The invention belongs to the technical field of biology, and discloses a high-throughput screening method based on glucose dehydrogenase coupled carbonyl reductase. The method comprises the followingsteps: Step I, building a high-throughput system applicable to all oxidordeuctases capable of directly or indirectly consuming NAD(P)H to generate NAD(P)<+> by adopting a glucose dehydrogenase coupling method; Step II, esterifying gluconic acid under an acidic condition to generate glucolactone; Step III, generating reaction between the glucolactone and hydroxylamine alkali to generate hydroxamicacid; and Step IV, generating reaction between the hydroxamic acid and ferric trichloride to generate a colored complex, wherein the complex has a characteristic absorption peak and can be quantifiedthrough a spectrophotometric colorimetry. The high-throughput screening method has the characteristics of high precision, broad spectrum, wide detection concentration range, low cost and the like. Thehigh-throughput screening method is easy to operate, has a low requirement for the operation skill of an experimenter, and does not need a large number of pre-experiments.