催化学报

催化学报 ›› 2008, Vol. 29 ›› Issue (6): 537-541.

• 研究论文 • 上一篇    下一篇

呼吸缺陷型面包酵母催化β-羰基酯的不对称还原

倪宏亮,姚善泾   

  1. 浙江大学化学工程与生物工程学系, 浙江杭州 310027
  • 收稿日期:2008-06-25 出版日期:2008-06-25 发布日期:2012-04-28

Asymmetric Reduction of β-Carbonyl Ester with Respiratory-Deficient Mutants of Baker’s Yeast

NI Hongliang, YAO Shanjing*   

  1. Department of Chemical and Biochemical Engineering, Zhejiang University, Hangzhou 310027, Zhejiang, China
  • Received:2008-06-25 Online:2008-06-25 Published:2012-04-28

PDF

1079

被引次数

4

摘要： 以4-氯-3-羰基丁酸乙酯为模型底物,葡萄糖为碳源,对呼吸缺陷型酵母催化不对称还原合成(S)-4-氯-3-羟基丁酸乙酯的反应进行了研究. 通过在254 nm下紫外诱变,经2,3,5-氯化三苯四氮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. 并对呼吸缺陷型酵母催化特性和线粒体复合体Ⅰ, Ⅰ+Ⅲ, Ⅱ+Ⅲ和Ⅳ的活性分别进行了测定. 结果表明,呼吸缺陷型酵母催化底物转化率在49%～75%,产物收率在31%～69%,对映体过量值最高可达72%, 比正常酵母提高了18%. 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, 4-氯-3-羰基丁酸乙酯, 不对称还原, 线粒体复合体, 紫外诱变

Abstract: Asymmetric synthesis of ethyl (S)-4-choloro-3-hydroxybutanoate was studied using ethyl 4-chloro-3-oxobutanoate as the model substrate and glucose as the carbon source. Six respiratory deficient (RD) mutants of natural baker’s yeast were induced by ultraviolet radiation at 254 nm and were identified by 2,3,5-triphenyltetrazolium chloride plate and carbon sources without sugar. The characteristics of catalytic asymmetric reduction and the activity of four mitochondria complexes, Ⅰ, Ⅰ+Ⅲ, Ⅱ+Ⅲ, and Ⅳ, in the electron transfer chain of RD mutants were detected. The results showed that the substrate conversions catalyzed by RD mutants were49%-75%, and the yields were31%-68%, which were lower than that of natural baker’s yeast. The enantiomeric excess (ee) of RD mutants increased to 72%, being 18% higher than that of natural baker’s yeast. The activities ofⅠ-Ⅳmitochondrial complexes in RD mutants were all lower than that in natural baker’s yeast to various degrees.

Key words: respiratory deficient baker’s yeast, ethyl 4-chloro-3-oxobutanoate, asymmetric reduction, mitochondria complex, ultraviolet mutagenesis