Academic literature on the topic 'Soybean protein hydrolysates'

Hydrolysates were prepared from soybean, walnut, and peanut protein by papain, respectively. Their amino acid compositions and molecular weight distributions, the effects of various hydrolysates on H2O2-induced injury PC12 cells, and cognition of mice were investigated, respectively. Results showed that the three hydrolysates were dominated by the peptides with 1–3 KDa with large amount of neurotrophic amino acids. All the hydrolysates exhibited much stronger inhibitory activity against H2O2-induced toxicity than cerebrolysin, and soy protein hydrolysate showed the highest activity. Moreover, the hydrolysates also could reduce the rate of nonviable apoptotic cells at the concentration of 2 mg/mL. The test of animal’s cognition indicated that three hydrolysates could present partly better effect of improving recurred memory ability of normal mice and consolidated memory ability of anisodine-treated mice than piracetam. Therefore, soybean, walnut, and peanut protein hydrolysates were recommended as a potential food raw material for prevention or treatment of neurodegenerative disorders.

Soybean (Glycine max) and soy milk residue (okara) are protein-rich materials. Soybean possesses the highest protein content among different types of beans (protein content of soybean varies from 40–42 %). Soy milk residue, a by-product of the soy milk manufacturing industry, contains approximately 27 % protein (by dry weight). A number of recent studies have investigated the improvement of functional properties of protein contained in soybean and okara by fermentation or by the use of proteolytic enzymes. The aim of this study was to evaluate the antioxidant activities of soybean and okara hydrolysates obtained by the fermentation with Aspergillus oryzae or by using proteolytic enzymes (neutrase and flavourzyme). DPPH radical scavenging assay was used to determine the antioxidant activities of hydrolysates. The concentration of peptides required to scavenge DPPH radical by 50 % (IC50 value) was used to evaluate the antioxidant activity of peptides produced obtained from hydrolysates. The results showed that when fermented with A. oryzae, the okara hydrolysate had higher antioxidant activity than the soybean hydrolysate, with IC50 values of 0.447 mg/ml and 3.95 mg/ml, respectively. The hydrolyzed okara obtained from hydrolysis using Neutrase had higher antioxidant activity than the one obtained from hydrolysis using Flavourzyme, with IC50 values of0.200 mg/ml and 0.407 mg/ml, respectively. Different peptide fractions obtained from the hydrolysates using cut-off membrane (10 kDa, 3 kDa and 1 kDa) possessed different antioxidant activities. The < 1 kDa peptide fraction exhibited the highest antioxidant activity with an IC50 value of 0.158 mg/ml.

Vegetable proteins are potential low-cost alternatives to solve the protein deficiency of the world population. A protein extracted from a mixture of soybean meal and maize germ was developed to offer more protein alternatives with high nutritional value. In this study, physicochemical, functional, and nutritional characteristics of isolates and hydrolysates of soybean and counterparts extracted from a soybean meal-maize germ were compared. The isolate and hydrolysate of the soybean-maize blend had a protein content of 93.9% and 73.6%, respectively. These protein mixtures contained 10% and 52% more solubility, 303.9%, and 22.7% more emulsifying capacity, 4.5% and 4.2% higher foam density and 36.3% and 1.2% more coagulation capacity compared to the soybean isolate and hydrolysate. Electrophoretic profiles of soybean-maize proteins showed four additional bands to the typical soybean pattern of 56, 55, 52 and 18 kDa, which could correspond to globulins and zeins from maize. The isolate extracted from the mixture of soybean meal and maize is a new alternative to provide the necessary amino acids for proper physical and mental development. Additionally, it has a high potential to be used as an ingredient by the food industry due to its excellent functionality and nutritional value.

Yoghurt strain Lactobacillus LBL-4 cultivated for 8–10 h at pH ~6.0 was investigated as a considerable food-grade source of intracellular aminopeptidase. Cell-free extract manifesting >200 AP U/l was obtained from cells harvested from 1 L culture media. Subtilisin-induced hydrolysates of casein, soybean isolate, and Scenedesmus cell protein with degree of hydrolysis 20–22% incubated at 45∘C for 10 h by 10 AP U/g peptides caused an enlarging of DH up to 40–42%, 46–48%, and 38–40% respectively. The DH increased rapidly during the first 4 h, but gel chromatography studies on BioGel P-2 showed significant changes occurred during 4–10 h of enzyme action when the DH increased gradually. After the digestion, the remained AP activity can be recovered by ultrafiltration (yield 40–50%). Scenedesmus protein hydrolysate with DH 20% was inoculated by Lactobacillus LBL-4 cells, and after 72 h cultivation the DH reached 32%. The protein hydrolysates (DH above 40%) obtained from casein and soybean isolate (high Q value) demonstrated a negligible bitterness while Scenedesmus protein hydrolysates (low Q value) after both treatments were free of bitterness.

Sport nutrition drinks are well-known in escalating athlete’s performance and endurance. These product developed from whey protein hydrolysates and soybean protein hydrolysates have already been recognized, however expansion from marine product is comparatively rare. Octopus (Octopus cyanea) widely acknowledged containing taurine and rich in amino acids is potential to be developed as ingredient for sport nutrition drink. The aims of this study were to create and characterize sport nutrition drinks based on marine peptides through Octopus protein hydrolyzate. Octopus protein hydrolysate has 77.78 ± 2.69% degree of hydrolysis and 751.02 ± 10.63 mg / 100g taurine. Sports nutrition drinks with the addition of 4% Octopus protein hydrolyzate was acceptable sensory panelists, and the serving size of 600 ml contained taurine 726.06 ± 0.82 mg and detected 17 types of amino acids.

<p>Abstract<br />Sport nutrition drinks are well-known in escalating athlete’s performance and endurance. These product developed from whey protein hydrolysates and soybean protein hydrolysates have already been recognized, however expansion from marine product is comparatively rare. Octopus (Octopus cyanea) widely acknowledged containing taurine and rich in amino acids is potential to be developed as ingredient for sport nutrition drink. The aims of this study were to create and characterize sport nutrition drinks based on marine peptides through Octopus protein hydrolyzate. Octopus protein hydrolysate has 77.78±2.69% degree of hydrolysis and 751.02±10.63 mg / 100g taurine. Sports nutrition drinks with the addition of 4% Octopus protein hydrolyzate was acceptable sensory panelists, and the serving size of 600 ml contained taurine 726.06±0.82 mg and detected 17 types of amino acids.</p>

The aim of this study was to analyze soybean proteins as sources of peptides likely to be bitter using fragmentomic and hybrid approaches involving in silico and in vitro studies. The bitterness of peptides (called parent peptides) was theoretically estimated based on the presence of bitter-tasting motifs, particularly those defined as bitter-tasting indicators. They were selected based on previously published multilinear stepwise regression results. Bioinformatic-assisted analyses covered the hydrolysis of five major soybean-originating protein sequences using bromelain, ficin, papain, and proteinase K. Verification of the results in experimental conditions included soy protein concentrate (SPC) hydrolysis, RP-HPLC (for monitoring the proteolysis), and identification of peptides using RP-HPLC-MS/MS. Discrepancies between in silico and in vitro results were observed when identifying parent peptide SPC hydrolysate samples. However, both analyses revealed that conglycinins were the most abundant sources of parent peptides likely to taste bitter. The compatibility percentage of the in silico and in vitro results was 3%. Nine parent peptides with the following sequences were identified in SPC hydrolysates: LSVISPK, DVLVIPLG, LIVILNG, NPFLFG, ISSTIV, PQMIIV, PFPSIL, DDFFL, and FFEITPEK (indicators are in bold). The fragmentomic idea of research might provide a supportive method for predicting the bitterness of hydrolysates. However, this statement needs to be confirmed experimentally.

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Financiadora de Estudos e Projetos
Diseases caused by Streptococcus pneumoniae are one of the main problems of public health in the world. The pneumococcal surface protein A(PspA) is a potential canditate as carrier in a conjugate vaccine against this bacteria. Considering the inherent high losses of the purification and conjugation steps, it is fundamental to adopt a strategy of cultivation and expression that allows the obtainance of large quantities of protein. Thus, the use of Escherichia coli as expression system as well as its cultivation in complex medium constitutes promising alternatives for reducing the cost and increasing the productivity of the process. The goal of this work was to study the influence of the temperature and cultivation medium composition over the production of a PspA belonging to clade 4 protein fragment (PspA4Pro) during rE coli cultivations, aiming at to evaluate the possibility of employing vegetable-based nitrogen sources (soybean protein hydrolisates) instead of the Triptona, an animal-derived nitrogen source. The experiments were carried out in both shakers and benchscale bioreactor, using a complex medium which contained glucose and glycerol as carbon sources, lactose as inducer and Soytone, Phytone or Triptone as nitrogen sources, besides yeast extract. Samples were collected during the experiments to follow the cell growth (measurements of absorbance, dry cell weight and permittivity signal from biomass sensors), the carbon sources consumption and the production of organic acids by HPLC analysis. The stability of the plasmid (agar plates with or without kanamycin) and the production of recombinant protein (Bradford and SDS-PAGE electrophoresis followed by densitometry) were also evaluated. Preliminary experiments were performed in shake flasks, incubated at 300rpm and 37ºC, employing both complex and defined media. The highest productivity was achieved in complex medium, with a 42% superior protein production. Subsequently, nine complementary experiments were conducted in shake flasks with complex medium, under the agitation of 300rpm and temperatures of 37ºC (growth phase) and 25, 31 or 37ºC (induction phase). The largest specific production of soluble PspA4Pro was verified at 25ºC, reaching, respectively, 209±6, 192±5mg/g dry cell mass for Phytone and Triptone, with final absorbance values (after 12h of induction) of 9.0±0.4 and 8.5±0.4. The best protein production for Soytone (124±4mg/g dry cell weight) was observed at 31ºC, yielding a final absorbance 8.0±0.4. From the results obtained in the preliminary tests, the nitrogen source Phytone was selected for experiments in bioreactor. Four batch cultures were conducted in bench-scale bioreactor (5L), containing a modified auto-induction complex medium (10g/L glucose, 60g/L glycerol and 20g/L lactose), being three of them with Phytone and one with Triptone, for comparison. The best results in terms of protein production (245±7mg of PspA4Pro soluble/g dry mass) were obtained in the presence of Phytone, corresponding to an increase of 16% towards the maximum value achieved in the cultivation with Triptone. These results demonstrate the potential of vegetable-based nutrients as alternatives to animal-derived nitrogen sources in complex media, contributing to adequate these media formulations to the current guidelines of good manufacturing practices.
Doenças causadas por Streptococcus pneumoniae constituem um dos principais problemas de saúde pública mundial. A proteína A de superfície de pneumococo (PspA) é candidata em potencial a ser carreadora em vacina conjugada contra essa bactéria. Considerando as altas perdas inerentes às etapas de purificação e conjugação da proteína, é fundamental adotar uma estratégia de cultivo e expressão que permita obter grandes quantidades de proteína. Nesse sentido, o emprego da bactéria Escherichia coli como sistema de expressão e o cultivo da mesma em meio complexo se apresentam como alternativas promissoras para redução do custo e aumento da produtividade do processo. O objetivo do presente trabalho foi estudar a influência da temperatura e da composição do meio de cultivo sobre a produção do fragmento da proteína PspA do clado 4 (PspA4Pro) em cultivos de rE. coli, visando avaliar a viabilidade de utilização de fontes de nitrogênio de origem vegetal (hidrolisados protéicos de soja) em substituição à Triptona, de origem animal. Os experimentos foram realizados em câmara incubadora e em biorreatores de bancada, utilizando meio complexo contendo glicose e glicerol e lactose como fontes de carbono, lactose como indutor e Soytone, Phytone ou Triptona como fontes de nitrogênio, além de extrato de levedura. Amostras foram coletadas ao longo dos experimentos para acompanhamento do crescimento celular (medida de absorbância, massa seca e permissividade por sensor de biomassa), do consumo das fontes de carbono e da produção de ácidos orgânicos por análises em cromatografia líquida de alto desempenho. A estabilidade do plasmídeo (plaqueamento em meio contendo ou não canamicina) e a produção de proteína recombinante (Bradford e eletroforese SDS-PAGE seguida por densitometria) também foram avaliadas. Experimentos preliminares foram realizados em frascos agitados e incubados a 300rpm e 37oC, empregando tanto o meio complexo como o definido. A maior produtividade foi obtida em meio complexo, a qual foi 42% superior a alcançada com meio definido. Em seguida, nove experimentos complementares foram conduzidos em frascos agitados em meio complexo sob agitação de 300rpm e à temperatura de 37ºC (fase de crescimento) e de 25, 31 ou 37ºC (fase de indução). Verificou-se que a temperatura de 25ºC proporcionou a maior produção específica de PspA4Pro solúvel, alcançando-se, respectivamente, 209±6, 192±5mg/g massa seca para o Phytone e para a Triptona, com absorbâncias finais (após 12h de indução) de 9,0±0,4 e 8,5±0,4. Já para o Soytone, a melhor produção de proteína (124±4mg/g massa seca) foi observada à temperatura de 31ºC, obtendo-se uma absorbância de final de 8,0±0,4. A partir dos resultados obtidos nos ensaios preliminares, a fonte de nitrogênio de origem vegetal Phytone foi selecionada para experimentos em biorreator. Quatro cultivos em batelada foram conduzidos em biorreator de bancada (5L), contendo meio complexo de autoindução modificado (10g/L glicose, 60g/L glicerol e 20g/L lactose), sendo 3 com Phytone e um com Triptona, para comparação. Os melhores resultados em termos de produção de proteína (245±7mg de PspA4Pro solúvel/g massa seca) foram obtidos na presença de Phytone, correspondendo a um aumento de 16% em relação ao valor máximo alcançado no cultivo com Triptona. Esses resultados comprovam o potencial dos nutrientes de origem vegetal como alternativa às fontes de nitrogênio de origem animal em meios complexos, contribuindo para adequar as formulações desses meios às atuais diretrizes de boas práticas de fabricação.

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Agência Nacional de Petróleo
Soybean hull is a lignocellulosic biomass that contains 38-51% of cellulose, which could be converted to ethanol. In addition, contains 9-14% of protein that can be hydrolyzed by endoproteases, releasing oligopeptides with nutritional applications. Although glycoside and peptide linkages can be hydrolyzed by acids, the cellulose molecules are more resistant than the protein and hemicellulose molecules. In this way, the acid treatment of the biomass would reduce its hemicellulose content, and the remnant cellulose into solid fraction would be more susceptible to hydrolytic enzymes. In this work, different routes of protein, hemicellulose, and lignin solubilization were evaluated, intending to obtain ethanol and soluble oligopeptides from the soybean hull. The protein was recovered as oligopeptides by hydrolysis of the lignocellulosic biomass using the commercial endoprotease Novo-Pro DR at 60oC, pH 9.0, 5 h, and different enzyme concentrations (1, 2, and 4%, m/m). A sequential hydrolysis using chymotrypsin and Novo-Pro DR, both at 1% (m/m) enzyme concentration, was also evaluated. The results showed that hydrolysis with 1% Novo-Pro DR allowed solubilization of 56.9% of protein from the soybean hulls. Nonetheless, at same temperature and pH, in absence of enzyme, was possible to solubilized 45.6% of the proteins. This solubilization is probably due to liberation of the physically aggregated units. The increase of endoprotease concentration from 1 to 2% increased the protein removal to 74%. However, the increase from 2 to 4% not increased significantly the protein solubilization. The use of chymotrypsin, an enzyme with high specificity and that work at mild conditions, allowed a solubilization of 44% protein. Nonetheless, the removal of lignin using chymotrypsin was higher than that using Novo-Pro DR. When in-nature soybean hull was hydrolyzed by acid or protease (1% Novo-Pro DR) followed by acid, the protein removal was around of 90%. The lignocellulosic biomass was hydrolyzed with 3% (v/v) H2SO4, solid:liquid ratio of 1:4, 120oC, and 20 min. 20 min. Carbohydrate analyses showed that the acid treatment allowed to hemicellulose removal around of 46.7% in the xylose form. The protein content of the soybean hull was almost totally solubilized during the acid hydrolysis, without significant loss of cellulose. On the contrary, large cellulose loss was observed during the acid hydrolysis of in-nature soybean hull. In this way, if it is intended to produce a protein hydrolysate containing controlled composition or ethanol from remnant solid fraction, is strongly recommended the previous enzymatic solubilization of the proteins. The chemical composition of the solid biomass after sequential hydrolyses with protease and acid showed cellulose content around of 49% for all samples. So, the biomass treated with 1% (m/m) Novo-Pro DR was saccharified with Acellerase 1500 at 50oC, pH 4.8, and enzyme/substrate ratio of 7 FPU/g of cellulose for 72 h. Under the same conditions, soybean hull in-nature, pretreated with acid, and pretreated with protease were submitted to cellulolytic hydrolyses. The cellulose-to-glucose conversion was around of 40% for the last two biomass. The increase of the enzymatic load to 20 FPU/g of cellulose allowed a cellulose conversion of 55% for biomass pretreated with 1% (m/m) Novo-Pro DR, followed by acid hydrolysis. The supplementation of the Acellarase 1500 with 120 IU of β-glucosidase and 1% (m/m) of pectinase not produced any increased in the cellulose conversion. The biomass was pretreated by organossolv method (50% ethanol, 170oC, and 1h) and saccharified with Acellerase 1500 under the same conditions described above. This procedure yielded a cellulose conversion of 52%, with less removal of hemicellulose. This result showed that lignin was causing greater steric hindrances to the enzymatic attack. The biomass pretreated with acid and with protease (1% Novo-Pro D) followed by acid yielded the same glucose-toethanol conversion, reaching an ethanol concentration around of 13 g/L.
A casca de soja, sendo um residuo lignocelulosico, contem celulose (38-51%) que pode ser convertida a etanol. Alem disso, contem 9-14% de proteinas, que uma vez hidrolisadas por endoproteases podem liberar de forma especifica oligopeptideos com aplicacoes nutricionais. Ligacoes glicosidicas e peptidicas podem ser rompidas por hidrolise acida, sendo hemicelulose mais susceptivel a que celulose. O ataque acido ao material permitiria assim reduzir o conteudo de hemicelulose da biomassa, tornando a celulose que permanece na fracao solida insoluvel mais acessivel as enzimas hidroliticas. Neste trabalho, foram estudadas diferentes rotas de solubilizacao de proteinas, hemicelulose lignina presentes na casca de soja, visando obtencao de etanol da fracao solida e oligopeptideos na fracao liquida. A recuperacao de proteinas na forma de peptideos foi feita hidrolisando-se a biomassa inicialmente com extrato comercial de endoprotease Novo-ProD, a 60oC, pH 9, por 5h, nas concentracoes enzimaticas de 1, 2 e 4% (m/m). Foi tambem testada, na sequencia da hidrolise com Novo-ProD1%, nova hidrolise com quimotripsina 1% (m/m). Os resultados mostraram que a hidrolise proteolitica com 1% de Novo-ProDpermitiu remocao de 56,9% da proteina presente na casca. Contudo, foi tambem verificado que e possivel remover 45,6% das proteinas nas mesmas condicoes, na ausencia de enzima, a pH9,0, possivelmente devido a liberacao de unidades agregadas apenas fisicamente. Um aumento da concentracao de endoproteases de 1% para 2% elevou a remocao para 74% de proteinas, nao se observando aumento significativo na extracao de proteinas aumentandose de 2 para 4%.. Com uso da enzima mais especifica, a quimotripsina, que opera em condicoes mais brandas, foi possivel remover 44% das proteinas, com uma maior remocao de lignina do material, comparando-se com Novo-ProD. Hidrolise acida de casca in natura ou sequencial a hidrolise com Novo-ProD1% sequencial permitiu atingir uma remocao total de aproximadamente 90%de proteinas O material solido remanescente e a casca in natura foram submetidos a hidrolise acida com H2SO4 3% (v/v), razao 1:4 (solido/liquido), 120oC, 20 min. Em todos os casos, as analises de carboidratos mostraram que foi possivel remover aproximadamente 46,7% de hemicelulose, maior parte na forma de xilose. Durante o tratamento acido ocorreu a remocao de quase toda a proteina da casca de soja, sem perda expressiva de celulose, o que se observou ocorrer na hidrolise acida da casca in natura. Assim, seja para obter-se um hidrolisado proteico de composicao controlada, seja para producao de etanol da fracao solida remanescente e recomendavel a remocao enzimatica previa de proteinas. A composicao quimica do material solido apos hidrolises proteolitica e acida sequenciais mostrou teores de celulose similares para todas as amostras (aproximadamente 49 %). Assim, o solido pre-tratado com 1% de Novo Pro-D, foi utilizado para a obtencao dos acucares fermentesciveis usando o complexo enzimatico Acellerase 1500 a 50oC, pH 4,8 e razao enzima/substrato de 7 FPU/g celulose durante 72 h. Nestas mesmas condicoes foram hidrolisadas as amostras de casca in natura pre-tratada com acido, a casca in natura e a casca apos hidrolise proteolitica. A conversao enzimatica de celulose em glicose foi em torno de 40%, tanto para as amostras pre-tratadas com Novo- ProD como para a amostra submetida so a hidrolise acida. Com um aumento de carga enzimatica para 20 FPU/g celulose foi possivel atingir uma conversao de 55% para amostras pre-tratadas com 1% de Novo-ProDe hidrolise acida sequenciais. A suplementacao do complexo enzimatico Acellerase 1500 com 120 UI de β-glicosidase e 1%(m/m) de pectinase nao produziu aumento na conversao enzimatica. A hidrolise da celulose proveniente de pre-tratado por organossolve usando etanol 50% a 170oC por 1 hora resultou em 52% de conversao com uma menor remocao de hemicelulose mostrando que a lignina estava causando o maior impedimento para o ataque enzimatico. A conversao de glicose em etanol foi similar para as amostras pre-tratadas por hidrolise acida e com as hidrolises proteoliticas (1% Novo-ProD) e acidas sequenciais chegando a uma concentracao aproximada de 13 g/L.

碩士
國立嘉義大學
生物農業科技學系研究所
106
Protein hydrolysate (SPH) and alginic acid (AlA) are biostimulants that can promote plant growth, enhance plant tolerance to stressful environment. AIA is a polysaccharide isolated from brown algae. Previous studies showed that AIA could promot plant growth, increase crop yield, and act as an elicitor to trigger defense response against pathogen infection. In this study, a SPH prepared by microbial fermentation and AlA were used to treat target plants. Our results showed that Arabidopsis treated with SPH and AlA increased root length and lateral numbers under osmotic stress. Treatment of SPH and AIA can increase root length, lateral numbers and decreased electrolyte leakage under cold stress. Treatment with SPH and AlA also promoted Arabidopsis growth. Tomato plant treated with SPH and AlA exhibited increased number of flowers in a single inflorescence, and increased GPX antioxidant enzyme activity and increased protein accumulation included CAT, AOX , LhcbI and PR3. Brassica (Brassica rapa chinensis, variety Brassica) treated with SPH and AlA exhibited lower H2O2 accumulation and malonaldehyde (MDA) content. In summary, our results showed that SPH combined with AlA can promote plant growth and antioxidant enzyme activity.

碩士
國立陽明大學
生物化學研究所
88
Protein can supply several physical properties on processing industries. Various peptides with low molecular weight were produced through protein hydrolysis. Production of bioactive peptides through microbial fermentation or protease hydrolysis is prevalent now. Besides having characteristics of easier digestion, better taste, and lower allergy, the resulted peptide products also contain functions in immunity, lipid metabolic, and blood pressure regulation etc. In order to develop and substantiate the production of bioactive peptides from soybean proteins, following four bioactive assay systems have been established in this study: (a). Inhibition-activity of angiotensin I converting enzyme (ACE). (b). Enhancement of activity through 3T3-L1 lipid cell differentiation. (c). Enhancement of the human absorption of calcium. This thesis is aimed to produce and purify the bioactive peptides from soybean protein through protease hydrolysis. The analysis of physiological activities was also useful in analyzing the peptides. The goal is to study the peptide composition and analyze the physiological activities by the systems mentioned above. In this thesis, the soybean protein was digested by using various protease on the market and the subtilisin YaB developed by protein engineering in this laboratory, and the resulted peptides were fractionated by gel filtration Fractogel HW-50(s) column chromatography and reverse phase HPLC. The rate of protease digestion and the optimum condition of hydrolysis process were studied. The bioactivity of each peptide fraction was assayed using these assay systems stated above. Peptides with molecular weight of 1,000 to 10,000 Da were produced from casein protease-digested soybean and purified by either gel filtration or anion exchange chromatography after Ca2+-ethanol precipitation. The peptides which contain high acidic amino acids ( aspartate, glutamate ), serine and threonine, or histidine are known to have high affinity to Ca2+. In this research, we have obtained such kind of peptides from trypsin and pepsin digested soybean. The peptide with high histidine content was shown to have high affinity for calcium binding. The physiological activities investigated in this project include the blood pressure regulation by ACE enzyme activity analysis, improving the lipid metabolism by 3T3-L1 lipid cells differentiation analysis. All these items are common concerns to the majority. The bioactive peptides developed in this project will be marked as the products containing physiological activities. The bioactive peptides were purified and the structure and action mechanism will be investigated. 縮寫-全文對照表 圖表目次 中文摘要 英文摘要 第一章 前言 第一節 蛋白質水解物之應用 第二節 黃豆蛋白質 第三節 Subtilisin YaB及其突變酵素G124A簡介 一、Subtilisin 二、Subtilisin YaB 三、Subtilisin YaB之酵素活性 四、Subtilisin YaB的應用價值 五、Subtilisin G124A 第四節 胜之生理活性 一、鈣吸收促進活性 二、脂肪細胞分化調節活性 三、ACE酵素抑制活性 第五節 研究構想與目的 第二章 實驗材料與方法 第一節 實驗材料 一、蛋白質來源 二、酵素來源 三、菌株來源、培養基及培養液 四、細胞株來源、培養基及培養液 五、管柱 六、藥品及儀器 第二節 實驗方法 一、Subtilisin YaB及G124A修飾型酵素之生產純化 二、蛋白質水解液之製備及酵素分解過程之探討 三、鈣結合性胜之生產純化 四、3T3-L1前驅脂肪細胞分化調節活性分析系統之建立 五、篩選具ACEI活性之胜 第三章 結果與討論 第一節 Subtilisin YaB 酵素及其突變酵素G124A之生產純化 一、Subtilisin YaB 之純化 二、G124A突變酵素之純化 第二節 酵素水解反應條件之探討及比較 一、Trypsin等八種蛋白對黃豆蛋白、酪蛋白分解率之比較 二、黃豆蛋白水解之探討結果及所產生胜分布之比較 三、蛋白水解液以 HW-50(s) 分子篩管柱層析分析結果 第三節 鈣吸收促進活性胜之生產純化 一、黃豆蛋白質之水解 二、鈣結合性胜之管柱層析分析及其胺基酸組成特性之比較 第四節 3T3-L1前驅脂肪細胞分化調節活性分析系統之建立 一、3T3-L1細胞經分化處理後之形態變化 二、3T3-L1脂肪細胞處理以各胜區分之變化比較 第五節 ACE酵素抑制活性之測定 第四章 參考文獻

碩士
國立嘉義大學
農業科技全英碩士學位學程
106
Abstract Reactive oxygen species (ROS) molecules can serve as signaling molecules or cause oxidative damage to the tissues that would depend on the delicate equilibrium between ROS production, and their scavenging activity to reduce ROS level. Biostimulants are natural compounds or microorganisms that can promote plant growth. Plant growth promoting rhizobacteria and soybean protein hydrolysate (SPH) are different kinds of biostimulants. In this study, Bacillus sp. WA strain and protein hydrolysate generated by the same bacterial species were used to treat Arabidopsis , bok choi, lettuce, and tomato plants. Our results showed that application of PGPR and soybean protein hydrolysate WA on Arabidopsis, bok choi, and lettuce significantly enhanced growth response in the vegetative phase, including shoot and root growth, fresh weight, chlorophyll content, chlorophyll fluorescence, soluble sugar, abiotic stress tolerance toward drought and heat stress exposure, and antioxidants activity such as catalase (CAT) and guaiacol peroxidase (GPX). Treatments of bacteria and SPH increased generative growth of tomato plant, fruit number, fruit ripening, and fruit fresh weight. Treated plant reduced nitrate accumulation, and concentrations of hydrogen peroxide (H2O2) and malondialdehyde (MDA) in the leaf tissues of bok choi and tomato plant; moreover, increased protein accumulation of SOD (superoxide dismutase), APX (L-ascorbate peroxidase), CAT (catalase); LhCb1 (LHCII type I chlorophyll a/b-binding protein), pathogenesis-related (PR) protein 3 (PR-3), cytosolic fructose-1,6-bisphosphatase (cFBPase), and NR (nitrate reductase). Those results suggest that WA strain or it protein hydrolysis product play important roles on plant growth and development. Keyword: PGPR Bacillus sp. WA); Soybean protein hydrolysate (SPH) by Bacillus sp. WA strain; antioxidant enzymes; abiotic stresses