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KMID : 0525720100150040203
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Journal of Chitin and Chitosan
2010 Volume.15 No. 4 p.203 ~ p.209
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Optimization for the Production of Chitinolytic and Proteolytic Enzymes from Pseudomonas fluorescens JK-0412
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Park Yong-Il
Seo Hyun-Hyo
Kim Woo-Jung
Ko Ju-Hee
Park Jae-Kweon
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Abstract
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The production of chitinolytic and proteolytic enzymes in submerged cultures of Pseudomonas fluorescens JK-0412 in media was investigated in experiment with pH, temperature and substrate concentration as the main parameters. The production of chitinolytic and proteolytic enzymes, named as CTNase and PRTase, the former has specific enzyme activity toward pNPGlcNAc mostly and the latter has specific enzyme activity toward skim-milk, was monitored under the influence at various concentrations of these substrates. The optimum conditions of pH, temperature and substrate concentration for the production of both enzymes was determined to be pH 8.0 and 37oC, and pH 8.0 and 45oC, respectively. The maximum production of CTNase was obtained when 1.0% dextran, 1.0% yeast extract and 1.0% MgSO4 were used as carbon, nitrogen and mineral sources in the absence of chitinous material. In addition, the maximum production of PRTase was obtained when 1.0% glucose, 1.0% ammonium sulfate, and 1.0% MgSO4 were used as carbon, nitrogen and mineral source, respectively. Furthermore, in the presence of flake chitin, chitosan, and colloidal chitin used as a sole carbon and nitrogen source, it was found that flake chitin was the most efficient elicitor of CTNase and PRTase after 3 and 4 day cultivation, respectively. Hence chitin powder is a suitable substrate for the production of both CTNase and PRTase to employ in an advanced biotechnological process, whereby several commercially applicable chitinous materials can be obtained from a waste product of the fishing industry.
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KEYWORD
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Chitin, chitinolytic enzyme, proteolytic enzyme, Pseudomonas fluorescens
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