?１５６４?化工进展２０１１年第３０卷
是ＮＡＤＰＨ【１１】。ＬｂＡＤＨ具有离度的立体专一性和
对映体选择性，可以还原带宥庞大镤｜ｌ链酶酮和酮酸
衍生物、历占二酮基酯类和环形酮等，生成相应的
（尺）．醇和（Ｒ）。羟基酸酯，且ｅ．ｅ．＞９９％；具有独
特的螽代苯乙醺逐瓤活性，且涵性较高Ｈ２】。Ｓｃｈｒｏｅｒ
等【ｌ习建立了双膜反应工艺，ＬｂＡＤＨ还原２，５．已二酮
生成（２Ｒ，５Ｒ）．己二醇，同时氧化ＮＡＤ（Ｐ）Ｈ生成
ＮＡＤ（Ｐ）＋，反应体系中加入异丙醚实现ＮＡＤ（Ｐ）Ｈ的再
生，产率达至ＴＪ＞１７０ｇ／（Ｌ?国，（２Ｒ，５Ｒ）．己二醇雳于合
成手性磷配体。值得一提的是ＬｂＡＤＨ能还原甲基．
卢萘基甲酮生成相应的（Ｒ）一醇，且ｅ．ｅ．》９９％。
ＬｂＡＤＨ对甲基。廖萘基甲酮的商活性，说暖该酶
对侧链庞大的酮类化合物的还原能力非常强，故
开展ＬｂＡＤＨ在绿色化学合成上的应用舆有重要
的意义。
厶ｋｅｆｉｒ的醇脱氢酶（ＬｋＡＤＨ）由７５９ｂｐ孩营
酸组成，编码２５２个氨基酸，属于短链脱氢酶，
ＮＡＤＰＨ依赖型。在大肠杆菌中重组表达的
ＬｋＡＤＨ能够还原多种脂肪族、芳香族、环形的酮
及掺。酾醮，生成褶应的（定），醇珏夺ｂｊ。Ｂｒａｄｓｈａｗ
等１１６１研究大肠杆菌重组的ＬｋＡＤＨ具有很宽的底
物特异性，催化多种类型的底物还原反应生成相
应的带芳香族的、嚣形酶、多繇酶帮脂臃族侧链
的手性醇，具有较高的产率和高度的对映体过量
（９４％－－一９９％）。利用Ｌ．ｋｅｆｉｒ全细胞催化，细胞内
的醇脱氢酶首先还原６一氯一３，５．二氧代己酸叔丁酯
的Ｃ，的酮基形成霞构型的手性羟基，进一步还原
ｃ５位酮基形成Ｓ构型的手性羟基，一锅法合成６。
氯一（３Ｒ，５ｓ）一二羟基己酸叔丁酯，在２２ｈ反应时间内
产率爻４７．５％和ｅ。ｅ。＞９９％，（３Ｒ，５ｓ）一二羟基已酸叔丁
酯为多种ＨＭＧ—ＣｏＡ还原酶抑制剂的合成提供手性
侧链（图１）【Ｊ门。
固定化细胞技术（ｉｍｍｏｂｉｌｉｚｅｄｃｅｌｌｓ）是将微生
物细脆蹇接作为酶原，进行催化反应，具霄多项优
ＯＯＯ
ｃｔ、儿人火∞。
ＯＨ０Ｏ
、人人儿蛐ＯＨＯＨＯ
ｃ１．．／１．．／ｋ．／Ｕ－ｏｍＨ
国｛两
图ｌＬ如归全细胞催化一锅法合成６－氯一（３Ｒ，５Ｓ）－
二羟基己酸叔丁醮点，如节省了酶分离纯化的时闻耪成本，多酶促反应，增加酶酶稳定性等。Ｔａｎ等疆ｓｊ幂｜ｊ用固定亿的Ｌｋｅｊ疗ｒ全细胞催化２，５．己二酮的还原生成（５尺）一羟基己烷一２一酮，ｅ．ｅ．＞９９％，在活塞流反应器中收率这８７∥＆?由，６天屠仍保持６８％麴残余活性。采用固定化细胞技术节约酶成本，具有应用的潜力和价值。
１．３赖氏蕊属的醇脱氢酶
赖氏菌属的Ｌｅｉｆｓｏｎｉａｓｐ。是Ｉｎｏｕｅ等从苯乙烯同化后的土壤样本，对约９００株菌株进行筛选所软得的能将前手性酮还原生成（Ｓ）．１．三氟苯乙醇的菌株。（ｓ）。ｌ一三氟苯乙醇是重要静医药中闻体瓣瑚。ＬｅｉｆｓｏｎｉａＡＤＨ的分子摄是１１０ｋＤ，同源四聚体，辅助因子是ＮＡＤＨ。ＬｅｉｆｓｏｎｉａＡＤＨ可还原一系列的醛帮酮，如：氯嚣耍｜霉、畚庚醺、２。辛鬻等前手性黧，对卤代的苯乙酮、甲氧苯乙酮、苯丁酮、环戊酮等有一定的活性，对酮酯如甲基丙酮酸、氧代丁酸盐等也有较离的活性（表２）。ＬｅｉｆｓｏｎｉａＡＤＨ催化反应具有高泼的立嚣选择性，三氟苯乙醺还原成（国．１一三氟苯乙醇，对映体过量大于９９％，且反应体系中加入异丙醇即可实现ＮＡＤＨ的再生。ＬｅｉｆｓｏｎｉａＡＤ珏还原苯乙醺、双乙醮基酮、长链烷基骥等生成＜Ｒ）。醇，且ｅ．ｅ＋＞９９％，有的底物转化率髅至高达１００％１２０－２Ｈ。带手性羟基的苯乙酮可在苯环的不同位置上进行卤代或氨基取代，产物可作为手性配钵帮医药中闻体溺于不对称合成，如耀于合成拟肾上腺素药，抗生素，ＨＩＶ－逆转录酶抑制剂等。
１．４囔热缴生物的醇脱氯酶
酶的稳定性，特别是耐有机溶剂的稳定性，在工业和医药行业的应用中起着十分重要的作用。嗜
表２ＬｅｉｆｓｏｎｉａｓＯ．ＡＤＨ的镁纯反应
底物产物相对活性／％转化率，％手性，ｅ．ｅ．，％
醇脱氢酶不对称还原制备手性醇的研究进展
作者：谌容， 王秋岩， 殷晓浦， 魏东芝， 谢恬， CHEN Rong， WANG Qiuyan， YIN Xiaopu， WEI Dongzhi， XIE Tian
作者单位：谌容,CHEN Rong(生物医药与健康研究中心，杭州师范大学，浙江杭州310012;生物反应器工程国家重点实验室，华东理工大学鲁华生物技术研究所，上海200237)， 王秋岩,殷晓浦,谢恬,WANG Qiuyan,YIN Xiaopu,XIE
Tian(生物医药与健康研究中心，杭州师范大学，浙江杭州310012)， 魏东芝,WEI Dongzhi(生物反应器工程国家
重点实验室，华东理工大学鲁华生物技术研究所，上海200237)
刊名：
化工进展
英文刊名：Chemical Industry and Engineering Progress
年，卷(期)：2011,30(7)
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引用本文格式：谌容.王秋岩.殷晓浦.魏东芝.谢恬.CHEN Rong.WANG Qiuyan.YIN Xiaopu.WEI Dongzhi.XIE Tian醇脱氢酶不对称还原制备手性醇的研究进展[期刊论文]-化工进展 2011(7)