Thèses : « Solid fermentation and submerged »

1

Kennedy, Max James. « The monitoring and evaluation of a solid substrate submerged culture fermentation ». Thesis, Massachusetts Institute of Technology, 1990. http://hdl.handle.net/1721.1/54965.

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2

Zhuang, Jun. « ECONOMIC ANALYSIS OF CELLULASE PRODUCTION BY CLOSTRIDIUM THERMOCELLUM IN SOLID STATE AND SUBMERGED FERMENTATION ». UKnowledge, 2004. http://uknowledge.uky.edu/gradschool_theses/170.

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Dependence on foreign oil remains a serious issue for the U.S. economy. Additionally, automobile emissions related to petroleum-based, fossil fuel has been cited as one source of environmental problems, such as global warming and reduced air quality. Using agricultural and forest biomass as a source for the biofuel ethanol industry, provides a partial solution by displacing some fossil fuels. However, the use of high cost enzymes as an input is a significant limitation for ethanol production.Economic analyses of cellulase enzyme production costs using solid state cultivation (SSC) are performed and compared to the traditional submerged fermentation (SmF) method. Results from this study indicate that the unit costs for the cellulase enzyme production are $15.67 per kilogram ($/kg) and $40.36/kg, for the SSC and SmF methods, respectively, while the market price for the cellulase enzyme is $36.00/kg. Profitability analysis and sensitivity analysis also provide positive results.Since these results indicate that the SSC method is economical, ethanol production costs may be reduced, with the potential to make ethanol a viable supplemental fuel source in light of current political, economic and environmental issues.

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3

Martins, Eduardo da Silva. « Purificação e caracterização bioquímica de poligalacturonases termoestáveis produzidas pelo fungo Thermoascus aurantiacus através de fermentação submersa e fermentação em estado sólido / ». Rio Claro : [s.n.], 2006. http://hdl.handle.net/11449/103964.

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Orientador: Eleni Gomes
Banca: Adalbeto Pessoa Júnior
Banca: Maria de Lourdes Teixeira de Moraes Polizeli
Banca: Márcia Regina Brochetto Braga
Banca: Gustavo Orlando Bonilla Rodriguez
Resumo: Pectinases termoestáveis apresentam características interessantes do ponto de vista da sua aplicação industrial, como alta estabilidade ao pH e à temperatura. Além disso, o tipo de processo fermentativo pode influenciar a produção e propriedades físico-químicas destas enzimas. A produção de poligalacturonase (PG) pelo fungo Thermoascus aurantiacus foi realizada em fermentação submersa (FSM) e em estado sólido (FES), usando substratos contendo pectina comercial ou subprodutos agro- industriais como fonte de carbono. A PG bruta obtida em FES apresentou atividade ótima a 65ºC e pH 5,0, com estabilidade na faixa de pH entre 4,0 e 5,0 e entre 7,5 e 8,5 e manteve 85% da atividade original quando incubada a 60ºC, por 1 hora. Em FSM, o melhor meio de cultivo foi a água amarela, com pH inicial de 5,5, após 5 dias de cultivo a 45ºC. A enzima em sua forma bruta apresentou temperatura ótima de 60ºC e pH ótimo de 5,0, maior estabilidade em pH ácido (3,0 a 4,5) e menor termoestabilidade, quando comparada com a obtida em FES, mantendo apenas 13% da atividade original quando incubada a 60ºC, por 1 hora. As enzimas foram purificadas utilizando-se cromatografias de filtração em gel e troca iônica. A PG purificada proveniente da FSM apresentou pH e temperatura ótimos de 5,5 e 60-65ºC, estabilidade em pH 5,0-5,5 e manteve, após 1 hora de incubação, 100% da atividade original até 50ºC. Resultados similares foram obtidos para a PG proveniente da FES. A PG de FES apresentou massa molar de 29,3 kDa, Km de 1,58 mg/mL e Vmáx de 1553,1 ? mol/min/mg, enquanto que a da FSM apresentou massa molar de 30,1 kDa, km de 1,46 mg/mL e Vmáx de 2433,3 ? mol/min/mg. Íons como Fe+3, Ca+2, e K+ praticamente não afetaram a atividade da enzima... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: Thermostable pectinases present important characteristics under the view of their industrial application, as their high stability to pH and temperature. Besides, the type of fermentative process used can affect the ir production and physical-chemical properties. The polygalacturonase (PG) production by the thermophilic fungus Thermoascus aurantiacus was carried out by submerged fermentation (SMF) and solid state fermentation (SSF) using substrates containing commercial pectin or agro- industrial residues as carbon sources. The crude PG from SSF presented optimum activity at 65ºC and pH 5.0, with stability at pH 4.0-5.0 and 7.5-8.5 and maintained 85% of its original activity at 60º C for 1 hour. In SMF the best cultivation medium was the liquid waste from juice extraction, with initial pH of 5.5, after 5 days of cultivation at 45ºC. The crude enzyme showed an optimum activity at 60ºC and pH 5.0, higher stability in acid ic pH (3.0 to 4.5) and was less thermostable when compared to that obtained in SSF, wich maintained only 13% of its original activity at 60ºC, for 1 hour. Purification of enzymes was carried out using filtration and ion-exchange chromatographies. The purified PG, from SMF, showed optimum pH and temperature of 5.5 and 60-65ºC, stability at pH 5.0-5.5 and preserved, after 1 hour incubation, 100% of its original activity at 50ºC. Similar results were obtained to PG from SSF. The PG obtained by SSF presented molar mass of 29.3 kDa, Km of 1.58 mg/ml and Vmáx of 1553.1 ? mol/min/mg, while that the enzyme from SMF presented molar mass of 30.1 kDa, km of 1.46 mg/ml and Vmáx of 2433.3 ? mol/min/mg. Ions such as Fe3+, Ca2+ and K+ practically did not affect the enzyme activity, while Mg2+, Mn2+ and Zn2+ decreased 7%, 75% and 50%... (Complete abstract, click electronic address below)
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Prevot, Vincent. « Comparaison de la production de complexes enzymatiques par fermentation en milieu solide et par fermentation en milieu liquide ». Thesis, Reims, 2013. http://www.theses.fr/2013REIMS009/document.

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La fermentation en milieu solide est un bioprocédé pouvant éventuellement être utilisé comme technologie de rupture pour diminuer le coût des biocatalyseurs utilisés dans la conversion de biomasse lignocellulosique comme le son de blé. La première partie de ces travaux de recherche a donc étudié le potentiel de cette technologie par rapport à celle de fermentation en milieu submergé lors d'une comparaison en application. Plusieurs tests de saccharification ont ainsi été réalisés sur différentes matières premières (cellulose, son de blé) et ont permis de montrer l'avantage différentiateur des biocatalyseurs produits par fermentation en milieu solide. Ensuite, la deuxième partie de ces travaux de recherche a porté sur l'étude des facteurs de la récalcitrance du son de blé à l'hydrolyse enzymatique. Deux principaux facteurs ont ainsi pu être démontrés : un facteur physique, lié à l'accessibilité des biocatalyseurs aux polysaccharides, et un facteur biochimique, lié à l'absence ou à la faible présence de certaines activités enzymatiques (féruloyl estérase,…) dans le complexe enzymatique de Trichoderma reesei Rut-C30. Cette étude a également permis d'identifier l'origine des différentes fractions glucidiques hydrolysées et de déterminer le potentiel glucidique actuellement hydrolysable à partir de cette biomasse. Enfin, la dernière partie de ces travaux de recherche a été consacrée à l'étude pratique d'un concept innovant de procédé permettant de favoriser la conversion des polysaccharides contenus dans le son de blé. Une levée de la barrière physique au transfert de masse et par conséquent une validation de ce concept a finalement pu être réalisée
Solid-state fermentation is a bioprocess that can optionally be used as disruptive technology to reduce the cost of biocatalysts used in the lignocellulosic biomass conversion like wheat bran. The first part of this research has explored the potential of this technology compared to submerged fermentation in an application comparison. Several saccharification tests have thus been carried on different feedstocks (cellulose, wheat bran) and have shown the differentiator advantage of biocatalysts produced by solid state fermentation. Then, the second part of this research has investigated the recalcitrance factors of wheat bran to enzymatic hydrolysis. Two main factors have thus been demonstrated: a physical factor, related to the accessibility of biocatalysts to the polysaccharides, and a biochemical factor, related to the absence or the low presence of some enzymatic activities (feruloyl esterase, ...) in the enzymatic complex of Trichoderma reesei Rut-C30. This study has also identified the origin of the various carbohydrate moieties hydrolyzed and has determined the carbohydrate potential currently releasable from this biomass. Finally, the last part of this research has been devoted to the practical study of an innovative concept of process to promote the conversion of polysaccharides in wheat bran. A removal of the physical barrier to mass transfer and therefore a validation of this concept has finally been achieved

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Martins, Eduardo da Silva [UNESP]. « Purificação e caracterização bioquímica de poligalacturonases termoestáveis produzidas pelo fungo Thermoascus aurantiacus através de fermentação submersa e fermentação em estado sólido ». Universidade Estadual Paulista (UNESP), 2006. http://hdl.handle.net/11449/103964.

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Pectinases termoestáveis apresentam características interessantes do ponto de vista da sua aplicação industrial, como alta estabilidade ao pH e à temperatura. Além disso, o tipo de processo fermentativo pode influenciar a produção e propriedades físico-químicas destas enzimas. A produção de poligalacturonase (PG) pelo fungo Thermoascus aurantiacus foi realizada em fermentação submersa (FSM) e em estado sólido (FES), usando substratos contendo pectina comercial ou subprodutos agro- industriais como fonte de carbono. A PG bruta obtida em FES apresentou atividade ótima a 65ºC e pH 5,0, com estabilidade na faixa de pH entre 4,0 e 5,0 e entre 7,5 e 8,5 e manteve 85% da atividade original quando incubada a 60ºC, por 1 hora. Em FSM, o melhor meio de cultivo foi a água amarela, com pH inicial de 5,5, após 5 dias de cultivo a 45ºC. A enzima em sua forma bruta apresentou temperatura ótima de 60ºC e pH ótimo de 5,0, maior estabilidade em pH ácido (3,0 a 4,5) e menor termoestabilidade, quando comparada com a obtida em FES, mantendo apenas 13% da atividade original quando incubada a 60ºC, por 1 hora. As enzimas foram purificadas utilizando-se cromatografias de filtração em gel e troca iônica. A PG purificada proveniente da FSM apresentou pH e temperatura ótimos de 5,5 e 60-65ºC, estabilidade em pH 5,0-5,5 e manteve, após 1 hora de incubação, 100% da atividade original até 50ºC. Resultados similares foram obtidos para a PG proveniente da FES. A PG de FES apresentou massa molar de 29,3 kDa, Km de 1,58 mg/mL e Vmáx de 1553,1 ? mol/min/mg, enquanto que a da FSM apresentou massa molar de 30,1 kDa, km de 1,46 mg/mL e Vmáx de 2433,3 ? mol/min/mg. Íons como Fe+3, Ca+2, e K+ praticamente não afetaram a atividade da enzima...
Thermostable pectinases present important characteristics under the view of their industrial application, as their high stability to pH and temperature. Besides, the type of fermentative process used can affect the ir production and physical-chemical properties. The polygalacturonase (PG) production by the thermophilic fungus Thermoascus aurantiacus was carried out by submerged fermentation (SMF) and solid state fermentation (SSF) using substrates containing commercial pectin or agro- industrial residues as carbon sources. The crude PG from SSF presented optimum activity at 65ºC and pH 5.0, with stability at pH 4.0-5.0 and 7.5-8.5 and maintained 85% of its original activity at 60º C for 1 hour. In SMF the best cultivation medium was the liquid waste from juice extraction, with initial pH of 5.5, after 5 days of cultivation at 45ºC. The crude enzyme showed an optimum activity at 60ºC and pH 5.0, higher stability in acid ic pH (3.0 to 4.5) and was less thermostable when compared to that obtained in SSF, wich maintained only 13% of its original activity at 60ºC, for 1 hour. Purification of enzymes was carried out using filtration and ion-exchange chromatographies. The purified PG, from SMF, showed optimum pH and temperature of 5.5 and 60-65ºC, stability at pH 5.0-5.5 and preserved, after 1 hour incubation, 100% of its original activity at 50ºC. Similar results were obtained to PG from SSF. The PG obtained by SSF presented molar mass of 29.3 kDa, Km of 1.58 mg/ml and Vmáx of 1553.1 ? mol/min/mg, while that the enzyme from SMF presented molar mass of 30.1 kDa, km of 1.46 mg/ml and Vmáx of 2433.3 ? mol/min/mg. Ions such as Fe3+, Ca2+ and K+ practically did not affect the enzyme activity, while Mg2+, Mn2+ and Zn2+ decreased 7%, 75% and 50%... (Complete abstract, click electronic address below)

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Ferrarezi, Ana Lúcia. « Seleção de fungos termofílicos para produção de lipase e aplicação na produção de biodiesel / ». Rio Claro : [s.n.], 2011. http://hdl.handle.net/11449/103950.

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Orientador: Eleni Gomes
Coorientador: Gustavo O. Bonilla-Rodriguez
Banca: Heizir Ferreira de Castro
Banca: Ernandes Benedito Pereira
Banca: Humberto Márcio Santos Milagre
Banca: Teresa Cristina Zangirolami
Resumo: As enzimas são catalisadores muito eficientes e de grande interesse na aplicação industrial. As lipases (E.C. 3.1.1.3) são serina-hidrolases que agem na hidrólise, esterificação e transesterificação de acilgliceróis de cadeia longa. Lipases microbianas têm sido amplamente usadas devido à sua especificidade. Na transesterificação moléculas de triacilglicerol reagem com um álcool na presença de um catalisador formando uma mistura de glicerol e ésteres de ácidos graxos. O biodiesel, definido como ésteres metílicos ou etílicos, tem atraído crescente interesse como uma fonte de energia renovável, substituindo o diesel produzido a partir de combustíveis fósseis. O presente trabalho tem como objetivo principal efetuar a prospecção de fungos termofílicos que apresentem produção significativa de lipase e, concomitantemente, atividade transesterificante. As linhagens foram selecionadas por detecção de atividade lipolítica em placas de ágar contendo Rodamina B, por fermentação submersa (FSm) e fermentação em estado sólido (FES). Foram testadas linhagens de fungos termofílicos da coleção do laboratório de Bioquímica e Microbiologia Aplicada, sendo Thermomucor indicae-seudaticae N31, Rhizomucor pusillus Myceliophtora sp F2.1.4, Myceliophtora sp M7.7, F2.1.1, F2.1.3 e Thermomyces lanuginosus TO-05 os que mostraram um maior potencial hidrolítico. Estudos adicionais avaliaram a produção de lipase através da modificação da fonte de componentes nutricionais e algumas propriedades físicas na produção de lipase por T. indicae-seudaricae N31 em FSm, e por R. pusillus e T. indicae-seudaticae N31 em FES. Os estudos dos processos fermentativos foram bem sucedidos, havendo um aumento de 16 vezes na produção de lipase de R. pusillus e de 36 vezes na lipase de T indicae-seudaticae... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: Enzymes are efficient catalysts and interesting for industrial applications. Lipases (EC 3.1.1.3) constitute a group of serine hydrolases that catalyze the hydrolysis, esterification and transesterification reactions of long chain acylglycerols. Microbial lipases have been widely used for biotechnological applications due to their specificity. In transesterification, molecule of a tryacylglicerol react with an alcohol in the presence of catalyst, producing a mixture of fatty esters and glycerol, Biodiesel, defined as methyl or ethyl fatty esters and has attracted considerable attention as a renewable source of energy, in substitution of fossil fuels. The main goal of the present work is the screening of thermophilic fungi that present outstanding lipase production and in parallel able to perform transesterification reaction. Strains were screened for lipase activity on agar plates containing Rhodamine B, for submerged fermentation (SmF) and solid state fermentation (SSF). The tested thermophilic strains were from the collections of the Biochemistry and Applied Microbiology Laboratory, where Thermomucor indicae-seudancae N31, Rhizomucor pusillus Myceliophtora sp F2.1.4, Myceliophtora sp M7.7, F2.1.1, F2.1.3 and Thermomyces lanuginosus TO-O5 had the highest lipase production. Additional studies attempted to improve lipase production by nutrient source modifications and physicals conditions in FmS by T. indicae-seudaticae N31 and in FSS by T. indicae-seudaticae N31 and R. pusillus. The fermentations studies were successful, with a 16 fold enhancement in lipase yield compared to the initial medim from lipase R. pusillus and 36 fold for the lipase T. indicae-seudaticae N31, both in FES. The lipase from T. indicae-seudaticae N31 cultured in SSF and SmF, exhibited maximum lipolytic activity at 40°C and stability for the pH range from 4 to 8. The enzyme produced by FmS presented maximum activity... (Complete abstract click electronic access below)
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Silva, Ronivaldo Rodrigues da. « Fermentação, purificação e caracterização da protease produzida pelo fungo Aspergillus fumigatus Fresenius / ». São José do Rio Preto : [s.n.], 2011. http://hdl.handle.net/11449/94802.

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Orientador: Hamilton Cabral
Banca: Rodrigo Simões Ribeiro Leite
Banca: Fabiana Fonseca Zanoelo
Resumo: A produção de proteases de origem microbiana depende das condições de cultivo e da diversidade bioquímica de cada espécie. Estudos comparativos entre fermentação em estado sólido (FES) e fermentação submersa (FSm) usando farelo de trigo e meio sintético, respectivamente, foram realizados para a determinação dos parâmetros de produção de proteases pelo fungo Aspergillus fumigatus Fresenius. A melhor produção de protease foi em FES no período de 96 horas utilizando farelo de trigo, temperatura de 30 ºC e 1x106 esporos/5g de substrato com 1.517 U/mL. Em FSm o pico de produção foi em pH 6,0, a 30ºC, 5x105 esporos/mL de meio no período de 72 e 96 horas em meio contendo 0,5 e 0,25% de caseína, respectivamente, ambos com 40 U/mL. Conforme a produtividade dos processos fermentativos, o extrato enzimático da FES foi utilizado para estudos de purificação e caracterização bioquímica. Neste estudo, a protease purificada apresentou atividade ótima em pH 7,5 e 50ºC, sendo inibida por Fenil-metil-sulfonil-fluoreto (PMSF) e mais intensamente por antipaína (1,6 µM). Sobre efeito de íons, foi observado modulação da atividade proteolítica, principalmente com inibição por AlCl3, cuja atividade proteolítica residual foi de 18% após incubação com este íon. Na presença de Ditiotreitol (DTT) e uréia houve diminuição da atividade proteolítica, apresentando atividades residuais de 63% em 200 mM de DTT e 10% com 5 M de uréia. Comparativamente, na concentração de 0,1% de cada surfactante estudado, notou-se redução da atividade proteolítica, sendo 97% em presença de Brometo de cetil-trimetil amônio (CTAB), 79% para 4 - (1,1,3,3 - Tetrametilbutil) fenil- polietileno glicol (Triton X-100), 55% com Polyoxyethylenesorbitan monolaurato (Tween-20) e completa redução da atividade (0%) em... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: The microbial protease production depends on growing conditions and the biochemical diversity of each species. Comparative studies between solid-state fermentation (SSF) and submerged fermentation (SmF) using wheat bran and synthetic medium, respectively, were performed to determine the optimum parameters for protease production by the fungus Aspergillus fumigatus Fresenius. The best protease production was in SSF within 96 hours using wheat bran, temperature 30°C and 1x106 spores/5g of substrate, with 1,517 U/mL. In SmF peak production was at pH 6.0 at 30°C, 5x105 spores/mL of media within 72 and 96 hours in medium containing 0.5 and 0.25% casein, respectively, with 40 U/mL. According to the productivity of the fermentative processes, enzymatic extract was used from SSF to study purification and biochemical characterization. In this study, purified protease showed optimum activity at pH 7.5 and 50°C, and inhibited by Phenylmethylsulfonyl fluoride (PMSF) and more intensely for antipain (1,6 µM). Concerning to the effect of ions, we observed modulation of the proteolytic activity, especially with inhibition by AlCl3, which residual activity was of 18 % after incubation with this ion. In the presence of Dithiothreitol (DTT) and ureia, we observed progressive decrease in proteolytic activity, presenting residual activities of 63% with 200 mM DTT, and 10% with 5 M ureia. Comparatively, in the concentration of 0.1% of each surfactant studied, there was a reduction in the proteolytic activity in 97% in presence of Cetyl trimethylammonium bromide (CTAB), 79% with 4-(1,1,3,3-Tetramethylbutyl)phenyl-polyethylene glycol (Triton X-100), 55% with Polyoxyethylenesorbitan monolaurate (Tween-20) and a complete inactivation in the presence of Sodium dodecyl sulfate... (Complete abstract click electronic access below)
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Habáníková, Kamila. « Využití odpadů rostlinného původu ». Master's thesis, Vysoké učení technické v Brně. Fakulta chemická, 2010. http://www.nusl.cz/ntk/nusl-216669.

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Production of cellulase and polygalacturonase by Aspergillus niger and Aureobasidium pullulans was studied in submerged (SmF) and solid state fermentation (SSF) systems. Substrates used in fermentation systems were mandarin peels and grape pomace. With Aspergillus niger used on grape pomace as a sole carbon source, cellulase production was detected after 72 hours in SSF and after 24 hours in SmF systems. The activity of cellulase per gram of substrate was higher in a submerged than in a solid state fermentation system. The longer time for higher polygalacturonase production was necessary in submerged fermentation systems and polygalacturonase activity was higher in SmF. The SSF fermentation with mandarin peels as a sole carbon source was similar, the highest detected activity of cellulase was determined after 72 hours. Different production of polygalacturonase was observed on mandarin peels in SmF systems. A comparison of enzyme productivities on grape pomace and mandarin peels showed that polygalacturonase activity per gram of substrate is highest in SmF system with mandarin peels as a sole carbon source. With Aureobasidium pullulans used on grape pomace as a sole carbon source, cellulase production was detected after 48 hours in SmF and SSF fermentation systems. The activity of cellulase per gram of substrate was higher in solid state system than in a submerged fermentation system. Longer time for higher polygalacturonase production was necessary in both fermentation systems. Polygalacturonase activity was higher in SmF. The SSF fermentation with mandarin peels as a sole carbon source was similar, the highest detected activity of cellulase was determined after 48 hours. Different production of polygalacturonase was observed on mandarin peels in SmF systems. A comparison of enzyme productivities on grape pomace and mandarin peels showed that polygalacturonase activity per gram of substrate is highest in SmF system with mandarin peels as a sole carbon source. For both systems and both substrates manganese-dependent peroxidase was detected for the first time. Differences in the enzyme synthesis by Aspergillus niger and Aureobasidium pullulans depend on both the substrates used as well as on the fermentation system.

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Ferrarezi, Ana Lúcia [UNESP]. « Seleção de fungos termofílicos para produção de lipase e aplicação na produção de biodiesel ». Universidade Estadual Paulista (UNESP), 2011. http://hdl.handle.net/11449/103950.

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
As enzimas são catalisadores muito eficientes e de grande interesse na aplicação industrial. As lipases (E.C. 3.1.1.3) são serina-hidrolases que agem na hidrólise, esterificação e transesterificação de acilgliceróis de cadeia longa. Lipases microbianas têm sido amplamente usadas devido à sua especificidade. Na transesterificação moléculas de triacilglicerol reagem com um álcool na presença de um catalisador formando uma mistura de glicerol e ésteres de ácidos graxos. O biodiesel, definido como ésteres metílicos ou etílicos, tem atraído crescente interesse como uma fonte de energia renovável, substituindo o diesel produzido a partir de combustíveis fósseis. O presente trabalho tem como objetivo principal efetuar a prospecção de fungos termofílicos que apresentem produção significativa de lipase e, concomitantemente, atividade transesterificante. As linhagens foram selecionadas por detecção de atividade lipolítica em placas de ágar contendo Rodamina B, por fermentação submersa (FSm) e fermentação em estado sólido (FES). Foram testadas linhagens de fungos termofílicos da coleção do laboratório de Bioquímica e Microbiologia Aplicada, sendo Thermomucor indicae-seudaticae N31, Rhizomucor pusillus Myceliophtora sp F2.1.4, Myceliophtora sp M7.7, F2.1.1, F2.1.3 e Thermomyces lanuginosus TO-05 os que mostraram um maior potencial hidrolítico. Estudos adicionais avaliaram a produção de lipase através da modificação da fonte de componentes nutricionais e algumas propriedades físicas na produção de lipase por T. indicae-seudaricae N31 em FSm, e por R. pusillus e T. indicae-seudaticae N31 em FES. Os estudos dos processos fermentativos foram bem sucedidos, havendo um aumento de 16 vezes na produção de lipase de R. pusillus e de 36 vezes na lipase de T indicae-seudaticae...
Enzymes are efficient catalysts and interesting for industrial applications. Lipases (EC 3.1.1.3) constitute a group of serine hydrolases that catalyze the hydrolysis, esterification and transesterification reactions of long chain acylglycerols. Microbial lipases have been widely used for biotechnological applications due to their specificity. In transesterification, molecule of a tryacylglicerol react with an alcohol in the presence of catalyst, producing a mixture of fatty esters and glycerol, Biodiesel, defined as methyl or ethyl fatty esters and has attracted considerable attention as a renewable source of energy, in substitution of fossil fuels. The main goal of the present work is the screening of thermophilic fungi that present outstanding lipase production and in parallel able to perform transesterification reaction. Strains were screened for lipase activity on agar plates containing Rhodamine B, for submerged fermentation (SmF) and solid state fermentation (SSF). The tested thermophilic strains were from the collections of the Biochemistry and Applied Microbiology Laboratory, where Thermomucor indicae-seudancae N31, Rhizomucor pusillus Myceliophtora sp F2.1.4, Myceliophtora sp M7.7, F2.1.1, F2.1.3 and Thermomyces lanuginosus TO-O5 had the highest lipase production. Additional studies attempted to improve lipase production by nutrient source modifications and physicals conditions in FmS by T. indicae-seudaticae N31 and in FSS by T. indicae-seudaticae N31 and R. pusillus. The fermentations studies were successful, with a 16 fold enhancement in lipase yield compared to the initial medim from lipase R. pusillus and 36 fold for the lipase T. indicae-seudaticae N31, both in FES. The lipase from T. indicae-seudaticae N31 cultured in SSF and SmF, exhibited maximum lipolytic activity at 40°C and stability for the pH range from 4 to 8. The enzyme produced by FmS presented maximum activity... (Complete abstract click electronic access below)

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Woods, Jeffrey. « The decrease in phenolic content of canola meal and the production of polyphenoloxidase in submerged and solid state fermentation using the white-rot fungus Trametes versicolor ». Thesis, National Library of Canada = Bibliothèque nationale du Canada, 1999. http://www.collectionscanada.ca/obj/s4/f2/dsk1/tape8/PQDD_0003/MQ46621.pdf.

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Klaic, Rodrigo. « PRODUÇÃO DE BIOHERBICIDA POR PROCESSOS FERMENTATIVOS A PARTIR DO FUNGO Phoma sp ». Universidade Federal de Santa Maria, 2014. http://repositorio.ufsm.br/handle/1/7982.

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Conselho Nacional de Desenvolvimento Científico e Tecnológico
Weeds are one of the major problems in the agricultural products cultivation and the main strategy of control is the use of chemical herbicides. Although is efficient, the herbicides bring direct and indirect consequences that overcome its benefits and this opened the way for bioherbicides. A significant barrier in the bioherbicide production is the development of an economically viable fermentation process. This way the aim of this work was to assess the production of a bioherbicide from the Phoma sp. fungus by submerged fermentation and solid-state fermentation, optimizing the fermentation medium in both cases and evaluating the herbicidal activity from the fermentation extracts in the control of the target plants. The culture medium was optimized and the cell growing was monitoring as response in the submerged fermentation. The optimum composition for the synthetic medium was glucose and peptone at 20 gL-1, yeast extract 7.5 gL-1 and an initial pH of 6.0. For the industrial medium, the ideal composition was 20 gL-1 of sucrose, 8 % CSL and initial pH of 6.0. Under these conditions, the maximum biomass obtained was 22 and 33 gL-1, for the synthetic and industrial medium, respectively. Therefore it was possible to obtain a 50 % higher biomass production in the industrial medium, reducing the cost of this fermentation medium. In the solid fermentation the composition of the culture medium was evaluated for obtaining the maximum herbicide action on target plants (cucumber). Bioassays were realized and the plant height, fresh weight, dry weight and phytotocixity mainly were the responses evaluated. The results obtained in the bioassays demonstrate that the bioherbicide presented activity towards the target plant and the intensity of the effect was influenced by the formulation of the fermentation medium. The optimized condition for bioherbicide production was moisture at 70 %, soybean bran of 30 wt% and CSL of 20 wt%, being possible to obtain a moderate injury, usually with recovery. The bioherbicide produced showed a mode of action based on the inhibition of carotenoids biosynthesis.
As plantas daninhas são um dos principais problemas no cultivo de produtos agrícolas e o principal método de controle é o uso de herbicidas químicos. Embora eficiente, os herbicidas trazem consequências diretas e indiretas que superam os benefícios em muitos casos, abrindo assim caminho para o desenvolvimento de bioherbicidas. Uma barreira significativa na produção de muitos bioherbicidas é o desenvolvimento de um processo economicamente viável, logo, os objetivos deste trabalho foram produzir um bioherbicida a partir do fungo Phoma sp. por fermentação submersa e fermentação em estado sólido, otimizar o meio de fermentação em ambos processos e avaliar a atividade herbicida dos extratos das fermentações no controle de plantas-teste. Na fermentação submersa foi otimizado o meio de cultivo e monitorado o crescimento celular como variável resposta. A composição otimizada do meio sintético foi 20 gL-1 de glicose e peptona, 7,5 gL-1 de extrato de levedura e pH inicial de 6.0. Para o meio industrial, a composição otimizada foi 20 gL-1 de sacarose, 8 % de água de maceração de milho (AMM) e pH inicial de 6.0. Nessas condições a máxima biomassa obtida foi de 22 e 33 gL-1, para o meio sintético e industrial, respectivamente, com uma produção de biomassa 50 % maior para o meio industrial e ainda possibilitou a redução do custo deste meio de fermentação. Na fermentação sólida foi otimizada a composição do meio de cultivo para obter a máxima ação do bioherbicida em plantas de pepino. Bioensaios foram realizados e o teste de Tukey mostrou que a altura da planta, massa verde da parte aérea, massa seca da parte aérea e principalmente os sinais de fitotoxidade foram significativos, entretanto, o comprimento do sistema radicular, a massa verde do sistema radicular, massa seca do sistema radicular e número de flores não apresentam resultados significativos. Os resultados obtidos nos bioensaios demonstraram que o bioherbicida apresenta atividade em plantas de pepino e a intensidade do efeito foi influenciado pela formulação do meio de fermentação. A condição otimizada para produção do bioherbicida foi 70 % de umidade, 30 % (p/v) de farelo de soja e 20 % (p/v) de AMM, nessas condições o nível de injúria obtida foi moderada com recuperação. O bioherbicida produzido mostrou um possível modo de ação baseado na inibição de biossíntese de carotenóides.

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Hu, Jinghua. « The production of phenolics-degrading enzymes in submerged and solid state fermentation and the decrease in phenolic content of canola meal using the white-rot fungus Pleurotus ostreatus ». Thesis, National Library of Canada = Bibliothèque nationale du Canada, 1999. http://www.collectionscanada.ca/obj/s4/f2/dsk1/tape8/PQDD_0001/MQ46584.pdf.

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Mascarin, Gabriel Moura. « Production by solid-state and liquid fermentation and formulation of virulent strains of the fungal entomopathogens Beauveria bassiana and Isaria fumosorosea against whiteflies ». Universidade de São Paulo, 2015. http://www.teses.usp.br/teses/disponiveis/11/11146/tde-28042015-111429/.

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Bemisia tabaci Gennadius (Hemiptera: Aleyrodidae) biotype B is a cosmopolitan, devastating insect pest due to their direct damages and transmission of plant viruses. Entomopathogenic fungi comprise the most diverse group of pathogens regulating arthropod pest populations in agroecosystems. Anamorphic fungal entomopathogens, including Beauveria bassiana, Isaria fumosorosea, and Lecanicillium spp., are among the main biocontrol agents of whitefly populations. Advances in research focusing on virulence, mass production, formulation, and storage stability of fungal propagules are imperative for the development of efficient mycopesticides toward whiteflies and other soft-bodied insects. Therefore, this study placed emphasis on screening for virulent fungal strains, enhancement of efficacy using nonionic surfactants in spray tank-mix, development of liquid culture conditions for rapid production and stabilization processes of single-yeast like cells known as blastospores. Firstly, we selected virulent strains of B. bassiana and I. fumosorosea displaying fastest speed of kill and inciting highest mortality levels of whitefly nymphs and adults along with their ability to produce high numbers of conidia on moistened parboiled rice. Secondly, insecticidal performance was enhanced by combining nonionic surfactants with spore suspensions rendering additive or synergistic effects. These surfactants also allowed reducing the volume application rate without altering fungal bioefficacy. Results from liquid fermentation studies using B. bassiana and I. fumosorosea revealed that appropriate amounts of inexpensive ingredients, such as cottonseed flour and glucose, are suitable for the rapid production of high yields of blastospores (3 days pre-culture and 2-3 days culture). The resultant blastospores of various strains survived well to desiccation and remained viable for more than one year under refrigeration. Moreover, these air-dried blastospores of both fungal species showed higher virulence against whitefly nymphs when compared with solid-substrate produced conidia. Optimized liquid culture production for B. bassiana blastospores was also achieved through the manipulation of oxygen rates and osmotic pressure in the liquid media. Furthermore, these blastospores produced in highly aerated and hyperosmotic liquid medium containing 140 g glucose L-1 were also more virulent to whitefly nymphs than those cells derived from low-osmotic medium amended with 40 g glucose L-1. These optimal conditions were also scaled up in 5-L bioreactor that yielded 1-2 × 1012 viable blastospores L-1 in 6 days at a cost of US$ 0.19 L-1. These blastospores were formulated with diatomaceous earth for air drying or for spray drying. Formulated blastospores of B. bassiana survived dehydration using both drying methods and showed improved shelf life when stored under vacuumpackaged at 4 °C rather than 28 °C. However, when these blastospores were actively packaged with dual action oxygen-moisture scavenging system, blastospores showed prolonged stability for up to 7 months at 28 °C and still remained virulent to whiteflies. Therefore, this low-cost production and stabilization method for the rapid production of shelf stable, virulent blastospores of B. bassiana and I. fumosorosea may expand the commercial use of mycopesticides for insect control in mainstream agriculture.
A mosca-branca, Bemisia tabaci Gennadius (Hemiptera: Aleyrodidae) biótipo B, é uma praga cosmopolita e devastadora devido aos prejuízos oriundos dos seus danos diretos e transmissão de vírus. Fungos entomopatogênicos compreendem um grupo diversificado, que desempenha ação importante na regulação de populações de praga em agroecossistemas. Fungos ascomicetos anamórficos, como Beauveria bassiana, Isaria fumosorosea e Lecanicillium spp., constituem relevantes agentes de biocontrole de moscas-brancas. Avanços na pesquisa focando virulência, produção massal, formulação e estabilização de propágulos fúngicos são fundamentais para o desenvolvimento de micopesticidas eficientes contra moscas-brancas e outros insetos. Desta forma, este trabalho objetivou selecionar isolados fúngicos virulentos à mosca-branca; aumentar a eficácia mediante uso de surfactants não-iônicos em suspensões conidiais; desenvolver meios de cultura para produção rápida e estável por fermentação líquida submersa de células leveduriformes conhecidas por blastosporos. Na primeira etapa, isolados virulentos de B. bassiana e I. fumosorosea foram selecionados pela rápida e elevada atividade inseticida a ninfas e adultos de mosca-branca, bem como alto rendimento de conídios em arroz parboilizado. A adição de surfactantes organosiliconados permitiu a redução do volume de calda aplicado com resultados aditivos ou sinérgicos de controle. Foi ainda verificado que altos rendimentos de blastosporos tanto de B. bassiana como I. fumosorosea foram obtidos em curto tempo de fermentação líquida (3 dias de précultivo e 2-3 dias de cultivo) usando nutrientes de baixo custo, como glucose e farelo de algodão. Esses blastosporos foram tolerantes à dessecação e mantiveram viabilidade por mais de um ano sob refrigeração (4 °C). Os blastosporos foram mais virulentos que conídios aéreos, o que coloca esta estrutura como a mais indicada como ingrediente ativo em bioinseticidas para moscas-brancas. Mediante manipulação nutricional e física do ambiente de fermentação, a produção de blastosporos de B. bassiana foi optimizada mediante aumento da aeração e pressão osmótica do meio líquido. Blastosporos produzidos em meio líquido altamente aerado e hiperosmótico (140 g glucose L-1) mostraram-se mais virulentos à mosca-branca em relação àqueles produzidos em meio hipo-osmótico (40 g glucose L-1). Esse processo foi reproduzido em escala piloto usando biorreator de 5 L resultando numa produção de 1-2 × 1012 blastosporos viáveis L-1 em apenas 3 dias a um custo de US$ 0,19 L-1. Blastosporos de B. bassiana formulados com terra de diatomáceas e secados em fluxo de ar contínuo, ou secados em spray dryer tiveram estabilidade extendida por até 8 meses a 4 °C e superior em relação a 28 °C. Durante empacotamento, o uso de sachês absorventes de oxigênio e umidade prolongou consideravelmente a viabilidade de blastosporos armazenados a 28 °C por até 7 meses sem afetar sua eficiência contra mosca-branca. Em suma, esses resultados demonstram a viabilidade técnica e econômica de produção de blastosporos virulentos de B. bassiana e I. fumosorosea, tolerantes à dessecação e estáveis durante armazenamento. Esta tecnologia é uma nova opção que pode contribuir para expansão comercial de bioinseticidas à base de fungos entomopatogênicos.

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Cruz, Filho Raimundo Felipe da. « Avaliação dos processos de produção de protease fibrinolitica por fermentação submersa, semi-sólida e extrativa utilizando uma espécie de bacilo da Amazônia ». Universidade Federal do Amazonas, 2013. http://tede.ufam.edu.br/handle/tede/3082.

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Conselho Nacional de Desenvolvimento Científico e Tecnológico
The fibrinolytic proteases degrade fibrin clots and therefore play an important role in the pharmaceutical industry as chemotherapeutic agents in the treatment of cardiovascular diseases. These biocatalysts were gradually discovered from plants, insects, earthworms, snakes and microorganisms (bacteria and fungi). Cardiovascular disease has been one of the leading causes of death in the world. A major cause of heart disease is the accumulation of fibrin in the arteries, causing thrombosis. According to the World Health Organization (WHO), about 17.5 million people will die of cardiovascular disease this year, and in 2030, this amount will be 23.6 million. Given the great potential of microbial biodiversity and the growing regional Amazon applicability of enzymes in the production of drugs, this research was conducted with the objective to (1) evaluate the growth of Bacillus stearothermophilus, (2) establish growth parameters associated with production of fibrinolytic proteases in submerged fermentation and (3) assess the effect of the extractive fermentation in the separation of these enzymes. In this study techniques of extractive fermentation and solid-state fermentation were employed. By submerged fermentation the following parameters were determined: Profile of growth of Bacillus stearothermophilus and the production of protease using 100 mL of the liquid medium [(g / L) 2 g KH2PO4, (NH4)2SO4 1 g, MgSO4 7H2O 0,1 g Na2HPO4 2H2O 0,9 g, yeast extract 1 g, distilled water 1000 ml] pH 7.2 supplemented with 0.5% gelatin in an 500 mL Erlenmeyer flask. The growth of the bacteria was determined at 610 nm, once every 2 hours for 36 hours. To determine the best conditions for the production of proteases were evaluated the influence of pH, stirring and temperature, age of inoculum and substrate concentration, the influence of natural sources of carbon (tapioca, arraruta and crueira), nitrogen sources and aeration. In the recovered extract was also performed a toxicity bioassay in Artemia salina and degradation tests in vitro of the blood clot by the fibrin plate method and the artificial clot degradation in tube. In addition, the partition coefficient (K), the purification factor (PF) and recovering the enzyme were determined. The solid-state fermentation was performed using as substrate 10g of manteiguinha bean [Vigna unguiculata (L.) Walp] with 60% humidity, pH 5.0 in a 250 ml Erlenmeyer flask. In extractive fermentation the best conditions were pH 5.0, 180 rpm and 25 °C in systems using PEG 1000 (g/mol-1) to 20% (w/w) and phosphate salts 15% (w/w) with K 1.05; FP 1.00; 152.54 Y. 34 mm halo in fibrin plate and partial degradation of the clot in tube. In the solid-state fermentation, the production of protease was 8.87 (U/mL), 23 mm of translucent halo in fibrin plate with total degradation of the blood clot in 24 hours. In this study, protease produced from Bacillus stearothermophilus by extractive fermentation and semi-solid fermentation was evaluated, showing in the optimum cultivation conditions that this microorganism presents physiology for industrial application in the production of the fibrinolytic protease
As proteases fibrinolíticas degradam coágulos de fibrina, por isso têm um importante papel na indústria farmacêutica como agentes quimioterapêuticos no tratamento de doenças cardiovasculares. Estes biocatalisadores foram descobertos gradualmente a partir de plantas, insetos, anelídeos, serpentes e micro-organismos (bactérias e fungos). Doenças cardiovasculares tem sido a principal causa de morte no mundo. Uma das principais causas de doenças cardíacas é o acúmulo de fibrina nas artérias, acarretando trombose. De acordo com Organização Mundial da Saúde (OMS), cerca de 17,5 milhões de pessoas morrerão este ano de doenças cardiovasculares, e em 2030, esse montante será de 23,6 milhões. Tendo em vista o grande potencial da biodiversidade microbiana regional Amazônica e a crescente aplicabilidade de enzimas na produção de medicamentos, esta pesquisa foi realizada com o objetivo de (1) avaliar o crescimento de Bacillus stearothermophilus, (2) estabelecer os parâmetros de crescimento associado a produção de proteases fibrinolíticas por fermentação submersa e (3) verifica o efeito da fermentação extrativa na separação dessas enzimas. Neste estudo foram empregados técnicas de fermentação extrativa e fermentação semi-sólida. Por fermentação submersa foram determinados os seguintes parâmetros: Perfil do crescimento de Bacillus stearothermophilus e a produção de protease utilizando 100mL do meio líquido [(g/L) KH2PO4 2g; (NH4)2SO4 1g; MgSO4 7H2O 0,1 g; Na2HPO4 2H2O 0,9 g; Extrato de Levedura 1 g; água destilada 1000mL] pH 7,2 suplementado com gelatina 0,5%, em frasco de Erlenmeyer de 500mL. O crescimento da bactéria foi determinado a 610nm, de 2 em 2 horas, durante 36 horas. Na Determinação das melhores condições para produção de proteases foram avaliados a influência do pH; agitação, temperatura, a idade do inóculo, da concentração do substrato, a influência das fontes de naturais de carbono (tapiocas, araruta e crueira), fontes de nitrogênio e aeração. No extrato recuperado foi realizado também bioensaio de toxicidade em Artemia salina e testes de degradação in vitro do coágulo sanguíneo pelos métodos da placa de fibrina e degradação do coagulo artificial em tubo. Foi determinado também o coeficiente de partição (K), o fator de purificação (FP) e a recuperação da enzima. A fermentação semi-sólida foi realizada utilizando como substrato 10g feijão manteiginha [Vigna unguiculata (L.) Walp], com 60% umidade, pH 5,0 em frasco Erlenmeyers de 250 mL. Na fermentação extrativa as melhores condições foram: pH 5,0; 180 rpm e 25 ºC, no sistemas utilizaram PEG 1000 (g/mol-1) a 20% (p/p) e sais fosfato a 15% (p/p) com K de 1,05; FP de 1,00; Y de 152,54. Halo de 34 mm na placa de fibrina e degradação parcial do coagulo em tubo. Na fermentação semi-sólida a produção de protease foi de 8,87 (U/mL), halo translucido de 23 mm em placa de fibrina com degradação total do coagulo de sangue em 24h. No presente estudo, protease de Bacillus stearothermophilus produzido em fermentação extrativa e fermentação semi-sólida foi avaliada, demonstrando nas condições ótimas de cultivo que este micro-organismo apresenta fisiologia para aplicações industriais na produção de protease fibrinolítica

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Silva, Ronivaldo Rodrigues da [UNESP]. « Fermentação, purificação e caracterização da protease produzida pelo fungo Aspergillus fumigatus Fresenius ». Universidade Estadual Paulista (UNESP), 2011. http://hdl.handle.net/11449/94802.

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A produção de proteases de origem microbiana depende das condições de cultivo e da diversidade bioquímica de cada espécie. Estudos comparativos entre fermentação em estado sólido (FES) e fermentação submersa (FSm) usando farelo de trigo e meio sintético, respectivamente, foram realizados para a determinação dos parâmetros de produção de proteases pelo fungo Aspergillus fumigatus Fresenius. A melhor produção de protease foi em FES no período de 96 horas utilizando farelo de trigo, temperatura de 30 ºC e 1x106 esporos/5g de substrato com 1.517 U/mL. Em FSm o pico de produção foi em pH 6,0, a 30ºC, 5x105 esporos/mL de meio no período de 72 e 96 horas em meio contendo 0,5 e 0,25% de caseína, respectivamente, ambos com 40 U/mL. Conforme a produtividade dos processos fermentativos, o extrato enzimático da FES foi utilizado para estudos de purificação e caracterização bioquímica. Neste estudo, a protease purificada apresentou atividade ótima em pH 7,5 e 50ºC, sendo inibida por Fenil-metil-sulfonil-fluoreto (PMSF) e mais intensamente por antipaína (1,6 µM). Sobre efeito de íons, foi observado modulação da atividade proteolítica, principalmente com inibição por AlCl3, cuja atividade proteolítica residual foi de 18% após incubação com este íon. Na presença de Ditiotreitol (DTT) e uréia houve diminuição da atividade proteolítica, apresentando atividades residuais de 63% em 200 mM de DTT e 10% com 5 M de uréia. Comparativamente, na concentração de 0,1% de cada surfactante estudado, notou-se redução da atividade proteolítica, sendo 97% em presença de Brometo de cetil-trimetil amônio (CTAB), 79% para 4 - (1,1,3,3 - Tetrametilbutil) fenil- polietileno glicol (Triton X-100), 55% com Polyoxyethylenesorbitan monolaurato (Tween-20) e completa redução da atividade (0%) em...
The microbial protease production depends on growing conditions and the biochemical diversity of each species. Comparative studies between solid-state fermentation (SSF) and submerged fermentation (SmF) using wheat bran and synthetic medium, respectively, were performed to determine the optimum parameters for protease production by the fungus Aspergillus fumigatus Fresenius. The best protease production was in SSF within 96 hours using wheat bran, temperature 30°C and 1x106 spores/5g of substrate, with 1,517 U/mL. In SmF peak production was at pH 6.0 at 30°C, 5x105 spores/mL of media within 72 and 96 hours in medium containing 0.5 and 0.25% casein, respectively, with 40 U/mL. According to the productivity of the fermentative processes, enzymatic extract was used from SSF to study purification and biochemical characterization. In this study, purified protease showed optimum activity at pH 7.5 and 50°C, and inhibited by Phenylmethylsulfonyl fluoride (PMSF) and more intensely for antipain (1,6 µM). Concerning to the effect of ions, we observed modulation of the proteolytic activity, especially with inhibition by AlCl3, which residual activity was of 18 % after incubation with this ion. In the presence of Dithiothreitol (DTT) and ureia, we observed progressive decrease in proteolytic activity, presenting residual activities of 63% with 200 mM DTT, and 10% with 5 M ureia. Comparatively, in the concentration of 0.1% of each surfactant studied, there was a reduction in the proteolytic activity in 97% in presence of Cetyl trimethylammonium bromide (CTAB), 79% with 4-(1,1,3,3-Tetramethylbutyl)phenyl-polyethylene glycol (Triton X-100), 55% with Polyoxyethylenesorbitan monolaurate (Tween-20) and a complete inactivation in the presence of Sodium dodecyl sulfate... (Complete abstract click electronic access below)

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Oliveira, Camila Fernanda Dias de. « Produção de pigmentos por Monascus ruber CCT 3802 a partir do xarope de maltose como substrato ». Universidade Federal de Goiás, 2017. http://repositorio.bc.ufg.br/tede/handle/tede/7711.

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Artificial dyes are commonly used in the food industry, both for their low cost and for ease of procurement. The consumer market, however, requires healthier products and a viable alternative would be the use of natural pigments. In addition to plants, flowers, fruits and animals, micro-organisms can be the source of this type of pigment, such as fungi, bacteria and microalgae. Therefore, the present study had as objective to produce pigments from the solid fermentation and submerged by filamentous fungus Monascus ruber CCT 3802 using maltose syrup as substrate. The effect of substrate concentration on maltose syrup and the influence of pH on pigment production was studied by evaluating the radial growth rate, pigment production and pigment properties by thermal stability. The main results demonstrate that the highest radial growth velocity was obtained from the plate containing 5 g L-1 maltose syrup 0.1053 mm h-1 , which corresponds to an increase of 71.70% when compared to the medium extract of Malt agar (MEA). In the submerged fermentation the concentration of 10 g L-1 of maltose obtained the highest absorbance (14.54 AU), lower biomass (4.65 g L-1 ) and greater dark red intensity. For pH determination, the highest radial growth rate was obtained when the fungus was cultivated at pH 6.5 and the production of yellow pigment was obtained at low pH (2.0 and 2.5) while the production of orange pigments was in the pH range (3.0 and 3.5) and the red pigment yield obtained when the fungus was grown at pH above 4.0. Thus, it can be concluded that maltose syrup and pH exerted a significant influence on both the radial growth rate and the production of pigments in submerged fermentation, showing that lower substrate concentrations favor higher amounts of red pigment and that associated with the culture in PH 6.5 favored the formation of red pigments.
Corantes artificiais são comumente utilizados na indústria de alimentos, tanto pelo seu baixo custo quanto pela facilidade de obtenção. O mercado consumidor, entretanto, requer produtos mais saudáveis e uma alternativa seria a utilização de pigmentos naturais. Além de plantas, flores, frutos e animais, micro-organismos podem ser fonte deste tipo de pigmento, como fungos, bactérias e microalgas. Portanto, o presente estudo teve como objetivo produzir pigmentos a partir da fermentação sólida e submersa pelo fungo filamentoso Monascus ruber CCT 3802 utilizando xarope de maltose como substrato. Foi estudado o efeito da concentração do substrato xarope de maltose e a influência do pH na produção de pigmentos, avaliando a velocidade de crescimento radial, a produção de pigmentos e as propriedades dos pigmentos mediante a estabilidade térmica. Os principais resultados demonstram que a maior velocidade de crescimento radial foi obtida da placa contendo 5 g L-1 de xarope de maltose 0,1053 mm h-1 , que corresponde ao um aumento de 71,70% quando comparado com o meio extrato de malte ágar (MEA). Na fermentação submersa a concentração de 10 g L-1 de maltose obteve a maior absorbância (14,54 UA), menor biomassa (4,65 g L-1 ) e maior intensidade da cor vermelho escuro. Já para determinação da influência do pH a velocidade de crescimento radial mais elevada foi obtida quando o fungo foi cultivado em pH 6,5. A obtenção dos pigmentos, amarelo, laranja e vermelho, deu-se nos intervalos de pH 2,0-2,5, 3,0-3,5 e acima de 4,0 respectivamente. Assim, pode-se concluir que a concentração de xarope de maltose e o pH exerceram influência significativa tanto na velocidade de crescimento radial quanto na produção de pigmentos em fermentação submersa, mostrando que menores concentrações de substrato favorecem maiores quantidades de pigmento vermelho e que associados ao cultivo em pH 6,5 favoreceram a formação de pigmentos vermelhos.

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Tonelotto, Mariana. « Produção de celulases, purificação e caracterização bioquímico-cinética da ß-Glicosidase produzida por fungo isolado da região amazônica ». Universidade Federal de São Carlos, 2012. https://repositorio.ufscar.br/handle/ufscar/7001.

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The selection of cellulase-producing fungi is one of the possible estrategies for obtaining necessary enzymes to hydrolyze the lignocellulosic material of plant biomass and thereby contribute to the viability of cellulosic ethanol production. The aim of this study was achive a screening of isolated fungi from the Amazon region to assess the production of enzymes related to plant biomass degradation, in order to select a line for production, purification and biochemical, kinetical and and structural biology characterizationof the ß-Galactosidase enzyme. Therefore, this work was undertaken in three stages, first of all it was performed a screening of 40 fungal strains isolated from the Amazon region through the cultivation in solid state fermentation (FES) at 35ºC for 240 hours, using as substrate wheat bran. It was evaluated the production of xylanase, endoglucanase, FPase, pectinase, ß-Glicosidase and total protein, and the fungi that stood out were: P6B2, the best producer of xylanase, P47C3 (Aspergillus niger), the best producer endoglucanase and ß-Glicosidase and P40B3, the best producer of FPase. These three fungi were selected for the second phase of this work for assessment in the production of xylanase, FPase, endoglucanase, ß-Glicosidase and total protein by submerged fermentation (FSm). The fermentation took place for 5 days at 30ºC and 200 rpm with a source of carbon: 1% of wheat bran washed and nutrient medium. The fungi P47C3, which was identified as Aspergillus niger, showed the best production of these enzymes, being selected for the third stage of this project. This last step involved the selection of an enzyme that has not been elucidated its structural biology. Given this fact, we carried out a study of selection of the medium, purification and biochemicalkinetical characterization of ß-Galactosidase. The Aspergillus niger (P47C3) was subjected to submerged for 5 days at 200 rpm at 30ºC. Purification occured in three steps using: ion exchange column SP-Sephadex C-50 and SP TSK-5PW column, and gelfiltration, with the resin Sephacryl S-200. The enzyme ß-Galactosidase showed a molecular weight of 125 kDa, being stable at pH 4,0, with anoptimum temperature of 55ºC. It was evaluated theKmap e Vmáxap of two substrates, PNPG and lactose, being: 2,204 mM-0,285 mM/min and 2,101 mM-0,75mM/min, respectively. The inhibition of hydrolasis of the substrate PNPG by ß-Galactosidase in the presence of galactose inhibitor product showed a Ki value of 5,01 mM. Finally, the ß-Galactosidase was subjected to crystallization conditions, the best conditions occurred in buffer 0,2M Tris- HCl, with the precipitation agent, 12% PEG 4000 at pH 8,6. Therefore, the unpublished protocol for purification of ß-Galactosidase was efficient and it is possible to crystallize this enzyme of isolated fungi from the Amazon region, which showed great potencial for the production of this enzyme and that the future can be used in industrial application and biotechnological innovations.
A seleção de fungos produtores de celulases é uma das possíveis estratégias para a obtenção das enzimas necessárias para hidrolisar o material lignocelulósico da biomassa vegetal e com isso contribuir para a viabilização da produção de etanol celulósico. O objetivo desse trabalho foi realizar um screening dos fungos isolados da região amazônica para a avaliação da produção de enzimas relacionadas à degradação da biomassa vegetal, a fim de selecionar uma linhagem para produção, purificação e caracterização bioquímica, cinética e biologia estrutural da enzima ß-Galactosidase. Dessa forma, esse trabalho foi realizado em três etapas, primeiramente foi realizado um screening de 40 linhagens fúngicas isoladas da região amazônica, através do cultivo em fermentação em estado sólido (FES), a 35°C, por 240 horas, utilizando como substrato o farelo de trigo. Avaliou-se a produção de xilanase, endoglucanase, FPase, pectinase, ßglicosidase e proteínas totais, sendo que os fungos que mais se destacaram foram o: P6B2, melhor produtor de xilanase, P47C3 (Aspergillus niger), melhor produtor de endoglucanase e ß-glicosidase e o P40B3, melhor produtor de FPase. Esses três fungos, foram selecionados para a segunda fase do trabalho para avaliação na produção de xilanase, FPase, endoglucanase, ß-glicosidase e proteínas Totais por fermentação submersa (FSm). A fermentação ocorreu por 5 dias, à 30ºC e 200 rpm tendo como fonte de carbono: 1% de farelo de trigo lavado e meio nutriente. O fungo P47C3, identificado como Aspergillus niger, apresentou melhor produção dessas enzimas, sendo selecionado para a terceira etapa desse projeto. Essa última etapa, envolveu a escolha de uma enzima que não estivesse sua biologia estrutural elucidada. Diante desse fato, realizou-se um estudo de seleção do meio de cultivo, purificação e caracterização bioquimico-cinética da ß-Galactosidase. O fungo Aspergillus niger (P47C3) foi submetido a fermentação submersa, durante 5 dias, à 200 rpm em 30ºC. A purificação ocorreu em três etapas utilizando: colunas de troca iônica SP - Sephadex C-50 e a coluna SP -TSK 5PW; e gel filtração, com a resina Sephacryl S-200. A enzima ß-Galactosidase apresentou uma massa molecular de 125 kDa, sendo estável em pH 4,0, e com temperatura ótima de 55ºC. Avaliou-se a Kmap e Vmáxap de dois substratos, o PNPG e a lactose, sendo: 2,204 mM - 0,285 mM/min e 2,101 mM 0,750 mM/min, respectivamente. A inibição da hidrólise do substrato PNPG pela ß-Galactosidase na presença do produto inibidor galactose apresentou um valor de Ki de 5,01 mM. Por fim, a ß-Galactosidase foi submetida a condições de cristalização, as melhores condições ocorreram em tampão 0,2M Tris-HCl, tendo como agente precipitante, PEG 4000 12% em pH 8,6. Portanto, o protocolo inédito de purificação da ß-Galactosidase foi eficiente, sendo possível cristalizar essa enzima do fungo isolado da região amazônica, o qual apresentou grande potencial para a produção dessa enzima e que futuramente possa ser utilizado em aplicações industriais e inovações biotecnológicas.

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Scheufele, Fabiano Bisinella. « Bioconversão de resíduos agroindustriais por micro-organismos do bioma amazônico produtores de enzimas lignocelulolíticas ». Universidade Estadual do Oeste do Parana, 2012. http://tede.unioeste.br:8080/tede/handle/tede/1906.

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior
Lignocellulosic biomass has high yields of cellulose which can be hydrolyzed to fermentable carbohydrates. Global generation of agro-industrial wastes grows simultaneously with the sector development resulting at the accumulation of lignocellulosic residues leading environmental pollution and loss of potential materials for the bioconversion to a wide range of high added value products, such as biofuels. Recently, the search of renewable sources of energy has grown, due to the depleting of fossil fuels, increasing the possibility at the conversion of the lignocellulosic biomass via hydrolytic enzymes. The aim of this work was evaluate cellulases production by lignocellulolytic fungi from the Amazonic biome aiming at the bioconversion of the agro-industrial residues. Submerged and solid-state fermentations were performed to select the microorganism with superior cellulase productive capacity. The influence of parameters such as pH, surfactant induction (Tween 80), aeration and agitation, besides the alkaline oxidative treatment of the sugarcane bagasse. Statistical design were carried out to estimate the influence of the moisture and the initial pH at cellulases production by solid-state fermentation. Trichoderma sp. and Aspergillus niger performed the best production of enzymes, where the highest yields of total cellulase were obtained by agitated submerged fermentation with sugarcane bagasse pretreated with H2O2 (1%) reaching 0.265 U.mL-1 (12.915 U.g-1) by Trichoderma sp. at the sugarcane bagasse, and 0.155 U.mL-1 (7.549 U.g-1) by Aspergillus niger. Through solid state fermentations with the pretreated sugarcane bagasse the influence of initial pH and the moisture were evaluated by statistical design. In the case of the Trichoderma sp. both parameters were significant at the cellulase production, as well as the synergistic interaction, within the confidence interval of 95%, yielding 0.167 U.mL-1 (2.695 U.g-1), at the pH 7.0 and 1:9 solid-liquid ratio. For Aspergillus niger only pH was significant and the cellulase content obtained was 0.098 U.mL-1 (1.695 U.g-1) at pH 7.0. Finally, a cellulase produced by Trichoderma sp. at solid state fermentation and a commercial enzyme were used at enzymatic hydrolysis tests. The parameters hydrolysis time, enzyme dilution, concentration of Tween 80 and solid-liquid ratio of sugarcane bagasse were evaluated. The significant variables were then optimized by a central composite rotational design. The strain of Trichoderma sp. from the Amazon biome showed potential at the cellulase production and the treated sugarcane bagasse was a fine substrate for the enzymatic production.
A biomassa lignocelulósica contêm altos teores de celulose e outros polissacarídeos em sua constituição química, podendo ser hidrolisados em açúcares fermentescíveis. A geração de resíduos agroindustriais anual tem crescido resultando no acúmulo de resíduos que contribuem para a poluição do meio ambiente e na perda de materiais que possuem potencial na bioconversão a produtos de alto valor agregado, como por exemplo, biocombustíveis. Recentemente, há a necessidade de fontes energéticas de origem renovável, devido à diminuição dos combustíveis fósseis, viabilizando a conversão das biomassas lignocelulósicas via enzimas hidrolíticas. O objetivo deste trabalho foi avaliar a produção de enzimas celulases por fungos lignocelulolíticos provenientes do bioma amazônico visando a bioconversão de resíduos agroindustriais. Diferentes fermentações foram realizadas, tanto em meio submerso quanto em estado sólido, através das quais selecionou-se os micro-organismos com melhor capacidade produtiva de celulases. Estudou-se a influência de parâmetros como pH, utilização de surfactante como indutor (Tween 80), aeração e agitação, além do tratamento alcalino oxidativo do bagaço de cana. Os micro-organismos que apresentaram melhor desempenho na produção das enzimas foram o Trichoderma sp. e o Aspergillus niger, sendo que os maiores níveis de celulase total foram obtidos por fermentação submersa nos ensaios agitados com bagaço de cana pré-tratado com H2O2 (1%), com 0,265 U.mL-1 (12,915 U.g-1) pelo Trichoderma sp., e 0,155 U.mL-1 (7,549 U.g-1) com Aspergillus niger. A partir de fermentações em estado sólido com o bagaço de cana pré-tratado avaliou-se a influência dos parâmetros pH inicial e umidade por planejamentos experimentais, verificando-se que para o Trichoderma sp. ambos os parâmetros, bem como a interação sinergética entre si, foram significativos dentro do intervalo de confiança de 95%, obtendo-se 0,167 U.mL-1 (2,695 U.g-1) no pH 7,0 e relação sólido-líquido 1:9. No caso do Aspergillus niger apenas o pH foi significativo e o teor de celulase obtido foi de 0,098 U.mL-1 (1,695 U.g-1) para um pH 7,0. Finalmente, a partir de uma celulase produzida por fermentação em estado sólido do Trichoderma sp. e uma enzima comercial foi realizada a avaliação da influência dos parâmetros tempo de hidrólise, diluição da enzima, concentração de Tween 80 e razão sólido-líquido do bagaço de cana sobre a hidrólise do mesmo. As variáveis significativas foram, posteriormente, otimizadas por um delineamento composto central rotacional. A cepa Trichoderma sp. proveniente do bioma amazônico apresentou potencial na produção de celulases e o o bagaço de cana submetido ao tratamento alcalino oxidativo apresentou-se como um bom substrato para a produção enzimática.

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Alper, Seda Harsa Şebnem. « Production and isolation of fungal chitosan by submerged fermentation/ ». [s.l.] : [s.n.], 2003. http://library.iyte.edu.tr/tezler/master/biyoteknoloji/T000249.rar.

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Schlepütz, Tino [Verfasser]. « Submerged vinegar fermentation in small scale culture systems / Tino Schlepütz ». Aachen : Hochschulbibliothek der Rheinisch-Westfälischen Technischen Hochschule Aachen, 2013. http://d-nb.info/1043612688/34.

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Oliveira, Simone Lopes do RÃgo de. « Optimization of the production of cellulase Melanoporia sp. submerged fermentation ». Universidade Federal do CearÃ, 2014. http://www.teses.ufc.br/tde_busca/arquivo.php?codArquivo=13646.

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Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico
Cellulases are enzyme complex composed of endoglucanases, exoglucanases and β-glucosidases with several biotechnological applications. However, their production cost is a major obstacle for its industrial application. About 40% of the total cellulase production cost is related to the culture medium used for the microorganism growth. In this context, efficient processes for cellulolytic enzyme production are of technical and economical interest. Thus, the present study aimed to optimize the production of cellulases by Melanoporia sp. using coconut shell powder as substrate in submerged fermentation. The influence of pH and temperature on the enzyme activity was evaluated by univariate experimental design. Then, the composition of the culture medium was sequentially optimized through Plaket -Burman followed by Central Composite experimental designs. The fermentation under optimized conditions was subsequently conducted in bioreactor to evaluate the influen ce of pH control and aeration on enzyme production. The stability of the enzyme was evaluated for 6 and 8 months at 4 ÂC and - 20 ÂC, respectively. The ability of the enzyme to hydrolyze coconut shell powder was evaluated at 65 ÂC and 80 ÂC using the crude enzyme extract produced by Melanoporia sp. The enzyme activity was determined by the quantification of reducing sugars using DNS method at pH 5.5 and 80 ÂC (optimum conditions). The composition of the culture medium which provided the highest enzyme yield was: 5 g/L of coconut shell; 15 g/L lactose; 3% tween 80; 1 g/L of KH2PO4 and 0.05 g/L FeSO4; pH 6.5 at 30ÂC for 72 hours. For batch enzyme production, the cult ure medium using non-delignified substrate, with pH controlled at 6.5, without aeration resulted in an increase of 90% in enzyme activity compared to the fermentation in a rotatory shaker. Under these conditions, the maximal enzyme production was obtained after 24 hours of fermentation. The crude enzyme extract produced by Melanoporia sp. was able to hydrolyze cellulose (coconut shell powder) efficiently, presenting industrial potential for the degradation of lignocellulosic residues. Unlike most of the cellulases produced by Trichoderma species, the strain reported as one of the best producers, the microorganism was capable of producing cellulases efficiently without the need of substrate pretreatment. Another feature of this enzyme complex is its high stability in the crude broth at-20ÂC e 4 ÂC
Celulases sÃo um complexo enzimÃtico constituÃdo por endoglucanases, exoglucanases e β-glicosidases com diversas aplicaÃÃes biotecnolÃgicas. No entanto, o elevado custo de produÃÃo dessas enzimas Ã o principal obstÃculo para sua aplicaÃÃo industrial. Estima-se que cerca de 40% do custo total de produÃÃo de celulases esteja relacionado ao meio de cultura utilizado para o crescimento do micro-organismo. Nesse contexto, Ã de fundamental importÃncia o desenvolvimento de processos para a produÃÃo de enzimas do complexo celulolÃtico que se mostrem tÃcnico e economicamente viÃveis. Diante do exposto, o presente estudo teve como objetivo avaliar a produÃÃo de celulases produzidas por Melanoporia sp. utilizando o pÃ da casca de coco como substrato em fermentaÃÃo submersa. A influÃncia dos parÃmetros pH e temperatura na determinaÃÃo da atividade da enzima foi avaliada atravÃs de planejamento experimental univariado. Em seguida, a composiÃÃo do meio de cultura foi otimizada atravÃs dos planejamentos experimentais Plaket-Burman e Composto Central. A fermentaÃÃo em condiÃÃes otimizadas foi posteriormente conduzida em fermentador para avaliar a influÃncia do controle de pH e oxigÃnio na produÃÃo da enzima. A estabilidade da enzima foi avaliada por 6 e 8 meses nas temperaturas de 4 ÂC e -20 ÂC, respectivamente. A capacidade das enzimas em hidrolisar o pÃ da casca do coco foi avaliada nas temperaturas de 65 ÂC e 80 ÂC utilizando o extrato enzimÃtico bruto produzido por Melanoporia sp. A atividade da enzima foi determinada atravÃs da quantificaÃÃo de aÃÃcares redutores pelo mÃtodo de DNS. O pH e a temperatura de determinaÃÃo da atividade enzimÃtica foram pH 5,5 e 80 ÂC, respectivamente. A composiÃÃo do meio de cultura que proporcionou o maior rendimento de produÃÃo da enzima foi: 5 g/L de casca de coco; 15 g/L de lactose; 3% de tween 80; 1 g/L de KH2PO4 e 0,05 g/L de FeSO4; pH 6,5 a 30 ÂC em 72 horas. Para a produÃÃo da enzima em fermentador, o meio de cultura utilizando substrato nÃo deslignificado, com controle do pH em 6,5, sem aeraÃÃo proporcionou um aumento de 90% na atividade da enzima, comparado Ã fermentaÃÃo em shaker. Nessas condiÃÃes, a mÃxima produÃÃo da enzima foi obtida apÃs 24 horas de fermentaÃÃo. O extrato enzimÃtico bruto produzido por Melanoporia sp. exibiu capacidade de hidrolisar celulose presente na casca de coco com eficiÃncia, apresentando potencial industrial para a degradaÃÃo de resÃduos lignocelulÃsicos. Diferentemente da maior parte das celulases produzidas por espÃcies de Trichoderma, micro-organismo reportado como bom produtor de enzimas celulolÃticas, o micro-organismo utilizado neste trabalho Ã capaz de produzir celulases de forma eficiente, sem necessidade de prÃ-tratamento do substrato. Outra caracterÃstica diferencial desta enzima Ã sua elevada estabilidade nas temperaturas de -20 ÂC e 4 ÂC no caldo bruto.

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Taurhesia, Shelly. « Exopolysachharide production by submerged culture of the fungus Sclerotium glucanicum ». Thesis, University of Strathclyde, 1992. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.319472.

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Abdul, Manan Musaalbakri. « Design aspects of solid state fermentation ». Thesis, University of Manchester, 2014. https://www.research.manchester.ac.uk/portal/en/theses/design-aspects-of-solid-state-fermentation(d64ea506-85ee-424f-9bca-531488e3e3c7).html.

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Solid state fermentation (SSF) refers to the microbial fermentation, which takes place in the absence or near absence of free water, thus being close to the natural environment to which the selected microorganisms, especially fungi, are naturally adapted. The current status of SSF research globally was discussed in terms of articles publication. This was followed by discussion of the advantages of SSF and the reason for interest in SSF as a notable bioprocessing technology to be investigated and compared to submerged fermentation (SmF) for the production of various added-value products. SSF also proved to be a potential technology to treat solid waste produced from food and agricultural industry and to provide environmental benefits with solid waste treatment. A summary was made of the attempts at using modern SSF technology for future biorefineries for the production of chemicals. Many works were carried out in the Satake Centre for Grain Process Engineering (SCGPE), University of Manchester, to prove the strategy of using SSF for the production of hydrolysate rich in nutrients for sequel microbial fermentation with or without adding any commercial nutrients. The research findings presented in this thesis are based on a series of SSF experiments carried out on systems varying in complexity from simple petri dishes to our own design of bioreactor systems. They were conducted to assess a solution for biomass estimation, enzymes production, and successful mass and heat transfer. A proper technique for inoculum transfer prior to the start of the fermentation process was developed. In SSF, estimation of biomass presents difficulties as generally the fungal mycelium penetrates deep and remains attached with the solid substrate particles. Although many promising methods are available, the evaluation of microbial growth in SSF may sometimes become laborious, impractical and inaccurate. Essentially, this remains another critical issue for monitoring growth. In these studies, measurement of colour changes during SSF are presented as one of the potential techniques that can be used to describe growth, complementary to monitoring metabolic activity measurement, such as CER, OUR and heat evolution, which is directly related to growth. For the growth of Aspergillus awamori and Aspergillus oryzae on wheat bran, soybean hulls and rapeseed meal, it was confirmed that colour production was directly proportional to fungal growth. This colourimetric technique was also proved to be a feasible approach for fungal biomass estimation in SmF. This new approach is an important complementation to the existing techniques especially for basic studies. The key finding is that the colourimetric technique demonstrated and provided information of higher quality than that obtained by visual observation or spores counting. The effect of aeration arrangements on moisture content, oxygen (O2), mass and heat transfer during SSF was investigated. A. awamori and A. oryzae were cultivated on wheat bran in newly designed four tray solid state bioreactor (SSB) systems. The new tray SSB systems were: (1) single circular tray SSB, (2) multi-stacked circular tray SSB, (3) Single rectangular tray SSB and (4) multi-square tray SSB. The purpose was to study the effect, on heat and water transfer, of operating variables, fermentation on the perforated base tray and internal moist air circulation under natural and forced aeration. Temperature, O2 and carbon dioxide were measured continuously on-line. Enzyme activity, moisture content and biomass were also measured. The results suggest that the air arrangements examined have a remarkable effect on the quantity of biomass produced using measurement of spores and enzymes production. The strategy presented in these studies allowed quantitative evaluation of the effect of forced internal moist air circulation on the removal of metabolic heat. With the proposed strategy, it was possible to maintain the bed temperatures at the optimum level for growth. However, the effect on moisture content was very different for the two fungi. It was found that the ability of A. oryzae to retain moisture was much higher than that of A. awamori. This is possibly due to the higher levels of chitin in A. oryzae. Greater spores and enzymes (glucoamylase, xylanase and cellulase) production was observed for A. awamori in multi-stacked circular tray and multi-square tray SSB systems compared to the conventional petri dishes and the other two systems. A. oryzae was excellent in producing protease in the same bioreactors. A direct technique of establishing a correlation between fungal growth and CER, OUR, heat evolved was proven successful in this work. The information obtained from CER and OUR led to the estimation of respiratory quotient (RQ). RQ describes the state of the fungal population in the tray SSB and gives an indication of fungal metabolic behaviour. RQ values < 1 were obtained from 38 experiments using four tray SSB systems for the two fungi. A kinetic model based on CO2 evolution instead of biomass concentration was examined in order to simplify the required experiments for kinetic model development. A Gompertz model was used to fit the integrated CO2 data and predict the quantity of CO2 evolution in all experiments. A correlation was found between the heat evolution and CER. The performances of tray SSB systems can be improved by constructing them as multi-trays. The multi-tray systems improved the mass transfer considerably compared with single tray systems. In addition, the multi-tray systems allowed precise measurement of the gradients of CO2, enzymes, spores and fungal biomass. In addition, the air arrangements using moistened air were successful in maintaining moisture content, adequate O2 supply and control of temperature, and hence, increased the productivity of both fungi. Overall A. awamori and A. oryzae have their own ability and performance to degrade and utilise the complex compositions contained in the solid substrate and fermentation conditions may lead to possible comparisons. In addition, multi-stacked circular tray and multi-square tray SSB systems demonstrated an excellent system for further investigations of mass transfer and possibly for large scale operation, though considerable optimisation work remains to be done, for example the height/diameter ratio and total number of trays should be optimised.

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Silva, Ellen Mae. « A gas-solid spouted bed bioreactor for solid state fermentation / ». The Ohio State University, 1997. http://rave.ohiolink.edu/etdc/view?acc_num=osu1487945320759412.

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Papagianni, Maria. « Morphology and citric acid production of 'Aspergillus niger' in submerged fermentation ». Thesis, University of Strathclyde, 1995. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.407845.

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Fazenda, Mariana L. « Submerged culture fermentation of the Basidiomycete fungus Ganoderma lucidum for biomass formation ». Thesis, University of Strathclyde, 2009. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.501792.

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The aim of the present study was to investigate a range of bioprocess strategies aimed at the achievement of maximum biomass yield in submerged cultivation of the Basidiomycete, Ganoderma lucidum. Although there had been previous studies into cultivation of G, lucidum, these had been almost exclusively centred round maximisation of the medically interesting polysaccharide, EPS. The present work is focused on the development of fermentation strategies to achieve this aim, which was a central interest of the sponsor. Additionally, to investigate the process physiology of these complex cultures to help improve the relatively poor, understanding of the bioprocessing of this Basidiomycete fungus and to understand the influence of process variables during submerged cultivation of G. lucidum on growth, polysaccharide production and substrate consumption.

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Robinson, Tim. « Solid state fermentation of dye-adsorbed agricultural residues ». Thesis, University of Ulster, 2002. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.274061.

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Lyons, Mark Pearse. « Optimisation of solid-state fermentation for enzyme production ». Thesis, Heriot-Watt University, 2007. http://hdl.handle.net/10399/2030.

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Wan, Mohtar Wan Abd A. Q. I. « Production and bioactivity of Ganoderma lucidum BCCM 31549 exopolysaccharide using submerged liquid fermentation ». Thesis, University of Strathclyde, 2016. http://oleg.lib.strath.ac.uk:80/R/?func=dbin-jump-full&object_id=26556.

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The RBF strategy has successfully produced fungal mycelial biomass and EPS in a very strictly regulated manner at high productivity rates compared to batch fermentation. The problematic lag phase and seed culture preparation were reduced in length; harvesting volume doubled, yield of product increased, and medium consumption was reduced in an RBF relative to batch. 80% broth replacement volume and transition phase were optimised. Dispersed mycelial filaments with ovoid-shaped pellets are the typical morphological characteristics associated with EPS production. N-limiting medium in an unbaffled 2.5-L bioreactor stimulated EPS formation during RBF compared to in baffled condition. The current study has managed to alter the molecule's hydrophobicity thus making it water-soluble as proved by compositional analysis and spectroscopy. The sulphated derivative of native glucan was identified as (1, 3)-(Sb(B-D-glucan. Sulphation was an effective approach to improve antibacterial, antifungal, antiproliferative and immunomodulatory (NO stimulation) activity of the sulphated (1,3)-(Sb(B-D-glucan or GS. GS maybe safe in in vitro trials due to its demonstrated lack of toxicity towards a normal human prostate cell line (PN2TA). GS also showed antimicrobial-antifungal-immunomodulatory activities derived from a single compound. Fungal cells tended to grow well in the porous structure of PUF cubes and the RBF using immobilised fungal cells was an efficient method for production of (Sb(B-glucan with a high yield. This study could be beneficial for other medicinal mushroom fermentation.

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Santos, Leandro Freire dos. « The pharmacological potential of the macromycetes (Cordyceps sinensis and Pleurotus ostreatus) cultived by submerged fermentation ». reponame:Repositório Institucional da UFPR, 2013. http://hdl.handle.net/1884/34767.

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Resumo: Este trabalho objetivou a avaliação das potencialidades farmacológicas dos macromicetos Cordyceps sinensis e Pleurotus ostreatus, obtidos por fermentação submersa, quanto à ação hipolipidêmica, antiaterosclerótica, e protetor da disfunção puberal em modelos hiperlipídicos de ratos machos Wistar, bem como a ação antitumoral do extrato aquoso de Cordyceps sinensis contra células do neuroblastoma humano. Para a instauração do modelo hiperlipídico, uma dieta comercial basal foi aditivada com gordura vegetal hidrogenada (6% p/p) e gordura de porco (14% p/p). Para as determinações bioquímicas, foram realizadas as dosagens do colesterol plasmático, lipoproteína de baixa densidade (LDL), atividade da aspartato transaminase, uréia e testosterona. A atividade antiaterosclerótica foi inferida pela determinação da ativação macrofágica: capacidade fagocítica, volume lisossomal e produção de íons (ânion superóxido, peróxido de hidrogênio e óxido nítrico). Os resultados indicaram que o Cordyceps sinensis diminuiu significativamente os parâmetros bioquímicos colesterol plasmático (37%), triglicerídeos (35%), LDL (40%), bem como normalizou os níveis de testosterona e restaurou a função hepática. Similarmente, o Pleurotus ostreatus também melhorou o perfil lipídico, bem como diminuiu a ativação macrofágica em aproximadamente 70%. Em adição, a ação antitumoral do Cordyceps sinensis contra células do neuroblastoma humano foi caracterizada por uma taxa de inibição de 20%. Sendo assim, estes cogumelos possivelmente não substituirão os tradicionais medicamentos já existentes para o tratamento destas patologias, mas poderão de forma valiosa complementá-los.

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Oliveira, Juliana de. « Poly(Lactic acid) production by conventional and microwave polymerization of lactic acid produced in submerged fermentation ». reponame:Repositório Institucional da UFPR, 2016. http://hdl.handle.net/1884/46421.

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Orientador : PhD. Luciana Porto de Souza Vandenberghe
Coorientadores : PhD. Carlos Ricardo Soccol e PhD. Sônia Faria Zawadzki
Tese (doutorado) - Universidade Federal do Paraná, Setor de Tecnologia, Programa de Pós-Graduação em Engenharia de Bioprocessos e Biotecnologia. Defesa: Curitiba, 09/06/2016
Inclui referências : f. 115-128
Área de concentração: Agroindústria e biocombustíveis
Resumo: Poli(ácido lático), poliéster, é um polímero biodegravável aplicado em produtos como embalagens, têxteis, médicos e farmacêuticos. Pode ser obtido a partir do monômero ácido lático (AL) por meio da reação de policondensação direta e pela polimerização por abertura de anel do lactídeo. O AL é um ácido orgânico que apresenta diversas aplicações principalmente na indústria alimentícia, assim como na indústria farmacêutica, química e de polímeros. A produção do AL por fermentação oferece vantagens tais como a produção do isômero opticamente puro. As necessidades nutricionais da bactéria aumentam o custo de produção do AL, portanto substratos alternativos tem sido estudados por apresentarem uma alternativa econômica para este processo. O objetivo deste trabalho foi a produção de ácido lático por Lactobacillus pentosus em fermentação submersa utilizando subproduto do processamento da batata e caldo de cana como substratos para a obtenção de poli(ácido lático). Estes sub-produtos porque possuem alta concentração de fonte de carbono e volumes significativos são gerados anualmente, o que justifica sua a re-utilização e valorização. O sub-produto do processamento da batata foi submetido a hidrólise ácida com o objetivo de converter o amido em glucose. A produção de AL foi otimizada utilizando etapas de planejamento experimental estatístico envolvendo a seleção de bactérias do gênero Lactobacillus, definição da composição do meio de cultivo e estudos de cinética em frascos de Erlenmeyer e biorreator do tipo tanque agitado. A produção de AL chegou a 150 g/L utilizando sub-produto do processamento da batata e 225 g/L utilizando caldo de cana em 96 horas de fermentação. O uso da célula inteira de levedura de panificação como fonte de nitrogênio e a condição de fermentação não estéril demostraram ser boas alternativas para um processo industrial de produção de AL. O processo de separação e recuperação do AL do caldo fermentado foi desenvolvido para obtenção da molécula purificada e estudos de polimerização com o monômero obtido. O processo desenvolvido consistiu no aquecimento do caldo fermentado seguido pela etapa de centrifugação. A etapa de clarificação foi realizada utilizando carvão ativado em pó seguida pela precipitação a baixa temperatura e acidificação do lactato de cálcio para conversão em ácido lático. O processo foi efetivo para remoção de contaminantes que estavam presentes no caldo fermentado. A concentração final de AL em solução aquosa foi de 416 g/L com um rendimento de 51%. Os estudos de polimerização foram desenvolvidos utilizando a técnica de policondensação direta do AL, por meio de dois diferentes sistemas de aquecimento, convencional e micro-ondas. Um polímero com massa molar de 6330 g/mol e 61% de rendimento foi obtido a partir de um AL comercial e utilizando o AL obtido por fermentação resultou em um polímero com massa molar de 2370 g/mol. O processo de aquecimento por micro-ondas proporcionou um maior rendimento, 79% e 76% para o AL comercial e obtido por fermentação, respectivamente. Porém, foi obtida menor massa molar que o processo convencional, 2070 para o AL comercial e 1450 para o AL obtido por fermentação. As propriedades físico-químicas do poli(ácido lático) demonstraram aplicação em encapsulamento de compostos bioativos e engenharia de tecido. As perspectivas de sequência de estudos são a aplicação em encapsulamento de moléculas, modificações do polímeros e desenvolvimento de compósitos. PALAVRAS CHAVE: Poli(ácido lático), sub-produto do processamento da batata, caldo de cana, policondensação
Abstract: Poly (lactic acid) (PLA) is a polyester, which has a predominant role as biodegradable plastic, that is applied in packaging, textile, medical and pharmaceutical products. It can be obtained from lactic acid by direct polycondensation and by ring-opening polymerization (ROP) of lactide. Lactic acid (LA) is an organic acid that presents diverse applications mostly in food industry, as well as in pharmaceutical, chemical industries and polymers. The production of LA by fermentation offers the advantage of producing optically high pure LA. Nutritional requirements of bacteria increase the cost of LA production so alternatives substrates have been studied to bring an economical alternative for this process. The aim of this work was the production of LA by Lactobacillus pentosus in submerged fermentation using potato processing waste and sugarcane juice as substrate in order to obtain poly(lactic acid). The fermentation process was developed using potato processing waste and sugarcane juice because of their high carbon source concentration. Important volumes of both sub-products were generated, which is another reason for their re-use and valorization. Potato processing waste was submitted to hydrolysis in order to convert starch to glucose. LA production by fermentation was optimized using, statistical experimental design approach steps of optimization involved the screening of bacteria of the genus Lactobacillus and definition of medium composition kinetics studies in Erlenmeyer flask and stirred tank reactor were also carried out. LA production reached 150 g/l using potato processing waste, it was and 225 g/l with sugar cane juice after 96 hours of fermentation. The use of baker's yeast as a source of nitrogen and nonsterile conditions demonstrated good alternatives for an industrial production process of LA. The separation and recovery process of LA from fermented broth was developed to obtain a purified molecule for further polymerization studies. The developed process consisted in heating the fermented broth, then a centrifugation step was conducted for removal of the cells and suspended solids. A clarification step was included with powered activated carbon with further precipitation at low temperature and acidification of calcium lactate to convert to LA. The process was effective for removal of contaminants that were present in the fermentation medium. Final concentration of LA in aqueous solution reached 416 g/l and a yield of 51%. Polymerization studies were then carried out using direct polycondensation of LA, that were carried out with two different heating systems, conventional and microwave heating. A polymer with 6330 g/mol of molecular weight and 61% of yield was obtained from commercial LA and using fermented LA resulted in 2370 g/mol. Microwave heating process provided a higher yield, 79% and 76% for commercial and fermented LA, respectively. Nevertheless, the molecular weight was lower than conventional process, 2070 for commercial LA and 1450 for fermented LA. Physicochemical properties of PLA demonstrated application in encapsulation of bioactive compounds and tissue engineering. Perspectives of sequence of the studies: application on encapsulation of molecules, modifications of polymer and development of composites. KEYWORDS: Poly(lactic acid); potato processing waste; sugarcane juice; polycondensation

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DIVATE, RUPESH-DATTATRAY, et 狄韋德. « Bioactivities of Xylaria Nigripes From Solid-state and Submerged Fermentation ». Thesis, 2016. http://ndltd.ncl.edu.tw/handle/43064345161585249617.

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博士
靜宜大學
食品營養學系
105
Xylaria nigripes (XN) is a medicinal fungus that was used traditionally as a diuretic, nerve tonic and for treating insomnia and trauma. In this dissertation, solid state fermentation (SSF) and submerged fermentation were attempted to produce XN with multi-bioactivities. This dissertation included four major objectives. The first and second objectives of this dissertation were to produce XN under SSF conditions with agriculture byproducts, including wheat bran (WB) and soy meal (SM), and edible whole grains as fermentation substances, respectively. The third objective was to produce XN under submerged fermentation and elucidated possible mechanism of neuroprotection effects of XN mycelium extract. The fourth objective was to evaluate anti-inflammatory and immunomodulatory activities of submerged fermented XN mycelium. When WB was used as the sole fermentation substrate under SSF conditions, the 70 % ethanol extract of XN-fermented substances obtained the highest antioxidant activities and anti-inflammation activities. When fermentation substrates contained equal amounts of WB and SM, the 70% ethanol extract of XN-fermented substances showed the highest effects against H2O2-induced damage in neuronal cells (PC12 cells). Both XN and residues of fermentation substrates may have contributed to the biological activities of XN-fermented substances. When whole grains, including brown rice, red beans, mung beans or soy beans, were used as fermentation substrate, the synergistic effects of bioactivities from XN mycelium and whole grains were obtained. Especially, XN-fermented red beans exhibited noteworthy activities in antioxidant, anti-inflammation and neuroprotection. When XN mycelium was produced by submerged fermentation, both XN water extract and 70% ethanol extract effectively protected PC12 cells against H2O2 - induced cell damage by inhibiting the release of lactate dehydrogenase, decreasing DNA damage, restoring mitochondrial membrane potential and arresting abnormal apoptosis through up-regulation of Bcl-2 and down-regulation of Bax and caspase-3. XN mycelium from submerged fermentation showed anti-inflammation activities by effectively reducing production of nitric oxide (NO), tissue necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in RAW264.7 cells and inhibiting cyclooxygenase-2 (COX-2) enzyme. Animal study using BALB/c mice demonstrated that the immunomodulatory activities of XN mycelium were achieved by enhancing splenocytes proliferation, cytokine inductions, and the cytotoxicity of splenic natural killer cells.

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KUO, CHUNG-YI, et 郭仲鎰. « Evaluate antioxidant capacity of solid and Submerged fermentation product from Inonotus obliquus ». Thesis, 2016. http://ndltd.ncl.edu.tw/handle/h3h9pd.

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碩士
南臺科技大學
生物科技系
104
The medicinal Inonotus obliquus (I. obliquus) is a white rot fungus, and it inhabits on birch trees in the birch forests of Russia, Eastern Europe, China, Japan. The sclerotia have been used as a folk medicine since the 16th century, and have exhibited various biological activities, including anti-tumor, anti-viral, anti-oxidation. In this study, four different fractions, hot water extraction (HWE), ethanol extraction (EE), hot water extraction followed by ethanol extraction (HW-EE) and gradient method of ethanol extraction, were evaluated for extraction of the total phenolic content from the solid state fermentation of I. obliquus. The change of the total phenolic content and the antioxidant capacity at the cultural period of the solid state and submerged fermentation were also investigated, respectively. The results showed that the total phenolic content increased with the cultural time of I. obliquus by solid-state fermentation using brown rice as culture medium. The highest of total phenolic content, 69.4 mg GAE/g, was obtained by gradient method of ethanol extraction at 120 days after inoculation. The EC50 concentration of ABTS, reducing power and OH radical scavenging activity were 3.84, 0.75and 3.63 mg/ml from the solid-state fermentation, respectively. The EC50 concentrations of ABTS, reducing power and OH radical scavenging activity was10.19, 3.2 and 5.16 mg/ml from the submerged fermentation is using maltose and himedia peptone, respectively. It showed the antioxidant capacities of the solid state and submerged fermentation of I. obliquus correlated with the total phenolic contents of extracts. The product of solid-state and submerged fermentation of I. obliquus has the potential to be the functional foods.

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Huang, Jia-You, et 黃嘉佑. « Solid-state and Submerged Cultivation and Fermentation Fluid Antioxidant Activity of Grifola frondosa ». Thesis, 2011. http://ndltd.ncl.edu.tw/handle/90570908753383082507.

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碩士
亞洲大學
生物科技學系碩士班
99
Grifola frondosa is one kind of medical and edible fungus, and it has been found lots of functions such as: anti-tumor, strengthen the immune system, lowering blood glucose, blood pressure and cholesterol, and weight-loss and other anti-diabetic effects. The international biological and medical community has been highly paid attention to G. frondosa recently. Due to the traditional cultivation of G. frondosa from the inoculation to produce fruiting bodies which takes a long time, the fermentation technology to produce mycelium and polysaccharides can not only shorten the training time, but also increase the quality and the yield stably. For the above reasons, the three purposes of this study are as follows. (1) To investigate the best carbon and nitrogen source of the five strains of G. frondosa by solid-state culture. (2) To study the production effects of G. frondosa mycelium and polysaccharide by different rpm and different initial pH value of submerged culture. (3) To analyze total polyphenol and flavonoid content and fermentation DPPH radical scavenging ability of the fermentation broth of G. frondosa. The first part of experimental results showed that best carbon source and the nitrogen source for the mycelium growth of five strains G. frondosa after 20 days cultivation were as follows: the 4% fructose as the carbon source and the 2% corn steep powder as the nitrogen source caused the mycelium growth of T1 strain reached 40.17 mm; the 3% glucose and the 1.5% corn steep powder let the mycelium growth of T2 strain reached 42 mm; the 4% fructose and the 1.5% yeast extract urged the mycelium growth of BT strain reached 42.83 mm; the 4% glucose and the 1.5% corn steep powder caused the mycelium growth of D-4 strain reached 41 mm; the 4% glucose and the 1.5% corn steep powder caused the mycelium growth of 5-T strain reached 42 mm. The second part of the study was designed to investigate the different rpm and initial pH value on the impact of submerged culture of G. frondosa. According to the experimental results, All strains (T1, T2, BT, D-4 ,5-T) in rpm 150 under cultivation, the highest yield of polysaccharide were 2.82±0.56, 2.11±0.13, 3.24± 0.71, 2.85±0.39, 2.06±1.23 g/L, but the mycelium and the final pH of no significant impact of the change. Part of the initial pH value, T1, T2 and BT strains of G. frondosa, produced respectively the highest mycelial yields as 1.83 g/L, 4.18 g/L and 3.16 g/L when the initial pH value was 5.5. In addition, D- 4 and 5-T strains, produced the highest mycelial yields when the initial pH values were 6.5 (3.83 g/L) and 4.5 (3.14 g/L). Furthermore, the maximum polysaccharide production of G. frondosa T1, T2, BT and 5-T strains were 8.39, 7.57, 5.56 and 5.07 g/L when the initial pH was 4.5. The D-4 strain presented the highest yield of polysaccharides (7.07 g/L) when the initial pH was 6.5. The third part of the antioxidant ability experiments indicated that the fermentation broth of T2 strain contained the highest total polyphenols (29.75 mg/g) and flavonoids (1.20 mg/g) above other strains under the cultivation conditions of 3% fructose and 1.5% corn steep powder at initial pH of 6.5. Moreover, T1, T2, BT, D-4, and 5-T strains presented the highest ability of scavenging DPPH radicals were 53.70%, 46.88%, 57.81%, 48.68% and 55.54%. Especially, G. frondosa 5-T strain presented the best results of EC50 within five strains was 23.67 mg/mL under the fermentation conditions of 4% glucose, 1.5% corn steep powder and initial pH of 4.5. Summarize the above results, higher concentrations (4~3%) of glucose and fructose as the carbon resources were suitable for the mycelia growth of all five strains of G. frondosa. Lower concentrations of corn steep powder (2~1.5%) as nitrogen resources were suitable for all strains of G. frondosa. The initial pH values (the case of weak acid) of G. frondosa fermentation were not significantly affected the mycelium and polysaccharide productions, but they had a little impact on the antioxidant capacity of G. frondosa. The relationship between different carbon-nitrogen resources and antioxidant abilities; or between different surfactants such as vegetable oil and polysaccharide production of G. frondosa can be further investigated. Based on the achievement of this study, the related information can be provided to the farmers for the solid-state of cultivation and the submerged culture in five different strains of G. frondosa. In the future, the fermentation fluid of G. frondosa can be applied for the development of functional health products to increases the industrial utilities and to promote the additional values of G. frondosa.

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Huang, Po-Hsin, et 黃伯欣. « Fructosyltransferase production by Aspergillus japonicus TU-26 in submerged and solid-state fermentation ». Thesis, 2008. http://ndltd.ncl.edu.tw/handle/83081296441044036298.

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碩士
大同大學
生物工程學系(所)
96
The production of fructosyltransferase (FTase) by Aspergillus japonicus TU-26 in submerged and solid-state fermentation was investigated. Submerged fermentation was carried out in a 5-L fermentor at 30 ℃, 1vvm, 500 rpm and pH 6, with a working volume of 2-3 L. The culture medium consisted of 1% yeast extract, 0.5% KH2PO4, 0.05% MgSO4•7H2O and sucrose of varied concentration. A maximum FTase activity of 359 U/mL was obtained at hour 24 when batch fermentation with 200 g/L of sucrose was performed. A maximum enzyme activity of 534 U/mL was achieved at hour 40 when 250 g/L of sucrose was used. As the fermentation was progressed, sucrose was converted to difructoside, fructo-oligosaccharides (FOS, including: trifructoside, tetrafructoside, 1-kestose, nystose and fructosyl nystose) , glucose and fructose. Eventually, all of the di- and oligo-saccharides were hydrolyzed into glucose and fructose. During the fermentation, FTase was continuously produced until both sucrose and FOS disappeared. In fed-batch fermentation, continuous sucrose-feeding to reach a final concentration of 200 g/L or 250 g/L in the first 24 hours was carried out. Maximum FTase activities of 463 U/mL and 615 U/mL at hour 36 and 52, respectively, were achieved. In solid-state fermentation, an omission test revealed that wheat bran, rice hull, yeast extract, sucrose, KH2PO4 and MgSO4•7H2O played important roles for the production of FTase by A. japonicus. During the solid-state fermentation, FTase production peaked at day 5. More wheat bran added resulted in more FTase produced. Solid-state fermentation and submerge fermentation were compared in FTase productivity based on that the cost of the solid culture medium in a 500-mL flask was equivalent to 100 mL of submerged culture medium. A flask of solid-state culture produced 42930 U of FTase, whereas 100 mL of submerged culture in the fed-batch fermentation with continuous sucrose-feeding gave 46310 U of FTase.

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Su, Cyuan-Sheng, et 蘇全生. « Study on the production and stability of nattokinase by solid state fermentation and submerged culture ». Thesis, 2005. http://ndltd.ncl.edu.tw/handle/56981589558555874234.

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碩士
國立嘉義大學
食品科學系碩士班
93
In this study, 65 Bacillus spp. had been isolated from Bacillus stock of Food Industry Research And Development Institute, soil, water, rice straw, Japanese commercial natto and other natto products. First, plasminogen-free fibrin plate method was used to pick out the high nattokinase production strains from 65 Bacillus spp.. Second, the optimal condition for cultivation of solid state fermentation and submerged culture for Bacillus strains was estabalished to increase nattokinase production. Than, crude enzyme of nattokinase was purified and identified the different recoveries identified and stability of nattokinase activity from each strains examined. The results indicated that strain number C13 which was isolated from Japanese commercial natto produced the highest nattokinase activity, 328.2 FU/g at 37℃ from 48 hours of culture. Explore sure of C13 strain to ultraviolet rays induced mutation and 46 mutant strains were. Mutant CM13 could produced the highest nattokinase activity 421.2 FU/g. Soybean was the optimal medium for CM13 mutant solid state fermentation. The optimal cultivation condition for inoculum size, relative humidity, temperature and time were 1.0 mL (107~108 cell/mL), 90%, 37℃ and 32 hours, respectively The highest activity of nattokinase achieved was 469.2 FU/g. Using submerged culture, CM13 mutant gave the activity of nattokinase approximately 70.3 FU/mL when incubated at 37℃ for 32 hours. At 60℃ or 37℃ the nattokinase from solid substrate fermentation was more stable than from submerged culture. The recovery of nattokinase purified from solid substrate fermentation reached 63.0%, much higher than 39.8% from submerged culture. This is was because of the γ-poly-glutamic acid (γ-PGA) which produced from natto during the process of cultivation. γ-PGA increased the stability of nattokinase, and the recovery was also enhanced after purification. Adding 0.1% γ-PGA to in crude enzyme from submerged fermentation increased the stability and recovery of nattokinase sigificantly.

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SU, GUO-CHIH, et 蘇國智. « Effects of solid and liquid submerged fermentation on the formation of β-carotene in Rhodotorula glutinis ». Thesis, 2016. http://ndltd.ncl.edu.tw/handle/fsjxsd.

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碩士
東海大學
化學工程與材料工程學系
104
Rhodotorula glutinis (BCRC 22360) is an oleaginous yeast that can accumulate high content of total lipid. In addition for total lipids production, R. glutinis was well known as having high content of β-carotene, which is a natural antioxidants to protect cells from being damaged by free radical through the interruption of lipid peroxidation chain reaction. Even though, the high cost of commercializing biosiesel production has been a thorny issue by using the microbial oils as the feedstock. In this study, the effects of liquid submerged fermentation on the cell growth and β-carotene production was investigated by using crude glycerol as carbon source. The corn is used as the substrate to reduce the cost for the extraction and the purification process. The effects of medium ingredients and environmental cultivation conditions on the growth of cell and the content of β-carotene in the shaker flask experiments.It was found that under the condition of 65 % content of water, the particle sizes which less than 0.85 mm, and cultivated for 96 hours can obtain the best consequence that the amount of fermentates of β-carotene reach 2.45 mg/kg. By dicing the substrate after sterilizing can enhance β-carotene from 2.45 to 2.96 mg/kg fermentates significantly. By adding 20 g/L of surcrose and 5 g/L of ammonium sulfate can enhance β-carotene from 2.96 to 3.67 mg/kg fermentates significantly. The effects on the growth of cell and β-carotene were evaluated by adding different volume of subsrtate in a 2 L rotary fermenter, powdering during the period of cultivating and methods of ventilation. While adding 200 g substrates in the 2 L rotary fermenter, the fermentates of β-carotene we obtained was 4.14 mg/kg and it fitted the economic benefits. The content of β-carotene was increased significantly from 4.14 to 6.389 mg/kg with the saturated gas pass into periodicity. The effects of C/N ratio and adding inducer in the medium on the growth of cell and β-carotene in the shaker flask by cultivating in the liquid fermentation were evaluated. The result demonstrated that a monoterpene ethanol solution did not increase the content of β-carotene significantly. Additionly, the growth of the cell was suppressed. However, it was found that adding 2.5 % palm oil can promote the growth of cell and β-carotene, which could obtain 11.5 g/L of biomass and 0.33 mg/g of β-carotene. In C/N ratio experiment, it seems that the increase of C/N ratio can increase the cells growth and β-carotene content. It can get 8.2 g/L of biomass and 0.26 mg/g of β-carotene by using 60 g/L crude glycerol and 1 g/L ammonium sulfate. The experiment of adding 2.5 % palm oil was discussed that scaled up by using the 5 L stirred tank reactor and 5 L airlift. It was found that adding palm oil can promote the growth of cell significantly, which could obtaine 29.8 g/L of biomass and 0.35 mg/g of β-carotene by using 5 L stirred tank reactor. While getting 24.5 g/L of biomass and 0.29 mg/g of β-carotene by using 5 L airlift. Comparing the two reactors, 5 L stirred tank reactor was better than another one. The experiment of changing C/N ratio was discussed that scaled up by using the 50 L airlift. The fed batch operation with the disposable feeding could enhance the lipid content from 41 to 60 %, but not promote for synthesizing the β-carotene, which could obtain 0.2 mg/g of β-carotene.

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38

Chen, Hang-Ming, et 陳漢銘. « The research of using the distiller'' grain for the fruit body culture of Ganoderma lucidum in solid state fermentation and thin stillage for the mycelial culture in submerged culture ». Thesis, 1998. http://ndltd.ncl.edu.tw/handle/44403414590720410467.

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碩士
東海大學
化學工程學系研究所
86
The main objective of this research is to investigate the feasibility of using the stillage grain for the fruit body culture of Ganoderma lucidum in solid state fermentation and thin stillage for the mycelial culture in submerged culture. The solid state fermentations were carried out in test tubes or propylene bags. The additions of NH4H2PO4 and CaCO3 were demon-strated to be in favor of the mycelial growth. The mixtures of wood chip (80%) and stillage grain (20%) at the moisture level of 60% were deter-mined to be optimal for the fruiting body culture in propylene bags. The compositions of fruiting body were also analyzed and compared with the control. Liquid cultures were performed in shaken flasks or fermenters. Different operating modes, including repeated batch and continuity, have been tested. In a repeated-batch culture using a 5-liter fermenter the concentration of polysaccharide reached 18.26mg/dL and the yields were estimated to be around 30-50mg/g glucose. In a continuous operation using a 12-liter fermenter the polysaccharide concentration and the yield were 13.5 mg/dL and 22 mg/g glucose at dilution rate (0.005hr-1). Con-tinuous production of polysaccharide was proved to be feasible.-1 -aThe research of using the distiller'' grain for the fruit body culture of Ganoderma lucidum in solid state fermentation and thin stillage for the mycelial culture in submerged culture

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39

Rühl, Martin [Verfasser]. « Laccases and other ligninolytic enzymes of the basidiomycetes Coprinopsis cinerea and Pleurotus ostreatus : submerged and solid state fermentation, morphological studies of liquid cultures and characterisation of new laccases / submitted by Martin Rühl ». 2009. http://d-nb.info/1002426499/34.

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40

Meysing, Daniel. « Investigations of Biomass Pretreatment and Submerged Fixed-bed Fermentation ». Thesis, 2011. http://hdl.handle.net/1969.1/ETD-TAMU-2011-12-10295.

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To improve the MixAlco process and biomass pretreatment, five studies were conducted. Three studies related to fermentation, whereas the other two investigated the effectiveness of shock tube pretreatment (STP) coupled with oxidative lime pretreatment (OLP). In the first study, the constant-selectivity assumption used in the continuum particle distribution model (CPDM) was determined to be invalid. During a 32-day batch fermentation, selectivity increased from 0.10 to 0.40 g acid/g non-acid volatile solid (NAVS) digested. Future revisions to CPDM should incorporate a non-constant selectivity term. In the second study, a revised procedure was developed to provide a more accurate determination of moisture content. Conventional drying at 105 degrees C allowed product acids to vaporize with water, which introduced errors. Using the revised procedure, calcium hydroxide or sodium hydroxide was added to samples at a concentration of 0.01 g base/g sample, which retained acids in the sample. The mass of additional retained material closely matched that of the additional retained acid. Three related studies involving biomass pretreatment were performed. In the first, recommended parameters for pretreating sugarcane bagasse with OLP and STP were determined. Recommended OLP parameters were 130 degrees C, 6.9-bar O2, and 2-h duration. The effects of solids concentration, liquid fill volume, particle size, type of shotgun shell, number of shocks, and pretreatment order were investigated. Liquid fill volume, particle size, type of shotgun shell, and pretreatment order were significant variables, whereas solids concentration and number of shocks were not. Recommended OLP parameters were used as a basis for an additional experiment. To simulate industrial-scale pile fermentation, fixed-bed batch fermentation of OLP + STP sugarcane bagasse was performed in 1-L PVC fermentors. Rubber mulch was used as a structural support material to prevent filter plugging, which had been reported in previous work. After 42 d, acid concentration reached 8 g/L with yield approximately 0.1 g acid/g NAVS fed. Poor fermentation performance was caused by short solid-liquid contact time and poor pH control. A third biomass pretreatment experiment investigated the potential of pretreated corn stover as a potential ruminant feed. Five samples (raw, OLP, STP, OLP + STP, and STP + OLP) were analyzed for composition and in vitro digestibility. STP followed by OLP increased neutral detergent fiber (NDF) digestibility from 49.3 to 79.0 g NDF digested/100 g NDF fed. On an organic matter basis, STP + OLP corn stover plus water-soluble extractives had a total digestible nutrients (TDN) of 74.9, nearly reaching corn grain at 88.1.

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Chiu, Chia-Hung, et 邱嘉鴻. « CoQ10 production by Rhodobacter sphaeroides in the submerged fermentation ». Thesis, 2007. http://ndltd.ncl.edu.tw/handle/42919512935777354392.

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碩士
東海大學
化學工程學系
95
CoQ10 is an intermediate component on the respiratory chain and also regarded as a kind of natural antioxidants. It is suggested as a health-supplemental material for many kinds of disease. The main objective of this research is using purple non-sulfur bacteria Rhodobacter sphaeroides to develop a submerged fermentation process for CoQ10 production. Firstly, the effects of addition amino acid and screening strategies in flask test were discussed. Then we use the tranditional 5 L agitation fermenter to find the optimize dissolve oxygen concentration and the test the possibility of CoQ10 production in the airlift fermenter. . The results of amino acid adding experiments show 1g/L Alanine can increase as high as 20 % of CoQ10 concentration and sodium azide added at 45 mg/l can be used a good screeing chemicals for finding high CoQ10 production strain. The anaerobic-light culture had a highest CoQ10 content 21.35 mg CoQ10/g DCW, but the maximum CoQ10 concentration of 23.73 mg /L was observed in the micoaerobic environment. The similar CoQ10 concentration was obtained from the results of airlift fermenter. The CoQ10 concentration 35.41 mg/l obtained in the airlift batch fermented with molasses as the carbon source. The final production of CoQ10 concentration reached 49.37 mg/L using feed strategy.

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42

Kao, Pao-Min, et 高博敏. « Production of Chitinase from Verticillium lecanii Using Submerged Fermentation ». Thesis, 2002. http://ndltd.ncl.edu.tw/handle/53565617178065744501.

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碩士
朝陽科技大學
應用化學系碩士班
90
Verticillium lecanii is an entomopathgenic fungus, which is also a major chitinase production organism. The main purpose of this research was to improve the productivity of chitinase from V. lecanii by submerged fermentation. The optimized culture medium obtained from response surface methodology (RSM) at the shaker-flask experiments was: 4.52% (w/v) maltose, 1.79% marine peptone extract, 0.41% shrimp powder and 0.3% isolated soy protein. The activity of chitinase was 18.3 mU/mL in such culture medium by controlling culture volume at 200 mL, shaker speed at 150 rpm, temperature at 24°C. Based on the result, we scaled up the operation to 5-L stirred-tank bioreactor and 30-L airlift bioreactor, and designed a series of experiments that changed different agitation rate, pH, and aeration rate etc. The highest activity of chitinase was observed as 16.9 mU/mL under the optimal cultivation conditions of temperature at 24°C, aeration rate at 0.6 vvm, pH 4, and agitation rate at 150 rpm, with 5-L stirred-tank bioreactor, and 19.9 mU/mL under the optimal cultivation conditions by using 24 mesh net-tube, temperature at 24°C, aeration rate at 0.9 vvm, and pH 4, with 30-L airlift bioreactor.

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Jech-Wei, Chen, et 陳志威. « Research on the quantification and submerged fermentation of destruxins ». Thesis, 1998. http://ndltd.ncl.edu.tw/handle/60086393681406413063.

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碩士
國立東華大學
生物技術研究所
86
Destruxins isolated from the entomogeneous fungi Metarhizium anisopliae isof a great potential for use as biopesticide. A processing scheme is described for preparative separation of destruxins produced from Metarhizium anisopliae fermentation broth. At the beginning, the fermentation broth was extracted with methylene dichloride. Extract was then further purified using ionexchange chromatography, silica gel chromatography, and semi-preparative HPLC chromatography. Quality of the semi-preparative products was unique. Over 85% purity (based on HPLC chromatograms) was achieved for either destru-xin A (DA) or destruxin B (DB). Determination of destruxins in the fermenta-tion broth could bedone by filtering the sample through a 0.22 mm membrane disk, and followed byinjecting into the HPLC directly. Selection of optimal medium for the production of DB was conducted by using different combination of carbon/nitrogen sources. The highest DB production (70 mg/L, after 13 days cultivation) was detected by using the combination of maltose and peptone among the three different carbon/nitrogen sources. In addition, production of DB could be enhanced by supplement of balanine in the fermentation medium. Consequently, the response surface methodology (RSM) was applied to shake-flask cultures of M. aniso products was unique. Over 85% purity (based on HPLC chromatograms) was achieved for either destruxin A (DA) or destruxin B (DB). Determination of destruxinsin the fermentation broth could be done by filtering the sample through a 0.22 mm membrane disk, and followed by injecting into the HPLC directly.A submerged fermentation production of destruxins by M. anisopliae was carried out in a5-L stirred-tank bioreactor (STR) by using maltose (3%)/peptone (0.5%) mediumat 28 *C, aeration 0.3vvm and stirred at 150 rpm. Maximal concentration of 20 mg/L DA and 223.5 mg/L DB were obtained after 15 days cultivation.

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Yang, Ya-ting, et 楊雅婷. « Production of Fructosyltransferase by Submerged Fermentation of Aspergillus japonicus ». Thesis, 2008. http://ndltd.ncl.edu.tw/handle/41351973392810525350.

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碩士
大同大學
生物工程學系(所)
96
Production of fructosyltransferase(FTase)by submerged fermentation of Aspergillus japonicus BCRC 93007 was investigated. After 4x107 spores of A. japonicus were inoculated into 200 mL of culture medium,the fermentation was carried out in a 500-mL Erlenmeyer flask in a 30°C rotary shaker at 200 rpm for more than 96 hours.The culture media consisted of 2 % yeast extract,20 % sucrose,0.5 % KH2PO4,0.05 % MgSO4•7H2O,and 1 % each of the following ingredients:carboxymethyl cellulose ( CMC ),starch,wheat bran and soybean meal. The resulted productivities of FTase were compared. When high viscosity CMC was added to the culture medium,maximum FTase at 593 U / mL was obtained after 120 hours of fermentation.The productivities of FTase resulted from these culture additives,from the highest to the lowest,in the following sequence: high viscosity CMC,starch,low viscosity CMC,wheat bran and soybean meal.It implicated that the culture medium of A. japonicus with higher viscosity makes mycelia more dispersed,and thus more FTase is produced.

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45

LIU, CHI-HSIEN, et 劉繼賢. « Production of B-Fructofuranosidase by submerged fermentation of Aspergillus japonicus ». Thesis, 1992. http://ndltd.ncl.edu.tw/handle/56879980592461306747.

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46

yu, wu yung, et 巫永裕. « Submerged cultural fermentation for chlamydospore of Trichoderma virens using winery waste ». Thesis, 2001. http://ndltd.ncl.edu.tw/handle/82789602204211895994.

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碩士
大葉大學
食品工程研究所
89
ABSTRACT The main purpose of this study was to investigate the influence of different additives on the production of mycelium and chlamydospore in submerged culture of Trichoderma virens incubated with thin stillage. On the cell growth, the pH4 was found to be better than other pH condition and the 80 % of thin stillage as the main media was also found the higher cell dry weight（7.2 g/L）after three days cultivation. The effect of addition of carbon source was shown that the 3 % glucose have the best growth of cell in the thin stillage. On the other hand, the effect of addition of nitrogen source was not shown significant difference of cell growth between the organic nitrogen and inorganic nitrogen . In the aspect of chlamydospore the 60 % thin stillage at pH4 was found higher spore production 1.69×107 chlamydospore/mL after 6 days cell cultivation.The medium contains V8 juice and other additives such as Tween 80 and glycerol can increase the production of chlamydospore. In same manner of addition of carbon source or nitrogen source in thin stillage 1 % glucose addition was found to produce higher chlamydospore than other additive sugars. However, the addition of organic or inorganic nitrogen source was not significant increase the spore production. After two or three days' cultivation, the concentration of glucose in the broth was increased to 8 % by addition concentrated sugar solution. The mycelia was found become thinner and without any increase of spore production.

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Lin, Jr-Hui, et 林志輝. « Production of Agaricus blazei Murrill and its Polysaccharide by Submerged Fermentation ». Thesis, 2004. http://ndltd.ncl.edu.tw/handle/92866903764204943493.

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碩士
國立臺灣大學
微生物與生化學研究所
92
Agaricus blazei Murrill is an edible mushroom originally used as folk remedy and food in Brazil. Over the last decade, A. blazei Murrill has been well studied and now developing as a novel functional food in Japan, Korea, China and Taiwan. The argument raised from morphological difference between A. blazei Murrill and A. blazei Heinem makes species-specific PCR is necessary before conduct subjects into test. In this study, 14 A. blazei Murrill strains was isolated from fruiting bodies, identified by species-specific PCR, established phylogenic tree by RAPD (random amplified polymorphic DNA) and optimized submerged fermentation by shaking flask fermentation. A new staining method that uses acridine orange to stain fungal mycelium and nucleus was established. A. blazei Murill mycelium has no clamp connection, with multi-nuclear(most are 4) cells. The phylogenic tree generated from RAPDs showed three groups and two distinct A. blazei Murrill strain M152 and A321 were isolated from all other strains. The optimized medium composition was 5% malt extract, 0.1% yeast extract and 0.5% peptone, pH 6. The optimized condition for submerged fermentation of A. blazei Murill was 27℃, 200mL optimal medium in 500mL hinton flask, incubated at 90 rpm for 3days then 105 rpm for 5days. The highest productivity of mycelium was 10.83±0.24g cell dry weight/L medium (A. blazei Murrill isolate M72). The highest yield of polysaccharide crude extract was 3.03±0.05 %, or 0.251±0.004 g/L medium(A. blazei Murrill isolate M152). All polysaccharide crude extracts’protein content was below 1.63% and total sugar content between 82.27∼99.14％. Polysaccharide crude extracts mainly consist of four different molecular weight components: 274.1kDa、32.7kDa、7.5kDa and 2.1 kDa. The 2.1 kDa portion should be a b-(1,3)-glucan.

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48

Murthy, Ramana M. V. « Fungal proteinase production by solid state fermentation ». Thesis, 1995. http://hdl.handle.net/2009/1436.

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49

Kang, Chih-Hsiung, et 康智雄. « Solid-state fermentation of various Bacillus species ». Thesis, 2016. http://ndltd.ncl.edu.tw/handle/06407269405967626629.

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碩士
國立屏東科技大學
食品科學國際碩士學位學程
104
Nowadays, antibiotics are widely used in livestock for increasing feed conversion efficacy and reducing costs. However, abuses of antibiotics have caused water and environment pollutions, as well as development of drug resistant pathogens, which become a major threat of human health. Researching alternative methods to reduce the use of antibiotics has become an important task or challenge in animal husbandry. Using probiotics to replace antibiotics in feeds is one of fensible strategies. People have drawn a great attention to the use of Bacillus spp. in animal feed. The present study investigated solid-state fermentation of Bacillus licheniformis, B. subtilis and B. subtilis var. natto using different agricultural by-products as fermentation substrates to make high cell density products as animal feed supplements. The investigation was initially studied on a lab scale and later was scaled up to a pilot production. The experimental results showed that rice husk to soybean meal on 2:1 ratio is suitable for use as medium formulation for cultivation of B. licheniformis, B. subtilis and B. subtilis var. natto. Sterilization time of 3 hours was required to kill all the heat resistant spores in the fermentation substrates. Additionally, glucose has to be sterilized separately from other ingredients in order to avoid Mallard reaction, which was found to inhibit the growth of Bacilli in this study. The optimized medium formulation and solid state fermentation process can produce fermentation products containing bacteria spores up to 10 to the power of 10 CFU/g after 3 days fermentation, which is satisfactory for the commercial production purpose.

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Tsai, Ming-Jin, et 蔡銘璡. « Study on functional ingredient of Cordyceps militaris by submerged andsolid state fermentation ». Thesis, 2004. http://ndltd.ncl.edu.tw/handle/24311300387759150688.

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碩士
大葉大學
生物產業科技學系碩士班
92
Pupa-Cordyceps is a famous traditional Chinese medicine, also known as “North Cordyceps” in flok in Chinese. It originates from infected larva or pupa with fruiting body by fungus Cordyceps militaris in nature. Natural Pupa-Cordyceps contains numerous functional ingredients, and possessing to nourish a body, repairing a lung, benefiting the kidney. Therefore it is called ”National Treasures”. Due to the occurrence of natural Pupa-Cordyceps is limited by the factors of environment and the climate, it is potential in viewing of the development of the mycelium and the bioactive ingredients of C. militaris using submerged fermentation. In the study, we investigated the effects of growth factors by using submerged and solid state fermentation on the production of the functional ingredients which including polysaccharide, cordycepin and adenosine. The results that was advantageous to use sucrose as a stimulator for polysaccharide production during submerged culture of C.militaris. Using 1 % corn steep powder can assist in liquid ferment. We found that cotton plug (using 4.5 g cotton as the plug in flask bottle neck with R.D 16 mm) had better cell concentration and polysaccharide production than flask with others. Using cotton plug transferring rubber plug had better the productions of extracellular cordycepin and adenosine. However, when plant oils and different surfactant was added into the medium, the mycelial concentration increased. Among that, the maximum of extracellular cordycepin content was obtained from 1 % peanut oil in medium. The maximum of intracellular adenosine content appeared 1 % olive oil adding into medium. The maximum of extracellular adenosine and intracellular cordycepin was obtained from 1 %Tween 20 in medium. In the solid state fermentation, it was advantageous to use cotton plug and maltose as a stimulator for cell growth. Solid state enhanced the biomass accumulation by using 1 % Tween 80.

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