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Isolation of a Psychrotrophic Exiguobacterium sp. SKPB5 (MTCC 7803) and Characterization of Its Alkaline Protease

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Abstract

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°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 β-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.

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Acknowledgments

The authors are grateful to Dr. P.S. Ahuja, Director, Institute of Himalayan Bioresource Technology, Palampur, for valuable suggestions and encouragement during the experimental study. We also thank Harkeerat Atwal and Prashant Mohanpuria for extending help in various experiments. Council of Scientific and Industrial Research (CSIR) is duly acknowledged for providing financial support in the form of internal grants to Institute of Himalayan Bioresource Technology (IHBT). This IHBT communication number for this study is 642.

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Authors and Affiliations

  1. Hill Area Tea Science Division, Institute of Himalayan Bioresource technology (CSIR), Palampur, 176061, India

    Ramesh C. Kasana

  2. Biotechnology Division, Institute of Himalayan Bioresource technology (CSIR), Palampur, 176061, India

    Sudesh K. Yadav

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  1. Ramesh C. Kasana
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  2. Sudesh K. Yadav
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Correspondence to Sudesh K. Yadav.

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Kasana, R.C., Yadav, S.K. Isolation of a Psychrotrophic Exiguobacterium sp. SKPB5 (MTCC 7803) and Characterization of Its Alkaline Protease. Curr Microbiol 54, 224–229 (2007). https://doi.org/10.1007/s00284-006-0402-1

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  • DOI: https://doi.org/10.1007/s00284-006-0402-1

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