Academic literature on the topic 'Separate Hydrolysis and Fermentation'

The purpose of this study to obtain the best reducing sugars using acid solvent H2SO4 and HCl to be used as a substrate in fermentation processes to produce bioethanol. The research phase includes the preparation of raw materials Sargassum sp., the processing of acid hydrolysis used a solvent H2SO4 and HCl. Hydrolysis then fermented for five days for the production of etanol. Hydrolysis using acid solvent H2SO4 obtained the best acid concentration of 2% with the result of reducing sugars 82,62 g/L, whereas using HCl acid solvent obtained the best acid concentration of 2% with the result of reducing sugars 74,79 g/L. Fermented for 120 hours to produce ethanol each H2SO4 2 ml and 3 ml HCl.

The fractionation of sugarcane bagasse using formic acid allowed removing lignin and hemicellulose, obtaining a material containing up to 90 % cellulose. The material can be easily hydrolyzed into glucose to serve as materials to produce high value added products such as biofuel, chemicals, pharmaceuticals, food additives, and the likes. The hydrolysate of fractionated bagasse was easily fermented with a (ethanol) fermentation yield attained 91.08 ± 2.02 %, showing no significant inhibition to the yeast in the hydrolysate. In this study, a process of simultaneous hydrolysis and fermentation (SSF) was performed to convert fractionated sugarcane bagasse at 20 % consistency to ethanol. The process with 6h pre-hydrolysis at 50 0C then SSF at 37 0C could attain a high ethanol concentration of 82.46 ± 3.42 g/L in the fermentation with the ethanol recovery yield of 81.66±1.88%; which was15.37 ± 1.06 % higher than that of the separate hydrolysis and fermentation (SHF) process (70.78 ± 0.25 %). In addition, in the SSF, the process time was shorten to 4 days instead of 7 days in the SHF.

This study compares the effects of pre- and post-hydrothermal treatment of source- separated organics (SSO) on solubilization of particulate organics and acidogenic fermentation for volatile fatty acids (VFAs) production. The overall COD solubilization and solids removal efficiencies from both schemes were comparable. However, the pre-hydrolysis of SSO followed by acidogenic fermentation resulted in a relatively higher VFA yield of 433 mg/g VSS, which was 18% higher than that of a process scheme with a post-hydrolysis of dewatered solids from the fermentation process. Regarding the composition of VFA, the dominance of acetate and butyrate was comparable in both process schemes, while propionate concentration considerably increased in the process with pre-hydrolysis of SSO. The microbial community results showed that the relative abundance of Firmicutes increased substantially in the fermentation of pretreated SSO, indicating that there might be different metabolic pathways for production of VFAs in fermentation process operated with pre-treated SSO. The possible reason might be that the abundance of soluble organic matters due to pre-hydrolysis might stimulate the growth of more kinetically efficient fermentative bacteria as indicated by the increase in Firmicutes percentage.

The production of single- and mixed-sugar streams and their conversion to bioproducts were studied, using sulfite pulping streams as feedstocks. Sulfite pulp, sludge, and spent sulfite liquor are concurrently generated alongside of bleached pulp, and the pulping process renders pretreatment of solid streams unnecessary. Streams were converted separately; however, due to their low production volume, solid and liquid streams were also combined as a means to increase the quantity of starting feedstock. Spent sulfite liquor, comprising mostly monomeric hexose and pentose sugars, was directly fermented to ethanol and xylitol with Candida guilliermondii. Single-sugar streams were generated through hydrolysis of pulp and sludge in water, followed by fermentation to ethanol with Saccharomyces cerevisiae. Mixed-sugar streams were generated through both separate hydrolysis and fermentation and simultaneous saccharification and fermentation of pulp and sludge in spent sulfite liquor using S. cerevisiae. The best single-sugar source was obtained by hydrolysis of pulp in water, which produced 78.8 g/L of glucose after 96 h. The glucose concentration from hydrolysis of sludge in water was lower (33.5 g/L). Both of these streams were easily converted to ethanol, with yields of 77.8% and 76.2%, respectively. Hydrolyzability of solids was the limiting factor in separate hydrolysis and fermentation conversion of pulp and sludge in water, but hydrolyzability of sludge was not affected when mixed with spent sulfite liquor.

Bioethanol from lignocellulose is expected to be the most likely fuel alternative in the near future. SEKAB E-Technology in Örnsköldsvik, Sweden develops the technology of the 2nd generation ethanol production; to produce ethanol from lignocellulosic raw material. The objective of this master’s thesis was to achieve a better knowledge of the potential and limitations of separate hydrolysis and fermentation (SHF) as a process concept for the 2nd generation ethanol production. The effects of enzyme concentration, temperature and pH on the glucose concentration in the enzymatic hydrolysis were investigated for pretreated spruce at 10% DM using a multiple factor design. Enzyme concentration and temperature showed significant effects on the glucose concentration, while pH had no significant effect on the concentration in the tested interval of pH 4.5-5.5. To obtain the maximum glucose concentration (46.4 g/l) for a residence time of 48 h, the optimal settings within the studied parameter window are a temperature of 45.7⁰C and enzyme concentration of 15 FPU/g substrate. However, a higher enzyme concentration would probably further increase the glucose concentration. If enzymatic hydrolysis should be performed for very short residence times, e.g. 6 h, the temperature should be 48.1⁰C to obtain maximum glucose concentration. The efficiency of the enzymes was inhibited when additional glucose was supplied to the slurry prior to enzymatic hydrolysis. It could be concluded that end product inhibition by glucose occurs and results in a distinct decrease in glucose conversion. No clear conclusions could be drawn according to different techniques for slurry and enzymes, i.e. batch and fed-batch, in the enzymatic hydrolysis process. Investigations of the fermentability of the hydrolysate revealed that the fermentation step in SHF is problematic. Inhibition of the yeast decrease the fermentation efficiency and it is therefore difficult to achieve the 4% ethanol limit. Residence time for enzymatic hydrolysis (48 h) and fermentation (24 h) need to be prolonged to achieve a sufficient SHF process. However, short processing times are a key parameter to an economically viable industrial process and to prolong the residence times should therefore not be seen as a desirable alternative. SHF as a process alternative in an industrial bioethanol plant has both potential and limitations. The main advantage is the possibility to separately optimize the process steps, especially to be able to run the enzymatic hydrolysis at an optimal temperature. Although, it is important to include all the process steps in the optimization work. The fermentation difficulties together with the end product inhibition are two limitations of the SHF process that have to be improved before SHF is a preferable alternative in a large scale bioethanol plant.

AgÃncia Nacional do PetrÃleo
CoordenaÃÃo de AperfeÃoamento de Pessoal de NÃvel Superior
In this work, the ethanol production from cashew bagasse was studied after acid followed by alkali pretreatment (CAB-OH) using the Separate Hydrolysis and Fermentation (SHF) and Simultaneous Saccharification and Fermentation (SSF) processes. In SHF process, the hydrolysate obtained from enzymatic hydrolysis of CAB-OH was used as carbon source for fermentation with different strains of Saccharomyces (S. cerevisiae CCA008, S. cerevisiae 01, S. cerevisiae 02 and Saccharomyces sp. 1238), Kluyveromyces (K. marxianus CCA510, CE025 and ATCC36907) and Hanseniaspora sp. GPBio03. The bioprocess was conducted at 30 ÂC and 50 g.L-1 initial glucose concentration. The K. marxianus ATCC36907 achieved ethanol concentration of 20 g.L-1 with consumption of all glucose in the hydrolysate. Similar results were obtained with Saccharomyces strains and higher ethanol concentration (23.43 g.L-1) was obtained by Saccharomyces sp. 1238. The maximum ethanol concentration of 24.54 g.L-1 was achieved by Hanseniaspora sp. GPBio03. Focused on further studies using SSF process, it was evaluated the temperature influence of thermotolerant yeast K. marxianus ATCC36907 in glucose and enzymatic hydrolysate from CAB-OH. The results showed that the temperature (30, 35, 40, 45 and 50 ÂC) did not affect the values of YE/G (0.45 to 0.46 gethanol/gglucose) using glucose as substrate. Moreover, the ethanol yields obtained with enzymatic hydrolysate were slightly influenced by temperature, 0.39 and 0.43 gethanol/gglucose were obtained at 30 and 40 ÂC, respectively. Based on this, the SSF of CAB-OH and K. marxianus ATCC36907 was conducted at 40 ÂC with cellulases from Celluclast 1.5L at 15 FPU/gcellulose. The highest ethanol concentration (24.90 Â 0.89 g.L-1) was obtained with 76h of fermentation with 0.33 g.L-1.h-1, 0.34 gethanol/gglucose and 66.3% of productivity, YʹE/G and of ethanol efficiency, respectively. In enzymatic hydrolysis studies, the cellulase NS 22074 at 30 FPU/gcellulose without cellobiases supplementation resulted in glucose yield of 93.77 Â 2.72% which is promising for studies of SSF with this enzyme complex. The temperature (40, 42 , 45 and 50 ÂC) influence in SSF process using microcrystalline cellulose, in contrast with SHF results, higher ethanol concentration, 19.86 Â 0.32 g.L-1, was obtained at 40 ÂC. The SSF using CAB-OH, 30 FPU/gcellulose cellulases NS 22074 at 40 ÂC showed higher ethanol concentration of 37.35 Â 0.64 g.L-1 at 80h, with productivity of 0.46 g.L-1.h-1. In this condition, there was an increase of YʹE/G from 0.34 to 0.49 gethanol/gglucose and the ethanol efficiency from 66.3% to 95.59% when compared to results obtained with SSF using Celluclast 1.5L. Based on the results of efficiency and ethanol yield (YʹE/G), the cashew apple bagasse showed as lignocelulose feedstock promising material for second generation ethanol production by SSF process using the yeast K. marxianus ATCC36907 and NS 22074 cellulases complex.
Nesse trabalho, estudou-se a produÃÃo de etanol de bagaÃo de caju apÃs prÃ-tratamento Ãcido seguido de Ãlcali (CAB-OH) atravÃs dos processos de FermentaÃÃo e HidrÃlise Separadas (SHF) e FermentaÃÃo e HidrÃlise SimultÃneas (SSF). No processo SHF, o hidrolisado obtido da hidrÃlise enzimÃtica de CAB-OH foi submetido à etapa de fermentaÃÃo com diferentes linhagens de Saccharomyces (S. cerevisiae CCA008, Saccharomyces sp. 1238, S. cerevisiae 01, S. cerevisiae 02), Kluyveromyces (K. marxianus CCA510, CE025 e ATCC36907) e Hanseniaspora sp. GPBio03. A fermentaÃÃo do hidrolisado foi conduzida a 30 ÂC com concentraÃÃo inicial de glicose de 50 g.L-1. ApÃs o screening de leveduras, a linhagem de K. marxianus ATCC36907 destacou-se com maior concentraÃÃo de etanol de 20 g.L-1 com consumo de toda glicose no hidrolisado. Resultados similares foram obtidos com Saccharomyces sp. 1238 e com a levedura isolada do caju (Hanseniaspora sp. GPBio03) com maiores concentraÃÃes de etanol de 22,41 g.L-1 e 24,54 g.L-1, respectivamente. Com o propÃsito de estudos posteriores de SSF, avaliou-se a influÃncia da temperatura da levedura termotolerante K. marxianus ATCC36907 em glicose PA e hidrolisado enzimÃtico de CAB-OH. Os resultados mostraram que para a glicose PA, a variaÃÃo da temperatura (30, 35, 40, 45 e 50 ÂC) nÃo influenciou nos valores de conversÃo de glicose em etanol (YE/G) obtendo-se valores na faixa de 0,45-0,46 getanol/gglicose. Por outro lado, os resultados de YE/G em hidrolisado enzimÃtico foram ligeiramente influenciados pela temperatura, obtendo-se 0,39 getanol/gglicose a 30ÂC e 0,43 getanol/gglicose a 40 ÂC. Em seguida, realizou-se a SSF de CAB-OH com K. marxianus ATCC36907 a 40 ÂC e celulases de Celluclast 1.5L a 15 FPU/gcelulose. A maior concentraÃÃo de etanol (24,90  0,89 g.L-1) foi obtida em 76h de fermentaÃÃo com produtividade de 0,33 g.L-1.h-1, conversÃo de glicose em etanol (YʹE/G) de 0,34 e eficiÃncia de produÃÃo de etanol de 66,3%. Contudo, visando aumentar a produÃÃo de etanol em estudos posteriores de SSF, realizou-se o estudo de hidrÃlise enzimÃtica com outros complexos de celulases (NS 22074) e celobiases (NS 50010). Os resultados de hidrÃlise enzimÃtica mostraram que a atividade de celulases NS 22074 a 30 FPU/gcelulose sem suplementaÃÃo de celobiase resultou no rendimento de glicose de 93,77  2,72% sendo resultado promissor para estudos de SSF com esse complexo enzimÃtico. Nos ensaios de SSF com celulases do complexo NS 22074, inicialmente realizou-se o estudo da temperatura (40, 42, 45 e 50 ÂC) com K. marxianus ATCC36907 utilizando celulose microcristalina; e, em contrapartida com os resultados SHF, na temperatura de 40 ÂC foi obtida a maior concentraÃÃo de etanol de 19,86  0,32 g.L-1, em 72h de fermentaÃÃo. Diante desses resultados, realizou-se o processo de SSF de CAB-OH nas seguintes condiÃÃes: 40 ÂC de temperatura e 30 FPU/gcelulose do complexo de celulases NS 22074. A maior concentraÃÃo de etanol (37,35  0,64 g.L-1) foi obtida em 80h de fermentaÃÃo, com produtividade de 0,46 g.L-1.h-1. Diante desses resultados, observa-se que a mudanÃa do complexo enzimÃtico de Celluclast 1.5L para NS 22074 proporcionou o aumento no valor de YʹE/G de 0,34 getanol/gglicose para 0,49 getanol/gglicose e no rendimento de etanol de 66,3% para 95,59%, o que torna o bagaÃo de caju prÃ-tratado promissor como matÃria-prima para produÃÃo de etanol de segunda geraÃÃo por processo SSF utilizando a levedura K. marxianus ATCC36907.

The diploma thesis is focused on production possibilities of bioethanol from waste paper by yeast Kluyveromyces marxianus. Waste cardboard was used as a potential substrate for bioethanol production. Several methods for cardboard preparation were introduced and compared as well as methods of fermentation. Simultaneous sacharification and fermentation and separate hydrolysis and fermentation of preprepared cardboard paper were performed in different pH buffer (4,8-7). Simultaneous sacharification and fermentation was held at a temperature of 45°C. Hydrolysis in separate hydrolysis and fermentation was performed at 50°C and fermentation at 25°C. Procedures outputs were obtained by sampling in specific time intervals and samples were analyzed by HPLC for presence and concentration glucose and ethanol. The results of the analysis have shown that the highest concentration of glucose produced by enzymatic hydrolysis was achieved by using microwaves, 2% H2SO4 and 2% NaOH pretreated paperboard at pH 4,8. The highest yield of ethanol was obtained by separate hydrolysis and fermentation of pulp pretreated by microwaves, 2% H2SO4 and 2% NaOH in pH 5,4 buffer. The method SHF proved to be more effective for the production of ethanol than SSF.

Thesis (MScEng)--Stellenbosch University, 2012
ENGLISH ABSTRACT: Through many years of research, a number of production schemes have been developed for converting lignocellulosic biomass into transport fuels. These technologies have been assessed through a number of techno-economic studies for application in a particular context in terms of the technical and economic feasibility. However, previous studies using these methods have tended to lack vigour in various aspects. Either the energy efficiency of the processes were not maximised through adequate heat integration, or a competing technology which existed was not considered. From an economic perspective, the financial models would often lack the vigour to account for the risk and uncertainty that is inherent in the market prices of the commodities. This phenomenon is especially relevant for the biofuel industry that faces the full fledge of uncertainties experienced by the agricultural sector and the energy sector. Furthermore, from an environmental perspective, the techno-economic studies had often ignored the environmental impacts that are associated with biofuel production. Thus, a comparative study could have favoured an option due to its economic feasibility, while it could have had serious environmental consequences. The aim of this study was to address these issues in a South African context, where biofuels could be produced from sugarcane bagasse. The first step would be to modify an existing simulation model for a bioethanol scenario that operates with a Separate Hydrolysis and Fermentation (SHF process) configuration into a second processing scenario that operates with a Simultaneous Saccharification and Fermentation (SSF process) configuration using reliable experimental data. The second step was to ensure that the maximum energy efficiency of each scenario was realised by carrying out pinch point analysis as a heat integration step. In contrast to these biological models is the thermochemical model that converts bagasse to gasoline and diesel via gasification, Fischer-Tropsch synthesis and refining (GFT process). While there were no significant advances in technology concerning this type of process, the energy efficiency was to be maximised with pinch point analysis. The GFT process obtained the highest energy efficiency of 50.6%. Without the affects of pinch point technology, the efficiency dropped to 46%, which thus emphasises the importance of heat integration. The SSF had an efficiency of 42.8%, which was superior to that of the SHF at 39.3%. This resulted from a higher conversion of biomass to ethanol in the SSF scenario. Comparing the SHF model to an identical model found in literature that did not have pinch point retrofits, this study showed lower efficiency. This arose because the previous study did not account for the energy demands of the cold utility systems such as the cooling tower operation, which has been shown in this study to account for 40% of the electrical energy needs. The economic viability of all three processes was assessed with Monte Carlo Simulations to account for the risks that the fluctuations in commodity prices and financial indices pose. This was accomplished by projecting the fluctuations of these parameters from samples of a historical database that has been transformed into a probability distribution function. The consequences were measured in terms of the Net Present Value (NPV) and Internal Rate of Return (IRR) for a large number of simulations. The results of these variables were aggregated and were then assessed by testing the probability that the NPV<0, and that the IRR recedes below the interest rate of 12.64%. The investment was thus deemed unfeasible if these probabilities were greater than 20%. Both biological models were deemed profitable in terms of this standard. The probabilities were 13% for the SSF and 14% for the SHF. The GFT process however was deemed completely unfeasible because the probability that the NPV<0 was 78%. Given that the GFT process had the highest energy efficiency, this result arises mainly because the capital investment of 140,000USD/MWHHV of biomass energy input is to enormous for any payback to be expected. The environmental footprint of each process was measured using Life Cycle Assessments (LCAs). LCAs are a scientifically intricate way of quantifying and qualifying the effects of a product or process within a specified boundary. The impacts are assessed on a range of environmental issues, such as Global Warming, Acidification, Eutrophication and Human toxicity. Furthermore, if the project under concern has multiple output products, then the impacts are distributed between the output products in proportion to the revenue that each generates. The impacts were either relative to the flow of feedstock, which was 600MW of bagasse, or to the functional unit, which was the amount of fuel required to power a standard vehicle for a distance of 1 kilometre. In either case, the GFT scenario was the least burdening on the environmental. This was expected because the GFT process had the highest energy efficiency and the process itself lacked the use of processing chemicals. Relative to the feedstock flow, the SSF was the most environmentally burdening scenario due to the intensive use of processing chemicals. Relative to the functional unit, the SHF was the most severe due to its low energy efficiency. Thus, the following conclusions were drawn from the study:  The GFT is the most energy and environmentally efficient process, but it showed no sign of economic feasibility. iv  There is no significant difference in the economic and environmental evaluation of the SSF and SHF process, even though the SSF is considered to be a newer and more efficient process. The major cause of this is because the setup of the SSF model was not optimised.
AFRIKAANSE OPSOMMING: Deur baie jare van navorsing is ‘n aantal produksie-skemas vir die omskakeling van lignosellulose biomassa na vloeibarebrandstof ontwikkel. Hierdie tegnologië is geassesseer ten opsigte van die tegniese en ekonomiese haalbaarheid deur middel van tegno-ekonomiese studies in bepaalde tekste. Tog het hierdie vorige studies besliste beperkings gehad. Of die energie-doeltreffendheid van die proses is nie gemaksimeer deur voldoende hitte-integrasie nie, of 'n mededingende tegnologie wat bestaan is nie oorweeg nie. Vanuit 'n ekonomiese perspektief, was die finansiële modelle dikwels nie die omvattend genoeg om rekening te hou met die risiko en onsekerheid wat inherent is in die markpryse van die kommoditeite nie. Hierdie verskynsel is veral relevant vir die biobrandstof bedryf wat die volle omvang van onsekerhede ervaar waaraan die landbousektor en die energiesektoronderhewig is. Verder het die tegno-ekonomiese studies dikwels die omgewingsimpakte wat verband hou met biobrandstofproduksie geïgnoreer. Dus kon ‘n opsie deur die ekonomiese haalbaarheid bevoordeel word, ten spyte van die ernstige omgewingsimpakte wat dit kon inhou. Die doel van hierdie studie was om hierdie kwessies aan te spreek in 'n Suid-Afrikaanse konteks, waar biobrandstof uit suikerriet bagasse geproduseer kan word. Die eerste stap was om 'n bestaande simulasiemodel vir 'n bio-scenario wat met Afsonderlike Hidroliese en Fermentasie (SHF proses) stappe werk, te modifiseer vir 'n tweede verwerking scenario wat met 'n gelyktydige Versuikering en Fermentasie (SSF proses) konfigurasie werk. Die verandering is gedoen deur die gebruik van betroubare eksperimentele data. Die tweede stap was om te verseker dat elke scenario die maksimum energie-doeltreffendheid het, deur 'n hitte-integrasie stap, wat gebruik maak van “pinch-point” analise. In teenstelling met hierdie biologiese modelle, is daar die thermochemiese roete waar petrol en diesel van bagasse vervaardig word via vergassing, Fischer-Tropsch-sintese en rafinering (GFT proses). Daar was geen betekenisvolle vooruitgang in tegnologie vir hierdie proses nie, maar die energie-doeltreffendheid is gemaksimeer word deur energie-integrasie. Die GFT proses toon die hoogste energie-doeltreffendheid van 50,6%. Sonder die invloed van energie-integrasie het die doeltreffendheid gedaal tot 46%, wat dus die belangrikheid van hitte-integrasie beklemtoon. Die SSF het 'n effektiwiteit van 42,8% gehad, wat beter was as dié 39,3% van die SHF opsie. Hierdie hoër effektiwiteit wasas gevolg van die hoër omskakeling van biomassa na etanol in die SSF scenario. Die energie doeltreffendheid vir die SHF-model was laer as met 'n identiese model (sonder energie-integrasie) wat in die literatuur gevind wat is. Dit het ontstaan omdat die vorige studie nie 'n volledig voorsiening gemaak het met die energie-eise van die verkillingstelselsnie, wat tot 40% van die elektriese energie behoeftes kan uitmaak. Die ekonomiese lewensvatbaarheid van al drie prosesse is bepaal met Monte Carlo simulasies om die risiko's wat die fluktuasies in kommoditeitspryse en finansiële indekse inhou, in berekening te bring. Hierdie is bereik deur die projeksie van die fluktuasies van hierdie parameters aan die hand van 'n historiese databasis wat omskep is in 'n waarskynlikheid verspreiding funksie. Die gevolge is gemeet in terme van die netto huidige waarde (NHW) en Interne Opbrengskoers (IOK) vir 'n groot aantal simulasies. Die resultate van hierdie veranderlikes is saamgevoeg en daarna, deur die toets van die waarskynlikheid dat die NPV <0, en dat die IRR laer as die rentekoers van 12,64% daal, beoordeel. Die belegging is dus nie realiseerbaar geag as die waarskynlikhede meer as 20% was nie. Beide biologieseprosesse kan as winsgewend beskou word in terme van bostaande norme. Die waarskynlikhede was 13% vir die SSF en 14% vir die SHF. Aangesien die NHW van die GFT-proses onder 0 met ‘n waarskynlikheid van 78% is, is die opsie as nie-winsgewend beskou. Gegewe dat die GFT-proses die hoogste energie-doeltreffendheid het, is die resultaat hoofsaaklik omdat die kapitale belegging van 140,000 USD / MWHHV-biomassa energie-inset te groot is, om enige terugbetaling te verwag. Die omgewingsvoetspoor van elke proses is bepaal deur die gebruik van Lewens Siklus Analises (“Life Cycle Assessments”) (LCAS). LCAS is 'n wetenskaplike metodeom die effek van ‘n produk of proses binne bepaalde grense beide kwalitatief en kwantitatief te bepaal. Die impakte word beoordeel vir 'n verskeidenheid van omgewingskwessies, soos aardverwarming, versuring, eutrofikasie en menslike toksisiteit. Voorts, indien die projek onder die saak verskeie afvoer produkte het, word die impakte tussen die afvoer produkte verdeel, in verhouding tot die inkomste wat elkeen genereer. Die impak was met of relatief tot die vloei van roumateriaal (600MW van bagasse), of tot die funksionele eenheid, wat die hoeveelheid van brandstof is om 'n standaard voertuig aan te dryf oor 'n afstand van 1 kilometer. In al die gevalle het die GFT scenario die laagste belading op die omgewing geplaas. Hierdie is te verwagte omdat die GFT proses die hoogste energie-doeltreffendheid het en die proses self nie enige addisionele chemikalieë vereis nie. Relatief tot die roumateriaal vloei, het die SSF die grootse belading op die omgewing geplaas as gevolg van die intensiewe gebruik van verwerkte chemikalieë. Relatief tot die funksionele eenheid, was die SHF die swakste as gevolg van sy lae energie-doeltreffendheid.

Research about resource recovery from complex waste streams is getting an increased scientific attention since valuable resources can be produced by sustainable biological means. In anaerobic degradation processes, resources such as volatile fatty acids (VFAs) and biogas are highly coveted. One of the key parameters affecting the yield of resources is the hydrolytic efficiency in the waste stream by hydrolytic bacteria. The aim of this study was to examine how bioaugmentation can be implemented as a strategy to enhance hydrolysis in complex waste streams. In pursuit of this aim, three selected species of hydrolytic bacteria, Bacteroides thetaiotaomicron, Bacteroides amylophilus and Bacteroides ruminicola were inoculated both in pure culture combinations and bioaugmented with granular sludge as mixed culture in reactors. The studied waste stream was food waste mixed with primary sludge collected from Henriksdals wastewater treatment plant at Stockholm, Sweden.  The highest hydrolytic efficiency (90%) was reached by the pure culture fermented reactor inoculated with Bacteroides thetaiotaomicron and Bacteroides ruminicola. This efficiency was measured at day 10 after reactor set-up. Among the bioaugmented reactors, highest hydrolytic activity (66%) was achieved by the reactor inoculated with Bacteroides thetaiotaomicron and it was measured at day 10. The increase in hydrolytic efficiency for bioaugmented reactors was slower compared to pure culture fermented reactors and the most probable reason to that is due to competition amongst introduced species and pre-existing mixed culture in granular seed sludge.
Mer uppmärksamhet riktas till forskning kring resursåtervinning från komplexa avfallsströmmar eftersom värdefulla resurser kan produceras genom mer hållbara biologiska tillvägagångssätt. I anaeroba nedbrytningsprocesser är produkter såsom flyktiga fettsyror (VFAs) och biogas mycket eftertraktade. En av huvudparametrarna som påverkar utbytet av återvunna resurser är den hydrolytiska effektiviteten i avfallsströmmen av hydrolytiska bakterier. Syftet med studien var att undersöka hur bioaugmentering kan implementeras som strategi för att förstärka hydrolys i komplexa avfallsströmmar. Därav utfördes fermentering med tre valda hydrolytiska bakterier, Bacteroides thetaiotaomicron, Bacteroides amylophilus och Bacteroides ruminicola både i renkultur och bioaugmenterat med granulärt slam som mixad kultur i reaktorer. Avfallsströmmen som studerades var matavfall mixat med primärt slam hämtat från Henriksdals vattenreningsverk i Stockholm, Sverige.  Högsta hydrolytiska effektivitet (90%) uppnåddes för reaktorn inokulerat med Bacteroides thetaiotaomicron och Bacteroides ruminicola i renkultur. Denna effektivitet uppmättes dag 10 efter reaktorerna sattes upp. För de bioaugmenterade reaktorerna så uppnåddes högsta hydrolytiska effektivitet (66%) dag 10 av reaktorn inokulerat med Bacteroides thetaiotaomicron. Ökningen i hydrolytisk effektivitet var långsammare för de bioaugmenterade reaktorerna jämfört med reaktorerna med renkultur. Den mest sannolika förklaringen till det är tävling om näringsämnen och vitaminer mellan introducerade bakterier och de bakterier som redan existerar i det granulära slammet.

Due to the depletion of petroleum reserves and environmental concerns, bioethanol has been identified as an alternative fuel to petrol. Bioethanol is a fuel of bio-origin derived from renewable biomass. Starch and sugar containing materials are the primary sources of carbon for bioethanol production. Starch is firstly hydrolysed into simple sugars which are later fermented to bioethanol using Saccharomyces cerevisiae (S. cerevisiae). The fermentation of sugars to bioethanol is however limited by inhibition of S. cerevisiae by the major product of the process, bioethanol. The challenge is thus in keeping the bioethanol concentration at levels which are not harmful to the fermenting organism. Keeping bioethanol concentration low in the broth will provide a suitable environment for yeast to grow and thus increase the overall production. Currently bioethanol producers use high water dilution rates to keep the bioethanol concentrations in the broth low enough so that yeast is not harmed. This excess water has to be removed in the downstream process, which is expensive. The use of excessive amounts of water in the fermentation can be avoided by continual removal of bioethanol from the broth. During this investigation the experimental conditions for the hydrolysis process were determined. A pH of 5.5 was determined as the best pH for Termamyl SC at 95°C with a pH of 5.0 for Spirizyme Fuel at 55°C during the liquefaction and the saccharification step, respectively. During the fermentation process the influence of yeast concentration on bioethanol production was investigated by varying the yeast concentration between 2 g.L-1 and 7 g.L-1. A yeast concentration of 5 g.L-1 produced the highest bioethanol yield of 0.48 g.g-1 after 48 hours of fermentation using S. cerevisiae. Later during the investigation a coupled fermentation/pervaporation system was employed in a batch system for continual removal of bioethanol in the fermentation broth in a process called simultaneous fermentation and separation (SFS). Through the continuous removal of bioethanol from the fermentation broth, the bioethanol concentration in the broth was kept low enough so that it was not harmful to the fermenting organism but the overall fermentation yield was not improved. Pervaporation is a membrane separation process used to separate azeotropic mixtures such as bioethanol and water. It is highly efficient, cost effective and uses less energy than distillation. During the SFS process a bioethanol yield of 0.22 g.g-1 was obtained. The SFS process yield for bioethanol was low compared to 0.45 g.g-1 of the traditional batch fermentation process. The lower overall bioethanol yield obtained in the SFS process could be attributed to only the supernatant being used in the SFS process and not the entire fermentation broth as in the traditional process. The results from this study proved that the SFS process was less efficient compared to the traditional batch fermentation process with respect to the bioethanol yield, but that the fermentation could be carried out without the necessity for additional process water.
Thesis (M.Sc. Engineering Sciences (Chemical and Minerals Engineering))--North-West University, Potchefstroom Campus, 2010.

Efficient processing of cassava roots by wet milling requires overcoming challenges associated with disaggregation of the starch-containing parenchyma cells. These cells entrap starch granules and hinder their release during wet milling. Steeping of ground cassava in 0.75% (w/v) NaOH in combination with wet milling was investigated to determine whether and how dilute NaOH modifies cassava cell walls. Gas chromatography (GC) data of cell wall constituent sugar composition and Fourier transform infrared (FTIR) data showed that NaOH steeping caused solubilisation of the cell wall pectin fraction. FTIR and wide-angle x-ray scattering (WAXS) spectroscopy indicated that NaOH steeping combined with fine (500 ?m opening screen size) wet milling reduced cellulose crystallinity. Dilute NaOH steeping also produced pits (micropores) through the cell wall structure as shown by scanning electron microscopy (SEM). The micropores seemed to have weakened the cell walls, as revealed by increased cellular disaggregation as viewed by light microscopy. Disaggregation of cassava root cells was associated with a reduction in large (diameter > 250 ?m) residue particle size in the bagasse and consequently more starch yield. Thus, it seems that mechanistically, dilute NaOH solubilisation of alkaline-soluble pectin weakens the cell walls of starch-containing cassava root parenchyma cells. Weakening of cassava cell walls with a combination of biological (14 day endogenous fermentation under microaerophilic conditions) and dilute alkaline pre-treatment (0.75% NaOH steeping) was investigated in an attempt to further increase starch yield by wet milling. However, the combined pre-treatment resulted in approx. 11.8% more starch yield, slightly less than the 12.3% increase obtained by using endogenous fermentation alone. The absence of an additive effect was probably because although endogenous fermentation (retting) and dilute NaOH steeping weakened cassava cell walls through different mechanisms (hydrolysis/solubilisation of pectin), the resultant loss in pectin cohesiveness was similar. Solid state fermentation of ground cassava using various alkaliphilic Bacillus spp. starter cultures separately and in combination was also investigated to determine their extracellular hydrolytic enzyme induced weakening effects on parenchyma cell walls. GC and FTIR data indicated that fermentation with Bacillus akibai + endogenous microflora (EM), B. cellulosilyticus + EM, B. hemicellulosilyticus + EM and B. spp. in combination + EM caused reduction in cell wall pectin, xyloglucan and cellulose contents. Cell wall solubilisation/hydrolysis seemed to have primarily involved the amorphous constituents, as indicated by an increase in cellulose crystallinity by WAXS spectroscopy. Enzyme assay and SEM indicated that Bacillus spp. extracellular cellulase and polygalacturonase weakened the cell walls through formation of micropores and possible rupturing of cellulose microfibril structures. These modifications seemed to have aided disaggregation of the cassava parenchyma cells and consequently liberation of more starch granules as indicated by light microscopy. Fermentation with B. akibai + EM, B. cellulosilyticus + EM, B. hemicellulosilyticus + EM and B. spp. in combination + EM also resulted in less large (diameter > 250 ?m) residue particle size in the bagasse and consequently higher starch yield. Thus, dilute NaOH steeping and fermentation with alkaliphilic Bacillus spp. starter cultures are techniques capable of improving the effectiveness of wet milling in disintegrating cassava cell walls. However, with regard to the demand for environmentally cleaner production, potential utilisation of alkaliphilic Bacillus spp. and more specifically Bacillus cellulosilyticus appears more promising.
Thesis (PhD)--University of Pretoria, 2017.
Food Science
PhD
Unrestricted

Thesis (PhD)--Stellenbosch University, 2014.
ENGLISH ABSTRACT: The development of ―energycane‖ varieties of sugarcane for ethanol production is underway, targeting the use of both sugar juice (first generation ethanol) and bagasse (second generation ethanol). Nevertheless, identification of the preferred varieties represents the biggest challenge to the development of energycane due to large number of samples produced during breeding. In the present study, dilute acid pretreatment, enzymatic hydrolysis and fermentation processes were used to evaluate the processability of bagasse (fibrous residue generated after juice sugar extraction) from different varieties of sugarcane to select preferred varieties with the properties of improving combined ethanol yield (ethanol from juice and bagasse) per hectare. The impact of variety selection on combined ethanol yield (ethanol from juice and bagasse) per hectare was also assessed. In the first part of this study, 115 varieties of sugarcane originated from classical breeding and precision breeding (genetic engineering) were screened based on agronomic data and experimental data from biochemical processes (dilute acid pretreatment and enzymatic hydrolysis) applied to the bagasse fraction of each variety. The results showed wide variations in the chemical composition of bagasse between the varieties. Structural carbohydrates and lignin content ranged from 66.6 to 77.6% dry matter (DM) and 14.4 to 23.1% DM, respectively. The majority of precision breeding varieties showed higher arabinoxylan, lower lignin and lower ash content than most of classical breeding varieties. Combined sugar yield from the bagasse after pretreatment and enzymatic hydrolysis also varied significantly among the varieties. Up to 27.9 g/100g (dry bagasse) difference in combined sugar yield was observed. Combined sugar yield was inversely correlated with lignin as well as ash content, but it correlated positively with structural carbohydrates content. Total potential ethanol yields per hectare, calculated based on cane yield, soluble and non-soluble sugar content also differed significantly among the varieties (8,602−18,244 L/ha). Potential ethanol from bagasse contributed approximately one third of the total potential ethanol yield. Interestingly, some of the varieties had combined properties of high potential ethanol yield per hectare and improved bagasse convertibility. Thus, six varieties (3 from each breeding technology) were selected as preferred varieties for further investigation. To enhance sugar yield from bagasse, optimisation of pretreatment was conducted on the selected varieties. Industrial bagasse was included for comparison purposes. The pretreatment optimisation was based on maximising combined sugar yield from the combined pretreatment-hydrolysis process. A central composite design (CCD) was applied to investigate the effects of temperature, acid concentration and residence time on the responses and was later used to determine the maximum combined sugar yield. Pretreatment optimisation was conducted at gram scale (22.9 ml reactor) and at bench scale (1000 ml reactor). Significant differences in sugar yields (xylose, glucose, and combined sugar) between the varieties were observed. The combined sugar yields from the best performing varieties and industrial bagasse at optimal pretreatment-hydrolysis conditions differed by up to 34.1% and 33% at gram and bench scale, respectively. A high ratio of carbohydrates to lignin and low ash contents increased the release of sugar from the substrates. At mild pretreatment conditions, the differences in bioconversion efficiency between varieties were greater than at severe conditions. This observation suggests that under less severe conditions the conversion efficiency was largely determined by the properties of the biomass. Furthermore, it was demonstrated that the pretreatment conditions with temperature ranged from 184 to 200 °C and varying residence time to provide a severity factor between 3.51 and 3.96 was observed to be the area in common where 95% of maximum combined sugar yield could be obtained. Simultaneous Saccharification and Fermentation (SSF) was performed on the unwashed pressed-slurry from bagasse pretreatment at conditions for maximum combined sugar yield at bench scale. Batch and fed-batch SSF feeding strategy at different solid loadings and enzyme dosages were used aiming to reach an ethanol concentration of at least 40 g/L. The results revealed significant improvement in overall ethanol yield after SSF for the selected varieties (84.5–85.6%) compared to industrial bagasse (74.8%). The maximum ethanol concentration from the best performing varieties was 48.6−51.3 g/l and for poor performing varieties was 37.1−38.3 g/l. Ethanol concentration in the fermentation broth was inversely correlated with lignin content and the ratio of xylose to arabinose, but it showed positive correlation with glucose yield from pretreatment-enzymatic hydrolysis. The overall assessment of the varieties showed greater improvement in combined ethanol yields per hectare (71.1–90.7%) for the best performing varieties with respect to industrial sugarcane. The performance in terms of ethanol yields of selected varieties from a number harvest years was evaluated. The results showed considerable variations in ethanol yields across harvests. The results showed that the best variety in terms combined ethanol yield was not maintained across harvests. The differences in ethanol yields were greater among the varieties than across the harvests. Prolonged severe drought significantly affected the ethnol yields of all varieties represented by lower and intermediate lignin content for cane yield compared to that which had highest lignin content. However, carbohydrates content in the bagasse and sugar yield/recovery between the harvest years did not change for the most of the varieties. In summary, the present study provides evidence of the impact of cultivar selection and pretreatment optimisation in increasing conversion efficiency of bagasse. The results demonstrate that varieties with lower lignin and ash content, as well as highly substituted xylan resulted in higher sugar and ethanol yields. These results suggest that lower process requirements can be achieved without adversely affecting juice ethanol and cane yield per hectare. Nonetheless, an attempt to reduce lignin content in the bagasse, to reduce processing requirements for ethanol production, can also target the improvement of crop tolerance toward severe drought conditions.
AFRIKAANSE OPSOMMING: Die ontwikkeling van ―energie-riet‖ rasse vir etanol produksie is goed op dreef, waar beide die sap (eerste generasie etanol) en die bagasse (tweede generasie etanol) geteiken word. Die groot aantal monsters wat tydens teling geproduseer word, bied egter die grootste uitdaging vir die identifisering van nuwe rasse ten einde energie-riet te ontwikkel. In die huidige studie is verdunde suurvoorbehandeling, ensiematiese hidrolise en fermentasie-prosesse gebruik om die verwerkbaarheid van bagasse (veselagtige residu gegenereer na sap suiker ekstraksie) van verskillende suikerrietrasse te evalueer om nuwe variëteite te selekteer wat eienskappe van verbeterde gekombineerde etanolopbrengs (etanol van sap en bagasse) per hektaar toon. Die impak van variëteit-seleksie op gekombineerde etanol opbrengs (etanol van sap en bagasse) per hektaar is ook beoordeel. In die eerste deel van hierdie studie het uit ‗n siftingsproses van 115 suikerriet rasse bestaan wat deur klassieke en presisie (geneties gemodifiseerde) teling gegenereer is. Die sifting was op agronomiese data gebaseer, asook op data van verdunde suur voorafbehandeling en ensimatiese hidrolise eksperimente wat op die bagasse fraksie van elke ras uitgevoer is. Die resultate het op groot variasie in die chemiese samestelling van die bagasse van verskillende rasse gedui. Die strukturele koolhidrate het tussen 66.6 en 77.6% droë massa (DM) gewissel, terwyl die lignien inhoud ‗n variasie van 14.4 en 23.1% DM getoon het. Verder het meeste van die presisie-teling variëteite ‗n hoër arabinoxilaan, maar ‗n laer lignien en as-inhoud as meeste van die klassieke teling rasse gehad. Die gekombineerde suikeropbrengs (GSO) van die bagasse na voorafbehandeling en ensimatiese hidrolise het ook beduidend tussen rasse gewissel, waar ‗n verskil van tot 27.9 g/100g (droë bagasse) waargeneem is. Daar was ‗n omgekeerde korrelasie tussen die gekombineerde suikeropbrengs en die lignien en as-inhoud gewees, maar die opbrengs het ‗n sterk positiewe korrelasie met die strukturele koolhidrate getoon. Die totale potensiële etanol opbrengs per hektaar wat vanaf die suikerriet se oplosbare en nie-oplosbare suikerinhoud bereken is, het ook beduidend tussen rasse verskil (8,602−18,244 L/ha), waar die potensiële etanol opbrengs van die bagasse gedeelte ongeveer een derde van die totale potensiële etanol opbrengs beslaan het. Interessante bevindinge het op sommige rasse met gekombineerde eienskappe van hoë potensiële opbrengs per hektaar asook ‗n hoë omskakelingsvermoë gedui. Derhalwe is ses variëteite (drie van elke telingstegnologie) as voorkeurvariëteite vir verdere studie gekies. Om die etanol opbrengs vanaf die bagasse te verbeter was voorafbehandeling van die voorkeurvariëteite geoptimeer, en waar industriële bagasse vir vergelykingsdoeleindes ingesluit was. Vir die optimering was dit ten doel gestel om die gekombineerde suikeropbrengs van die gekombineerde voorafbehandeling-hidrolise proses te maksimeer. ‗n Sentrale saamgestelde ontwerp (SSO) is gebruik om die effek van temperatuur, suurkonsentrasie en residensietyd op die responsveranderlikes vas te stel wat uiteindelik gebruik is om die maksimum gekombineerde suikeropbrengs te bepaal. Die optimering van die voorafbehandeling is op gram-skaal in ‗n 22.9 ml reaktor, asook op bank-skaal in ‗n 1000 ml reaktor uitgevoer. Beduidende verskille in die suikeropbrengs (xilose, glukose en gekombineerde suiker) is tussen die voorkeurrasse waargeneem. Tussen die rasse wat die beste gevaar het, asook die industriële bagasse, het die gekombineerde suikeropbrengs by optimale voorafbehandeling-hidrolise toestande onderskeidelik met tot 34.1% en 33% op gram-skaal en bank-skaal gevarieer. ‗n Hoë verhouding van koolhidrate tot lignien, asook ‗n lae as-inhoud het tot ‗n toename in die vrystelling van suiker uit die substraat gelei. By matige voorafbehandelingstoestande was die verskille in omskakelingseffektiwiteit tussen rasse groter as onder hewige toestande, wat daarop gedui het dat omskakelingseffektiwiteit grotendeels deur die eienskappe van die biomassa bepaal is. Verder is daar ook gedemonstreer dat die voorbehandelingsomstandighede met temperatuur tussen 184 en 200ºC en verandering van die residensietyd om 'n hewigheidsfaktor van tussen 3.51 en 3.96 te verskaf, 'n gemeenskaplike area gelewer het waar 95% van maksimum gekombineer suiker opbrengs (GSO) verkry kon word. Gelyktydige versuikering en fermentasie (GVF) is na voorafbehandeling op ongewaste, gepersde bagasse substraat by toestande vir die maksimum gekombineerde suikeropbrengs op bank-skaal uitgevoer. Bondel en voerbondel SSF voerstrategie by verskillende vaste ladings en ensiemdoserings is gebruik om 'n etanol konsentrasie van ten minste 40 g/L te bereik. Ná GVF was die algehele etanol opbrengs vir die voorkeurvariëteite (84.5–85.6%) beduidend beter relatief tot die industriële bagasse (74.8%). Die maksimum etanol opbrengs na SSF van die rasse met die beste prestasie was 48.6-51.3 g/L en 37.1-38.3 g/L vir rasse wat swak presteer het. Die etanol konsentrasie in die fermentasiesop was omgekeerd met lignien en die verhouding van xilose tot arabinose gekorreleer, maar was duidelik positief met die glukose opbrengs vanaf voorafbehandeling-hidrolise gekorreleer. ‗n Algemene assessering het op ‗n duidelike verbetering van die voorkeurvariëteite in terme van gekombineerde etanol opbrengs per hektaar gedui (71.1–90.7%), relatief tot die industriële suikerriet. Die prestasie in terme van etanol opbrengs van geselekteerde variëteite is oor 'n reeks oesjare ge-evalueer. Die resultate het aansienlike variasies in etanol opbrengs oor oesjare getoon. Die resultate het gewys dat die beste variëteite in terme van gekombineerde etanol opbrengs nie volhou is oor oeste nie. Die verskille in etanol opbrengste tussen variëteite was groter as die verskille oor oesjare. Verlengde ernstige droogte het die etanol opbrengs van alle variëteite met laer en intermediere lignien inhoud vir rietopbrengs aansienlik beinvloed, in vergelyking met dié wat die hoogste lignien inhoud gehad het. Die koolhidraatinhoud in die bagasse en suiker opbrengs/lewering tussen die oesjare het vir die meeste variëteite egter nie gewissel nie. Ter opsomming, die huidige studie verskaf bewyse van die impak van kultivarseleksie en voorbehandelings optimisering op die verhoging van die omskakelings-doeltreffendheid van bagasse. Die resultate wys dat variëteite met laer lignien- en asinhoud, en hoogs-gesubstitueerde xilaan hoër suiker- en etanol opbrengs gelewer het. Hierdie resultate stel voor dat verminderde voorbehandelingsvereistes bereik kan word sonder om die sap etanol en rietopbrengs per hektar te benadeel. Nieteenstaande, 'n poging om die lignien inhoud van die bagasse te verminder om die verwerkingsvereistes vir etanolproduksie te verminder, kan ook die verbetering van gewas-toleransie tov ernstige droogte-toestande teiken.

The fields of biodiesel and bioethanol research and development have largely developed independently of one another. Opportunities exist for greater integration of these processes that may result in decreased costs of production for both fuels.To that end, this work addresses the use of the starches and glycerol from processed algal biomass as substrates for fermentation by the yeasts Saccharomyces cerevisiae and Pachysolen tannophilus, respectively. Ethanol producers commonly employ the former yeast for ethanol production and include the latter yeast among candidate species for cellulosic ethanol production.A simple 95% ethanol extraction at 70°C followed by sulfuric acid hydrolysis at 121°C and 2 atm proved a sufficient pretreatment for S. cerevisiae fermentation of starch from Chlamydomonas reinhardtii mutant cw15. The maximum rate of ethanol production was observed as 14 mL/g-h and a maximum concentration of 0.9±0.01% (m/v) was observed by 28 hours. Some starch appeared invulnerable to hydrolysis.P. tannophilus fermentation of glycerol, both independently and among mixed substrates, was likewise demonstrated. It was found that glucose consumption preceded that of glycerol and xylose, but that the latter two substrates were consumed concurrently. Under aerobic, batch conditions, the maximum specific growth rate of the species on a 2% glycerol substrate was observed as 0.04/hr and the yield coefficient for conversion of glycerol to ethanol was 0.07 g/g. While the maximum observed concentration of ethanol in the glycerol-only fermentation was 0.1% m/v, that in mixed media containing 2% each glucose, xylose, and glycerol was 1.5%.Also investigated here was the flocculation of a mutant species of the algae C. reinhardtii by a combination of methanol and calcium. Algae harvest is typically an energy-intensive process, but the technique demonstrated here is not. Complete flocculation of cells was observed with only 5 minutes of mixing and less than 10 minutes of settling using 12 mM CaCl₂ and 4.6% methanol. Ethanol was observed to operate in the same capacity, intimating another area in which yeast bioethanol and algal biodiesel processes might enable one another. During growth, either an inhibitor of flocculation was produced or a facilitator was consumed.

During organic material degradation in oxic environments, electrons from organic material, the electron donor, are transferred to oxygen, the electron acceptor, during aerobic respiration. Other compounds, such as nitrate, iron, sulfate, and carbon dioxide, take the place of oxygen during anaerobic respiration in anoxic environments. The order in which these compounds are used by bacteria and archaea (only a few eukaryotes are capable of anaerobic respiration) is set by thermodynamics. However, concentrations and chemical state also determine the relative importance of electron acceptors in organic carbon oxidation. Oxygen is most important in the biosphere, while sulfate dominates in marine systems, and carbon dioxide in environments with low sulfate concentrations. Nitrate respiration is important in the nitrogen cycle but not in organic material degradation because of low nitrate concentrations. Organic material is degraded and oxidized by a complex consortium of organisms, the anaerobic food chain, in which the by-products from physiological types of organisms becomes the starting material of another. The consortium consists of biopolymer hydrolysis, fermentation, hydrogen gas production, and the reduction of either sulfate or carbon dioxide. The by-product of sulfate reduction, sulfide and other reduced sulfur compounds, is oxidized back eventually to sulfate by either non-phototrophic, chemolithotrophic organisms or by phototrophic microbes. The by-product of another main form of anaerobic respiration, carbon dioxide reduction, is methane, which is produced only by specific archaea. Methane is degraded aerobically by bacteria and anaerobically by some archaea, sometimes in a consortium with sulfate-reducing bacteria. Cultivation-independent approaches focusing on 16S rRNA genes and a methane-related gene (mcrA) have been instrumental in understanding these consortia because the microbes remain uncultivated to date. The chapter ends with some discussion about the few eukaryotes able to reproduce without oxygen. In addition to their ecological roles, anaerobic protists provide clues about the evolution of primitive eukaryotes.

Processing of biomass for obtaining of liquid ethanol, platform chemicals and solid biofuel, is topical biorefinery schema intensively developing. Acid hydrolysis, separated hydrolysis and fermentation, and simultaneous saccharification and fermentation are integrated with other treatments for the advanced technology development. The aim of this work was estimation of effect of softwood and wheat straw hydrolysis type on fuel characteristics of rich-in-lignin residues with emphasis on content and component composition of ash and feedstock. Elemental Analysis; Atomic absorption spectroscopy; calorimetric method; Klason lignin determination were used. Laboratory scale pellet mill KAHL 14-175 and original small pilot-scale gasifier were used for study of residues granulation ability and combustion behavior of pellets obtained. The ash content in softwood residues slightly increase but does not exceed 1%, for wheat straw residues it is >14% (0,2% and 10% for feedstock, correspondingly). The ashing temperature of 650oC is experimentally established as optimal. Detection of ash component allows to foreseen possible contamination connected with materials of the devices and chemicals used in the technological stream. Combustion mechanism of solid residues differs from that of feedstock by increasing of ratio duration of glue combustion to flame combustion steps, that is more characteristic for coal. The efficiency of biomass combustion was regulated by changing the ratio of primary and secondary air supply. Direct correlation established between higher heating value and Klason lignin content for samples, allows to recommend these analyses for evaluation of biomass potential as a fuel. Residues under study meet the requirements of EU Standard CN/TS 335.