一种胆固醇氧化酶高产菌株筛选方法的建立

A Novel Screening Method for Isolating High Cholesterol Oxidase Producing Strain

摘要:

用亚硝基胍(1 mg/mL)与超声波(200 W, 50 kHz)复合诱变的方法, 对产胆固醇氧化酶短杆菌Brevibacterium sp. DGCDC-82进行诱变处理, 得到一株桔红色突变株其产胆固醇氧化酶能力提高140%, 酶活达到1.24 U/mL, 又用同样的方法对桔红色突变株进行回复突变处理, 得到一株白色回复突变株和一株淡粉色回复突变株, 两株回复突变株产胆固醇氧化酶的能力又明显下降, 酶活分别为0.17 U/mL和0.69 U/mL。说明短杆菌Brevibacterium sp.产胆固醇氧化酶能力与其产红色素成正相关偶联关系, 这种相关性模型的建立可以作为以后诱变或定向进化研究的筛子。

Abstract:

The cholesterol oxidase producing strain Brevibacterium sp. DGCDC-82 was treated with NTG (1 mg/mL) under ultrasonicztion(200 W, 50 kHz). A red mutant named Brevibacterium sp. DGCCN-25 showed higher and stable production of cholesterol oxidase was obtained. The enzyme activity was increased by 140%, it is 1.24 U/mL.Then dealed with DGCCN-25 using the same method, two revertants were obtained, one was white and the other was rose pink. The enzyme activity of two revertants was obvious decrease, they are 0.17 U/mL and 0.69 U/mL.The results showed the positive correlation between COD acticity and red pigment producing by Brevibacterium sp.. The relativity model can be used as a method of screening for mutation and directed evolution.