Amidase from Bacillus sp. APB-6 with very good acyltransferase activity was purified to homogeneity with a purification fold of 3.68 and 53.20% enzyme yield. The purified protein's subunit molecular mass was determined approximately 42 kDa. Hyperactivity of the enzyme was observed at pH 7.5 (150 mM, potassium-phosphate buffer) and 50 °C of incubation. An enhancement in activity up to 42% was recorded with ethylenediaminetetraacetic acid and dithiothreitol. The kinetic parameter K_m values for substrates: acetamide and hydroxylamine-hydrochloride were 73.0 and 153 mM, respectively. Further, the V_max for acyltransferase activity was 1667 U/mg of protein and the K_i for acetamide was calculated as 37.0 mM. The enzyme showed tolerance to various organic solvents (10%, v/v) and worked well in the biphasic reaction medium. The acyltransferase activity in presence of solvents i.e. biphasic medium may prove highly favorable for the transformation of hydrophobic amides, which otherwise is not possible in simple aqueous phase.

中文翻译：

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芽孢杆菌属的耐溶剂酰基转移酶。APB-6：纯化和表征。

来自芽孢杆菌属的酰胺酶。将具有非常好的酰基转移酶活性的APB-6纯化至均一，纯化倍数为3.68和53.20％的酶产率。确定纯化的蛋白质的亚基分子量约为42 kDa。在pH 7.5（150 mM，磷酸钾缓冲液）和50°C的孵育条件下观察到该酶的过度活跃。乙二胺四乙酸和二硫苏糖醇的活性提高到42％。底物：乙酰胺和羟胺盐酸盐的动力学参数K _m值分别为73.0和153 mM。此外，酰基转移酶活性的V _max为1667 U / mg蛋白质，K _i乙酰胺的计算值是37.0 mM。该酶显示出对各种有机溶剂的耐受性（10％，v / v），并且在双相反应介质中表现良好。在溶剂即双相介质的存在下，酰基转移酶的活性可能证明对疏水性酰胺的转化非常有利，否则在简单的水相中是不可能的。