克隆了来自于枯草芽孢杆菌的羰基还原酶基因IolS和葡萄糖脱氢酶基因GDH,采用Ni-NTA镍亲和层析柱对重组蛋白IolS进行纯化,并对纯酶进行了酶学性质研究.结果表明,该羰基还原酶的最适温度和pH值分别为30℃和6.0；在40℃以下具有较好的热稳定性；在pH 5.5-7.0的偏酸性范围内能保持75％以上的酶活.采用三种策略构建了IolS和GDH的共表达重组质粒,结果发现,采用双启动子的重组质粒能够实现羰基还原酶IolS的高效表达,粗酶液中的IolS和GDH的比酶活均达到1.5 U/mg.运用该重组菌对10 g/L的OPBE进行不对称还原,反应15h后,底物转化率大于99％,产物(R)-2-羟基-4-苯基丁酸乙酯的ee值达到99.5％.
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