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Stepwise application of affinity chromatography on bacitracin-silochrome, gel filtration on Acrylex P-10, rechromatography on bacitracin-Sepharose 4B and gel filtration on Sephadex G-15, a homogeneous metalloproteinase (M(r) = 35,000 Da) has been isolated from the cultural filtrate of B. megaterium strain 599. The amino acid composition and N-terminal sequence (20 amino acids) of the enzyme have been determined. The proteinase is not inhibited by diisopropyl-fluorophosphate, is inhibited by o-phenanthroline, EDTA, and Zn2+, and is activated by Co2+. The enzyme has a peak activity at 60-65 degrees C. The maximum of the enzymatic activity after hydrolysis of synthetic substrates is at pH 6.5-7.0. The enzyme is stable at pH 7.0-9.0 and retains its stability at 45-60 C for several hours. In acid media the enzyme undergoes irreversible inactivation. The dependence of kcat/Km on pH points to the involvement of an ionogenic group with pKa 7.5 in the catalytic act, most probably of the imidazole group of histidine. The metalloproteinase hydrolyzes synthetic peptide substrates at the bonds formed by the amino groups of hydrophobic amino acids-Phe, Leu, Ile and Val.
I P Morozova, and
G G Chestukhina, and
M E Bormatova, and
M Iu Gololobov, and
N M Ivanova, and
E N Lysogorskaia, and
I Iu Filippova, and
O M Khodova, and
E A Timokhina, and
V M Stepanov
January 2014,
PloS one,
I P Morozova, and
G G Chestukhina, and
M E Bormatova, and
M Iu Gololobov, and
N M Ivanova, and
E N Lysogorskaia, and
I Iu Filippova, and
O M Khodova, and
E A Timokhina, and
V M Stepanov
January 1975,
The Journal of general virology,
I P Morozova, and
G G Chestukhina, and
M E Bormatova, and
M Iu Gololobov, and
N M Ivanova, and
E N Lysogorskaia, and
I Iu Filippova, and
O M Khodova, and
E A Timokhina, and
V M Stepanov
January 1987,
Microbiology and immunology,
I P Morozova, and
G G Chestukhina, and
M E Bormatova, and
M Iu Gololobov, and
N M Ivanova, and
E N Lysogorskaia, and
I Iu Filippova, and
O M Khodova, and
E A Timokhina, and
V M Stepanov
September 1977,
The Journal of general virology,
I P Morozova, and
G G Chestukhina, and
M E Bormatova, and
M Iu Gololobov, and
N M Ivanova, and
E N Lysogorskaia, and
I Iu Filippova, and
O M Khodova, and
E A Timokhina, and
V M Stepanov
July 1983,
Hoppe-Seyler's Zeitschrift fur physiologische Chemie,
I P Morozova, and
G G Chestukhina, and
M E Bormatova, and
M Iu Gololobov, and
N M Ivanova, and
E N Lysogorskaia, and
I Iu Filippova, and
O M Khodova, and
E A Timokhina, and
V M Stepanov
September 1980,
Journal of bacteriology,
I P Morozova, and
G G Chestukhina, and
M E Bormatova, and
M Iu Gololobov, and
N M Ivanova, and
E N Lysogorskaia, and
I Iu Filippova, and
O M Khodova, and
E A Timokhina, and
V M Stepanov
March 1966,
The Biochemical journal,
I P Morozova, and
G G Chestukhina, and
M E Bormatova, and
M Iu Gololobov, and
N M Ivanova, and
E N Lysogorskaia, and
I Iu Filippova, and
O M Khodova, and
E A Timokhina, and
V M Stepanov
January 1990,
Folia microbiologica,
I P Morozova, and
G G Chestukhina, and
M E Bormatova, and
M Iu Gololobov, and
N M Ivanova, and
E N Lysogorskaia, and
I Iu Filippova, and
O M Khodova, and
E A Timokhina, and
V M Stepanov
March 1990,
Journal of general microbiology,
I P Morozova, and
G G Chestukhina, and
M E Bormatova, and
M Iu Gololobov, and
N M Ivanova, and
E N Lysogorskaia, and
I Iu Filippova, and
O M Khodova, and
E A Timokhina, and
V M Stepanov
January 1986,
Journal of bacteriology,
