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  • Title: Synthesis of calf prochymosin (prorennin) in Escherichia coli.
    Author: Emtage JS, Angal S, Doel MT, Harris TJ, Jenkins B, Lilley G, Lowe PA.
    Journal: Proc Natl Acad Sci U S A; 1983 Jun; 80(12):3671-5. PubMed ID: 6304731.
    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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