Isolation of a psychrotrophic Exiguobacterium sp. SKPB5 (MTCC 7803) and characterization of its alkaline protease.
Out of nine psychrotrophic bacterial strains isolated from cold environments of the Western Himalayas, SKPB5 was selected for protease purification and characterization because it had the largest zone of clearance on plate assay. On the basis of the phenotypic and biochemical characterization and 16S rRNA gene-sequencing studies, isolate was identified as Exiguobacterium sp. SKPB5. The protease was purified near to homogeneity with a purification fold of 7.1, and its molecular weight was determined to be 36 kDa. The enzyme exhibited maximum stability at 50 degrees C and an optimal pH of 8.0. Metal ions Mg2+, Ca2+, Zn2+, and Mn2+ enhanced the enzyme activity, whereas Cu2+ had no effect. Phenylmethanesulfonyl fluoride and ethylenediaminetetraacetic acid did not show any effect on the activity of the enzyme, whereas a 20% increase in activity was observed when it was incubated in presence of reducing agents such as beta-mercaptoethanol and dithiothreitol. This suggests that the protease isolated from psychrotrophic Exiguobacterium sp. SKPB5 belongs to the cysteine family. The results highlight the relevance of unexplored microbes from cold environments of Western Himalayas for the isolation of protease enzymes active at wide range of temperature and pH.
Kasana
Ramesh C
RC
Hill Area Tea Science Division, Institute of Himalayan Bioresource technology (CSIR), Palampur 176061, India.
Yadav
Sudesh K
SK
D001406:Bacillaceae / Q000201:enzymology* / Q000302:isolation & purification* / Q000502:physiology; D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D015373:Bacterial Typing Techniques; D003080:Cold Temperature; D003546:Cysteine Endopeptidases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D004229:Dithiothreitol / Q000494:pharmacology; D010450:Endopeptidases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D004783:Environmental Microbiology; D020536:Enzyme Activators / Q000494:pharmacology; D004795:Enzyme Stability; D020459:Genes, rRNA; D006863:Hydrogen-Ion Concentration; D007194:India; D008623:Mercaptoethanol / Q000494:pharmacology; D008670:Metals / Q000494:pharmacology; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010802:Phylogeny; D012329:RNA, Bacterial / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid; D013696:Temperature
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D020536:Enzyme Activators; D008670:Metals; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S; D008623:Mercaptoethanol; D010450:Endopeptidases; D003546:Cysteine Endopeptidases; C087822:alkaline protease; D004229:Dithiothreitol