哈茨木霉高产几丁质酶菌株的筛选及产酶条件研究  

Screening of high chitinase-producing stains from Trichoderma harzianum and conditions for enzyme production

作　　者：王红梅[1] 蒋立科[1] 陶芳[1] 魏练平[1] 岳永德[2] 

机构地区：[1]安徽农业大学生命科学学院,安徽合肥230036 [2]国际竹藤网络中心,北京100102

出　　处：《浙江大学学报（农业与生命科学版）》2010年第2期146-152,共7页Journal of Zhejiang University:Agriculture and Life Sciences

基　　金：国家高技术研究发展计划"863"资助项目(2003AA241171)

摘　　要：通过氮离子注入法诱变哈茨木霉(Trichoderma harzianum)野生菌,再以透明圈法对平板培养菌进行筛选,用DNS法测定液体培养菌的酶活,从中筛选出1株产几丁质酶能力高的哈茨木霉菌株H-13,并通过单因素试验和正交试验优化该目标菌株的发酵条件.单因素试验结果表明,添加1.0%蛋白胨时产酶量最高,1.0%胶体几丁质对菌体产几丁质酶的诱导性最强,初始pH 5.5、培养96 h时产酶量较高.正交试验表明,H-13以1.0%蛋白胨为氮源、0.5%胶体几丁质为碳源,在初始pH 5.0的培养基中培养96 h时,发酵液中的几丁质酶活可达到731.69 U?mL-1,比同样条件下的野生菌株酶活力提高1倍.Trichoderma harzianum wild-type strains were implanted by N＋ to produce mutagenesis. A mutant stain H-13 with higher activity of chitinase was obtained by screening the mutants with transparent zone, and analyzing chitinase activity by DNS method. Single-factor design and orthogonal test design were used for optimizing cultural conditions of chitinase production from mutant strain H-13. Single-factor experimentation showed that the media added 1.0% peptone led to highest chitinase production, and 1.0% colloidal chitin was the best media for inducing chitinase. The mutant strain showed higher enzymatic activity in this media with initial pH 5.5 for 96 h of culture. Orthogonal test showed that chitinase activity of strain H-13 could reach 731.69·mL^-1 cultured in the optimal medium with 1.0~ peptone, 0.5% colloidal chitin and initial pH 5.0 for 96 h, which was one fold increase compared to the wild strain enzyme vigor.

关 键 词：哈茨木霉 几丁质酶 氮离子注入 产酶条件 

分 类 号：Q939.9[生物学—微生物学]