Synthesis of Calf Prochymosin (prorennin) in Escherichia Coli

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1983 Jun 1

PMID 6304731

Citations 25

Authors

J S Emtage

S Angal

M T Doel

T J Harris

B Jenkins

G Lilley

P A Lowe

Affiliations

Soon will be listed here.

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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