Enzymatic saccharification of cellulose using an ionic liquid [Amim] [COOH] pretreatment

In this study we used l-allyl-3-methyl imidazole formate （[Amim][COOH]） as ionic liquid to pre-treat the cellulose and determined the rate of polymerization and enzymatic hydrolysis. The results showed that pretreatment with （[Amim][COOH]） significantly decreased the cellulose polymerization. As the pretreatment temperature went up, the enzymatic hydrolysis rate was first increased and then decreased The maximal enzymatic hydrolysis rate was achieved when the pretreatment temperature was 90 ℃. Under the ultrasonic condition, the initial rate of enzmatic hydrolysis for the ionic liquid-treated cellulose was up to 11.10 gL-1h-1, which was 33% increase compared to the untreated cellulose. Scanning Electronic Microscopy （SEM） and Fourier Transform Infrared-Raman Spectroscop （FT-IR） analysis showed that ionic liquidtreated cellulose started to depolymerize. In addition, the cr3＇stallinity of the cellulose was significantly decreased after pretreatment with ionic liquid.

Biorefinery of bacterial cellulose from rice straw:enhanced enzymatic saccharification by ionic liquid pretreatment

The pretreatment of rice straw is often used to enhance the hydrolysis. 1-allyl-3-methylimidazolium chloride （ [ AMIM ] C1） is a kind of low viscous, nontoxic and recyclable ionic liquid. It was used to treat rice straw and improve the enzymatic hydrolysis of rice straw in this study. The factors influencing the pretreatment were as follows： the dosage of rice straw in [ AMIM ] Cl, crush mesh of rice straw, pretreatment temperature and time. After the pretreatment with a 3 % （the weight ratio of rice straw to ionic liquid） rice straw dosage in [AMIM]Cl at 110 ℃ for 1 h, the yield of reducing sugar of regenerated rice straw by 33 U/mL cellulase hydrolysis was 53.3 %, which was two times higher than that of un-treated rice straw （23.7 % ）. More researches regarding straw biorefinery to bacterial cellulose are being performed in the lab and prospective results will be published in near future.

Effects of Chemical Pretreatment on Enzymatic Saccharification and Fermentation of Forages

[ Objective] To study the effect of pretreatment with chemical substances on enzymatic saccharification of affalfa （ Medic, ago sativa L. ）, sorghum hybrid sudan grass [ Sorghum bicolor （ L. ） Moench x Sorghum sudanese （Piper） Stapf], erect milkvetch （Astraga/us adsurgens Pall. ） and pearl millet （ Pennisetum americanum （ L. ） Leeke）. [ Method ] The forages were pretreated with sulfuric acid at different concentration, and then the content of cellulose, hemicellulose, and lignin were detected and compared with that before pretreatment. The concentration of glucose and ethanol after different fermentation time was also determined. [ Result] After pretreatment, the content of cellulose increased, while that of hemicel- lulose and lignin decreased. After treatment with 1.0% （W/V） sulfuric acid, the four kinds of forages all had the highest concentration of ethanol in the citric acid-sodium citrate buffer system （pH 4.8）. Dudng fermentation process, the concentration of glucose and ethanol first increased and then declined, peaking respectively at 24 h and 48 h post fermentation. [Condusion] Pretreatment promotes the enzymatic saccharification and fermen- tation of alfalfa, sorehum hvbrid sudan orass. Dead millet, and erect milkvetch, and their enerov performance decreases in order.

Effect of various aromatic compounds with different functional groups on enzymatic hydrolysis of microcrystalline cellulose and alkaline pretreated wheat straw

Low molecular aromatic compounds are detrimental to the enzymatic hydrolysis of lignocellu-lose.However,the specific role of their functional groups remains unclear.Here,a series of nine aromatic compounds as additives were tested to understand their effect on the hydrolysis yield of microcrystalline cellulose(MCC)and alkaline pretreated wheat straw.Based on the results,the inhibition of aldehyde groups on MCC was greater than that of carboxyl groups,whereas for the alkaline pretreated wheat straw case,the inhibitory effect of aldehyde groups was lower than that of carboxyl groups.Increased methoxyl groups of aromatic compounds reduced the inhibitory ef-fect on enzymatic hydrolysis of both substrates.Stronger inhibition of aromatic compounds on MCC hydrolysis was detected in comparison with the alkaline pretreated wheat straw,indicating that the substrate lignin can offset the inhibition to a certain extent.Among all aromatic com-pounds,syringaldehyde with one aldehyde group and two methoxyl groups improved the glucan conversion of the alkaline pretreated wheat straw.

Conversion of Sugarcane Shoots and Leaves into Reducing Sugars by Pretreatment and Enzymatic Hydrolysis

Sugarcane shoots and leaves consist of 38% cellulose, 30.6% hemicellulose and 12.8% lignin on dry solid （DS） basis and have the potential to serve as low cost feedstocks for ethanol production. The pretreatment and enzymatic hydrolysis conditions include particle size, alkali （NaOH）/dilute acid （H2SO4） pretreatment, chemical and substrate concentrations, temperature, autoclaving time for pretreatment, enzyme concentration, pH and temperature for hydrolysis varied were evaluated for conversion of sugarcane shoots and leaves cellulose and hemicellulose to reducing sugar. The optimum conditions were accomplished by using 14% w/v DS of 0-10 mm sugarcane shoots and leaves in particle size, pretreated with 1.5% w/v of dilute sulfuric acid at 121℃, 15 lbs/in2 for 15 min and enzymatic saccharification using 40 FPU/g DS cellulose at 50℃ and pH 5, After incubating at 160 rpm for 12 hrs, 59 g/L or 386,38 mg/g DS of reducing sugar and 50.69% saccharification were obtained.

Research on Hydrolysis and Saccharification of Corn Stover

In this paper three methods(dilute acid pretreatment, aqueous ammonia/dilute acid pretreatment and alkaline pretreatment) were used to study the hydrolysis of corn stover and characteristics of each method were compared. The results showed that the lignin removal rate was 71.8% when the corn stover was treated with a caustic soda solution containing 1.5% of NaOH, at a temperature of 75 ℃ for 90 min with an initial solid-liquid ratio of 1:8(w/v). Hydrolysis yield of the NaOH pretreated sample reached 78.5%, which was much higher than other control groups. These results are useful for evaluation of pretreatment technologies, and identification of key factors that limit cellulose hydrolysis, and can also serve as a basis for designing and screening appropriate pretreatment technologies.

Effect of Different Pretreatment Methods on Enzymatic Saccharification and Ethanol Production from Sugarcane Shoots and Leaves

Sugarcane shoots and leaves consist of 35.2% cellulose, 23.43% hemicellulose, 12.6% lignin and 6.59% ash on dry solid （DS） basis and have the potential to serve as low cost feedstocks for ethanol production. To improve the enzymatic digestibility of these biomass and bioethanol production, three pretreatment methods had been investigated and compared, including： （1） 2% w/v NaOH solution autoclaving pretreatment; （2） 2% w/v H2SO4 solution autoclaving pretreatment and （3） two steps of 2% w/v NaOH solution autoclaving followed by 2% w/v H2SO4 solution autoclaving pretreatment. Among them, the best result for ethanol production was obtained when 15 g DS of sugarcane shoots and leaves was pretreated by using two step of 2% w/v NaOH solution autoclaving followed by 2% w/v H2SO4 solution autoclaving. The highest ethanol concentration 30.40 g/L （92.65% in fermentation efficiency） was obtained from reducing sugar 89.25 g/L at 48 h. Moreover, the washing step of solid residue after pretreatment could reduce furfural and hydroxymethylfurfural （HMF） in all pretreatment methods when compared to unwashing solid residue after pretreatment.

Optimization on Pretreatment and Enzymatic Hydrolysis of Sugarcane Trash for Ethanol Production

The present study was conducted for the optimization of pretreatment and enzymatic hydrolysis of lignocellulosic biomass （sugarcane trash）, which is a renewable resource for the production of bioethanol. The pretreatment and enzymatic hydrolysis conditions including alkali （NaOH）/dilute acid （H2SO4）, substrate and chemical concentration for pretreatment, enzyme dosage, pH, temperature and substrate concentration for hydrolysis were varied and evaluated for sugar and ethanol production at the end. The optimum condition was accomplished using 15% w/v DS of 0-2 mm sugarcane trash in size of particle. It was pretreated with two steps of 2% w/v NaOH autoclaving followed by 2% w/v H2SO4 autoclaving with washing step after pretreatment. An enzymatic hydrolysis was then performed using 15% w/v DS pretreated substrate, hydrolyzed with cellulase 50 filter paper unit （FPU）/g DS at 50 ℃ and pH 5. After incubating at 160 r for 48 h, 117.16 g/L reducing sugar was obtained. The achieved sugar after enzymatic hydrolysis was finally fermented to ethanol by Saccharomyces cerevisiae TISTR 5596, with concentration of 48.17 g/L ethanol or yield 0.509 g/g reducing sugars which was equal to 99.81% of theoretical yield.

Optimal Method for Production of Amorphous Cellulose with Increased Enzymatic Digestibility

In this paper,a simple and cheap method for producing of amorphous cellulose was studied by treating the initial cellulosic material(MCC and waste paper)with a cold solvent,such as aqueous solution of 7%NaOH/12%Urea,at the various ratios of the solvent to cellulose(v/w)(R).If was found that after treatment of cellulose materials with the solvent at R≥5,a completely amorphous cellulose(AC)is formed.Due to high digestibility,the AC with concentration of 50 g/L is converted to glucose almost completely for 48 h under the action of cellulolytic enzyme CTec-3 with a dose of 30 mg/g solid sample.Such sample can be used as an amorphous standard in the study of crystallinity degree and enzymatic hydrolysis of various types of cellulose and lignocellulose.It was found that enzymatic saccharification is most advantageous to carry out at elevated concentrations of AC,150 g/L.Due to high cost of MCC,it is preferable to use a cheap cellulose raw material,such as mixed waste paper(MWP),for the commercial production of AC and glucose.The resulting glucose can find application in biotechnology as a promising nutrient for various microorganisms.

3D-Printed PLA Filaments Reinforced with Nanofibrillated Cellulose

In the current study poly(lactic acid)PLA composites with a 3 wt%and 5 wt%of nanofibrillated cellulose(NFC)were produced by 3D-printing method.An enzymatic pretreatment coupled with mechanical fibrillation in a twin screw extruder was used to produce high consistency NFC.Scanning electron microscopy(SEM)equipped with Fibermetric software,FASEP fiber length distribution analysis,Furrier transform infrared spectroscopy(FT-IR),thermogravimetric analysis(TGA),tensile tests,impact tests and differential scanning calorimetry were used to characterize NFC and PLA/NFC composites.The results of the fiber length and width measurements together with the results of the SEM analysis showed that enzymatic hydrolysis coupled with a twin screw extrusion could effectively reduce the diameter and length of cellulose fibers.The produced NFC consisted of microand nanosized fibers entangled in a characteristic 3D-network.Based on the FT-IR analysis,no new bonds were formed during the enzymatic hydrolysis or fibrillation process.The TGA analysis confirmed that produced NFC can be used in hightemperature extrusion processing without NFC degradation.During the PLA/NFC composites preparation the NFC agglomerates were formed,which negatively influenced PLA/NFC composites impact properties.The slightly improved tensile strength and elastic modulus were reported for all composites when compared to the neat PLA.The elongation at break was not affected by the NFC addition.No significant differences in thermal stability were detectable among composites nor in comparation with the neat PLA.However,the crystallinity degree of the composite containing 5 wt%NFC was increased in respect to the neat PLA.

Enzymatic hydrolysis of cellulose materials treated with ionic liquid [BMIM] Cl

A new cellulose solvent ionic liquid 1-butyl-3-methylimidazolium chloride ([BMIM]Cl) was used to treat wheat straw and steam-exploded wheat straw (SEWS) in order to improve the enzymatic hy- drolysis rates, while the water was used as the con- trol. The enzymatic hydrolysis results showed that the hydrolysis rates of materials treated with [BMIM]Cl were improved. The hydrolysis rate of treated wheat straw could reach 70.37% and the SEWS could be completely hydrolyzed, while hydrolysis rates of the wheat straw and SEWS treated with water were 42.78% and 68.78% under the same conditions, re- spectively. The FTIR analysis and polymerization degree measurement indicated that the hydrolysis rates improvement was attributed to the decrease of the polymerization degrees of cellulose and hemi- cellulose, the absolute crystallinity degree of cellu- lose and the increase of its reaction accessibility.

Ameliorated enzymatic saccharification of corn stover with a novel modified alkali pretreatment

Enzymatic saccharification/hydrolysis is one of the key steps for the bioconversion of lignocelluloses into sustainable biofuels.In this work,corn stover was pretreated with a novel modified alkali process(NaOH+anthraquinone(AQ)+sodium lignosulfonate(SLS)),and then enzymatically hydrolyzed with an enzyme cocktail(cellulase(Celluclast 1.5L),β-glucosidase(Novozyme 188)and xylanase(from thermomyceslanuginosus))in the pH range of 4.0-6.5.It was found that the suitable pH for the enzymatic saccharification process to achieve a high glucan yield was between 4.2 and 5.7,while the appropriate pH to obtain a high xylan yield was in the range of 4.0-4.7.The best pH for the enzymatic saccharification process was found to be 4.4 in terms of the final total sugar yield,as xylanase worked most efficiently in the pH range of 4.0-4.7,under the conditions in the study.The addition of xylanase in the enzymatic saccharification process could hydrolyze xylan in the substrates and reduce the nonspecific binding of cellulase,thus improving the total sugar yields.

基于磺化碳基固体酸的纤维素水解糖化研究进展

对磺化碳基固体酸制备原料及制备方法进行系统介绍,分析磺化碳基固体酸物理化学结构与其催化纤维素水解的构效关系,讨论碳基固体酸失活原因及分离回收方法。针对磺化碳基固体酸与纤维素间的传质限制,总结归纳先“降结晶、再水解”的纤维素高效水解策略。最后对磺化碳基固体酸的未来研究重点和前景进行展望,以期为促进磺化碳基固体酸在木质纤维素糖平台构建中的创新发展提供科学参考。

甘蔗渣的碱催化甘油水溶液预处理

建立了一种木质纤维素的碱催化甘油水溶液预处理方法以期选择性分离组分和提高底物的可酶解性。实验确定了碱催化甘油水溶液预处理甘蔗渣的条件为:温度180℃、NaOH添加量7%、反应时间45 min以及甘油水溶液浓度80%。在该条件下,甘蔗渣的纤维素和半纤维素保留率分别为93.0%和83.4%,而木质素脱除率接近80%,达到了理想的组分分离效果。预处理后底物(20 g/L)在CTec2酶载量10 FPU/g干基下水解72 h酶解率为79.6%,表明预处理后底物具有较好的可酶解性。利用现代分析技术解析了甘蔗渣预处理前后的组成结构演变规律,初步探明了碱催化甘油水溶液预处理有效提升生物质原料可酶解性的原因。

白酒糟中纤维素酶解工艺优化研究

为实现白酒糟资源的综合利用,探究了白酒糟渣中纤维素酶解的最佳工艺参数。以可发酵糖得率为评价指标,通过单因素实验探究了预处理条件(固液比、硫酸体积分数、处理温度、处理时间)及酶解条件(酶添加量)对酒糟中纤维素酶解效果的影响,在此基础上用神经网络分析结合遗传算法进行进一步优化。结果表明,白酒糟纤维素的最佳酶解工艺参数为:固液比为1∶10、硫酸体积分数为7%、温度为170℃、时间为40 min、酶添加量为23 FPU/g。此条件下酒糟纤维素酶解得到的可发酵糖总量为(19.77±0.56) g/L,得率为88.97%。以该酶解液作为碳源进行丁醇发酵,溶剂(丁醇、乙醇、异丙醇和丙酮)总产量为1.56 g/L。

Process development for producing a food-grade glucose solution from rice straws

In this paper, processes for producing a food-grade glucose solution through enzymatic hydrolysis of celluloserich solids obtained from rice straws are presented. The rice straws were pretreated by acid-catalyzed steam explosion, and the reaction efficiency, toxicity control, and process economic feasibility were studied. Mass transfer resistance to the hydrolysis reaction was reduced by grinding with glass beads. A higher glucose concentration could be obtained by feeding more cellulose in the hydrolysis reaction; however, this also resulted in the production of undesired byproducts. Thus, a soaking process for the cellulose solids in water was developed to effectively reduce the generation of byproducts in the hydrolysis reaction. The resulting food-grade glucose solution can provide 414 kilocalories per liter, and could be used during a food-shortage crisis in the future.The current production cost is estimated to be 0.82 USD·L^(-1).

纤维素酶酶解苇浆纤维微观结构和结晶结构的变化

采用红外光谱(IR)、X射线衍射分析和扫描电镜对在纤维素酶解进程中苇浆纤维素大分子的结构及纤维形态的变化进行了研究。研究结果表明,纤维素酶酶解进程中,纤维素大分子的晶型没有改变,但结晶度呈现周期性变化,微晶尺寸略有降低。在酶解初期,纤维素酶不仅作用于纤维素无定形区,也开始作用在纤维素结晶区表面;在酶解中、后期,纤维素酶对纤维素结晶区和无定形区的作用呈现周期性变化。纤维素酶解过程中纤维表面呈现周期性“剥皮”现象,并且在纤维表面出现孔洞和沟槽,在孔洞处纤维易断裂,导致纤维长度和粗度的降低。

蒸汽爆破预处理对玉米秸秆化学组成及纤维结构特性的影响

研究了蒸汽爆破预处理对玉米秸秆化学组成变化及纤维素酶水解得率的影响,并采用扫描电镜(SEM)、X射线衍射仪(XRD)和红外光谱(IR)对玉米秸秆纤维结构特性进行了分析。结果表明,蒸汽爆破预处理对纤维素影响不显著,半纤维素含量大幅度降低,木质素含量也有所降低。纤维素酶水解得率随着爆破压力的增大和维压时间的延长而增加,当蒸汽压力为1.6MPa,维压时间9 min时所得固体渣经纤维素酶水解,得率最大为75.76%。蒸汽爆破预处理后玉米秸秆纤维表面和细胞壁受到不同程度的破坏,表面积增大,孔洞增加,纤维素的结晶度降低,有利于纤维素酶水解作用的进行。蒸汽爆破是一种有效的木质纤维预处理方法。

化学预处理提高酒糟生物质酶解糖化效果

为促进酒糟生物质的酶解糖化,筛选适宜的预处理方法,以脱除木质素,提高综纤维素(纤维素和半纤维素之和)保留率为目标,研究比较了酸-超声波耦合(ultrasound-assisted acid pretreatment,UAAP)、液氨(pretreatment by soaking in aqueous ammonia,PSAA)、碱性双氧水(alkaline hydrogen peroxide pretreatment,AHPP)和酸性亚硫酸氢盐(bisulfite pretreatment,BP)4种预处理法对酒糟化学组分、结构特性和酶解得率的影响。结果表明,与其余3种方法相比,BP法处理后酒糟的纤维素和半纤维素保留率最高,分别为84.59%和84.87%,即综纤维素保留率为84.68%。与未处理酒糟(unpretreatment,UP)相比,4种方法预处理后酒糟的综纤维素酶解得率分别提高了49.12%(酸-超声波,UAAP)、55.48%(液氨,PASS)、92.79%(碱性双氧水,AHPP)和99.15%(酸性亚硫酸氢盐,BP),其中BP法对酒糟酶解糖化的促进作用最有效。扫描电镜(scanning electron microscope,SEM)和X-衍射(X-ray differaction,XRD)结果显示,酒糟经不同方法预处理后表观结构发生了明显变化,木质纤维网络结构遭到破坏,表面呈现无规则或形状各异的膨松状态,沟壑明显,孔隙率增加,比表面积增大,有利于提高水解酶的可及性。化学组分和结构特性的变化说明酒糟的酶解得率与综纤维素的保留、木质素的去除、表面微观形貌变化以及纤维素结晶度等因素直接相关。总之,酸性亚硫酸氢盐(BP)法是适用于酒糟生物质糖化预处理的一种有效可行方法。