ABSTRACT
The characteristics of Bacillus pumilus subtilisin-like protease expression were investigated. Potential binding sites for the transcription factor DegU~P have been identified in the regulatory region of the enzyme gene, which have been optimized for interaction with the regulatory protein. The expression of the extracellular subtilisin-like protease modified gene has been examined. It was established that the modification of one of the sites has resulted in increased expression of the proteinase in 2 times. It is concluded that the optimization of the promoter led to increased expression of subtilisin-like protease.
Subject(s)
Bacillus/enzymology , Peptide Hydrolases/biosynthesis , Serine Endopeptidases/biosynthesis , Subtilisin/biosynthesis , Amino Acid Sequence , Gene Expression Regulation, Enzymologic , Peptide Hydrolases/chemistry , Promoter Regions, Genetic , Serine Endopeptidases/chemistry , Serine Endopeptidases/metabolism , Substrate Specificity , Subtilisin/chemistryABSTRACT
Bacillus pumilus 3-19 glutamylendopeptidase has been isolated from culture liquid of Bacillus subtilis recombinant strain on different growth stages: growth retardation (early enzyme) and stationary phase (late enzyme). The effect of purified proteinase of different growth stages on insulin beta-chain, protein and oligopeptide substrates has been studied. Comparative study of physicochemical properties of early and late proteinases was carried out. Two protein fractions were different in catalytic characteristics and demonstrated different sensitivity to the presence of metal cations.