粗纤维素酶液对微晶纤维素酶解的影响因素

doi:10.16085/j.issn.1000-6613.2016.09.019

化工进展 ›› 2016, Vol. 35 ›› Issue (09): 2766-2774.DOI: 10.16085/j.issn.1000-6613.2016.09.019

粗纤维素酶液对微晶纤维素酶解的影响因素

沙如意^1,2,3,, Edwin Menledy Gbor JR^1,2,3, 楼坚^1,2,3, 蔡成岗^1,2,3, 毛建卫^1,2,3, 刘士旺^1,2,3

1 浙江科技学院生物与化学工程学院/轻工学院, 浙江杭州 310023;
2 浙江省农产品化学与生物加工技术重点实验室, 浙江杭州 310023;
3 浙江省农业生物资源生化制造协同创新中心, 浙江杭州 310023

收稿日期:2016-04-27 修回日期:2016-05-12 出版日期:2016-09-05 发布日期:2016-09-05
通讯作者: 毛建卫,教授,研究领域为农业生物资源利用。E-mail:zjhzmjw@163.com。
作者简介:沙如意(1982-),男,博士研究生。
基金资助:
国家级星火计划（2014GA690036、2015GA690057、2015GA700079）、浙江省重点研发计划（2015C02031）、浙江省教育厅科研项目（Y201327551、Y201224264）及浙江科技学院科研启动基金（F501103C01）项目。

Influence factors of microcrystalline cellulose hydrolysis by crude cellulase solution

SHA Ruyi^1,2,3,, MENLEDY Gbor JR Edwin^1,2,3, LOU Jian^1,2,3, CAI Chenggang^1,2,3, MAO Jianwei^1,2,3, LIU Shiwang^1,2,3

1 School of Biological and Chemical Engineering/School of Light Industry, Zhejiang University of Science and Technology, Hangzhou 310023, Zhejiang, China;
2 Zhejiang Provincial Key Lab for Chemical and Biological Processing Technology of Farm Products, Hangzhou 310023, Zhejiang, China;
3 Zhejiang Collaborative Innovation Center of Chemical and Biological Manufacturing for Agricultural Biological Resources, Hangzhou 310023, Zhejiang, China

Received:2016-04-27 Revised:2016-05-12 Online:2016-09-05 Published:2016-09-05

摘要/Abstract

摘要： 栗生灰黑孔菌多被用于产漆酶的研究，很少有利用其纤维素酶的报道。为了降低在纤维素水解中纤维素酶的使用成本，利用栗生灰黑孔菌发酵制备的粗纤维素酶液，以微晶纤维素为底物模型，研究粗纤维素酶液水解微晶纤维素的最佳pH、温度和最佳表面活性剂助剂种类及浓度，并对不同表面活性剂存在条件下的纤维素酶解动力学、紫外和荧光光谱进行了研究。结果表明，粗纤维素酶水解微晶纤维素的最佳条件为pH 4.8，温度50℃，最佳表面活性剂助剂为吐温80，添加剂量为1.12mg/g底物；吐温80的添加可提高粗纤维素酶解的最大反应速度常数V_max，降低米氏常数K_m；表面活性剂改变了纤维素酶的紫外和荧光最大吸收峰，酰胺Ⅰ带和酰胺Ⅲ带的谱峰，可能通过与纤维素酶中的氨基酸残基发生反应影响了纤维素酶的结构，进而影响了微晶纤维素的水解反应。该研究为进一步降低纤维素水解成本提供了理论指导。

关键词: 纤维素酶, 微晶纤维素, 表面活性剂, 生物催化, 生物质

Abstract: Melanoporia castanea was often used as laccase-producing strain,but little study was reported about the production of cellulase by this strain. In order to reduce the cost of cellulase in the hydrolysis of cellulose,the crude cellulase solution prepared by the fermentation of Melanoporia castanea. Then microcrystalline cellulose(MCC) was used as a model substrate,and the conditions of microcrystalline cellulose hydrolysis by crude cellulase solution was optimized,including pH,temperature,and type and concentration of surface active agents. Then crude cellulase solution hydrolysis kinetics,and ultraviolet and fluorescence spectrum were studied with the surfactant agents. The results showed that the optimum hydrolysis conditions of microcrystalline cellulose with crude cellulase solution were pH 4.8,temperature 50℃,and surfactant Tween-80 at dosage of 1.12mg/g substrate. It demonstrated that Tween-80 addition could improve the maximum reaction velocity constants V_max of crude cellulase solution and reduce Michaelis constant(K_m). Both maximum absorbance peaks of ultraviolet and fluorescence were changed after addition of surfactants,with amideⅠand Ⅲ bands infrared spectrum changed,showing that the aromatic amino acid residues in cellulose were reacted with surfactants and then influenced the hydrolysis of microcrystalline cellulose. The study provides a theoretical guide for further reducing the cost of cellulose hydrolysis.

Key words: cellulase, microcrystalline cellulose, surfactants, biocatalysis, biomass

中图分类号:

TS201.2

沙如意, Edwin Menledy Gbor JR, 楼坚, 蔡成岗, 毛建卫, 刘士旺. 粗纤维素酶液对微晶纤维素酶解的影响因素[J]. 化工进展, 2016, 35(09): 2766-2774.

SHA Ruyi, MENLEDY Gbor JR Edwin, LOU Jian, CAI Chenggang, MAO Jianwei, LIU Shiwang. Influence factors of microcrystalline cellulose hydrolysis by crude cellulase solution[J]. Chemical Industry and Engineering Progree, 2016, 35(09): 2766-2774.

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粗纤维素酶液对微晶纤维素酶解的影响因素

Influence factors of microcrystalline cellulose hydrolysis by crude cellulase solution

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