title: Expression studies of Bacillus licheniformis chitin deacetylase in E. coli Rosetta cells.
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.
authors.affiliation: Department of Biotechnology, Manipal Institute of Technology, Manipal University, Karnataka 576104, India. Electronic address: ritu.raval@manipal.edu.
authors.forename: Ritu
authors.initials: R
authors.lastname: Raval
authors.affiliation: Department of Biotechnology, Manipal Institute of Technology, Manipal University, Karnataka 576104, India.
authors.forename: Robin
authors.initials: R
authors.lastname: Simsa
authors.affiliation: Department of Chemical Engineering, National Institute of Technology Surathkal, Karnataka 575025, India.
authors.forename: Keyur
authors.initials: K
authors.lastname: Raval
mesh_terms: D000581:Amidohydrolases / Q000235:genetics* / Q000378:metabolism; D000069976:Bacillus licheniformis / Q000201:enzymology* / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D005990:Glycerol / Q000378:metabolism
chemical_list: D000581:Amidohydrolases; C019866:chitin deacetylase; D005990:Glycerol
keywords: Chitin deacetylase; Expression; Glycerol; Purification; Recombinant E. coli