摘要
【目的】建立里氏木霉(Trichoderma reesei)高产突变菌株的快速筛选方法,选育出高产内切葡聚糖酶的突变株。【方法】对里氏木霉T306菌株的初筛培养基进行优化,建立快速筛选方法;通过紫外诱变手段选育内切葡聚糖酶高产突变菌株,并对突变菌株的产酶培养基进行优化。【结果】在初筛培养基中添加浓度为0.1%(W/V)的乳糖、蛋白胨及脱氧胆酸钠有利于菌株的筛选。诱变后筛选出菌落形态发生明显变化的内切葡聚糖酶高产突变株0516,其羧甲基纤维素酶活力(CMC酶)较出发菌株提高了38.9%。其产酶培养基经优化后,得到最适碳、氮源分别为:乳糖1.50%、硫酸铵0.14%、尿素0.05%、蛋白胨0.10%,优化后CMC酶活力达64.2 U/mL,较优化前提高了2.3倍。【结论】建立了里氏木霉高产突变菌株的快速筛选方法,通过紫外诱变育种获得了产内切葡聚糖酶能力高且遗传稳定的突变株0516。
[Objective] To establish the rapid screening method of mutant strain of Trichoderma reesei and to screen out mutants with high yield endoglucanase. [Methods] The screening medium of Trichoderma reesei T306 was optimized to establish the rapid screening method, and the mutant strain with high-production endoglucanase was screened through UV mutation. Furthermore, the fermentation medium of mutant strain was optimized. [Results] Addition of 0.1% （W/V） lactose, peptone and sodium deoxycholate was beneficial for the screening of mutants. The mutant strain 0516 was obtained, which had higher production of endoglucanase and obviously varied in colony morphology. The CMCase activity of the mutant was increased by 38.9% compared with original strain. The optimum carbon source and nitrogen source of culture medium were as follows： lactose 1.50%, ammonium sulfate 0.14%, urea 0.05%, and peptone 0.10%. Under these conditions, the CMCase activity of mutant 0516 reached 64.2 U/mL, which was 2.3 times higher than before. [Conclusion] A method for rapidly screening mutant of Trichoderma reesei with high production endoglucanase was established, and mutant 0516 with higher production of endoglucanase and genetic stability was obtained after UV mutation.
出处
《微生物学通报》
CAS
CSCD
北大核心
2013年第8期1448-1456,共9页
Microbiology China
基金
福建省发改委生物技术开发专项项目(No.闽发改高技【2011】1598号)
福州市科技计划项目(No.榕科【2012】78号)
2013年海洋公益性行业科研专项项目(No.201305015)
