反应详情
EQUATION
反应方程式
REACTANTS
反应物
PRODUCTS
生成物
PROCEDURE
实验过程
The asymmetric reduction of ethyl 2-oxocyclohexanecarboxylate was carried out by using this reductase (2). Namely, 4 ml of the reaction mixture containing the reductase (2) (2U), NADPH (0.8 μmol), ethyl 2-oxocyclohexanecarboxylate (100 μmol, 17 mg), phosphate buffer solution (pH 7.0, 400 μmol), glucose dehydrogenase (the same enzyme as that used in Example 4; 2U) and glucose (100,μmol) was incubated at 30° C. with gentle shaking to promote the reaction. After 4 hour-incubation, the reaction mixture was extracted according to the same manner as in Example 3. The extract was analyzed by gas chromatography described in Example 1. As a result, the yield of the cis isomer was 17.0 mg (reduction yield >99%), and neither the remaining substrate nor the trans isomer was detected. On the other hand, the extract above was distilled off according to the the same manner as in Example 3 to obtain the reduction product, ethyl 2-hydroxycyclohexanecarboxylate. The resulting reduction product was analyzed by high performance liquid chromatography described in Example 1. As a result, only the cis 2 isomer was detected and no other isomers were detected.
WORKUP
后处理
- customwas incubated at 30° C.
- customthe reaction
- extractionthe reaction mixture was extracted
- extractionOn the other hand, the extract
- distillationabove was distilled off