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光学纯的3-奎宁醇是一种重要的手性医药中间体,目前已报道的通过酶催化3-奎宁酮的不对称还原几乎都是得到(R)-3-奎宁醇.在前期的工作中我们筛选获得一株能催化还原得到(S)-3-奎宁醇的红串红球菌(Rhodococcus erythropolis WY1406),其ee值达到99%.本研究从该红串红球菌中克隆得到6个3-奎宁酮还原酶基因,并将其转入到大肠杆菌BL21(DE3)中诱导表达,利用粗酶液检测它们催化3-奎宁酮还原的活性及立体选择性.结果发现其中两种酶Re QR-13和Re QR-25催化3-奎宁酮还原生成(S)-3-奎宁醇,ee值大于99%.此外还发现了能催化生成(S)-3-奎宁醇的ee值分别为83%、46%和57%的奎宁酮还原酶Re QR-18、Re QR-27和Re QR-28.其中表达Re QR-25的整细胞催化5 g/L的底物3-奎宁酮还原,转化率达93%.本研究为合成光学纯的(S)-3-奎宁醇提供了一种新的方法.
Optically pure 3-quinuclidin is an important chiral pharmaceutical intermediate. Up to now, it has been reported that (R) -3-quinuclidin is almost always obtained by asymmetric reduction of 3-quinuclidin by an enzyme. In the previous work, we screened Rhodococcus erythropolis WY1406 which can be catalytically reduced to obtain 99% ee value.This study was cloned from Rhodococcus erythropolis Six 3-quinone reductase genes were obtained and transformed into E. coli BL21 (DE3) to induce expression, and their activity in catalyzing the reduction of 3-quinone and their stereoselectivity were detected by using a crude enzyme solution. Two of the enzymes, Re QR-13 and Re QR-25, catalyzed the reduction of 3-quinuclidine to (S) -3-quinuclidin with an ee value of over 99%. In addition, The ee values of quinine were 83%, 46% and 57% of the quinone reductase Re QR-18, Re QR-27 and Re QR-28, respectively. The whole cells expressing Re QR- L of the substrate 3-quinone reduction, conversion rate of 93% .This study provides a new method for the synthesis of optically pure (S) -3-quinucl alcohol.