Biochemical and Molecular Insights into Bile Salt Hydrolase in the Gastrointestinal Microflora - A Review -

  • Kim, Geun-Bae (Department of Food Science and Agricultural Chemistry, McGill University) ;
  • Lee, Byong H. (Department of Food Science and Agricultural Chemistry, McGill University)
  • Received : 2005.04.11
  • Accepted : 2005.05.07
  • Published : 2005.10.01


Bile salt deconjugation is the most biologically significant reaction among the bacterial alterations of bile acids in the gastrointestinal tract of human and animal. The responsible enzyme, bile salt hydrolase (BSH), catalyzes the hydrolysis of glycineand/or taurine-conjugated bile salts into amino acid residues and deconjugated bile acids. Herein we review current knowledge on the distribution of BSH activity among various microorganisms with respect to their biochemical and molecular characteristics. The proposed physiological impact of BSH activity on the host animal as well as on the BSH-producing bacterial cells is discussed. BSH activity of the probiotic strains is examined on the basis of BSH hypothesis, which was proposed to explain cholesterol-lowering effects of probiotics. Finally, the potential applications of BSH research are briefly discussed.


Bile Salt Hydrolase (BSH);Gut Microbes;Probiotics;Cholesterol Lowering;Genetic Marker


  1. Anderson, J. W. and S. E. Gilliland. 1999. Effect of fermented milk (yogurt) containing Lactobacillus acidophilus L1 on serum cholesterol in hypercholesterolemic humans. J. Am. College Nutr. 18:43-50.
  2. Baron, S. F. and P. B. Hylemon. 1997. Biotransformation of bile acids, cholesterol, and steroid hormones. In: Gastrointestinal Microbiology, Vol. I, Gastrointestinal Ecosystems and Fermentations, (Ed. R. I. Mackie and B. A. White), International Thomson Publ., New York, pp. 470-510.
  3. Canzi, E., E. Maconi, F. Aragozzini and A. Ferrari. 1989. Cooperative 3-epimeization of chenodoexycholic acid by Clostridium innocuum and Eubacterium lentum. Curr. Microbiol. 18:97-104.
  4. Chikai, T., H. Nakao and K. Uchida. 1987. Deconjugation of bile acids by human intestinal bacteria implanted in germ-free rats. Lipids 22:669-671.
  5. Coleman, J. P. and L. L. Hudson. 1995. Cloning and characterization of a conjugated bile acid hydrolase gene from Clostridium perfringens. Appl. Environ. Microbiol. 61:2514-2520.
  6. De Smet, I., L. van Horde, N. De Saeyer, M. vande Woewtyne, and W. Verstraete. 1994. In vitro study of bile salt hydrolase (BSH) activity of BSH isogenic Lactobacillus plantarum 80 strains and estimation of cholesterol lowering through enhanced BSH activity. Microbial Ecol. Health Dis. 7:315-329.
  7. Drasar, B. S. and M. J. Hill. 1974. Human intestinal flora. Academic Press, New York, pp. 103-123.
  8. Elkins, C. A., S. A. Moser and D. C. Savage. 2001. Genes encoding bile salt hydrolases and conjugated bile salt transporters in Lactobacillus johnsonii 100-100 and other Lactobacillus species. Microbiol. 147:3404-3412.
  9. Grill, J. P., F. Schneider, J. Crociani and J. Ballongue. 1995. Purification and characterization of conjugated bile salt hydrolase from Bifidobacterium longum BB536. Appl. Environ. Microbiol. 61:2577-2582.
  10. Gustafsson, B. E., T. Midtvedt and A. Norman. 1966. Isolated fecal microorganisms capable of $7{\alpha}$-dehydroxylating bile acids. J. Exp. Med. 123:413-432.
  11. Kawamoto, K., I. Horibe and K. Uchida. 1989. Purification and characterization of a new hydrolase for conjugated bile acids, chenodeoxycholyltaurine, from Bacteroides vulgatus. J. Biochem. 106:1049-1053.
  12. Kleerebezem, M., J. Boekhorst, R. van Kranenburg, D. Molenaar, O. P. Kuipers, R. Leer, R. Tarchini, S. A. Peters, H. M. Sandbrink, M. W. E. J. Fiers, W. Stiekema, R. M. Klein Lankhorst, P. A. Bron, S. M. Hoffer, M. N. Nierop Groot, R. Kerkhoven, M. de Vries, B. Ursing, W. M. de Vos and R. J. Siezen. 2003. Complete genome sequence of Lactobacillus plantarum WCFS1. Proc. Natl. Acad. Sci. USA. 100:1990-1995.
  13. Lack, L. and I. M. Weiner. 1966. Intestinal bile salt transport: structure-activity relationship and other properties. Am. J. Physiol. 210:1142-1152.
  14. Lundeen, S. and D. C. Savage. 1990. Characterization and purification of bile salt hydrolase from Lactobacillus sp. strain 100-100. J. Bacteriol. 172:4171-4177.
  15. Midtvedt, T. 1974. Microbial bile acid transformation. Am. J. Clin. Nutr. 27:1341-1348.
  16. Moore, W. E. C. and L. V. Holdeman. 1974. Human fecal flora: the normal flora of 20 Japanese-Hawaiians. Appl. Microbiol. 27:961-979.
  17. NCBI Microbial Genomes Annotation Project. 2003a. Lactobacillus gasseri Lgas_3, whole genome shotgun sequence. Direct submission to the GenBank/EMBL/DDBJ. Accession number NZ_AAAO02000005.
  18. Paulsen I. T., L. Banerjei, G. S. Myers, K. E. Nelson, R. Seshadri, T. D. Read, D. E. Fouts, J. A. Eisen, S. R. Gill, J. F. Heidelberg, H. Tettelin, R. J. Dodson, L. Umayam, L. Brinkac, M. Beanan, S. Daugherty, R. T. DeBoy, S. Durkin, J. Kolonay, R. Madupu, W. Nelson, J. Vamathevan, B. Tran, J. Upton, T. Hansen, J. Shetty, H. Khouri, T. Utterback, D. Radune and C. M. Fraser. 2003. Role of mobile DNA in the evolution of vancomycinresistant Enterococcus faecalis. Sci. 299:2071-2074.
  19. Pridmore, R. D., B. Berger, F. Desiere, D. Vilanova, C. Barretto, A. C. Pittet, M. C. Zwahlen, M. Rouvet, E. Altermann, R. Barrangou, B. Mollet, A. Mercenier, T. Klaenhammer, F. Arigoni and M. A. Schell. 2004. The genome sequence of the probiotic intestinal bacterium Lactobacillus johnsonii NCC 533. Proc. Natl. Acad. Sci. USA. 101:2512-2517.
  20. Stellwag, E. J. and P. B. Hylemon. 1976. Purification and characterization of bile salt hydrolase from Bacteroides fragilis subsp. fragilis. Biochim. Biophys. Acta 452:165-176.
  21. Tannock, G. W., J. M. Bateup and H. F. Jenkinson. 1997. Effect of sodium taurocholate on the in vitro growth of lactobacilli. Microbiol. Ecol. 33:163-167.
  22. Wijaya, A., A. Herrmann, H. Abriouel, I. Specht, N. M. K. Yousif, W. H. Holzapfel and C. M. A. P. Franz. 2003. Enterococcus faecium bile salt hydrolase (bsh) gene, complete cds. Direct submission to the Genbank/EMBL/DDBJ. Accession number AY260046.
  23. Corzo, G. 1997. Bile salt deconjugation by three strains of Lactobacillus acidophilus and characterization of their bile salt hydrolase. Doctoral Thesis, Oklahoma State University, USA.
  24. Tanaka, H., H. Hashiba, J. Kok and I. Mierau. 2000. Bile salt hydrolase of Bifidobacterium longum-Biochemical and genetic characterization. Appl. Environ. Microbiol. 66:2502-2512.
  25. du Toit, M., C. M. A. P. Franz, L. M. T. Dicks, U. Schillinger, P. Haberer, B. Warlies, F. Ahrens and W. H. Holzapfel. 1998. Characterization and selection of probiotic lactobacilli for a preliminary minipig feeding trial and their effect on serum cholesterol levels, faeces pH and faeces moisture content. Int. J. Food Microbiol. 40:93-104.
  26. Mallory, A., F. Kern, J. Smith and D. Savage. 1973. Patterns of bile acids and microflora in the human small intestine. I. Bile acids. Gastroenterol. 64:26-33.
  27. De Smet, I., P. De Boever and W. Verstraete. 1998. Cholesterol lowering in pigs through enhanced bacterial bile salt hydrolase activity. Br. J. Nutr. 79:185-194.
  28. Schell, M. A., M. Karmirantzou, B. Snel, D. Vilanova, B. Berger, G. Pessi, M. C. Zwahlen, F. Desiere, P. Bork, M. Delley, R. D. Pridmore and F. Arigoni. 2002. The genome sequence of Bifidobacterium longum reflects its adaptation to the human gastrointestinal tract. Proc. Natl. Acad. Sci. USA. 99:14422-144227.
  29. Shimizu, T., K. Ohtani, H. Hirakawa, K. Ohshima, A. Yamashita, T. Shiba, N. Ogasawara, M. Hattori, S. Kuhara and H. Hayashi. 2002. Complete genome sequence of Clostridium perfringens, an anaerobic flesh-eater. Proc. Natl. Acad. Sci. USA. 99:996-1001.
  30. Northfield, T. G. and I. McColl. 1973. Postprandial concentrations of free and conjugated bile acids sown the length of the normal human small intestine. Gut. 14:513-518.
  31. Sherwood, L. G. and S. Tabaqchali. 1969. Bacteria, bile, and the small bowel. Gut 10:963-972.
  32. Aries, V. and M. J. Hill. 1970a. Degradation of steroids by intestinal bacteria. I. Deconjugation of bile salts. Biochim. Biophys. Acta 202:526-534.
  33. Gilliland, S. E. and M. L. Speck. 1977. Deconjugation of bile acids by intestinal lactobacilli. Appl. Environ. Microbiol. 33:15-18.
  34. Leer, R. J., H. Christiaens, W. Verstraete, L. Peters, M. Posno and P. H. Pouwels. 1993. Gene disruption in Lactobacillus plantarum strain 80 by site-specific recombination: isolation of a mutant strain deficient in conjugated bile salt hydrolase activity. Mol. Gen. Genet. 239:269-272.
  35. Aries, V. and M. J. Hill. 1970b. Degradation of steroids by intestinal bacteria. II. Enzymes catalyzing the oxidoreduction of the $3{\alpha}$-, $7{\alpha}$-, and $12{\alpha}$-hydroxyl groups in cholic and the dehydroxylation of the $7{\alpha}$-hydroxyl group. Biochim. Biophys. Acta 202:535-543.
  36. Marteau, P., M. F. Gerhardt, A. Myara, E. Bouvier, F. Trivin and J. C. Rambaud. 1995. Metabolosm of bile salts by alimentary bacteria during transit in the human small intestine. Microbial Ecol. Health Dis. 8:151-157.
  37. Savage, D. C. and S. A. Moser. 1999. Lactobacillus acidophilus putative bile salt hydrolase operon, complete sequence. Direct submission to the Genbank/EMBL/DDBJ. Accession no. AF091248
  38. Batta, A. K., G. Salen, R. Arora, S. Shefer, M. Batta and A. Person. 1990. Side chain conjugation prevents bacterial 7-dehydroxylation of bile acids. J. Biol. Chem. 265:10925-10928.
  39. Masuda, N. 1981. Deconjugation of bile salts by Bacteroides and Clostridium. Microbiol. Immunol. 25:1-11.
  40. Russell, W. M. and T. R. Klaenhammer. 2001. Identification and cloning of gusA, encoding a new beta-glucuronidase from Lactobacillus gasseri ADH. Appl. Environ. Microbiol. 67:1253-1261.
  41. Savage, D. C., S. G. Lundeen and L. T. O’Connor. 1977. Microbial ecology of the gastrointestinal tract. Ann. Rev. Microbiol. 31:107-133.
  42. Christiaens, H., R. J. Leer, P. H. Pouwels and W. Verstraete. 1992. Cloning and expression of a conjugated bile acid hydrolase gene from Lactobacillus plantarum by using a direct plate assay. Appl. Environ. Microbiol. 58:3792-3798.
  43. Corzo, G. and S. E. Gilliland. 1999b. Measurement of bile salt hydrolase activity from Lactobacillus acidophilus based on disappearance of conjugated bile salts. J. Dairy Sci. 82:466-471.
  44. Hofmann, A. F. and K. J. Mysels. 1992. Bile acid solubility and precipitation in vitro and in vivo: the role of conjugation, pH, and $Ca^{2+}$ ions. J. Lipid Res. 33:617-626.
  45. Kobayashi, H. 1985. A proton-translocating ATPase regulates pH of the bacterial cytoplasm. J. Biol. Chem. 260:72-76.
  46. Tannock, G. W., M. P. Dashkevicz and S. D. Feighner. 1989. Lactobacilli and bile salt hydrolase in the murine intestinal tract. Appl. Environ. Microbiol. 55:1848-1851.
  47. Cole, C. B. and R. Fuller. 1984. Bile acid deconjugation and attachment of chicken gut bacteria: their possible role in growth depression. Br. Poult. Sci. 25:227-231.
  48. Batta, A. K., G. Salen and S. Shefer. 1984. Substrate specificity of cholylglycine hydrolase for the hydrolysis of bile acid conjugates. J. Biol. Chem. 259:15035-15039.
  49. Dashkevicz, M. P. and S. D. Feighner. 1989. Development of a differential medium for bile salt hydrolase-active Lactobacillus spp. Appl. Environ. Microbiol. 55:11-16.
  50. Salminen, S., E. Isolauri and E. Salminen. 1996. Clinical uses of probiotics for stabilizing the gut mucosal barrier: Successful strains for future challenges. Antonie van Leeuwenhoek 70:347-358.
  51. Dean, M., C. Cervellati, E. Casanova, M. Squerzanti, V. Lanzara, A. Medici, P. P. de Laureto and C. M. Bergamini. 2002. Characterization of cholylglycine hydrolase from a bileadapted strain of Xanthomonas maltophilia and its application for quantitative hydrolysis of conjugated bile salts. Appl. Environ. Microbiol. 68:3126-3128.
  52. Dussurget, O., D. Cabanes, P. Dehoux, M. Lecuit, C. Buchrieser, P. Glaser and P. Cossart. 2002. Listeria monocytogenes bile salt hydrolase is a virulence factor involved in the intestinal and hepatic phases of listeriosis. Mol. Microbiol. 45:1095-1106.
  53. Saavedra, L., M. P. Taranto, F. Sesma and G. F. de Valdez. 2003. Homemade traditional cheeses for the isolation of probiotic Enterococcus faecium strains. Int. J. Food Microbiol. 88:241-245.
  54. De Smet, I., L. van Horde, M. van de Woestyne, H. Christiaens, and W. Verstraete. 1995. Significance of bile salt hydrolytic activities of lactobacilli. J. Appl. Bacteriol. 79:292- 301.
  55. van Faassen, A., J. Bol, W. van Dokkum, N. A. Pikaar, T. Ockhuizen and R. J. J. Hermus. 1987. Bile acids, neutral steroids and bacteria in faeces as affected by a mixted, a lactoovovegetarian and a vegan diet. Am. J. Clin. Nutr. 46:962-967.
  56. Corzo, G. and S. E. Gilliland. 1999a. Bile salt hydrolase activity of three strains of Lactobacillus acidophilus. J. Dairy Sci. 82:472-480.
  57. Moser, S. A. and D. C. Savage. 2001. Bile salt hydrolase activity and resistance to toxicity of conjugated bile salts are unrelated properties in lactobacilli. Appl. Environ. Microbiol. 67:3476-3480.
  58. Tanaka, H., K. Doesburg, T. Iwasaki and I. Mierau. 1999. Screening of lactic acid bacteria for bile salt hydrolase activity. J. Dairy Sci. 82:2530-2535.
  59. Tanida, N., Y. Hikdsa, Y. Shimoyama and K. D. R. Setchell. 1984. Comparison of faecal bile acid profiles between patients with adenomatous polyps of the large bowel and healthy subjects in Japan. Gut 25:824-832.
  60. Gopal-Srivastava, R. and P. B. Hylemon. 1988. Purification and characterization of conjugated bile salt hydrolase from Clostridium perfringens. J. Lipid. Res. 29:1079-1085.
  61. Bateup, J. M., M. A. McConnell, H. F. Jenkinson and G. W. Tannock. 1995. Comparison of Lactobacillus strains with respect to bile salt hydrolase activity, colonization of the gastrointestinal tract, and growth rate of the murine host. Appl. Environ. Microbiol. 61:1147-1149.
  62. Rodriguez, E., J. L. Arques, R. Rodriguez, M. Nunez and M. Medina. 2003. Reuterin production by lactobacilli isolated from pig faeces and evaluation of probiotic traits. Lett. Appl. Microbiol. 37:259-263.
  63. van der Meer, R., D. S. M. L. Termont and H. T. de Vries. 1991. Differential effects of calcium ions and calcium phosphate on cytotoxicity of bile acids. Am. J. Physiol. 260:G142-147.
  64. Van Eldere, J., J. Robben, G. de Pauw, R. Merckx and H. Eyssen. 1988. Isolation and identification of intestinal steroiddesulfating bacteria from rats and humans. Appl. Environ. Microbiol. 54:2112-2117.
  65. Elkins, C. A. and D. C. Savage. 1998. Identification of genes encoding conjugated bile salt hydrolase and transport in Lactobacillus johnsonii 100-100. J. Bacteriol. 180:4344-4349.
  66. Franz, C. M. A. P., I. Sepcht, P. Haberer and W. H. Holzapfel. 2001. Bile salt hydrolase activity of enterococci isolated form food: Screening and quantitative determination. J. Food Prot. 64:725-729.
  67. Hofmann, A. F. 1977. The enterohepatic circulation of bile acids in man. Clin. Gastroenterol. 6:3-24.
  68. Nair, P. P., M. Gordon and J. Reback. 1967. The enzymatic cleavage of the carbon-nitrogen bond in $3{\alpha}$, $7{\alpha}$, $12{\alpha}$-trihydroxy-$5{\beta}$-cholan-24-oylglycine. J. Biol. Chem. 242:7-11.
  69. Farkkila, M. and T. A. Miettinen. 1990. Lipid metabolism in bile acid malabsorption. Ann. Med. 22:5-13.

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