Production and characterization of a novel extracellular metalloproteinase by a newly isolated moderate halophile, Halobacillus sp. LY6
Language English Country United States Media print-electronic
Document type Journal Article
- MeSH
- Sodium Chloride metabolism MeSH
- Phylogeny MeSH
- Halobacillus classification enzymology genetics MeSH
- Cations, Divalent MeSH
- Hydrogen-Ion Concentration MeSH
- Culture Media chemistry MeSH
- Matrix Metalloproteinases biosynthesis isolation & purification metabolism MeSH
- Molecular Weight MeSH
- RNA, Ribosomal, 16S genetics MeSH
- Temperature MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- Sodium Chloride MeSH
- Cations, Divalent MeSH
- Culture Media MeSH
- Matrix Metalloproteinases MeSH
- RNA, Ribosomal, 16S MeSH
A moderately halophilic bacterium LY6 with high proteolytic activity was isolated. Biochemical and physiological characterization, along with 16S rDNA sequence analysis placed the isolate in the genus Halobacillus. The salinity of the culture medium strongly influenced the proteinase production of LY6. Maximum enzyme production was observed in the medium containing 5% Na(2)SO(4) or 10% NaCl. Proteinase production was synchronized with bacterial growth and reached a maximum level during the mid-stationary phase. Enzyme purification was carried out by a simple approach including a combination of ammonium sulfate precipitation and Sephacryl S-100 gel filtration chromatography. SDS-PAGE and gelatin zymography analysis revealed it was a monomer with high molecular weight of 69 kDa. Optimal proteinase activity was obtained at pH 10.0, 40°C, and 10% NaCl. It was high active over broad temperature (30-80°C), pH (6.0-12.0), and NaCl concentration (0-25%) ranges, indicating its thermostable, alkali-stable, and halotolerant nature. Moreover, the enzyme activity was markedly enhanced by Ca(2+) and Cu(2+), but strongly inhibited by EDTA, PAO, and DEPC, indicating that it probably was a metalloproteinase with cysteine and histidine residues located in its active site.
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