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Expression studies of Bacillus licheniformis chitin deacetylase in E. coli Rosetta cells

Author:
Raval, Ritu, Simsa, Robin, Raval, Keyur
Source:
International journal of biological macromolecules 2017
ISSN:
0141-8130
Subject:
Bacillus licheniformis, Escherichia coli, N-acetylglucosamine, additives, bioenergy industry, biomass, biopolymers, cellulose, chitin, chitin deacetylase, chitosan, genes, glycerol, metagenomics, soil, wastes
Abstract:
Chitin, the biopolymer of the N-acetylglucosamine, is the most abundant biopolymer on the planet after cellulose. However owing to its crystalline nature, its deacetylated derivative; chitosan is industrially more potent. This conversion on an enzymatic scale can be made using chitin deacetylase. The metagenomics library constructed from the soil exposed to chitin and chitosan yielded chitin modifying enzymes, one of them being chitin deacetylase (CDA) utilized for the present study. The gene was amplified and expressed using the pET 22b vector in E. coli Rosetta cells. The effect of two additives; chitin and glycerol on the CDA activity were studied. The inclusion of glycerol in the medium improved the biomass by 50% from the initial value of 1.25g/l to 2.5g/l. The activity of CDA increased from 90μmol/min/ml to 343μmol/min/ml. The CDA activity reported in the present paper is the highest observed for any strain. The addition of glycerol to the media not only helped improve the yield of the chitin deacetylase but also imparted value addition to the waste of the biofuel industry.
Agid:
5638066