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Characterisation of a detergent-stable alkaline protease from a novel thermophilic strain Paenibacillus tezpurensis sp. nov. AS-S24-II

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Abstract

An alkaline-protease-producing bacterial strain (AS-S24-II) isolated from a soil sample in Assam is a Gram-stain-positive, catalase-positive, endospore-forming rod and grows at temperatures ranging from 30 °C to 60 °C and salinity ranging from 0% to 7% (w/v) NaCl. Phenotypic characterisation, chemotaxonomic properties, presence of Paenibacillus-specific signature sequences, and ribotyping data suggested that the strain AS-S24-II represents a novel species of the genus Paenibacillus, for which the name Paenibacillus tezpurensis sp. nov. (MTCC 8959) is proposed. Phylogenetic analysis revealed that P. lentimorbus strain DNG-14 and P. lentimorbus strain DNG-16 represent the closest phylogenetic neighbour of this novel strain. Alkaline protease production (598 × 103 U l−1) by P. tezpurensis sp. nov. in SmF was optimised by response surface method. A laundry-detergent-stable, Ca2+-independent, 43-kDa molecular weight alkaline serine protease from this strain was purified with a 1.7-fold increase in specific activity. The purified protease displayed optimum activity at pH 9.5 and 45–50 °C temperature range and exhibited a significant stability and compatibility with surfactants and most of the tested commercial laundry detergents at room temperature. Further, the protease improved the wash performance of detergents, thus demonstrating its feasibility for inclusion in laundry detergent formulations.

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Acknowledgements

Authors acknowledge the help extended by the Institute of Microbial Technology (IMTECH), Chandigarh, for bacterial cellular fatty acid analysis and Delhi University for 16S rDNA sequencing. Mr. S. K. Rai and Mr. J. K. Roy were recipients of Senior Research Fellowship from the Department of Biotechnology, New Delhi and Institutional fellowship from Tezpur University, respectively. Authors acknowledge the technical assistance received from Ms. P. Bhuyan, T.U. This work was supported by financial assistance from the Department of Biotechnology, Ministry of Science and Technology, New Delhi.

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Authors and Affiliations

  1. Department of Molecular Biology and Biotechnology, School of Science and Technology, Tezpur University, Tezpur, 784 028, Assam, India

    Sudhir K. Rai, Jetendra K. Roy & Ashis K. Mukherjee

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  1. Sudhir K. Rai
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  2. Jetendra K. Roy
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  3. Ashis K. Mukherjee
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Correspondence to Ashis K. Mukherjee.

Additional information

The 16S rDNA sequence to GenBank and the accession number are bankit1191281 FJ804507, and the strain is deposited in Microbial Type Culture Collection (MTCC 8959).

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Rai, S.K., Roy, J.K. & Mukherjee, A.K. Characterisation of a detergent-stable alkaline protease from a novel thermophilic strain Paenibacillus tezpurensis sp. nov. AS-S24-II. Appl Microbiol Biotechnol 85, 1437–1450 (2010). https://doi.org/10.1007/s00253-009-2145-y

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  • DOI: https://doi.org/10.1007/s00253-009-2145-y

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