手性药物中间体R扁桃酸的不对称合成及扁桃酸脱氢酶特性研讨.pdfVIP

M039 手性药物中间体R(-)-扁桃酸的不对称合成 及扁桃酸脱氢酶特性研究 严芬 黄雅燕 郭养浩* （福州大学生物科学与工程学院，福建 福州 350002 ） 摘 要 本工作从酵母、乳酸菌、粪链球菌、假丝酵母等 4 个种属 18 种菌株中筛选出的对苯乙 酮酸具有较高的转化活性和光学选择性的酵母 sp.strain by1，经高浓度底物苯乙酮酸和高浓度产 物扁桃酸的驯化，进一步筛选得到 by1.1b 。分别考察了pH 值、反应温度、能源供体（葡萄糖） 浓度、底物浓度等因素对扁桃酸得率和对映体过量值的影响。在本实验的最适条件下，生物转化 18 h，底物转化率可达 97.0%，产物扁桃酸的得率可达 96.1%，产物(R−扁桃酸)e.e.值为 98.3 ％。 从增殖酵母中提取扁桃酸脱氢酶（粗酶）。该酶与酵母醇脱氢酶和马肝醇脱氢酶的底物特异性差 别较大。本文测定并比较了所提取的扁桃酸脱氢酶(粗酶)对不同极性、不同取代基团的羰基化合 物的催化活性。 关键词 酵母 R-扁桃酸 扁桃酸脱氢酶 底物特异性 ASYMMETRIC BIOSYNTHESIS OF INTERMEDIATE OF CHIRAL DRUGDS——R (-)-MANDELIC ACID BY SACCHAROMYCES CEREVISIAE YAN Fen HUANG Yayan and GUO Yanghao (College of biological science and technology , Fuzhou University, Fuzhou 350002,China) Abstract Saccharomyces cerevisiae sp. strain BY1 from 18 strains of Saccharomyces cerevisiae, Lactoballius, Streptococcus faecalis, Candida albicans was screened for bioreducing phenylglyoxilic acid to R-(-)-mandelic acid. After BY1 being mutagenized and demosticated, BY1.1b was obtained which showed high bioreduction activity and opticselectivity in higher concentrations of phenylglyoxilic acid and mandelic acid.Effects of pH，reaction temperature ， energy source(glucose) concentration，substrate concentration on the yield and enantiometric excess of the mandelic acid were examined respectively..Under the optimal conditions, 18 h after inoculation, the conversion of substrate of 97.0% and the yield of mandelic acid of 96.1% and enantiomeric excess value of R-(-)-mandelic acid of 98.3% could been achieved. Mandelic acid dehydrogenase which was isolated from proliferous cells of Saccharomyces cerevisiae has distinct difference with yeast alcohol dehydrogen