食品与生物技术学报2024,Vol.43Issue(9) :163-172.DOI:10.12441/spyswjs.20240224004

多酶一步法还原石胆酸制备熊去氧胆酸

Multi-Enzymatic One-Pot Reduction of Lithocholic Acid to Ursodeoxycholic Acid

吕迪 席志文 解修兵 徐岩 张文池 张荣珍
食品与生物技术学报2024,Vol.43Issue(9) :163-172.DOI:10.12441/spyswjs.20240224004

多酶一步法还原石胆酸制备熊去氧胆酸

Multi-Enzymatic One-Pot Reduction of Lithocholic Acid to Ursodeoxycholic Acid

吕迪 1席志文 1解修兵 1徐岩 1张文池 2张荣珍1
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作者信息

  • 1. 江南大学工业生物技术教育部重点实验室,江苏 无锡 214122
  • 2. 约翰·霍普金斯大学医学院,美国马里兰州 巴尔的摩21205
  • 折叠

摘要

熊去氧胆酸是一种重要的治疗性胆汁酸,特别是在治疗胆囊炎和胆固醇型胆结石方面有重要应用.利用大肠杆菌全细胞系统中的P450单加氧酶CYP107D1(OleP),在石胆酸7β-位置区域进行选择性和立体选择性羟基化反应产生熊去氧胆酸.一步法生物合成熊去氧胆酸可以避免残酷的活熊胆汁提取和低效率的化学合成.通过对OleP的突变体F84Q/S240A/V291G(OlePs)与氧化还原伴侣蛋白的共表达组合设计,构建了一种高效合成熊去氧胆酸的重组表达菌株Escherichia coli BL21/PDR-PDX-SD-AS-OlePs.基于重组菌株E.coli BL21/PDR-PDX-SD-AS-OlePs的全细胞催化反应,熊去氧胆酸产量显著提升,产率为88.28%,比未共表达氧化还原伴侣时的产率提升73.96%.该研究结果在一步法还原石胆酸制备熊去氧胆酸方面取得进展,为以生物废料石胆酸为底物制备熊去氧胆酸开辟了新的合成途径.

Abstract

Ursodeoxycholic acid is an important therapeutic bile acid,especially in the treatment of cholecystitis and the prevention of cholesterol-type gallstones.The authors produced ursodeoxycholic acid through regioselective and stereoselective hydroxylation at the 7β-position of lithocholic acid using the P450 monooxygenase CYP107D1(OleP)in E.coli whole-cell system.Ursodeoxycholic acid was biosynthesized using an efficient one-step method,avoiding the cruel practice of extracting bile from live bear and the inefficiencies of chemical synthesis.Through the design of the co-expression combination of the mutant F84Q/S240A/V291G(OlePs)of OleP and the redox chaperone,a recombinant expression strain Escherichia coli BL21/PDR-PDX-SD-AS-OlePs was developed.Utilizing this recombinant strain,the whole-cell catalytic reaction of E.coli BL21/PDR-PDX-SD-AS-OlePs led to a substantial increase in a significantly increased yield of ursodeoxycholic acid yield at 88.28%,which was 73.96%higher than that of without the co-expression of redox chaperone.The author has made progress in the preparation of ursodeoxycholic acid through one-step reduction of lithocholic acid,offering possibilities for the synthesis of ursodeoxycholic acid from biowaste lithocholic acid.

关键词

P450单加氧酶/理性设计/石胆酸/熊去氧胆酸/全细胞一步催化

Key words

P450 monooxygenase/rational design/lithocholic acid/ursodeoxycholic acid/whole-cell one-step catalysis

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出版年

2024
食品与生物技术学报
江南大学

食品与生物技术学报

CSTPCD北大核心
影响因子:0.674
ISSN:1673-1689
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