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254nm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.
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-triphenyhe-trazolium 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 were 49 %-75 %, and the yields were 31%-68 %, which were lower than that of natural baker＇s yeast. The enan-tiomeric excess （ee） of RD mutants increased to 72% , being 18% higher than that of natural baker＇s yeast. The activities of Ⅰ-Ⅳ mitochondrial complexes in RD mutants were all lower than that in natural baker＇s yeast to various degrees.
出处
《催化学报》
SCIE
CAS
CSCD
北大核心
2008年第6期537-541,共5页
基金
高等学校博士学科点专项科研基金(20050335131)
