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PMID: 6304731 Published · ppublish English Journal Article

Synthesis of calf prochymosin (prorennin) in Escherichia coli.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 80 ·No. 12 ·1983-06-00 ·Pages 3671-5

Emtage JS, Angal S, Doel MT, Harris TJ, Jenkins B, Lilley G, Lowe PA

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.

MeSH Terms
Animals Base Sequence Cattle Chymosin/genetics,metabolism Cloning, Molecular DNA/metabolism DNA Restriction Enzymes Enzyme Precursors/genetics,metabolism Escherichia coli/genetics Plasmids RNA, Messenger/genetics
Chemicals
Enzyme Precursors RNA, Messenger DNA DNA Restriction Enzymes prorennin Chymosin
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Emtage J S
Angal S
Doel M T
Harris T J
Jenkins B
Lilley G
Lowe P A
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.80.12.3671
Published
1983-06-00
Pages
3671-5
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC394112
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.80.12.3671 PMC Full Text View journal details
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