Chemical characteristics and ethanol fermentation of the cellulose component in autohydrolyzed bagasse

The chemical characteristics, enzymatic saccharification, and ethanol fermentation of autohydrolyzed lignocellulosic material that was exposed to steam explosion were investigated using bagasse as the sample. The effects of the steam explosion on the change in pH, organic acids production, degrees of polymerization and crystallinity of the cellulose component, and the amount of extractive components in the autohydrolyzated bagasse were examined. The steam explosion decreased the degree of polymerzation up to about 700 but increased the degree of crystallinity and the micelle width of the cellulose component in the bagasse. The steam explosion, at a pressure of 2.55 MPa for 3 mins, was the most effective for the delignification of bagasse. 40 g/L of glucose and 20 g/L of xylose were produced from 100 g/L of the autohydrolyzed bagasse by the enzymatic saccharification using mixed cellulases, acucelase and meicelase. The maximum ethanol concentration, 20 g/L, was obtained from the enzymatic hydrolyzate of 100 g/L of the autohydrolyzed bagasse by the ethanol fermentation usingPichia stipitis CBS 5773; the ethanol yield from sugars was 0.33 g/g sugars.     

Characterization of the steam-exploded spent Shiitake mushroom medium and its efficient conversion to ethanol

Spent Shiitake mushroom medium was subjected to steam explosion followed by simultaneous saccharification and fermentation (SSF) using Meicelase and Saccahromyces cerevisiae AM12. Water extraction of the medium exposed to steam at 20 atm for 5 min enhanced the saccharification rate by about 20% compared to steam-exploded medium before water extraction and resulted in the production of 23.8 g/l ethanol from a substrate concentration of 100 g/l. This corresponded to 87.6% of the theoretical ethanol yield, i.e., 15.9 g ethanol was obtained from 100 g of spent Shiitake mushroom medium. Spent Shiitake mushroom medium subjected to steam explosion and then water extraction appears to be a candidate for efficient bioconversion to ethanol.     

沙柳原料高浓度底物酶解发酵乙醇工艺

为了提高沙柳生物转化过程的经济可行性,考察了沙柳原料经过蒸爆、超微粉碎+稀酸、超微粉碎+稀碱预处理后高浓度底物补料酶解的效果,并对其高浓度水解糖液进行了乙醇发酵。结果表明:蒸爆处理法水解效果最好,通过补料酶解,底物质量分数可以达到30%,酶解液中总糖质量浓度达到132 g/L,葡萄糖质量浓度105 g/L;超微粉碎+稀酸预处理原料底物质量分数可以达到22%,酶解液中总糖质量浓度达到123 g/L,葡萄糖质量浓度73 g/L;超微粉碎+稀碱预处理原料底物质量分数可以达到22%,酶解液中总糖质量浓度133 g/L,葡萄糖质量浓度77 g/L。3种预处理使沙柳原料的酶解糖液都可以较好地被酿酒酵母利用发酵产乙醇,蒸爆处理原料的酶解糖液乙醇发酵效果最好,乙醇质量浓度达到47 g/L。     

木薯纤维素乙醇发酵的纤维素酶成本评价

木薯中的纤维素成分约占木薯干重的10％(W/W).文中以木薯燃料乙醇生产的木薯纤维素酒渣为原料,从纤维素酶成本角度评估了三种利用木薯纤维素组分发酵生产乙醇的方法,包括木薯纤维素酒渣的直接糖化和乙醇发酵、木薯纤维素酒渣预处理后的糖化与乙醇发酵、木薯乙醇发酵中同步淀粉与纤维素糖化以及乙醇发酵.结果表明,前两种方法的纤维素利用效率不高,酶成本分别达到13602、11659元/吨乙醇.第三种方法,即在木薯乙醇发酵过程同时加入糖化酶和纤维素酶,进行同步淀粉与纤维素糖化,进而进行乙醇发酵,木薯纤维素乙醇的收益最高.发酵结束时的乙醇浓度从101.5g/L提高到107.0g/L,纤维素酶成本为3 589元/吨乙醇.此方法利用木薯纤维素与木薯淀粉同时进行,不会带来额外的设备及操作投入,酶成本低于产品乙醇价格,可实现盈利,因此第三种方法为木薯纤维用于乙醇发酵的最适方法,本研究结果将为木薯乙醇产业深度利用木薯纤维提供依据.     

湿氧化爆破稻秆的同步糖化发酵条件研究

探讨了木质纤维素经过湿氧化爆破后在同步糖化发酵过程中酵母产乙醇的基本规律.采用单因素方法对湿氧化爆破条件、酶系组成和添加量以及预酶解时间和温度进行了优化.不同湿氧化爆破预处理条件下的稻秆对同步糖化发酵工艺的影响较大,在预处理温度160 ℃,进氧压力为4×105 Pa,碱用量为6%(w/w),反应时间为20 min的条件...     

Immobilization of co-culture of Saccharomyces cerevisiae and Scheffersomyces stipitis in sodium alginate for bioethanol production using hydrolysate of apple pomace under separate hydrolysis and fermentation

Apple pomace as a substrate for bioethanol production is interesting due to its abundance and sustainable availability in varied states like Himachal Pradesh (H.P.), Jammu and Kashmir, Uttarakhand and Arunachal Pradesh, India. In the current study, apple pomace which is the main fruit industrial waste of H.P. was evaluated as feedstock for bioethanol production by the process of enzymatic saccharification using multiple carbohydrases. Microwave pretreatment of the apple pomace resulted in the efficient removal of lignin and crystalline structure of cellulose fibre. The enzymatic saccharification of the pretreated biomass was done by optimizing parameters for maximal saccharification leads to production of 27.50?mg/g of reduce, ng sugar. An enhanced ethanol yield of 44.46?g/l and fermentation efficiency of 58% by immobilized co-culture of Saccharomyces cerevisiae MTCC 3089 and Scheffersomyces stipitis NCIM 3498 under SHF as compared to fermentation performed with free yeast cells, i.e. 34.46?g/l of ethanol and 45% of fermentation efficiency.     

Ethanol production from corn cob pretreated by the ammonia steeping process using genetically engineered yeast

Summary A new and effective pretreatment process for biomass conversion involves the steeping of biomass in 2.9 M NH₄OH. This resulted in the removing about 80–90% of the lignin along with almost all the acetate from cellulosic residues. Based on dry cellulose from corn cob, a high glucose yield of 92% was obtained after enzymatic saccharification of cellulose fraction. By using a genetically engineered, xylosefermenting Saccharomyces 1400(pLNH33) in the batch fermentation of a glucose-xylose mixture from corn cob, an ethanol concentration of 47 g/L was obtained within 36 h with 84% yield. In addition, an ethanol concentration of 45 g/L was obtained within 48 h with 86% yield using simultaneous saccharification-fermentation process.     

Dilute acid pretreatment, enzymatic saccharification, and fermentation of rice hulls to ethanol

Rice hulls, a complex lignocellulosic material with high lignin (15.38 +/- 0.2%) and ash (18.71 +/- 0.01%) content, contain 35.62 +/- 0.12% cellulose and 11.96 +/- 0.73% hemicellulose and has the potential to serve as a low-cost feedstock for production of ethanol. Dilute H2SO4 pretreatments at varied temperature (120-190 degrees C) and enzymatic saccharification (45 degrees C, pH 5.0) were evaluated for conversion of rice hull cellulose and hemicellulose to monomeric sugars. The maximum yield of monomeric sugars from rice hulls (15%, w/v) by dilute H2SO4 (1.0%, v/v) pretreatment and enzymatic saccharification (45 degrees C, pH 5.0, 72 h) using cellulase, beta-glucosidase, xylanase, esterase, and Tween 20 was 287 +/- 3 mg/g (60% yield based on total carbohydrate content). Under this condition, no furfural and hydroxymethyl furfural were produced. The yield of ethanol per L by the mixed sugar utilizing recombinant Escherichia colistrain FBR 5 from rice hull hydrolyzate containing 43.6 +/- 3.0 g fermentable sugars (glucose, 18.2 +/- 1.4 g; xylose, 21.4 +/- 1.1 g; arabinose, 2.4 +/- 0.3 g; galactose, 1.6 +/- 0.2 g) was 18.7 +/- 0.6 g (0.43 +/- 0.02 g/g sugars obtained; 0.13 +/- 0.01 g/g rice hulls) at pH 6.5 and 35 degrees C. Detoxification of the acid- and enzyme-treated rice hull hydrolyzate by overliming (pH 10.5, 90 degrees C, 30 min) reduced the time required for maximum ethanol production (17 +/- 0.2 g from 42.0 +/- 0.7 g sugars per L) by the E. coli strain from 64 to 39 h in the case of separate hydrolysis and fermentation and increased the maximum ethanol yield (per L) from 7.1 +/- 2.3 g in 140 h to 9.1 +/- 0.7 g in 112 h in the case of simultaneous saccharification and fermentation.     

Ethanol production from artificial domestic household waste solubilized by steam explosion

Solubilization of domestic household waste through steam explosion with subsequent ethanol production by the microbial saccharification and fermentation of the exploded product was studied. The effects of steam explosion on the changes of the density, viscosity, pH, and amounts of extractive components in artificial household waste were determined. The composition of artificial waste used was similar to leftover waste discharged from a typical home in Japan. Consecutive microbial saccharification and fermentation, and simultaneous microbial saccharification and fermentation of the steam-exploded product were attempted usingAspergillus awamori, Trichoderma viride, andSaccharomyces cerevisiae, the ethanol yields of each process were compared. The highest ethanol yield was obtained with simultaneous microbial saccharification and fermentation of exploded product at a steam pressure of 2 MPa and a steaming time of 3 min.     

Ethanol from lignocellulosic materials by a simultaneous saccharification and fermentation process (SFS) with Kluyveromyces marxianus CECT 10875

Simultaneous saccharification and fermentation (SSF) process for ethanol production from various lignocellulosic woody (poplar and eucalyptus) and herbaceous (Sorghum sp. bagasse, wheat straw and Brassica carinata residue) materials has been assayed using the thermotolerant yeast strain Kluyveromyces marxianus CECT 10875. Biomass samples were previously treated in a steam explosion pilot plant to provide pretreated biomass with increased cellulose content relative to untreated materials and to enhance cellulase accessibility. SSF experiments were performed in laboratory conditions at 42 °C, 10% (w/v) substrate concentration and 15 FPU/g substrate of commercial cellulase. The results indicate that it is possible to reach SSF yields in the range of 50–72% of the maximum theoretical SSF yield, based on the glucose available in pretreated materials, in 72–82 h. Maximum ethanol contents from 16 to 19 g/l were obtained in fermentation media, depending on the material tested.     

Evaluation of nanoparticle-immobilized cellulase for improved ethanol yield in simultaneous saccharification and fermentation reactions

Ethanol yields were 2.1 (P = 0.06) to 2.3 (P = 0.01) times higher in simultaneous saccharification and fermentation (SSF) reactions of microcrystalline cellulose when cellulase was physisorbed on silica nanoparticles compared to enzyme in solution. In SSF reactions, cellulose is hydrolyzed to glucose by cellulase while yeast simultaneously ferments glucose to ethanol. The 35°C temperature and the presence of ethanol in SSF reactions are not optimal conditions for cellulase. Immobilization onto solid supports can stabilize the enzyme and promote activity at non-optimum reaction conditions. Mock SSF reactions that did not contain yeast were used to measure saccharification products and identify the mechanism for the improved ethanol yield using immobilized cellulase. Cellulase adsorbed to 40 nm silica nanoparticles produced 1.6 times (P = 0.01) more glucose than cellulase in solution in 96 h at pH 4.8 and 35°C. There was no significant accumulation (<250 μg) of soluble cellooligomers in either the solution or immobilized enzyme reactions. This suggests that the mechanism for the immobilized enzyme's improved glucose yield compared to solution enzyme is the increased conversion of insoluble cellulose hydrolysis products to soluble cellooligomers at 35°C and in the presence of ethanol. The results show that silica-immobilized cellulase can be used to produce increased ethanol yields in the conversion of lignocellulosic materials by SSF.     

Effect of inhibitors formed during wheat straw pretreatment on ethanol fermentation by Pichia stipitis

The inhibitory effect of the main inhibitors (acetic acid, furfural and 5-hydroxymethylfurfural) formed during steam explosion of wheat straw was studied through ethanol fermentations of model substrates and hydrolysates from wheat straw by Pichia stipitis. Experimental results showed that an increase in acetic acid concentration led to a reduction in ethanol productivity and complete inhibition was observed at 3.5 g/L. Furfural produced a delay on sugar consumption rates with increasing concentration and HMF did not exert a significant effect. Fermentations of the whole slurry from steam exploded wheat straw were completely inhibited by a synergistic effect due to the presence of 1.5 g/L acetic acid, 0.15 g/L furfural and 0.05 g/L HMF together with solid fraction. When using only the solid fraction from steam explosion, hydrolysates presented 0.5 g/L of acetic acid, whose fermentations have submitted promising results, providing an ethanol yield of 0.45 g ethanol/g sugars and the final ethanol concentration reached was 12.2 g/L (10.9 g ethanol/100 g DM).     

Evaluation of steam explosion pre-treatment for enzymatic hydrolysis of sunflower stalks

Sunflower stalks, a largely available and cheap agricultural residue lacking of economic alternatives, were subjected to steam explosion pre-treatment, the objective being to optimize pre-treatment temperature in the range 180-230°C. Enzymatic hydrolysis performed on the pre-treated solids by a cellulolytic complex (Celluclast 1.5L) and analysis of filtrates were used to select the best pre-treatment temperature. Temperature selection was based on the susceptibility to enzymatic hydrolysis of the cellulose residue and both the cellulose recovery in the solid and the hemicellulose-derived sugars recoveries in the filtrate. After 96h of enzymatic action, a maximum hydrolysis yield of 72% was attained in the water-insoluble fiber obtained after pre-treatment at 220°C, corresponding to a glucose concentration of 43.7g/L in hydrolysis media. Taking into account both cellulose recovery and hydrolysis yield, the maximum value of glucose yield referred to unpretreated raw material was also found when using steam pre-treated sunflower stalks at 220°C, obtaining 16.7g of glucose from 100g of raw material. With regard to the filtrate analysis, most of the hemicellulosic-derived sugars released during the steam pre-treatment were in oligomeric form, the highest recovery being obtained at 210°C pre-treatment temperature. Moreover, the utilisation of hemicellulosic-derived sugars as a fermentation substrate would improve the overall bioconversion of sunflower stalks into fuel ethanol.     

The effects of four different pretreatments on enzymatic hydrolysis of sweet sorghum bagasse

Four pretreatment processes including ionic liquids, steam explosion, lime, and dilute acid were used for enzymatic hydrolysis of sweet sorghum bagasse. Compared with the other three pretreatment approaches, steam-explosion pretreatment showed the greatest improvement on enzymatic hydrolysis of the bagasse. The maximum conversion of cellulose and the concentration of glucose obtained from enzymatic hydrolysis of steam explosion bagasse reached 70% and 25 g/L, respectively, which were both 2.5 times higher than those of the control (27% and 11 g/L). The results based on the analysis of SEM photos, FTIR, XRD and NMR detection suggested that both the reduction of crystallite size of cellulose and cellulose degradation from the Iα and Iβ to the Fibril surface cellulose and amorphous cellulose were critical for enzymatic hydrolysis. These pretreatments disrupted the crystal structure of cellulose and increased the available surface area, which made the cellulose better accessible for enzymatic hydrolysis.     

Cellulosic ethanol production on temperature-shift simultaneous saccharification and fermentation using the thermostable yeast Kluyveromyces marxianus CHY1612

In cellulosic ethanol production, use of simultaneous saccharification and fermentation (SSF) has been suggested as the favorable strategy to reduce process costs. Although SSF has many advantages, a significant discrepancy still exists between the appropriate temperature for saccharification (45-50 °C) and fermentation (30-35 °C). In the present study, the potential of temperature-shift as a tool for SSF optimization for bioethanol production from cellulosic biomass was examined. Cellulosic ethanol production of the temperature-shift SSF (TS-SSF) from 16 w/v% biomass increased from 22.2 g/L to 34.3 g/L following a temperature shift from 45 to 35 °C compared with the constant temperature of 45 °C. The glucose conversion yield and ethanol production yield in the TS-SSF were 89.3% and 90.6%, respectively. At higher biomass loading (18 w/v%), ethanol production increased to 40.2 g/L with temperature-shift time within 24 h. These results demonstrated that the temperature-shift process enhances the saccharification ratio and the ethanol production yield in SSF, and the temperature-shift time for TS-SSF process can be changed according to the fermentation condition within 24 h.     

Steam pretreatment of H(2)SO(4)-impregnated Salix for the production of bioethanol

In the bioconversion of lignocellulosic materials to ethanol, pretreatment of the material prior to enzymatic hydrolysis is essential to obtain high overall yields of sugar and ethanol. In this study, steam pretreatment of fast-growing Salix impregnated with sulfuric acid has been investigated by varying the temperature (180-210 degrees C), the residence time (4, 8 or 12 min), and the acid concentration (0.25% or 0.5% (w/w) H(2)SO(4)). High sugar recoveries were obtained after pretreatment, and the highest yields of glucose and xylose after the subsequent enzymatic hydrolysis step were 92% and 86% of the theoretical, respectively, based on the glucan and xylan contents of the raw material. The most favorable pretreatment conditions regarding the overall sugar yield were 200 degrees C for either 4 or 8 min using 0.5% sulfuric acid, both resulting in a total of 55.6g glucose and xylose per 100g dry raw material. Simultaneous saccharification and fermentation experiments were performed on the pretreated slurries at an initial water-insoluble content of 5%, using ordinary baker's yeast. An overall theoretical ethanol yield of 79%, based on the glucan and mannan content in the raw material, was obtained.     

Biosaccharification of cellulosic biomass in immiscible solvent–water mixtures

The enzymatic hydrolysis of wheat straw was carried out in bi-phasic media prepared with acetate esters and Na-acetate buffer. The volume percentage of the organic chemicals was 75%. The biomass was pretreated in a steam explosion plant at 217°C and for 3 min. A cellulase complex from commercial source was utilised and the experiments were run at 45°C and at constant enzyme to biomass weight ratio (0.06). Biomass loadings ranged from 6.25 to 100 g per litre of reactor. The amount of glucose formed per litre of reactor and hour and the glucose yield (grams of product per gram of biomass) were close to the values attained in pure buffer. The glucose concentration in the aqueous phase was in bi-phasic media much higher than in pure buffer and reached the value of 146 g l_H2O⁻¹ during 72 h of saccharification. The results were poorly dependent on the physical–chemical properties of the solvents. Nevertheless, butyl acetate could be slightly preferred to propyl and i-amyl acetate. The use of bi-phasic media did not require stirring rate higher than in pure buffer. The presence of acetate ester traces did not alter markedly the production of ethanol in the fermentation stage, but determined the extension of the lag phase.     

白腐真菌Pleurotus sajor-caju预处理对红麻秸秆发酵乙醇的影响

为研究微生物法预处理对红麻秸秆中木质素的降解及后续的红麻纤维素酶促糖化和发酵效率的影响,将白腐真菌Pleurotus sajor-caju接种在红麻秸秆培养基上固态培养,对红麻秸秆进行预处理。经P. sajor-caju培养25~35 d后,有效转化红麻秸秆中的木质素,转化率最高可达50.20％,并提高红麻纤维素的酶促水解效率,糖化率达69.33％~78.64％,与对照组相比提高了3.5~4.1倍。以微生物法预处理后的红麻秸秆样品为底物的同步糖化发酵实验表明,发酵72 h,发酵液中乙醇浓度达到18.35~     

树干毕赤酵母和酿酒酵母混合糖发酵产乙醇

以树干毕赤酵母和酿酒酵母为发酵菌株,酸性蒸汽爆破玉米秸秆预水解液和纯糖模拟液为C源,采用固定化酵母细胞的方法,研究了酸爆玉米秸秆预水解液初始pH、N源种类及其浓度、3种发酵模式对树干毕赤酵母戊糖发酵的影响。结果表明:玉米秸秆预水解液适合发酵的初始pH范围为6.0～7.0;1.0 g/L的(NH4)2SO4作为N源,在40 g/L葡萄糖和25 g/L木糖培养基中发酵24 h,糖利用率达到99.47%,乙醇质量浓度为24.72 g/L,优于尿素和蛋白胨作为N源;3种模式的发酵体系中,以游离树干毕赤酵母和固定化酿酒酵母发酵性能最好,糖利用率和乙醇得率分别为99.43%和96.39%。     

Cellulose degradation and glucose accumulation byClostridium thermocellum ATCC 27405 under different cultural conditions

Effect of various cultural parameters on cellulose degradation, glucose accumulation and ethanol production byClostridium thermocellum ATCC 27405 were investigated. Optimum pH values for glucose accumulation and ethanol production were determined as 7 and 10, respectively. Highest amount of ethanol (0.92 g/l) was obtained from the culture which contains 10 g urea/l with 34.5% decrease in glucose accumulation. Addition of 100 mM phosphate to the medium increased ethanol production while cellulose degradation and sugar accumulation decreased by 34 and 99%, respectively. Among minerals tested, Mg⁺² was found to be the most important element which affects cellulose degradation. When the medium contained no Mg⁺², residual cellulose concentration was 4.3 g cellulose/l. When the cultural parameters were optimised, glucose accumulation started at early days of fermentation and glucose concentration was 60% higher than that of the control at the 10^th day of fermentation.