酿酒酵母烟酰胺核苷激酶的异源表达及其酶学性质分析  

Heterologous Expression and Enzymatic Properties of Nicotinamide Nucleoside Kinase from Saccharomyces cerevisiae

作　　者：何建菊 刘欣欣 史红玲[1] 冉璐妮 王贤 张英君[1] 唐存多[1,2,3] HE Jianju;LIU Xinxin;SHI Hongling;RAN Luni;WANG Xian;ZHANG Yingjun;TANG Cunduo(College of Life Science and Agricultural Engineering,Nanyang Normal University,Nanyang 473061,China;Postdoctoral Innovation Practice Base,She Dian Lao Jiu Co.Ltd.,Nanyang 473300,China;College of Food Science and Technology,Henan Agricultural University,Zhengzhou 450002,China)

机构地区：[1]南阳师范学院生命科学与农业工程学院,河南南阳473061 [2]赊店老酒股份有限公司博士后创新实践基地,河南南阳473300 [3]河南农业大学食品科学技术学院,河南郑州450002

出　　处：《食品科学》2023年第16期192-197,共6页Food Science

基　　金：国家自然科学基金青年科学基金项目(31900916);河南省高校科技创新人才项目(21HASTIT041);河南省中央引导地方科技发展资金项目(Z20221343035)。

摘　　要：从酿酒酵母中克隆烟酰胺核苷激酶(nicotinamide riboside kinase,NRK)Sc NRK1的编码基因,借助p ET28a质粒在大肠杆菌中实现了可溶性表达。结果表明,发酵液中酶活力为14.75 IU/m L,纯化后的比活力为2252.59 IU/mg。此外,Sc NRK1的动力学参数也明显优于其他已报道的NRK,在烟酰胺单核苷酸的酶促合成中具有更大的优势。In this study,the nicotinamide riboside kinase 1 gene(ScNRK1)was cloned from Saccharomyces cerevisiae,and its soluble expression in Escherichia coli was achieved using pET28a plasmid.The enzyme activity in the fermentation broth was 14.75 IU/mL,and the specific activity the purified enzyme was 2252.59 IU/mg.In addition,the kinetic parameters of ScNRK1 were significantly higher than those of other reported nicotinamide nucleoside kinases,so ScNRK1 could be more advantageous in the enzymatic synthesis of nicotinamide mononucleotide(NMN).

关 键 词：烟酰胺核苷激酶 β-烟酰胺核苷酸 表达 鉴定 生物催化 

分 类 号：Q814.9[生物学—生物工程]