中度嗜盐菌Martelella sp.AD-3降解菲过程中水杨酸-5-羟化酶的酶学性质  

Characterization of salicylate 5-hydroxylase for phenanthrene degradation using moderately halophilic Martelella sp. AD-3

作　　者：董斐[1] 崔长征[1] 冯天才[1] 冯耀宇[1] 刘勇弟[1] 

机构地区：[1]华东理工大学资源与环境工程学院、国家环境保护化工过程环境风险评价与控制重点实验室,上海200237

出　　处：《微生物学报》2012年第8期985-993,共9页Acta Microbiologica Sinica

基　　金：中央高校基本科研业务费专项资金(WB1114031);高等学校博士学科点专项科研基金(20110074130002);环境模拟与污染控制国家重点联合实验室专项经费(11K02ESPCT)~~

摘　　要：【目的】研究中度嗜盐菌Martelella sp.AD-3在降解菲过程中水杨酸-5-羟化酶的活性与菲降解效率的关系及其酶学性质。【方法】通过HPLC分析菲的降解效率和AD-3菌粗酶液催化水杨酸的产物,根据NADH在340 nm处的吸光度变化计算水杨酸-5-羟化酶的活性。【结果】水杨酸-5-羟化酶是一种诱导酶,在AD-3菌的对数生长期和稳定初期时活性较高,酶活力大小与该菌对菲的降解速率基本一致。在菲浓度为200 mg/L、生长盐度为3%、pH为9.0的培养条件下,AD-3菌株表达的水杨酸-5-羟化酶的活力最高,为132.8 nmol/(min.mg)。水杨酸-5-羟化酶催化水杨酸降解时的最适温度、pH和盐度分别为30℃、7.5和3%,酶的最大反应速率为200 nmol/(min.mg)、米氏常数Km为8.7μmol/L。【结论】AD-3菌在降解菲的过程中表达水杨酸-5-羟化酶,该酶的活性与菲降解速率具有相关性。[Objective] To identify the relationship between salicylate 5-hydroxylase activity and the degradation rate of phenanthrene by moderately halophilic Martelella sp.AD-3,and to investigate their enzymatic characteristics.[Methods] The products resulted from degradation of salicylic acid using the crude enzyme produced by AD-3 was analyzed using HPLC analysis,and salicylate 5-hydroxylase activities at different conditions was calculated according to the change of NADH absorbance at 340 nm.[Results] The salicylate 5-hydroxylase activity was higher during log-phase and early stationary phase of AD-3,and the enzyme activity was in consistent with phenanthrene degradation rate.The enzyme production was induced by phenanthrene and salicylic acid.The highest enzyme activity,132.8 nmol/（min·mg）,from strain AD-3 was obtained under the phenanthrene concentration of 200 mg/L,3% salinity and a pH of 9.0.The optimum conditions for degradation of salicylic acid using this enzyme were 30℃,pH 7.5,and salinity 3%.The Vmax and Km for the enzyme were 200 nmol/（min·mg） and 8.7 μmol/L,respectively.[Conclusion] The salicylate 5-hydroxylase was produced in the degradation of phenanthrene in AD-3.The enzyme activity was related to phenanthrene degradation rate.

关 键 词：菲 水杨酸-5-羟化酶 酶活 盐度 

分 类 号：X172[环境科学与工程—环境科学]