食品科学2018，Vol.39Issue(6)：69-74,6.DOI:10.7506/spkx1002-6630-201806012
蜡样芽孢杆菌Bacillus cereus LJ01中亚硝酸盐还原酶的基因克隆、表达和纯化
Cloning, Expression and Purification of the Nitrite Reductase Gene from Bacillus cereus LJ01
摘要
Abstract
A nitrite reductase (NiR) gene was cloned from Bacillus cereus LJ01 and expressed in Escherichia coli.The recombinant enzyme was purified by His-Tag nickel affinity chromatography (Ni Sepharose 6 Fast Flow) and DEAE Sepharose Fast Flow ion exchange chromatography for characterization.The results showed that the recombinant NiR gene contained a 1 623 bp-length ORF encoding 540 amino acids,and the molecular weight was about 67 ku.Iron and copper ions were simultaneously present in the enzyme and their contents were 51.0 and 184.5 mg/kg,respectively.The results of circular dichroism showed that the helix structure accounted for the largest proportion of the recombinant NiR.关键词
亚硝酸盐还原酶/蜡样芽孢杆菌/基因克隆/蛋白表达/工程菌Key words
nitrite reductase/Bacillus cereus/gene cloning/protein expression/engineered strain分类
生物科学引用本文复制引用
陈思敏,罗彤晖,费永涛,吴健锋,刘冬梅..蜡样芽孢杆菌Bacillus cereus LJ01中亚硝酸盐还原酶的基因克隆、表达和纯化[J].食品科学,2018,39(6):69-74,6.基金项目
国家自然科学基金青年科学基金项目(31101254) （31101254）
广东省自然科学基金项目(S2011010005679) （S2011010005679）
广东省科技攻关项目(2014A020208019 （）
2013B020312002) （）
