Pseudomonas fluorescens JK-0412가 생산하는 키틴 분해 효소와 단백질 분해효소의 생산성을 다양한 기질을 이용하여 실험조사 하였다. 각각 키틴 분해효소와 단백질 분해효소 (CTNase와 PRTase로서 명명)의 생산에 여러 가지 기질의 특이성을 검토하였다. 전자는 주로, pNP-GlcNAc에 기질 특이성을 보이는 CTNase, 그리고, 후자는 skim-milk에 기질특이 성을 갖는 PRTase로서 두 효소의 최적 생산 조건은 각각 pH8.0와 37℃였으며, 또한 1.0% dextran, 1.0% yeast extract, 1.0% MgSO4를 각각 carbon, nitrogen, and mineral 원으로서 이용했을 때, CTNase의 최대 생산성을 얻을 수 있었다. 또한 각각, 1.0% glucose, 1.0% ammonium sulfate, 1.0% MgSO4를 carbon, nitrogen, 및 mineral 원으로서 이용했을 때, PRTase의 최대 생산성을 얻을 수 있었다. 특히, 유일한 carbon과 nitrogen 원으로 chitin powder, chitosan, colloidal chitin 과 dextran을 기질로 이용하여 12일 동안 배양하여 효소활성을 측정한 결과, CTNase 생산에 있어 기질로 powder chitin를 사용하였을 경우 3일 이후 효소활성이 확인되었으며, PRTase 활성이 측정되기 시작한 4일 이후의 PRTase 생산에 있어서도 powder chitin이 가장 적합한 기질로 확인되었다. 따라서 상업적으로 적용할 수 있는 chitin은 수산가공 부산물로부터 얻을 수 있을 수 생물소재로서 CTNase와 PRTase 생산을 위한 생물 공학적인 공정에 이용가치가 매우 높다.

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 37 o C, and pH 8.0 and 45 o C, 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.