生物加工过程2023，Vol.21Issue(6)：632-641,10.DOI:10.3969/j.issn.1672-3678.2023.06.006
多酶—锅法合成硫酸基供体3'-磷酸腺苷-5'-磷酸硫酸(PAPS)
One-pot biosynthesis of sulfate group donor 3'-phosphoadenosine-5'-phosphosulfate(PAPS)through catalysis with multiple enzymes
摘要
Abstract
The application of two enzymes,ATP sulfatase(ATPS)from Kluyveromyces lactis and APS kinase(APSK)from Penicillium chrysogenum,could readily convert adenine nucleoside triphosphate(ATP)to 3'-phosphoadenosine-5'-phosphosulfate(PAPS).More importantly,the addition of inorganic pyrophosphorylase(PPase)from Pseudomonas aeruginosa could hydrolyze the corresponding by-product PPi generated in this enzymatic process,which assisted the reaction to proceed in the direction of PAPS biosynthesis.In this study,three recombinant plasmids pET-28a-sumo-ATPS,pET-28a-APSK,and pET-28a-PPase were constructed to express the corresponding proteins in E.coli.Through the investigation of enzymatic properties and the optimization of reaction conditions,the cumulative yield of PAPS was greatly improved to 14.47 g/L.This one-pot enzymatic process can potentially offer an alternative approach for the production of PAPS at relatively low cost.关键词
3'-磷酸腺苷-5'-磷酸硫酸/ATP硫酸化酶/APS激酶/无机焦磷酸水解酶Key words
3'-phosphoadenosine-5'-phosphosulfate/ATP sulfurylase/APS kinase/inorganic pyrophosphohydrolase分类
生物科学引用本文复制引用
岳继恒,储梦萍,储建林,何冰芳..多酶—锅法合成硫酸基供体3'-磷酸腺苷-5'-磷酸硫酸(PAPS)[J].生物加工过程,2023,21(6):632-641,10.基金项目
国家重点研发计划(2018YFA0902000) （2018YFA0902000）
江苏省自然科学基金面上项目(BK20181379) （BK20181379）
