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对一株产肉碱脱水酶的菌株用溴化乙锭进行诱变 ,并经肉碱选择性培养基进行筛选 ,获得了一株高活性酶活的菌株 ,并进行了休止细胞转化巴豆甜菜碱为L 肉碱的反应条件的研究。实验确定休止细胞的最适反应温度为 30℃～ 32℃ ,pH8.1、0 .0 1mol L磷酸缓冲液 ,时间 8.5～ 10h。最适底物浓度为 6 0mg ml,最适休止细胞浓度为 6 0mg ml(湿重 ) ,产率 4 0 % (摩尔比 )。建立了休止细胞酶反应的表观米氏方程 ,其中Vm =0 .0 0 12 2 5mol (min·15mg菌体 ) ,Km =0 .0 4 0 114mol L。
A strain of carnitine-producing dehydratase was mutagenized with ethidium bromide and screened by a carnitine-selective medium to obtain a highly active enzyme-producing strain, and the resting cells were transformed with crotylbetaine Study on the Reaction Conditions for L Carnitine. The optimum reaction temperature of the resting cells was determined to be 30 ℃ ~ 32 ℃, pH8.1,0.0 1mol L phosphate buffer for 8.5 ~ 10h. The optimal substrate concentration was 60 mg, the optimal resting cell concentration was 60 mg ml (wet weight) and the yield was 40% (molar ratio). The established Mie equation of resting cell enzyme reaction was established, where Vm = 0.012 2 5mol (min · 15mg cells), Km = 0.014 0 114mol L.