Comparison of bioethanol production of simultaneous saccharification & fermentation and separation hydrolysis & fermentation from cellulose-rich barley straw

Cellulose rich barley straw, which has a glucan content of 62.5%, followed by dilute acid pretreatment, was converted to bioethanol by simultaneous saccharification and fermentation (SSF). The optimum fractionation conditions for barley straw were an acid concentration of 1% (w/v), a reaction temperature of 158 °C and a reaction time of 15 min. The maximum saccharification of glucan in the fractionated barley straw was 70.8% in 72 h at 60 FPU/gglucan, while the maximum digestibility of the untreated straw was only 18.9%. With 6% content WIS (water insoluble solid) for the fractionated barley straw, 70.5 and 83.2% of the saccharification yield were in SHF and SSF (representing with glucose equivalent), respectively, and a final ethanol concentration of 18.46 g/L was obtained under the optimized SSF conditions: 34 °C with 15 FPU/g-glucan enzyme loading and 1 g dry yeast cells/L. The results demonstrate that the SSF process is more effective than SHF for bioethanol production by around 18%.     

Different process configurations for bioethanol production from pretreated olive pruning biomass

BACKGROUND: In Mediterranean countries, olive tree pruning provides a widely available renewable agricultural residue with, currently, no industrial application. This residue could provide feedstock for the bioethanol industry. In the present study, olive tree pruning biomass pretreated with both ‘liquid hot water’ and ‘dilute‐sulfuric acid’ was tested as a substrate for ethanol production. Three different process configurations, separate hydrolysis and fermentation (SHF), simultaneous saccharification, fermentation and prehydrolysis (PSSF), and simultaneous saccharification and fermentation (SSF), were compared at different water‐insoluble solids concentrations. RESULTS: High ethanol concentration of about 3.7% (v/v) was obtained by separate hydrolysis and fermentation or prehydrolysis and simultaneous saccharification and fermentation of liquid hot water pretreated at 23% (w/w) substrate loading. CONCLUSION: The nature of the pretreated residue allows high substrate concentration (≥17% w/w) to be used in the enzymatic hydrolysis step. Substrate loading of 17% DM has been shown to provide a compromise between hydrolysis efficiency and glucose concentrations for the same enzyme/substrate ratio. Prehydrolysis prior to simultaneous saccharification and fermentation facilitated SSF performance at high substrate loading on liquid hot water pretreated olive pruning residue. This effect was not observed with dilute‐acid pretreated substrate. Copyright © 2011 Society of Chemical Industry     

Bioethanol production from wheat straw by the thermotolerant yeast Kluyveromyces marxianus CECT 10875 in a simultaneous saccharification and fermentation fed-batch process

Solid content in the simultaneous saccharification and fermentation (SSF) broth should be as high as possible in order to reach higher ethanol concentration. In this work, several feeding strategies for ethanol production from steam-exploded wheat straw by Kluyveromyces marxianus CECT 10875 have been studied with the aim of obtaining higher ethanol concentrations. Previous fermentability tests as well as SSF processes showed the difficulty of using the slurry for ethanol production under the studied conditions. Notwithstanding, fed-batch SSF processes with water-insoluble solids (WIS) fraction resulted in better configuration, reaching the highest ethanol concentration (36.2 g/L) with an initial WIS content of 10% (w/v) and 4% (w/v) of substrate addition at 12 h, which meant 20% more ethanol when compared with batch SSF.     

高底物浓度纤维乙醇同步糖化发酵工艺的比较

引言日益加剧的能源危机和环境污染,正迫使人们寻求新的可再生替代能源。纤维乙醇作为一种重要的生物质替代能源,经过近40多年的发展,已经具备了实现工业化生产的潜力。为了进一步降低纤     

蒸汽爆破麦草同步糖化发酵转化乙醇的研究

近年来对木质生物资源同步糖化发酵转化乙醇的研究较多,但是,麦草同步糖化发酵转化乙醇的最佳工艺条件还未确定。文中采用正交试验设计的方法,对在混合酶(纤维素酶Celluclast 1.5 1,β-葡萄糖苷酶Novozym 188)与酿酒酵母菌作用下,稀硫酸催化的蒸汽爆破麦草原料同步糖化发酵转化乙醇的工艺条件进行研究,详细讨论了反应温度、底物质量浓度、发酵液pH值、纤维素酶浓度对乙醇质量浓度和得率的影响。结果表明,工艺条件对乙醇质量浓度和得率的影响程度由高到低依次为:底物质量浓度、纤维素酶浓度、发酵液pH值、反应温度。最佳工艺条件为反应温度35℃,底物质量浓度100 g/L,发酵液pH值5.0,纤维素酶浓度30 FPU/g。在此条件下,随着反应时间的延长,乙醇质量浓度持续上升。反应72 h后,乙醇质量浓度和得率分别达到22.7 g/L和65.8%。     

Study of simultaneous saccharification and fermentation for steam exploded wheat straw to ethanol

Although simultaneous saccharification and fermentation (SSF) has been investigated extensively, the optimum condition for SSF of wheat straw has not yet been determined. Dilute sulfuric acid impregnated and steam explosion pretreated wheat straw was used as a substrate for the production of ethanol by SSF through orthogonal experiment design in this study. Cellulase mixture (Celluclast 1.5 l and ?-glucosidase Novozym 188) were adopted in combination with the yeast Saccharomyces cerevisiae AS2.1. The effects of reaction temperature, substrate concentration, initial fermentation liquid pH value and enzyme loading were evaluated and the SSF conditions were optimized. The ranking, from high to low, of influential extent of the SSF affecting factors to ethanol concentration and yield was substrate concentration, enzyme loading, initial fermentation liquid pH value and reaction temperature, respectively. The optimal SSF conditions were: reaction temperature, 35°C; substrate concentration, 100 g·L^-1; initial fermentation liquid pH, 5.0; enzyme loading, 30 FPU·g^-1. Under these conditions, the ethanol concentration increased with reaction time, and after 72 h, ethanol was obtained in 65.8% yield with a concentration of 22.7 g·L^-1.     

Production of fuel ethanol from softwood by simultaneous saccharification and fermentation at high dry matter content

BACKGROUND: The production of bio‐ethanol from softwood is considered a promising alternative to fossil fuels in Sweden. In order to make fuel ethanol economically competitive with fossil fuels, it is important to reduce the production cost, which can be done by increasing the dry matter content of the fermentation medium, thus reducing the energy demand in the final distillation of the fermentation broth. Running simultaneous saccharification and fermentation (SSF) at higher dry matter content has, however, been found to decrease the ethanol yield. RESULTS: The use of different stirrer types and stirring speeds in the present study has shown to have an influence on the final ethanol yield in SSF with 10% water‐insoluble solids (WIS). Also, higher concentration of pretreatment hydrolysate, i.e., with increased inhibitor concentration, at the same WIS resulted in a decreased ethanol yield. However, despite stirring problems and high inhibitor concentration, ethanol was produced at 12% WIS with an ethanol yield in the SSF step of 81% of the theoretical based on the content of fermentable sugars in the fermentor. CONCLUSION: The decrease in ethanol yield in SSF at high dry matter content has been shown to be a combined effect of increased mass transfer resistance and increased inhibitor concentration in the fermentation broth. Copyright © 2008 Society of Chemical Industry     

Tween80对稻草水解及同步糖化与发酵产乳酸的影响

在生物转化纤维原料产乳酸的过程中,酶解纤维原料产还原糖是限速步骤。为了获得较高的产物产率,需较高的酶用量,这使大规模酶解废弃纤维原料的成本很高。对吐温80在酶解稻草纤维素产糖,以及耐高温乳酸菌同步糖化发酵稻草产乳酸过程中的作用进行了考察。初步结果表明,吐温80加入可使保持同等程度的水解率所需的酶用量降低,添加0.2 g/g底物的吐温80到酶用量10 FPU/g体系,水解120 h的糖产率为292.2 mg/g,比不加表面活性剂体系的糖产率增加了11%;添加0.7 g/L的吐温80进行同步糖化与发酵72 h,能使乳酸产量提高24.2%。     

木质纤维生物质同步糖化发酵(SSF)生产乙醇的研究进展

综述了有关木质纤维生物质原料同步糖化发酵生产乙醇的最新研究进展和未来发展方向:同步糖化发酵是一种用于从木质纤维原料生产乙醇的工艺过程,此工艺的优点是酶水解与发酵同时进行,可以减少最终产物对酶水解的抑制作用,并减少投资成本,是最具发展潜力和优势的工艺之一。近年来在优化预处理工艺、降低纤维素酶成本以及己糖戊糖协同发酵等方面的研究都取得了长足的进步,其中以小麦秸秆为原料进行同步糖化发酵所得到的乙醇浓度接近40g/L。     

Pretreatment of barley husk for bioethanol production

This paper reports on the optimization of steam pretreatment of barley husk for high pentose and hexose recovery in the subsequent enzymatic hydrolysis step, as well as high ethanol yield, following simultaneous saccharification and fermentation. The parameters optimized in the steam pretreatment step were residence time (5–15 min), temperature (190–215 °C), and concentration of the acid catalyst (0 or 0.5% H₂SO₄). A microwave oven was employed for screening of the optimal conditions to obtain the highest sugar yield following combined pretreatment and enzymatic hydrolysis. The final optimization of the pretreatment prior to enzymatic hydrolysis was performed on a larger scale, in a steam pretreatment unit. Simultaneous saccharification and fermentation was carried out following steam pretreatment on 5 and 10% dry matter steam‐pretreated slurries. Fermentability tests were performed to determine the effect of by‐products (ie furfural and 5‐hydroxymethyl furfural) in the bioconversion of glucose to ethanol by baker's yeast. The maximum glucose yield, 88% of the theoretical, was obtained following steam pretreatment with 0.5% H₂SO₄ at 200 °C for 10 min. Under these conditions, a sugar to ethanol conversion of 81% was attained in simultaneous saccharification and fermentation. Copyright © 2004 Society of Chemical Industry     

An approach to optimization of enzymatic hydrolysis from sugarcane bagasse based on organosolv pretreatment

BACKGROUND: The organosolv pretreatment followed by enzymatic hydrolysis of the pretreated material and subsequent fermentation of the hydrolysate produced, was the strategy used for ethanol production from sugarcane bagasse. The effect of different operational variables affecting the pretreatment (the catalyst type and its concentration, and the pretreatment time) and enzymatic hydrolysis stage (substrate concentration, cellulase loading, addition of xylanase and Tween 20, and the cellulase/β‐glucosidase ratio), were investigated. RESULTS: The best values of glucose concentration (28.8 g L^?1) and yield (25.1 g per 100 g dry matter) were obtained when the material was pretreated with 1.25% (w/w) H₂SO₄ for 60 min, and subsequently hydrolyzed using 10% (w/v) substrate concentration in a reaction medium supplemented with xylanase (300 UI g^?1) and Tween 20 (2.5% w/w). Fermentation of the broth obtained under these optimum conditions by Saccharomyces cerevisiae resulted in an ethanol yield of 92.8% based on the theoretical yield, after 24 h. CONCLUSION: Organosolv pretreatment of sugarcane bagasse under soft conditions, and subsequent enzymatic hydrolysis of the pretreated material with a cellulolytic system supplemented with xylanase and Tween 20, is a suitable procedure to obtain a glucose rich hydrolysate efficiently fermentable to ethanol by Sacharomyces cerevisiae yeasts. Copyright © 2010 Society of Chemical Industry     

A comprehensive ligninolytic pre-treatment approach from lignocellulose green biotechnology to produce bio-ethanol

In the present study, we developed a novel ligninolytic enzymes based pre-treatment method for lignocellulosic wheat straw to depolymerize lignin and expose the cellulose polymers to produce bio-ethanol. Wheat straw was pre-treated with ligninolytic enzymes extract produced from Ganoderma lucidum under optimum solid state fermentation conditions. The pre-treated biomass was further subjected to the enzymatic hydrolysis by the crude unprocessed cellulases (β-1,4 endoglucanase, 53.5 ± 1.24 U/mL; β-1,4 exoglucanase, 41.3 ± 1.31 U/mL; β-1,4 glucosidase, 46.8 ± 1.43 U/mL; and xylanase 39 ± 2.2 U/mL) produced by Trichoderma harzaianum. Under optimal conditions for enzymatic saccharification, 10% (w/v) of pre-treated biomass was hydrolyzed completely and converted to 72.5 and 2.4 g/L of glucose and xylose, respectively. Initial time screening Sequential Saccharification and Fermentation (SSF) of the concentrated enzymatic hydrolyzate (10%, w/v) by Saccharomyces cerevisiae produced 22.6 g/L ethanol in a fermented medium after 72 h of temperature controlled incubation at 37 °C. For maximum ethanol production, different physical and nutritional parameters like pH, temperature, substrate level and inoculum sizes were optimized. Under optimal conditions ethanol production of 33.5 g/L was obtained from ligninolytic treated residual (wheat straw) biomass.     

麦秸秆的氢氧化钙预处理及酶解试验研究

采用氢氧化钙对麦秸秆进行预处理,以酶解还原糖得率为目的,分别优化预处理及酶解条件。结果表明,氢氧化钙预处理麦秸秆的最佳条件是:Ca(OH)2添加量为0.06g/g(对秸秆),固液比为1:10,在120℃下反应时间为2h;最佳酶解条件是:温度50℃,pH4.8,纤维素酶17FPU/g(对秸秆),木聚糖酶160IU/g,在添加0.15g/g(对秸秆)Tween80条件下,酶解液中还原糖质量浓度为62.32g/L,酶解还原糖得率达85.23%。     

Study of simultaneous saccharification and fermentation for steam exploded wheat straw to ethanol

Although simultaneous saccharification and fermentation (SSF) has been investigated extensively, the optimum condition for SSF of wheat straw has not yet been determined. Dilute sulfuric acid impregnated and steam explosion pretreated wheat straw was used as a substrate for the production of ethanol by SSF through orthogonal experiment design in this study. Cellulase mixture (Celluclast 1.5 l and β-glucosidase Novozym 188) were adopted in combination with the yeast Saccharomyces cerevisiae AS2.1. The effects of reaction temperature, substrate concentration, initial fermentation liquid pH value and enzyme loading were evaluated and the SSF conditions were optimized. The ranking, from high to low, of influential extent of the SSF affecting factors to ethanol concentration and yield was substrate concentration, enzyme loading, initial fermentation liquid pH value and reaction temperature, respectively. The optimal SSF conditions were: reaction temperature, 35°C; substrate concentration, 100 g·L⁻¹; initial fermentation liquid pH, 5.0; enzyme loading, 30 FPU·g⁻¹. Under these conditions, the ethanol concentration increased with reaction time, and after 72 h, ethanol was obtained in 65.8% yield with a concentration of 22.7 g·L⁻¹. __________ Translated from Chemical Engineering (China), 2007, 35(12): 42–45 [译自: 化学工程]     

FED-BATCH SIMULTANEOUS SACCHARIFICATION AND FERMENTATION OF MICROWAVE/ACID/ALKALI/H2O2 PRETREATED RICE STRAW FOR PRODUCTION OF ETHANOL

The batch simultaneous saccharification and fermentation (SSF) of microwave/acid/alkali/H₂O₂ pretreated rice straw to ethanol was optimized using cellulase from Trichoderma reesei and Saccharomyces cerevisiae YC-097 cells prior to the fed-batch SSF studies. The batch SSF optima were 10% w/v substrate, 40°C, 15 mg cellulase/g substrate, initial pH 5.3, and 72 hours. Under the optimum conditions the ethanol concentration and its yield were 29.1 g/L and 61.3% respectively. Based on the optimal batch SSF, the fed-batch SSF was investigated and its operation parameters were optimized. Under its optimal conditions the ethanol concentration reached 57.3 g/L, while its productivity and yield were only slightly less than those in the batch SSF. This suggests that fed-batch SSF is a potential operation mode for effective ethanol production from microwave/acid/alkali/H₂O₂ pretreated rice straw.     

Enzymatic hydrolysis and fermentation of lime pretreated wheat straw to ethanol

BACKGROUND: The objective of this work is to develop an efficient pretreatment method that can help enzymes break down the complex carbohydrates present in wheat straw to sugars, and to then ferment of all these sugars to ethanol. RESULTS: The yield of sugars from wheat straw (8.6%, w/v) by lime pretreatment (100 mg g^?1 straw, 121 °C, 1 h) and enzymatic hydrolysis (45 °C, pH 5.0, 120 h) using a cocktail of three commercial enzyme preparations (cellulase, β‐glucosidase, and xylanase) at the dose level of 0.15 mL of each enzyme preparation g^?1 straw was 568 ± 13 mg g^?1 (82% yield). The concentration of ethanol from lime pretreated enzyme saccharified wheat straw (78 g) hydrolyzate by recombinant Escherichia coli strain FBR5 at pH 6.5 and 35 °C in 24 h was 22.5 ± 0.6 g L^?1 with a yield of 0.50 g g^?1 available sugars (0.29 g g^?1 straw). The ethanol concentration was 20.6 ± 0.4 g L^?1 with a yield of 0.26 g g^?1 straw in the case of simultaneous saccharification and fermentation by the E. coli strain at pH 6.0 and 35 °C in 72 h. CONCLUSION: The results are important in choosing a suitable pretreatment option for developing bioprocess technologies for conversion of wheat straw to fuel ethanol. Copyright © 2007 Society of Chemical Industry     

变温调控对汽爆秸秆发酵产氢的影响

在以汽爆秸秆为原料,利用丁酸梭菌(Clostridium butyricum)同步糖化发酵产氢的过程中,存在着纤维素酶解和微生物发酵的最适温度不一致的矛盾,影响底物的转化和微生物利用。在发酵开始 10 h 和 30 h 的时候,分别进行两次变温调控,首先迅速升温至 45℃,保持 2 h 后又迅速将温度降低回到丁酸梭菌的最适发酵温度 35℃。这一变温调控能够促进纤维素酶解,并有利于微生物保持高活性。与恒温发酵相比,两次变温的最终产氢量提高1.2倍,纤维素和半纤维素的转化率分别为 75%和 88%。     

木薯酒精渣的预处理及补料同步糖化发酵制取乙醇

木薯酒精渣的处置是制约木薯燃料乙醇大规模产业化的问题之一。本文立足于探索木薯酒精渣利用途径,分析了木薯酒精渣的主要成分,对比了氨水、氢氧化钠、氨水组合稀硫酸3种预处理方式对于木薯酒精渣纤维素和木素含量及纤维素酶水解效率的影响,分析了处理前后木薯酒精渣的表面结构及纤维素结晶度,并以氨水处理后的木薯酒精渣为底物,进行了同步糖化发酵。结果表明,3种预处理方法中组合预处理能更好地增加纤维素含量和提高纤维素酶水解效率,与未处理原料相比,组合预处理后纤维素含量增加了111.26%,木素下降了35.05%,酶水解72h纤维素转化率从42.10%增加到61.71%。氨水预处理后,原料的木素含量降低,处理后木薯酒精渣的表面变得更加粗糙,纤维素结晶度有所增加,以氨水处理后的木薯酒精渣为底物进行分批补料同步糖化发酵,当初始底物浓度为100.0g/L,分别在20h、40h、60h进行补料至最终底物浓度为400.0g/L时,发酵120h乙醇浓度达到51.0g/L。     

Simultaneous saccharification and fermentation of alkaline-pretreated corn stover to ethanol using a recombinant yeast strain

Bio-ethanol converted from cheap and abundant lignocellulosic materials is a potential renewable resource to replace depleting fossil fuels. Simultaneous saccharification and fermentation (SSF) of alkaline-pretreated corn stover for the production of ethanol was investigated using a recombinant yeast strain Saccharomyces cerevisiae ZU-10. Low cellobiase activity in Trichoderma reesei cellulase resulted in cellobiose accumulation. Supplementing the simultaneous saccharification and fermentation system with cellobiase greatly reduced feedback inhibition caused by cellobiose to the cellulase reaction, thereby increased the ethanol yield. 12 h of enzymatic prehydrolysis at 50 °C prior to simultaneous saccharification and fermentation was found to have a negative effect on the overall ethanol yield. Glucose and xylose produced from alkaline-pretreated corn stover could be co-fermented to ethanol effectively by S. cerevisiae ZU-10. An ethanol concentration of 27.8 g/L and the corresponding ethanol yield on carbohydrate in substrate of 0.350 g/g were achieved within 72 h at 33 °C with 80 g/L of substrate and enzyme loadings of 20 filter paper activity units (FPU)/g substrate and 10 cellobiase units (CBU)/g substrate. The results are meaningful in co-conversion of cellulose and hemicellulose fraction of lignocellulosic materials to fuel ethanol.     

Production of bioethanol from corn meal hydrolyzates

The two-step enzymatic hydrolysis of corn meal by commercially available α-amylase and glucoamylase and further ethanol fermentation of the obtained hydrolyzates by Saccharomyces cerevisiae yeast was studied. The conditions of starch hydrolysis such as substrate and enzyme concentration and the time required for enzymatic action were optimized taking into account both the effects of hydrolysis and ethanol fermentation. The corn meal hydrolyzates obtained were good substrates for ethanol fermentation by S. cerevisiae. The yield of ethanol of more than 80% (w/w) of the theoretical was achieved with a satisfactory volumetric productivity P (g/l h). No shortage of fermentable sugars was observed during simultaneous hydrolysis and fermentation. In this process, the savings in energy by carrying out the saccharification step at lower temperature (32 °C) could be realized, as well as a reduction of the process time for 4 h.