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Expression of catalytically active radish 3‐hydroxy‐3‐methylglutaryl coenzyme A reductase in Escherichia coli
Author(s) -
Ferrer Albert,
Aparicio Cristina,
Nogués Núria,
Wettstein Annette,
Bach Thomas J.,
Boronat Albert
Publication year - 1990
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(90)81508-l
Subject(s) - escherichia coli , complementary dna , reductase , cytosol , biochemistry , enzyme , coenzyme a , cofactor , chemistry , microbiology and biotechnology , biology , gene
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.