土壤中腈水解酶产生菌的快速筛选.pdfVIP

Vol. 35 No. 4 ( ) 2009- 08 Journal of East China University of cience and Technology ( Natural cience Edition) 545 : 1006-3080( 2009) 04-0545-04 周冬杰, 欧阳立明, 许建和, 周鹏鹏, 潘 江 ( 华东理工大学生物反应器工程国家重点实验 , 生物催化研究 , 上海200237) : 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 : 腈水解酶; 苯酚-次氯酸盐比色法( Berthelot 比色法) ; 微孔板; 筛选 : Q939. 9; Q33 : A Rapid Screening of Nitrilase Producing Strains from Soil ZH O UDong-j ie, OUYA N GL i-ming, X UJian-he, ZH OUPeng-p eng, PA N J iang (Laboratory of iocatalysis, StateK ey Laboratory of ioreactor Engineering, East China University of Science and Technology , Shanghai 200237, China) Abstract: T o isolate nitrilase producing strains from soil, a rapid enzyme activity analysis method based on phenol-hypochlorite colorimetry ( Berthelot colorimetry) for the determination of ammonia w as established. U sing this method, 9 strains w ith relatively high nitrilase activity w ere obtained. The p-hydroxybenzonitride conversion rate of the 9 strains w ere also quantitatively determined by HPLC and the results of the tw o methods corresponded w ell. The highest conversion rate of one of the 9 strains w as 49. 9%. 16s rDNA sequence analysis show ed that the 9 strains belong to genus of Chryseobacterium, acillus, Stap hylococcus, Stenotrop homonas etc. Key words: nitrilase; phenol-hypochlorite colorimetry ( Berthelot colorimetry) ; micro plate; screening [ 2] ( Nitrilase) , ( Nitrile hydratas