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Alkaline protease from a non-toxigenic mangrove isolate of Vibrio sp. V26 with potential application in animal cell culture

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Abstract

Vibrio sp. V26 isolated from mangrove sediment showed 98 % similarity to 16S rRNA gene of Vibrio cholerae, V. mimicus, V. albensis and uncultured clones of Vibrio. Phenotypically also it resembled both V. cholerae and V. mimicus. Serogrouping, virulence associated gene profiling, hydrophobicity, and adherence pattern clearly pointed towards the non—toxigenic nature of Vibrio sp. V26. Purification and characterization of the enzyme revealed that it was moderately thermoactive, nonhemagglutinating alkaline metalloprotease with a molecular mass of 32 kDa. The application of alkaline protease from Vibrio sp. V26 (APV26) in sub culturing cell lines (HEp-2, HeLa and RTG-2) and dissociation of animal tissue (chick embryo) for primary cell culture were investigated. The time required for dissociation of cells as well as the viable cell yield obtained by while administering APV26 and trypsin were compared. Investigations revealed that the alkaline protease of Vibrio sp. V26 has the potential to be used in animal cell culture for subculturing cell lines and dissociation of animal tissue for the development of primary cell cultures, which has not been reported earlier among metalloproteases of Vibrios.

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Acknowledgments

This work was carried out with the financial assistance from the Department of Biotechnology, Government of India, under Programme Support in Marine Biotechnology (BT/PR4012/AAQ/03/204/2003).The first author thanks the University Grants Commission for Fellowship.

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Authors and Affiliations

  1. Department of Marine Biology, Microbiology and Biochemistry, School of Marine Sciences, Cochin University of Science and Technology, Fine Arts Avenue, Cochin, 682016, Kerala, India

    K. Manjusha & A. V. Saramma

  2. National Centre for Aquatic Animal Health, Cochin University of Science and Technology, Fine Arts Avenue, Cochin, 682016, Kerala, India

    P. Jayesh, Divya Jose, B. Sreelakshmi, P. Priyaja, Prem Gopinath & I. S. Bright Singh

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  1. K. Manjusha
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Correspondence to I. S. Bright Singh.

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Manjusha, K., Jayesh, P., Jose, D. et al. Alkaline protease from a non-toxigenic mangrove isolate of Vibrio sp. V26 with potential application in animal cell culture. Cytotechnology 65, 199–212 (2013). https://doi.org/10.1007/s10616-012-9472-z

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