METHOD OF PREPARING TAUROURSODEOXYCHOLIC ACID BY BIOTRANSFORMATION AND APPLICATION THEREOF

摘要

A method of preparing tauroursodeoxycholic acid by biotransformation and an application thereof, comprising genetic codon optimization, engineered bacteria construction, engineered bacteria cultivation, substrate transformation, and product preparation. Tauroursodeoxycholic acid is prepared by transforming a substrate by means of direct fermentation of engineered bacteria, and the substrate is taurochenodeoxycholic acid. The substrate may reach 250 g/L in concentration, the reaction time is short, the transformation rate of the substrate reaches 98% and above, and the obtained product reaches 99% and above in purity; cyclic regeneration of NAD⁺ in the reaction system helps greatly reduce the usage of coenzyme NAD⁺, the cost of enzyme catalysis reaction is reduced, and industrial amplification is benefited. Steroid dehydrogenase and coenzyme regeneration enzyme are connected by a flexible polypeptide sequence to construct a fusion protein polymer, binding distances to the substrate and the coenzyme are shorter, transformation reaction is more facilitated, the number of times of fermentations in industrial production is decreased, the process is simplified, and time costs and raw material costs are saved.