海洋科学2016，Vol.40Issue(5)：1-8,8.DOI:10.11759/hykx20151031001
南极低温降解菌Shewanella sp. NJ49羟化酶分离纯化及最适产酶条件研究
Separation and purification of hydroxylase from Antarctic psychrophilic bacteriumShewanella sp. NJ49 and optimum conditions for its enzyme production
摘要
Abstract
Hydroxylase is a key enzyme with respect to bacteria during hydrocarbon degradation. In this paper, we investigate the Antarctic psychrophilic bacteriumShewanella sp. NJ49. To obtain a single component of hydroxy-lase, we separated and purified hydroxylase from NJ49 in a series of steps, i.e., ammonium sulfate precipitation, ultrafiltration concentration, and column chromatography with Sephadex G-75 as the column media. In addition, we studied the relationships between the activity of NJ49 hydroxylase and environmental factors such as temperature, pH, surfactant, and salinity. We found that purified hydroxylase comprised subunitsα,β, andγ, with estimated molecular masses of 56, 45, and 36 kD, respectively. The optimum conditions for producing NJ49 hydroxylase were a temperature of 15℃, a pH of 7.2, and a salinity of 55‰. Different types of surfactants had different effects on enzyme activity. Although individual amphoteric, cationic, and nonionic surfactants did not inhibit enzyme activity, a mixture of anionic and nonionic surfactants, as well as a mixture of cationic and nonionic surfactants, improved the activity of the hydroxylase.关键词
Shewanella/羟化酶/分离纯化/环境因素Key words
Shewanella/hydroxylase/separation and purification/environmental factors分类
生物科学引用本文复制引用
刘芳明,王以斌,曲长凤,梁强,郑洲,缪锦来,肖天..南极低温降解菌Shewanella sp. NJ49羟化酶分离纯化及最适产酶条件研究[J].海洋科学,2016,40(5):1-8,8.基金项目
国家自然科学基金项目(31200097 （）
41576187) （）
中央级公益性科研院所基本科研业务费专项资金(2013G33 （）
2015G10) （）
国家自然科学基金-山东省联合资助开发项目(U1406402-5)@@@@National Science Foundation of China , No.31200097, No.41576187 （U1406402-5）
Basic science and technology research funds projects of central level Public Scientific Research Institute,2013G33,2015G10 （）
The Joint Funds of the National Natural Science Foundation of China and Shandong Province, U1406402-5。 （）
