3-甾酮-1-脱氢酶基因的表达及甾体转化分析

中国生物工程杂志

研究报告

3-甾酮-1-脱氢酶基因的表达及甾体转化分析

李玉路福平孙同毅刘逸寒杜连祥

天津科技大学生物工程学院天津科技大学生物工程学院天津科技大学生物工程学院天津科技大学生物工程学院天津科技大学生物工程学院

Expression of ksdD gene encoding 3-ketosteroid-△1-dehydrogenase from Arthrobacter simplex in Bacillus subtilis and analysis of steroid

全文: PDF HTML

摘要： 本文利用带有P43启动子的表达分泌载体pWB980，实现了简单节杆菌3-甾酮-1-脱氢酶在枯草芽孢杆菌中的表达，表达出的目的蛋白的分子量为55KDa。利用分光光度法检测得到胞内和胞外可溶性部分的酶活分别为110±0.5mU和15±0.6mU每毫克蛋白, 比出发菌株简单节杆菌提高了将近30倍。重组芽孢杆菌对甾体底物4-AD的转化率为45.3％，比出发菌株简单节杆菌提高了近10倍。利用枯草芽孢杆菌对甾体底物进行脱氢为甾体药物的生产开辟了一个新的途径。

Abstract: To improve 3-ketosteroid-△1-dehydrogenase(KSDH) activity and the transformation level for androst-4-ene-3,17-dione, 3-ketosteroid-△1-dehydrogenase gene(ksdD) from Arthrobacter Simplex was cloned into plasmid pWB980 and expressed in B. subtilis WB600 under the control of promoter P43. The molecular weight of expressed enzyme was about 55KDa by SDS-PAGE analysis. The activitities assayed by spectrophotometrical method of intracellular and extracellular soluble enzyme were 110±0.5mU and 15±0.6mU per milligram of protein respectively. The transformation rate of androst-4-ene-3,17-dione by the B. subtilis recombinant cells was 45.3%. Compared with Arthrobacter Simplex, the enzyme activity of KSDH expressed in B. subtilis was improved about 30 fold, and the transformation level of androst-4-ene-3,17-dione by the B. subtilis recombinant cells was improved about 10 fold. The recombinant B. subtilis cells used in biotransformation of steroids provided a new way for production steroid medicines.

Key words: expression steroid transformation. 3-ketosteroid-△1-dehydrogenase B. subtilis

收稿日期: 2006-06-29 出版日期: 2006-11-25

通讯作者: 李玉

	服务
	把本文推荐给朋友
	加入引用管理器
	E-mail Alert
	RSS
	作者相关文章
	路福平
	刘逸寒
	杜连祥
	孙同毅
	李玉

引用本文:

李玉,路福平,孙同毅,刘逸寒,杜连祥. 3-甾酮-1-脱氢酶基因的表达及甾体转化分析[J]. 中国生物工程杂志, .

. Expression of ksdD gene encoding 3-ketosteroid-△1-dehydrogenase from Arthrobacter simplex in Bacillus subtilis and analysis of steroid. China Biotechnology, .

链接本文:

https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2006/V26/I11/24

No related articles found!

Viewed

Full text

Abstract

Cited

Shared

Discussed