Purification and characterization of a subtilisin-like proteinases secreted in the stationary growth phase of Bacillus amyloliquefaciens H2

Proteinases secreted during the early and late stationary phases have been isolated from the culture liquid of Bacillus amyloliquefaciens H2 using CM-cellulose ion-exchange chromatography with subsequent FPLC on a Mono S column. Considering the character of hydrolysis of specific chromogenic substra...

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Bibliographic Details
Published in:Biochemistry (Moscow) 2007-04, Vol.72 (4), p.459-465
Main Authors: Balaban, N P, Malikova, L A, Mardanova, A M, Rudenskaya, G N, Sharipova, M R
Format: Article
Language:English
Subjects:
Bacillus (Bacteria)
Bacillus - enzymology
Bacillus - growth & development
Bacillus amyloliquefaciens
Bacteria
Cellulose
Electrophoresis, Polyacrylamide Gel
Enzyme Stability
Enzymes
Microbiology
Physiological aspects
Properties
Proteases
Substrate Specificity
Subtilisins - isolation & purification
Subtilisins - metabolism
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Summary:Proteinases secreted during the early and late stationary phases have been isolated from the culture liquid of Bacillus amyloliquefaciens H2 using CM-cellulose ion-exchange chromatography with subsequent FPLC on a Mono S column. Considering the character of hydrolysis of specific chromogenic substrates and the type of inhibition, these enzymes were identified as subtilisin-like proteinases. The molecular weight of both proteinases is 29 kD. The proteolytic activity of the proteinases secreted during the early and late stationary phases towards the synthetic substrate Z-Ala-Ala-Leu-pNA was maximal at pH 8.5 and 9.0, respectively. The maximal activity of both proteinases was observed at 37 degrees C, and the proteins were stable within the pH range of 7.2-9.5. The subtilisin-like proteinases from B. amyloliquefaciens were shown to catalyze synthesis of peptide bonds.
ISSN:0006-2979
1608-3040
0320-9725
DOI:10.1134/S0006297907040141