Isolation, Identification and Molecular Cloning of Chitinase Gene in Bacteria Isolated from Crustacean Wastes

Abstract

This research envisaged cloning of genes encoding chitinases in bacteria isolated from crustacean wastes and its transformation in E.coli by TOPO cloning method for complete sequencing of partial gene. Chitin and chitinolytic enzymes are gaining importance for their biotechnological applications. Particularly, chitinases are used in both agricultural, industrial applications and also in human health care. The chitinases are present in chitin-containing microorganisms, bacteria and plants with a diversity of roles, such as chitin metabolism in growing hyphae, defence mechanisms in response to pathogens and abiotic stress, in nutrition and parasitism. Apart from the application of chitinases as inhibitors and bio pesticides, the chitinases have been used for production of single cell protein for animal and aquaculture feed, for the isolation of fungal protoplasts, preparation of bioactive chito-oligosaccharides and for phytopathogen inhibition, reinforcement of plant defence. The success in employing chitinases for different aspects depends on the supply of highly active preparations at reasonable cost. Therefore, the understanding of biochemistry and genetics of chitinolytic enzymes, their phylogenetic relationships and methods of estimation will make them more useful in a variety of processes in near future.

Keywords: Chitin, bacterial chitinase, chitinase Gene, TOPO, TA cloning, sequence analysis