通过对苹果酸酶(ME)辅酶结合域L310、Q401、I404饱和位点突变库与辅酶烟酰胺腺嘌呤二核苷酸(NAD+)类似物库的高通量筛选,研究了苹果酸酶结合域位点对NAD+及其类似物(B1～B7)催化活性的影响.结果表明,突变后酶ME-Q401H/L404T对类似物B4的kcat/Km是野生型酶的50倍;突变后酶ME-L310M/Q401N对类似物B4的kcat/Km是野生型酶的16倍,对类似物B3的kcat/Km是野生型酶的5倍,因此通过对结合域定点突变,NAD+类似物的催化活性得到提高.
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