Production of NADP by Immobilized Brevibacterium ammoniagenes and ATP- regenerating System of Acetate Kinase

고정화 Brevibacterium ammoniagenes와 Acetate Kinase의 ATP생성계에 의한 NADP생산

For the conversion of WAD to NADP, Immobilized Brevibacterium ammoniagenes cells with NAD kinase was coupled with ATP-generating system by acetate kinase. The membrane permeability of B. ammoniagenes was improved by toluene treatment of cells. The toluene treated B. ammoniagenes cells were immobilized for stable enzyme activity. Partially purified acetate kinase was used in the reaction system. The optimum conditions for the efficient conversion of UAD to WADP by energy-coupled system were investigated. B. ammoniagenes cells treated with toluene for the Improvement of membrane permeability showed 4.5 fold improved permeability in the conversion of NAD to NADP compared with Intact cells. 3% k-carrageenan as the immobilization matrix of B. ammoniagenes showed the best efficiency for the conversion of NAD to NADP The optimum conditions for the WAR to WARP conversion reaction coupled nth ATP-generating system were 10mM acetylphosphate, 5mM ADP 200mM inorganic phosphate, 10mM MgCl2, 250mg/ml Immobilized cells, 49.3mUnit/ml acetate kinase, pH 7.5 and 37$^{\circ}C$. Under the optimum conditions, 72% of 5mM(340mg/ml ) NAD was converted to UADP In 12 hours.