The invention discloses a
carbonyl reductase directional modification method for synthesis of
atorvastatin intermediates, which belongs to the technical field of
enzyme gene engineering, and is characterized in that
carbonyl reductase genes are used as templates,
mutation primer pairs are designed, double-site
mutation is carried out by adopting an overlapping extension PCR (
polymerase chain reaction) method,
escherichia coli is transformed, IPTG (
isopropyl-beta-d-thiogalactoside) is used for inducing expression, and
mutant carbonyl reductase is prepared. The K205D / R237W
double mutation modified carbonyl
reductase improves the
enzyme activity, combines the synergistic effect of the
thiophene pyridine coenzyme analogue, improves the conversion rate of the
atorvastatin intermediate and the product purity, is significantly superior to the traditional
chemical reduction method, avoids the low-temperature and high-pressure conditions of the traditional chemical method, reduces the
energy consumption, and is suitable for industrial production. The use amount of an
organic solvent and the generation amount of byproducts are reduced, and the generation amount of
wastewater is reduced.