辅酶自循环再生系统高效合成熊去氧胆酸
Ursodeoxycholic acid was efficiently synthesized by coenzyme self-circulation regeneration system
胡青 1何建新 1宋士奎 2苏正定3
作者信息
- 1. 湖北工业大学生命科学与健康工程学院,湖北武汉 430068
- 2. 新疆师范大学生命科学学院,新疆乌鲁木齐 830017
- 3. 新疆大学药学院,新疆乌鲁木齐 830046
- 折叠
摘要
[目的]运用突变后的高活性7α-羟基类固醇脱氢酶(7α-HSDH)和7β-羟基类固醇脱氢酶(7β-HSDH)偶联成一组辅酶自循环再生催化系统,优化催化体系高效合成熊去氧胆酸(UDCA).[方法]通过人工优化合成沙丁梭菌(Clostridium absonum)DSM 599 的 7α-HSDH 和瘤胃球菌(Ruminococcus torques)ATCC 35915 的 7β-HSDH 突变型基因;对7α/β-HSDH诱导温度进行了梯度优化;以单位时间内鹅去氧胆酸(CDCA)的转化率为考察指标,筛选出葡萄糖脱氢酶(GDH)、7α/β-HSDH添加比例、温度、pH值、助溶剂对催化反应体系的影响.[结果]7α/β-HSDH最佳诱导温度为30 ℃,CDCA催化合成UDCA最佳工艺参数为:7α/β-HSDH添加比例为1∶1.5,温度为30℃,pH为8.0,助溶剂为0.4％的吐温80,在此催化反应体系CDCA转化率可达到90.8％.[结论]选用的高活性的7α-HSDH和7β-HSDH可以实现辅酶自循环再生,通过对催化体系的优化可经济高效的将CDCA催化合成UDCA.
Abstract
[Objective]To optimize the catalytic system for the efficient synthesis of ursodeoxycholic acid(UDCA)by using mutated highly active 7 α-hydroxysteroid dehydrogenase(7 α-HSDH)and 7 β-hydroxysteroid dehydrogenase(7 β-HSDH)coupled to form a set of coenzyme self-circulation regeneration catalytic systems.[Method]The 7α-HSDH mutant gene of Clostridium absonum DSM 599 and the 7 β-HSDH mutant gene of Ruminococcus torques ATCC 35915 were synthesized by arti-ficial optimization.The gradient optimization of 7α/β-HSDH induction temperature was carried out,and the conversion rate of chenodeoxycholic acid(CDCA)per unit time was used as the index to screen out the effects of glucose dehydrogenase(GDH),7α/β-HSDH addition ratio,temperature,pH and co-solvent on the catalytic reaction system.[Result]The optimal induction temperature of 7α/β-HSDH was 30 ℃,and the optimal process parameters for the synthesis of UDCA catalyzed by CDCA were as follows:the addition ratio of 7α/β-HSDH was 1:1.5,the temperature was 30 ℃,the pH value was 8.0,and the co-solvent was 0.4％Tween 80,and the conversion rate of CDCA in the catalytic reaction system could reach 90.8％.[Conclusion]The highly active 7α-HSDH and 7 β-HSDH can realize the self-recycling and regeneration of coenzymes,and the synthesis of UDCA can be catalyzed by CDCA cost-effectively through the optimization of the catalytic system.
关键词
7α-HSDH/7β-HSDH/辅酶自循环/CDCA/UDCA/催化体系/工艺优化/转化率Key words
7α-HSDH/7β-HSDH/coenzyme self-circulation/CDCA/UDCA/catalytic system/process optimization/conversion rate引用本文复制引用
出版年
2024
NETL
NSTL
万方数据