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从6株粪产碱杆菌中筛选出对邻氯扁桃腈具有较高芳基乙腈水解酶活性和中等对映选择性的Alcaligenes faecalis CGMCC 1.2006.研究了反应介质对Alcaligenes faecalis CGMCC 1.2006芳基乙腈水解酶活性和对映选择性的调控作用.反应介质中水溶性辅溶剂如甲醇的加入降低芳基乙腈水解酶活性,但显著提高对映选择性.而非离子表面活性剂的加入具有相反的作用.系统优化了底物浓度、细胞浓度、pH、温度和反应时间等反应条件对芳基乙腈水解酶活性和对映选择性的影响.在最优反应条件下,采用分批补料策略,(R)-邻氯扁桃酸的产量在22 h内达到32.2 g/L,产率82.4%,ee 93.1%.以邻氯苯甲醛和氰化钾为底物合成(R)-邻氯扁桃酸,收率89.5%,ee98.6%.
Alcaligenes faecalis CGMCC 1.2006 was selected from 6 Alcaligenes faecalis Alcaligenes faecalis which had higher aryl-acetohydrolase activity and moderate enantioselectivity to o-chloro-mandelic acid.The effects of reaction medium on Alcaligenes faecalis CGMCC 1.2006 aryl-acetonitrile hydrolase activity and Enantioselective regioselective addition of a water-soluble auxiliary solvent such as methanol to the reaction medium lowers arylacetonitrile hydrolase activity but significantly increases enantioselectivity, whereas non-ionic surfactant addition has the opposite effect. The effects of reaction conditions, such as substrate concentration, cell concentration, pH, temperature and reaction time, on the activity and enantioselectivity of arylacetonitrile hydrolase were studied.Under the optimal conditions, fed - batch strategy, (R) - The yield of o-chloromandelic acid reached 32.2 g / L within 22 h with the yield of 82.4% and ee of 93.1%. (R) -chloro-mandelic acid was synthesized from o-chlorobenzaldehyde and potassium cyanide with the yield of 89.5 %, ee98.6%.