一种产邻苯二酚菌株高通量筛选方法

目的：对产邻苯二酚菌株进行筛选。方法：采用前期筛得的产邻苯二酚菌为出发菌株,通过硫酸二乙酯诱变的方法使该菌株突变,同时建立96孔板培养和酶标仪检测方法对产邻苯二酚菌株进行高通量筛选。结果：硫酸二乙酯的终浓度为0.1%,诱变时间为8 min的条件下,突变菌致死率为84.5%,突变效果最好。筛选培养基中吸光值（495nm）和富集培养基中菌液浊度值（OD630）的加和值较大的突变菌株产邻苯二酚能力高。通过诱变和筛选得到的菌株,产邻苯二酚浓度可达0.87mg/ml,比出发菌株的提高了262.5%。经过形态学和生理生化反应,初步鉴定该菌株属于假单胞菌属（Pseudomonas sp.）。结论：硫酸二乙酯诱变和96孔板筛选的方法能以高通量方式快速筛选出产邻苯二酚菌株。

A Method of High Throughput Screen of Catechol-Producing Strain

Objective: Screen of strains producing catechol. Method: Strains producing catechol were screened using diethyl sulphate as mutageniz-ing reagent and 96 well plate screening method. Result: It showed that mutagenizing reagent content at ratio of 0.1 %, at treatment time of 8min were suitable conditions for mutagenesis. Under specific conditions, the lethal rate was 84.5% .The mutant strains which showed high absorbance value in 495nm in sceening fermentation medium and high OD_(630) in enrichment medium would be better candidate for catechol-producing. Though mutagenizing and plate selection, the catechol yield of the strian was 0.87rng/ml, increasing by 262.5% compared to the original one. The strain was preliminary identified as Pseudomonas sp. by means of morphological, physiological and biochemical identification. Conclusion: Catechol producing strains could rapidly screened by high throughout screening using diethyl sulphate as mutagenizing reagent and 96 well plate screening method.