Method for synthesizing UDCA (ursodesoxycholic acid) by catalyzing CA (cholic acid) by chemical cells
A cell and chemical technology, applied in the field of chemical cell catalysis of CA to synthesize UDCA, can solve the problems of expensive coenzyme, incomplete reaction, strict storage conditions, etc., and achieve simplified catalytic steps and catalytic conditions, simple reaction steps and conditions, and reaction conditions. mild effect
Inactive Publication Date: 2017-11-24
SUZHOU DUMEI BIOTECH CO LTD
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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-
Method used
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Embodiment 1-4
[0029] Example 1-4 is the construction of 3α-HSDH-7β-HSHD-GDH recombinant plasmid transformed into BL21 (DE3)
Embodiment 1
[0031] 3α-HSDH from Clostridium perfingens, 7β-HSDH from Bacteroides fragilis and GDH from Agrobacterium Tumefaciens were co-constructed. The gene was constructed on the plastid vector in a tandem manner, and then transformed into E. coli cells to obtain the recombinant strain SDM109.
Embodiment 2
[0033] 3α-HSDH from Eubacterium lentum, 7β-HSDH from Collinsella aerofaciens and GDH from Bacillus Anthracis were co-constructed. These three genes were chromosomally Integration, and then transformed into E. coli cells to obtain recombinant strain SDM109.
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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 ...
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Login to view more IPC IPC(8): C12P33/02
CPCC12P33/02
Inventor 陈车翘李国庆
Owner SUZHOU DUMEI BIOTECH CO LTD
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