Research Center for Biomedicine and Health, College of Life and Environmental Science, Hangzhou Normal University, Hangzhou 310012, China.
Bioresour Technol. 2012 Nov;123:690-4. doi: 10.1016/j.biortech.2012.07.060. Epub 2012 Jul 25.
The gene encoding a putative protein from Candida parapsilosis CDC317 (CPE) was cloned and overexpressed in Escherichia coli. The protein was amenable to overexpression in E. coli and constituted 35% of the total cell protein content. The optimal activity was determined at pH 5.5 and 40°C with the substrate 4-chloro-3-oxobutanoate ethyl ester (COBE). The optical purity of the product was over 99% enantiomeric excess for the (S)-isomer, and the molar conversion yield of the product was 91.1%. The apparent k(m) value for COBE was 0.19±0.01mM, which is an order of magnitude lower than that of other enzymes in the literature.
从近平滑假丝酵母 CDC317（CPE）中克隆并在大肠杆菌中过表达了一个假定蛋白的编码基因。该蛋白在大肠杆菌中易于过表达，占总细胞蛋白含量的 35%。最佳活性在 pH 5.5 和 40°C 下测定，底物为 4-氯-3-氧代丁酸乙酯（COBE）。产物的光学纯度对（S）-异构体的对映体过量超过 99%，产物的摩尔转化率为 91.1%。COBE 的表观 k(m) 值为 0.19±0.01mM，比文献中其他酶低一个数量级。
