复合诱变选育腈水合酶高产菌株  

Breeding of High-yield Strain of Nitrile Hydratase by Compound Mutation

作　　者：朱婷婷[1,2] 秦智[2] 孙晓君[2] 蒋忠平[2] 

机构地区：[1]新余高等专科学校太阳能科学与工程系,江西新余338000 [2]哈尔滨理工大学化学与环境工程学院,哈尔滨150040

出　　处：《安徽农业科学》2009年第4期1397-1399,共3页Journal of Anhui Agricultural Sciences

基　　金：黑龙江省自然科学基金项目(E2004-08);黑龙江省普通高等学校青年学术骨干支持计划项目(1153G008);哈尔滨市科技攻关项目(2004AA4CS049);哈尔滨市青年科学研究基金(2004AFQXJ041)资助

摘　　要：[目的]选择有效的方法选育稳定的腈水合酶高产菌株。[方法]以Rhodococcussp.HUST为出发菌株,采用紫外线和硫酸二乙酯对其进行复合诱变处理,选育出酶活力高的突变株进行培养并测定腈水合酶活性。[结果]菌株HUST复合诱变的条件为:出发菌株在距离为15cm时紫外诱变45s后再经1.1%的硫酸二乙酯诱变处理20m in,然后进行有利突变株的筛选,通过初筛、复筛和遗传稳定性试验,获得1株遗传性状稳定的腈水合酶高产菌株HUST-1,其酶活高达698.6U,比诱变前提高了68.3%。[结论]通过紫外线和硫酸二乙酯复合诱变能选育出有较高腈水合酶活力的菌株。[ Objective ] The aim was to select effective methods to breed the stable high-yield strains of nitrile hydratase. [ Method ] With Rhodococcus sp. HUST as the starting strain, the mutant with high enzyme activity were bred after HUST was treated by compound mutation with ultraviolet and diethyl sulfate mutation, and then its nitrile hydratase activity was detected. [Result] The compound mutation condition of HUST strain was that after the starting strain was mutated by ultraviolet for 45 s at the distance of 15 era, it was mutated by 1.1% diethyl sulfate for 20 min, then the available mutant was screened and a high-yield strain HUST-1 of nitrile hydratase with stable genetic characters was obtained through the primary screening, secondary screening and the genetic stability experiment. The enzyme activity of HUST-1 was 698.6 U, which was 68.3% higher than the starting strain. [ Conclusion] Compound mutation with ultraviolet and diethyl sulfate mutation could breed strains with higher nitrile hydratase activity.

关 键 词：复合诱变 腈水合酶 选育 

分 类 号：TQ920.1[轻工技术与工程—发酵工程]