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Cytotechnology

, Volume 65, Issue 2, pp 199–212 | Cite as

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

  • K. Manjusha
  • P. Jayesh
  • Divya Jose
  • B. Sreelakshmi
  • P. Priyaja
  • Prem Gopinath
  • A. V. Saramma
  • I. S. Bright SinghEmail author
Original Research

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.

Keywords

Alkaline protease Vibrio sp. Animal cell culture Primary cell culture Virulence genes Trypsin 

Notes

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.

Supplementary material

10616_2012_9472_MOESM1_ESM.doc (5.7 mb)
Supplementary material 1 (DOC 5794 kb)

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Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • K. Manjusha
    • 1
  • P. Jayesh
    • 2
  • Divya Jose
    • 2
  • B. Sreelakshmi
    • 2
  • P. Priyaja
    • 2
  • Prem Gopinath
    • 2
  • A. V. Saramma
    • 1
  • I. S. Bright Singh
    • 2
    Email author
  1. 1.Department of Marine Biology, Microbiology and Biochemistry, School of Marine SciencesCochin University of Science and TechnologyCochinIndia
  2. 2.National Centre for Aquatic Animal HealthCochin University of Science and TechnologyCochinIndia

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