Expression of catalytically active radish 3‐hydroxy‐3‐methylglutaryl coenzyme A reductase in Escherichia coli

FEBS Letters

Wettstein, Annette¹  Ferrer, Albert²  Bach, Thomas J.¹  Boronat, Albert²  Aparicio, Cristina²  Nogués, Núria² 

[1] Botanisches Institut II, Universität Karlsruhe, Kaiserstrasse 12, D-7500 Karlsruhe, FRG;Unitat de Bioquímica, Facultat de Farmacia, Universitat de Barcelona, 08028-Barcelona, Spain

关键词: HMG-CoA reductase;    Isoprenoid biosynthesis;    Cloned enzyme expression;    Rhaphanus sativus;    Escherichia coli;   

DOI  :  10.1016/0014-5793(90)81508-L

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Two fragments of a cDNA encoding radish 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) were cloned into the vector pET-8c and expressed in Escherichia coli. The large fragment, encoding both the membrane and the cytosolic domains, was expressed at low level, essentially as an insoluble protein without enzymatic activity. In contrast, the fragment encoding only the cytosolic domain was expressed at a high level in a catalytically active form. The amount of soluble active enzyme in cell-free extracts of E. coli dramatically increased when the temperature during the induction was lowered from 37°C to 22°C.

【 授权许可】

Unknown