食品科学2024，Vol.45Issue(24)：92-99,8.DOI:10.7506/spkx1002-6630-20240504-003
基于理性设计提高酿酒酵母烟酰胺核糖激酶1热稳定性
Rational Design for Improving the Thermostability of Saccharomyces cerevisiae Nicotinamide Riboside Kinase 1
摘要
Abstract
In order to improve the thermal stability of nicotinamide riboside kinase 1 from Saccharomyces cerevisiae(Sc NRK1),six single-point mutants of Sc NRK1 were virtually designed using computer-aided technology,and their expression using site-directed mutagenesis and enzymatic characterization were carried out.Out of these,three superior mutants were selected for a second round of combined mutagenesis.The results showed that after two rounds of mutagenesis,a mutant named T136P/S209A with significantly improved thermal stability and catalytic activity was obtained.Its optimal reaction temperature was increased to 45℃,and its half-life at 45℃was 48.98 min,which was 4.2 times as high as that of the wild-type ScNRK1.The specific enzyme activity of the purified mutant enzyme was 146.63 U/mg,which was 1.98 times as high as that of the wild type.This study is expected to provide new ideas for improving the thermal stability of enzymes through rational design,and provide a new enzyme source for efficient and low-cost production of nicotinamide mononucleotide.关键词
烟酰胺核糖激酶1/理性设计/定点突变/热稳定性/烟酰胺单核苷酸Key words
nicotinamide riboside kinase 1/rational design/site-directed mutagenesis/thermal stability/nicotinamide mononucleotide分类
生物科学引用本文复制引用
王瑶,沈太松,李思晨,史红玲,姚伦广,唐存多..基于理性设计提高酿酒酵母烟酰胺核糖激酶1热稳定性[J].食品科学,2024,45(24):92-99,8.基金项目
河南省青年科学家项目(225200810076) （225200810076）
