可高效降解甜高粱秸秆产糖的纤维素酶研究
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摘要
为评价实验室自制的纤维素酶对甜高粱秸秆纤维素的降解效果,以5种不同发酵工艺所产的5组纤维素酶和商品纤维素酶为研究对象,通过对各纤维素酶的滤纸酶活、分解微晶纤维素所产的还原糖量及对甜高粱秸秆中纤维素转化率的检测可知,在5组自制的纤维素酶中,纤维素酶Ⅱ的滤纸酶活最高(1 364.84 U/m L),但纤维素酶Ⅴ对微晶纤维素的分解能力最强,并在50℃,48 h时其对甜高粱秸秆纤维素的转化率最高,达到11.69%,表明纤维素酶Ⅴ可高效分解甜高粱生物质材料转化成可发酵的糖,在发酵生物乙醇和高附加值的化工产品方面极具潜力。
To evaluate the effect of lab-made cellulase on sweet sorghum straw cellulose degradation, the filter paper activity(FPA) of cellulase, the reducing sugar content of microcrystalline cellulose(MCC) decomposition and the cellulose conversion rate of sweet sorghum straw were detected,respectively, with the 5 batches of cellulase produced by 5 different of fermentation processes and commercial cellulase as research object. The results showed that the FPA of cellulase Ⅱ was the highest(1 364.84 U/ml) in the 5 lab-made cellulase, which the decomposition capability of cellulase V on microcrystalline cellulose was the strongest, and its conversion rate was the highest on sweet sorghum straw cellulose at 50 ℃ for 48 h, reaching11.69%. It indicated that cellulase V could effectively decompose sweet sorghum biomass materials into fermentable sugars, and had great potential for fermentation of bioethanol and high value-added chemical products.
To evaluate the effect of lab-made cellulase on sweet sorghum straw cellulose degradation, the filter paper activity(FPA) of cellulase, the reducing sugar content of microcrystalline cellulose(MCC) decomposition and the cellulose conversion rate of sweet sorghum straw were detected,respectively, with the 5 batches of cellulase produced by 5 different of fermentation processes and commercial cellulase as research object. The results showed that the FPA of cellulase Ⅱ was the highest(1 364.84 U/ml) in the 5 lab-made cellulase, which the decomposition capability of cellulase V on microcrystalline cellulose was the strongest, and its conversion rate was the highest on sweet sorghum straw cellulose at 50 ℃ for 48 h, reaching11.69%. It indicated that cellulase V could effectively decompose sweet sorghum biomass materials into fermentable sugars, and had great potential for fermentation of bioethanol and high value-added chemical products.
引文
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[2]THRAN D,SEIDENBERGER T.Global biomass potentials-resources,drivers and scenario results[J].Energ Sust Dev,2010(10):200-205.
[3]DEUTSCHMANN R,DEKKER R F.From plant biomass to bio-based chemicals:latest developments in xylan research[J].Biotechnol Adv,2012,30(6):1627-1640.
[4]SINNOTT M L.Catalytic mechanism of enzymic glycosyl transfer[J].Chem Rev,1990,90(7):1171-1202.
[5]JUTURU V,WU J C.Microbial cellulases:Engineering,production and applications[J].Renew Sust Ener Rev,2014,33(5):188-203.
[6]KUMAR R,SINGH S,SINGH O V.Bioconversion of lignocellulosic biomass:biochemical and molecular perspectives[J].J Ind Microbiol Biotechnol,2008,35(5):377-391.
[7]彭良才.论中国生物能源发展的根本出路[J].华中农业大学学报,2011,92(2):1-6.
[8]BHAT M K.Cellulases and related enzymes in biotechnology[J].Biotechnol Adv,2000,18(5):355-383
[9]GUPTA R,MEHTA G,DESWAL D,et al.Cellulases and their biotechnological applications[J].Biotechnol Environ Manage Res Recov,2013,12:89-106.
[10]BANSAL P,HALL M,REALFF M J,et al.Modeling cellulase kinetics on lignocellulosic substrates[J].Biotechnol Adv,2009,27(6):833-848.
[11]KUHAD C,GUPTA R,SINGH A.Microbial cellulases and their industrial applications[J].Enzyme Res,2011(2):1-9.
[12]姜伯玲.重离子辐照诱变选育纤维素酶高产菌株及其发酵工艺研究[D].北京:中国科学院大学,2016.
[13]姜伯玲,王曙阳,李文建,等.绿色木霉与黑曲霉混合发酵产纤维素酶的研究[J].中国酿造,2015,34(7):28-31.
[14]ZHAO K,XUE P J,GU G Y.Study on determination of reducing sugar content using 3,5-dinitrosalicylic acid method[J].Food Sci,2008,29(8):534-536.
[15]王义甫,何艳玲,孔令娜.饲用纤维素酶测定方法的探讨[J].饲料与畜牧,2008,67(1):35-38.
[16]张全国,张志萍,赵民善,等.秸秆制氢过程中纤维素酶酶活测定方法研究[J].热科学与技术,2011,10(1):128-132.
[17]DECKER S R,ADNEY W S,JENNINGS E,et al.Automated filter paper assay for determination of cellulose activity[J].Biotechnol Fuels Chem,2003,107(1):687-703.
[18]陆红佳,郑龙辉,刘雄.微晶纤维素在食品工业中的应用研究进展[J].食品与发酵工业,2011,37(3):141-144.
[19]BOMMARIUS A S,KATONA A,CHEBEN S E,et al.Cellulase kinetics as a function of cellulose pretreatment[J].Metab Eng,2008,10(6):370-381.
[20]杨胜.饲料分析及饲料质量检测技术[M].北京:中国农业大学出版社,1999:16-17.
[21]全国饲料工业标准化技术委员会.NY/T 1459—2007饲料中酸性洗涤纤维的测定[S].西安:农业部饲料质量监督检查测试中心,2007.
[22]全国饲料工业标准化技术委员会.GB/T 20805—2006饲料中酸性洗涤木质素(ADL)的测定[S].北京:中国标准出版社,2006.
[23]许富强,董妙音,王曙阳,等.甜高粱最佳青贮时期及延期对其营养成分的影响研究[J].饲料研究,2017(4):20-24.
[24]麻星星.天然纤维素酶法水解条件的研究[D].北京:北京化工大学,2010.
[25]中华人民共和国卫生部、中国国家标准化管理委员会.GB/T 5009.7—2008食品中还原糖的测定[S].北京:中国标准出版社,2008.
[26]WILSON D B,IRWIN D C.Genetics and properties of cellulases[M].New York:Springer Nature,1999,1-21.
[27]YE S,CHENG J.Hydrolysis of lignocellulosic materials for ethanol production:A review[J].Bioresource Technol,2003,34(1):1-11.
[28]MOOD S H,GOLFESHAN A H,TABATABAEI M,et al.Lignocellulosic biomass to bioethanol,a comprehensive review with a focus on pretreatment[J].Renew Sust Energ Rev,2013,27:77-93.