Method for synthesizing UDCA (ursodesoxycholic acid) by catalyzing CA (cholic acid) by chemical cells

Abstract

The invention provides a novel method for synthesizing UDCA (ursodesoxycholic acid) by catalyzing CA (cholic acid) by a whole-cell combined chemical method. According to the method, cholic acid is oxidized by the aid of sodium hypochlorite to obtain dehydrocholic acid, the dehydrocholic acid is catalyzed by the aid of a built Escherichia coli engineering bacterium cell to synthesize 12-keto-ursodesoxycholic acid, and the 12-keto-ursodesoxycholic acid is reduced into the ursodesoxycholic acid by a Wolff-Kishner method. The Escherichia coli engineering bacterium cell comprises a co-expression system of three foreign genes such as 7beta-hydroxysteroid dehydrogenase (7beta-HSDH), 3alpha-hydroxysteroid dehydrogenase (3alpha-HSDH) and coenzyme NADP (nicotinamide adenine dinucleotide phosphate) +regenerative glucose dehydrogenase (GDH). The method is cheap in chemical oxidation raw material, less in catalytic by-products and by-products, high in yield and cell catalyst yield and applicable to industrial production, and addition of coenzyme is omitted.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a method for synthesizing UDCA from chemical cell catalysis CA. Background technique [0002] Ursodeoxycholic acid (UDCA), namely 3α,7β-dihydroxy-5β-cholestane-24-acid, is a non-toxic hydrophilic cholic acid that can competitively inhibit toxic endogenous cholic acid Absorption in the ileum increases the secretion capacity of cholestatic hepatocytes by activating the signaling network composed of calcium ions and protein kinase C, and by activating fission-active protein kinases, reducing the concentration of endogenous hydrophobic cholic acid in blood and hepatocytes , to achieve the effect of anti-cholestasis. Clinically, it is mainly used to treat various gallstone diseases and various acute and chronic liver diseases. [0003] With the increasing domestic and foreign market demand of ursodeoxycholic acid, and the trend is increasing year by year, the synthesis of ursodeoxycholic ...

AI Technical Summary

Problems solved by technology

The purity of the final product is 98%, and the yield is 89.3%. Although it is greatly improved compared with the chemical method, there are also two problems, or the problems of this type of route. The first is that the chemical method oxidizes CDCA to 7-KLCA The process will have by-products that are not easy to control, such as impurities that are only oxidized at the 3-position and diketone impurities that are oxidized at the same time at the 3-position and 7-position, which increases the difficulty and cost of post-processing; the second is that there are many types of enzymes in the reduction process, and there are 3 types Enzyme and 1 coenzyme, the ratio of the enzyme needs to be well controlled, otherwise it will increase the cost or cause incomplete reaction, affecting the stability of the production batch, while the coenzyme is more expensive, the stability is not good, and the storage conditions are strict, etc.

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