工业微生物2025，Vol.55Issue(2)：19-21,3.DOI:10.3969/j.issn.1001-6678.2025.02.007
黄嘌呤氧化酶在微生物代谢中的调控机制与酶活性研究
The Regulatory Mechanism and Enzymatic Activity of Xanthine Oxidase in Microbial Metabolism
杨平1
作者信息
- 1. 常州卫生高等职业技术学校,江苏 常州 213000
- 折叠
摘要
Abstract
In this study,the regulatory mechanism of xanthine oxidase(XO)in the metabolism of Escherichia coli and its impact on uric acid metabolism were systematically explored.The XO gene was knocked out using CRISPR-Cas9 technology.The results of qRT-PCR indicated that the mRNA expression level in the XO gene knockout group was significantly decreased.In the overexpression experiment,by constructing the pET28a expression vector,the enzymatic activity of the XO overexpression strain was increased.Enzymatic activity determination and enzyme kinetics analysis demonstrated that the enzymatic activity of the R258A mutant was significantly enhanced,and the uric acid conversion rate was increased.However,the enzymatic activity of the E402Q mutant was significantly reduced.Through the research on enzymatic activity and enzyme kinetics,the crucial roles of the R258 and E402 sites in the catalytic mechanism of XO were disclosed.This study provided a theoretical basis for the application and optimization of xanthine oxidase in industrial microorganisms and layed a foundation for enhancing enzyme performance through directed mutation technology in the future.关键词
黄嘌呤氧化酶/大肠杆菌/酶活性/定点突变/基因工程Key words
xanthine oxidase/Escherichia coli/enzymatic activity/site-directed mutation/genetic engineering引用本文复制引用
杨平..黄嘌呤氧化酶在微生物代谢中的调控机制与酶活性研究[J].工业微生物,2025,55(2):19-21,3.
