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. 1983 Jun;80(12):3671-5.
doi: 10.1073/pnas.80.12.3671.

Synthesis of calf prochymosin (prorennin) in Escherichia coli

J S Emtage, S Angal, M T Doel, T J Harris, B Jenkins, G Lilley, P A Lowe

Synthesis of calf prochymosin (prorennin) in Escherichia coli

J S Emtage et al. Proc Natl Acad Sci U S A. 1983 Jun.

Abstract

A gene for calf prochymosin (prorennin) has been reconstructed from chemically synthesized oligodeoxyribonucleotides and cloned DNA copies of preprochymosin mRNA. This gene has been inserted into a bacterial expression plasmid containing the Escherichia coli tryptophan promoter and a bacterial ribosome binding site. Induction of transcription from the tryptophan promoter results in prochymosin synthesis at a level of up to 5% of total protein. The enzyme has been purified from bacteria by extraction with urea and chromatography on DEAE-cellulose and converted to enzymatically active chymosin by acidification and neutralization. Bacterially produced chymosin is as effective in clotting milk as the natural enzyme isolated from calf stomach.

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