土壤中腈水解酶产生菌的快速筛选  

Rapid Screening of Nitrilase Producing Strains from Soil

作　　者：周冬杰[1] 欧阳立明[1] 许建和[1] 周鹏鹏[1] 潘江[1] 

机构地区：[1]华东理工大学生物反应器工程国家重点实验室,生物催化研究室,上海200237

出　　处：《华东理工大学学报（自然科学版）》2009年第4期545-548,共4页Journal of East China University of Science and Technology

基　　金：上海市自然科学基金(07ZR14030)

摘　　要：为了从土壤中快速筛选具有腈水解能力的微生物,建立了以苯酚-次氯酸盐比色法(又名Berthelot比色法)检测96孔板反应产物中氨离子浓度为基础的腈水解酶酶活快速分析方法。用该法从土壤样本中筛选获得了9个酶活力较高的菌株。用HPLC方法定量检测9个菌株对对羟基苯甲腈的转化率,证明苯酚-次氯酸盐比色法微孔板分光光度计的测定结果与HPLC结果对应良好,转化率最高为49.9%。经16s rDNA鉴定,菌株分别属于金黄杆菌属、芽胞杆菌属、葡萄球菌属、寡养单胞菌属等。To isolate nitrilase producing strains from soil, a rapid enzyme activity analysis method based on phenol-hypochlorite colorimetry （Berthelot colorimetry） for the determination of ammonia was established. Using this method, 9 strains with relatively high nitrilase activity were obtained. The p-hydroxybenzonitride conversion rate of the 9 strains were also quantitatively determined by HPLC and the results of the two methods corresponded well. The highest conversion rate of one of the 9 strains was 49.9%. 16s rDNA sequence analysis showed that the 9 strains belong to genus of Chryseobacterium, Bacillus, Staphylococcus, Stenotrophomonas etc.

关 键 词：腈水解酶 苯酚-次氯酸盐比色法(Berthelot比色法) 微孔板 筛选 

分 类 号：Q939.9[生物学—微生物学]