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Large-scale production of ursodeoxycholic acid from chenodeoxycholic acid by engineering 7α- and 7β-hydroxysteroid dehydrogenase

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Abstract

7α- and 7β-hydroxysteroid dehydrogenases (HSDHs) are key biocatalysts for the biotransformation of ursodeoxycholic acid (UDCA) from chenodeoxycholic acid (CDCA). Various researches focused on heterogeneously expressed engineering enzymes to epimerize CDCA for UDCA, however not yet applied to further industrial application. In this work, we present the large-scale production of UDCA from CDCA by 7α- and 7β-HSDH enzymatic synthesis. Engineering enzymes were both successfully heterologous overexpressed in Escherichia coli BL21, and the effect of the enzymatic synthesis was investigated. The mass analysis (MS), IR spectrum, 1H NMR and 13C NMR were used to characterize the product. 500-L fermentor fermentation strategy producing a stable supply of HSDH and large-scale production process of UDCA in dozens kilogram class enabled industrial application.

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References

  1. Im E, Martinez JD (2004) Ursodeoxycholic acid (UDCA) can inhibit deoxycholic acid (DCA)-induced apoptosis via modulation of EGFR/Raf-1/ERK signaling in human colon cancer cells. J Nutr 134:483–486

    Article  CAS  PubMed  Google Scholar 

  2. Fieser LF, Herz JE, Klohs MW, Romero MA, Utne T (1952) Cathylation (carbethoxylation) of steroid alcohols. J Am Chem Soc 74:3309–3313

    Article  CAS  Google Scholar 

  3. Hu XZ, Liu AJ (2013) Method for producing ursodesoxycholic acid by using chenodeoxycholic acid as raw material. Chinese Patent CN102911235

  4. Tian H, Zhao HB, Cao XJ (2013) Catalytic transfer hydrogenation of 7-ketolithocholic acid to ursodeoxycholic acid with Raney nickel. J Ind Eng Chem 19:606–613

    Article  CAS  Google Scholar 

  5. Yuan XX, Ma XL, Cao XJ (2014) Preparation of ursodeoxycholic acid by direct electro-reduction of 7-ketolithocholic acid. Korean J Chem Eng 31:1276–1280

    Article  CAS  Google Scholar 

  6. Edenharder R, Knaflic T (1981) Epimerization of chenodeoxycholic acid to ursodeoxycholic acid by human intestinal lecithinase-lipase-negative Clostridia. J Lipid Res 22:652–658

    CAS  PubMed  Google Scholar 

  7. Lepercq P, Gérard P, Béguet F, Raibaud P, Grill JP, Relano P, Cayuela C, Juste C (2004) Epimerization of chenodeoxycholic acid to ursodeoxycholic acid by Clostridium baratii isolated from human feces. FEMS Microbiol Lett 235:65–72

    Article  CAS  PubMed  Google Scholar 

  8. Medici A, Pedrini P, Bianchini E, Fantin G, Guerrini A, Natalini B, Pellicciari R (2002) 7α-OH epimerisation of bile acids via oxidoreduction with Xanthomonas maltophilia. Steroids 67:51–56

    Article  CAS  PubMed  Google Scholar 

  9. Yoshimoto T, Higashi H, Kanatani A, Lin XS, Nagai H, Oyama H, Kurazono K, Tsuru D (1991) Cloning and sequencing of the 7alpha-hydroxysteroid dehydrogenase gene from Escherichia coli HB101 and characterization of the expressed enzyme. J Bacteriol 173:2173–2179

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Tanaka N, Nonaka T, Tanabe T, Yoshimoto T, Tsuru D, Mitsui Y (1996) Crystal structures of the binary and ternary complexes of 7 alpha-hydroxysteroid dehydrogenase from Escherichia coli. Biochemistry 35:7715–7730

    Article  CAS  PubMed  Google Scholar 

  11. Liu L, Aigner A, Schmid RD (2011) Identification, cloning, heterologous expression, and characterization of a NADPH-dependent 7β-hydroxysteroid dehydrogenase from Collinsella aerofaciens. Appl Microbiol Biotechnol 90:127–135

    Article  CAS  PubMed  Google Scholar 

  12. Bennett MJ, McKnight SL, Coleman JP (2003) Cloning and characterization of the NAD-dependent 7α-hydroxysteroid dehydrogenase from Bacteroides fragilis. Curr Microbiol 47:475–484

    Article  CAS  PubMed  Google Scholar 

  13. Ferrandi EE, Bertolesi GM, Polentini F, Negri A, Riva S, Monti D (2012) In search of sustainable chemical processes: cloning, recombinant expression, and functional characterization of the 7α- and 7β-hydroxysteroid dehydrogenases from Clostridium absonum. Appl Microbiol Biotechnol 95:1221–1233

    Article  CAS  PubMed  Google Scholar 

  14. Lee JY, Arai H, Nakamura Y, Fukiya S, Wada M, Yokota A (2013) Contribution of the 7β-hydroxysteroid dehydrogenase from Ruminococcus gnavus N53 to ursodeoxycholic acid formation in the human colon. J Lipid Res 54:3062–3069

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Zheng MM, Wang RF, Li CX, Xu JH (2015) Two-step enzymatic synthesis of ursodeoxycholic acid with a new 7β-hydroxysteroid dehydrogenase from Ruminococcus torques. Process Biochem 50:598–604

    Article  CAS  Google Scholar 

  16. Bakonyi D, Hummel W (2017) Cloning, expression, and biochemical characterization of a novel NADP+-dependent 7α-hydroxysteroid dehydrogenase from Clostridium difficile and its application for the oxidation of bile acids. Enzyme Microb Tech 99:16–24

    Article  CAS  Google Scholar 

  17. Zheng MM, Chen KC, Wang RF, Li H, Li CX, Xu JH (2017) Engineering 7β-hydroxysteroid dehydrogenase for enhanced ursodeoxycholic acid production by multiobjective directed evolution. J Agric Food Chem 65:1178–1185

    Article  CAS  PubMed  Google Scholar 

  18. Zheng MM, Chen FF, Li H, Li CX, Xu JH (2018) Continuous production of ursodeoxycholic acid by using two cascade reactors with co-immobilized enzymes. ChemBioChem 19:347–353

    Article  CAS  PubMed  Google Scholar 

  19. Pedrini P, Andreotti E, Guerrini A, Dean M, Fantin G, Giovannini PP (2006) Xanthomonas maltophilia CBS 897.97 as a source of new 7α-and 7β-hydroxysteroid dehydrogenases and cholylglycine hydrolase: improved biotransformations of bile acids. Steroids 71:189–198

    Article  CAS  PubMed  Google Scholar 

  20. Eggert T, Bakonyi D, Hummel W (2014) Enzymatic routes for the synthesis of ursodeoxycholic acid. J Biotechnol 191:11–21

    Article  CAS  PubMed  Google Scholar 

  21. Korz DJ, Rinas U, Hellmuth K, Sander EA, Deckwer WD (1995) Simple fed-batch technique for high cell density cultivation of Escherichia coli. J Biotechnol 39:59–65

    Article  CAS  PubMed  Google Scholar 

  22. Kole MM, Altosaar I (1985) Conversion of chenodeoxycholic acid to ursodeoxycholic acid by Clostridium absonum in culture and by immobilized cells. FEMS Microbiol Lett 28:69–72

    Article  CAS  Google Scholar 

  23. Bovara R, Canzi E, Carrea G, Pilotti A, Riva S (1993) Enzymatic α/β inversion of the C-7-hydroxyl of steroids. J Org Chem 58:499–501

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. Shaanxi Key Laboratory of Degradable Biomedical Materials, School of Chemical Engineering, Northwest University, Xi’an, 710069, China

    Xuan Zhang & Daidi Fan

  2. Department of Scientific Research and Development, Tsinghua University, Beijing, 100084, China

    Xiufu Hua

  3. Department of Chemical Engineering, Tsinghua University, Beijing, 100084, China

    Tian Zhang

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  1. Xuan Zhang
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  2. Daidi Fan
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  3. Xiufu Hua
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  4. Tian Zhang
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Correspondence to Daidi Fan or Xiufu Hua.

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Zhang, X., Fan, D., Hua, X. et al. Large-scale production of ursodeoxycholic acid from chenodeoxycholic acid by engineering 7α- and 7β-hydroxysteroid dehydrogenase. Bioprocess Biosyst Eng 42, 1537–1545 (2019). https://doi.org/10.1007/s00449-019-02151-4

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  • DOI: https://doi.org/10.1007/s00449-019-02151-4

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