Production and Characterization of Enzymes Involved in Chitin Catabolic Cascade from a Bacterial Strain Isolated from Soil

Abstract

The present study was aimed to isolate the bacteria producing chitinolytic enzymes including chitin deacetylase (CDA) from soil origin. About 24 bacteria were screened for their chitinolytic enzymes producing ability on the basis of colloidal chitin. Amongst one of the most potent isolates designated to JI-02 was further selected as the most prominent chitinolytic and CDAlike enzyme (CDA-X) producer based on the ability of deacetylation of an artificial substrate pNP-acetamilide. The maximum production of CDA-X in crude enzyme from JI-02 was observed in the presence of 1% colloidal chitin at 37°C and pH 6.4 after 120 h of incubation. Potent enzyme activity of CDA-X was determined toward pNP-acetamilide, demonstrating that optimal pH, temperature and ionic strength of the CDA-X were assessed to be 7.0, 52°C and 125 mM, respectively. Unexpectedly, Nacetylglucosamine (GlcNAc) was not fully converted by the action of CDA-X to glucosamine (GlcN) under the established conditions. Further study on enzyme activity toward chitin-oligosaccharides consisting of multi-N-acetylglucosamine (GlcNAc)n, n=2-5 may be necessary to elucidate the mode of action which requires the minimum size of (GlcNAc)n. However, our data suggest that CDA-X can convert chitin to chitosan with the maximum yield of approximately 0.08g/L during the fermentation of the strain in the presence of chitin. Furthermore, we found that crude enzyme showed the significant chitosanolytic activity. Taken together, our data suggest that the strain JI-02 may be a potential candidate as a particular strain for better understating of chitin catabolic pathway.