VOPROSY MEDITSINSKOI KHIMII (ISSN 0042-8809)

Isolation of B. licheniformis glutamylendopeptidase precursor and study of its in vitro processing

   
Serkina A.V.1, Bushueva A.M.1, Chestukhina G.G.1, Gumpert J.2, Hoischen Ch.2, Kujau M.2, Shevelev A.B.1

1. Stepanov Laboratory of Protein Chemistry, Federal Research Center GNIIGenetika
2. Research Group of Prokaryotic Cell Biology, Institute for Molecular Biotechnology, Beutenbergstr
PubMed Id: 11385993
Year: 2001  Volume: 47  Issue: 1  Pages: 111-122
A secretory system based on L-form cells of Proteus mirabilis was developed for production of native Bacillus licheniformis glutamylendopeptidase precursor never formerly available. The produced precursor was stable per se under physiological conditions and in presence of trypsin and glutamylendopeptidase from B. intermedius. Complete conversion of the precursor to the mature glutamylendopeptidase was performed by bacillar metalloproteases and subtilisin. The artificially processed glutamylendopeptidase was purified by affinity chromatography on bacitracin-sepharose. Native tertiary structure in the purified glutamylendopeptidase was confirmed by demonstrating its activity towards a specific chromogenous peptide substrate.
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Serkina A.V., Bushueva A.M., Chestukhina G.G., Gumpert J., Hoischen Ch., Kujau M., Shevelev A.B. (2001) Voprosy meditsinskoi khimii, 47(1), 111-122.
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