Lab of Biocatalysis, Hangzhou Normal University, 402-Building D, 1378 West Wenyi Road, Hangzhou 311121, China.
Biotechnol Lett. 2013 May;35(5):757-62. doi: 10.1007/s10529-013-1141-6. Epub 2013 Jan 22.
A novel aldo-keto reductase gene, Tm1743, from Thermotoga maritima was overexpressed in Escherichia coli. The enzyme displayed the highest activity at 90 °C and at pH 9. It retained 63 % of its activity after 15 h at 85 °C. The enzyme also could tolerate (up to 10 % v/v) acetonitrile, ethanol and 2-propanol with slightly increased activities. Methanol, DMSO and acetone decreased activity slightly. Furthermore, Tm1743 exhibited broad substrate specificity towards various keto esters, ketones and aldehydes, with relative activities ranging from 2 to 460 % compared to the control. Its optimum substrate, 2,2,2-trifluoroacetophenone, was asymmetrically reduced in a coupled NADPH-regeneration system with an enantioselectivity of 99.8 % and a conversion of 98 %.
一种新型的醛酮还原酶基因 Tm1743 来自于海洋栖热菌（Thermotoga maritima），在大肠杆菌中得到了过表达。该酶在 90°C 和 pH 值 9 时表现出最高的活性。在 85°C 下 15 小时后，它仍保持 63%的活性。该酶还能耐受（最高达 10%v/v）乙腈、乙醇和异丙醇，活性略有增加。甲醇、DMSO 和丙酮则略微降低了活性。此外，Tm1743 对各种酮酯、酮和醛具有广泛的底物特异性，与对照相比，相对活性范围为 2%至 460%。其最佳底物 2,2,2-三氟苯乙酮在 NADPH 再生系统中进行不对称还原，对映选择性为 99.8%，转化率为 98%。
