[Isolation of glutamylendopeptidase precursor from B. licheniformis and its processing in vitro].
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.
Serkina
A V
AV
Stepanov Laboratory of Protein Chemistry, Federal Research Center GNIIGenetika, 1st Dorozhny pr., 1, 113545 Moscow, Russia. serkina@rocketmail.com
Bushueva
A M
AM
Chestukhina
G G
GG
Gumpert
J
J
Hoischen
C
C
Kujau
M
M
Shevelev
A B
AB
D001407:Bacillus / Q000201:enzymology* / Q000235:genetics; D001483:Base Sequence; D002846:Chromatography, Affinity; D004789:Enzyme Activation; D004792:Enzyme Precursors / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D010957:Plasmids; D011499:Protein Processing, Post-Translational; D012697:Serine Endopeptidases / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism
D004792:Enzyme Precursors; D015345:Oligonucleotide Probes; D012697:Serine Endopeptidases; C028344:glutamyl endopeptidase