用呼吸缺陷型酵母催化4氯3羰基丁酸乙酯的不对称还原.pdfVIP

用呼吸缺陷型酵母催化 4-氯-3-羰基丁酸乙酯的不对称还原 倪宏亮，姚善泾* (浙江大学 化学工程与生物工程学系，浙江杭州 310027) 摘要：通过紫外诱变，经 2 ，3，5-氯化三苯四氯唑平板和非发酵型碳源培养基鉴定，得到 6 株面包 酵母的呼吸缺陷型变异株。考察了 6 株呼吸缺陷型酵母催化 4-氯-3-羰基丁酸乙酯的不对称还原反应， 分别测定了其中 3 株呼吸缺陷型酵母的线粒体复合体Ⅰ、Ⅰ+ Ⅲ、Ⅱ+ Ⅲ和Ⅳ的活性。结果表明，4- 氯-3-羰基丁酸乙酯的转化率在 49~75%之间，产物收率在 31~69%之间，对映体过量值最高可达 72%， 比正常酵母的54%提高了 18%；3 株突变株线粒体内复合体 I-Ⅳ的活性与正常酵母相比都有不同程度 的下降。 关键词：呼吸缺陷型；面包酵母；不对称还原；线粒体复合体；紫外诱变；4-氯-3-羰基丁酸乙酯 Asymmetric Reduction of Ethyl 4-chloro-3-oxobutanoate with Respiratory-Deficient Mutants of Baker’s Yeast NI Hongliang, YAO Shanjing (Department of Chemical and Biochemical Engineering, Zhejiang University, Hangzhou 310027, Zhejiang, China) Abstract ：Six respiratory deficient (RD) mutants of bakers yeast were induced with ultraviolet radiation, which were identified by 2,3,5-triphenyltetrazolium chloride plate and carbon sources without sugar. The characteristics of asymmetric reduction of ethyl 4-chloro-3-oxobutanoate catalysed by six RD mutants were studied. And activities of four complexes, Ⅰ, Ⅰ+Ⅲ, Ⅱ+Ⅲ and Ⅳ, in electron transfer chain of three RD mutants were detected separately. The results showed that the conversions of ethyl 4-chloro-3-oxobutanoate catalyzed by RD mutants were among 49%~75%, and yields were among 31%~68%, while enantiomeric excess (e.e.) of RD mutants increased to 72%, which was 18% higher than that of natural bakers yeast. The activities of 4 complexes in RD mutants were all lower than in normal yeast. Key words: Respiratory deficient; Baker’s yeast; Asymmetric reduction; Mitochondria complex; Ultraviolet mutagenesis; Ethyl 4-chloro-3-oxobutanoate 1. 引言