Abstract
A facultative anaerobic bacterium, Pseudomonas sp. strain Chol1, degrading cholate and other bile acids was isolated from soil. We investigated how strain Chol1 grew with cholate and whether growth was affected by the toxicity of this compound. Under anoxic conditions with nitrate as electron acceptor, strain Chol1 grew by transformation of cholate to 7,12-dihydroxy-1,4-androstadiene-3,17-dione (DHADD) as end product. Under oxic conditions, strain Chol1 grew by transformation of cholate to 3,7,12-trihydroxy-9,10-seco-1,3,5(10)-androstatriene-9,17-dione (THSATD), which accumulated in the culture supernatant before its further oxidation to CO2. Strain Chol1 converted DHADD into THSATD by an oxygenase-dependent reaction. Addition of cholate (≥10 mM) to cell suspensions of strain Chol1 caused a decrease of optical density and viable counts but aerobic growth with these toxic cholate concentrations was possible. Addition of CCCP or EDTA strongly increased the sensitivity of the cells to 10 mM cholate. EDTA also increased the sensitivity of the cells to DHADD and THSATD (≤1.7 mM). The toxicity of cholate and its degradation intermediates with a steroid structure indicates that strain Chol1 requires a strategy to minimize these toxic effects during growth with cholate. Apparently, the proton motive force and the outer membrane are necessary for protection against these toxic effects.
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References
Barnes PJ, Bilton RF, Mason AN, Fernandez F, Hill MJ (1975) The coupling of anaerobic steroid dehydrogenation to nitrate reduction in Pseudomonas N.C.I.B. 10590 and Clostridium paraputrificum. Biochem Soc Trans 3:299–301
Bortolini O, Medici A, Poli S (1997) Biotransformations on steroid nucleus of bile acids. Steroids 62:564–577
Colborn T (2004) Endocrine disruption overview: are males at risk? Adv Exp Med Biol 545:189–201
van der Geize R, Hessels GI, van Gerwen R, van der Meijden P, Dijkhuizen L (2002) Molecular and functional characterization of kshA and kshB, encoding two components of 3-ketosteroid 9alpha-hydroxylase, a class IA monooxygenase, in Rhodococcus erythropolis strain SQ1. Mol Microbiol 45:1007–1018
Gunn JS (2000) Mechanisms of bacterial resistance and response to bile. Microbes Infect 2:907–913
Hancock RE (1984) Alterations in outer membrane permeability. Annu Rev Microbiol 38:237–264
Hayakawa S (1982) Microbial transformation of bile acids. A unified scheme for bile acid degradation, and hydroxylation of bile acids. Z Allg Mikrobiol 22:309–326
Helenius A, Simons K (1975) Solubilization of membranes by detergents. Biochim Biophys Acta 415:29–79
Hesse M, Meier H, Zeeh B (1995) Spektroskopische Methoden in der organischen Chemie, 5th edn. Thieme, Stuttgart
Hoben HJ, Somasegaran P (1982) Comparison of the pour, spread, and drop plate methods for enumeration of Rhizobium spp. in inoculants made from presterilized peat. Appl Environ Microbiol 44:1246–1247
Horinouchi M, Hayashi T, Yamamoto T, Kudo T (2003) A new bacterial steroid degradation gene cluster in Comamonas testosteroni TA441 which consists of aromatic-compound degradation genes for seco-steroids and 3-ketosteroid dehydrogenase genes. Appl Environ Microbiol 69:4421–4430
Hylemon PB, Harder J (1998) Biotransformation of monoterpenes, bile acids, and other isoprenoids in anaerobic ecosystems. FEMS Microbiol Rev 22:475–488
Jenkins RL, Wilson EM, Angus RA, Howell WM, Kirk M (2003) Androstenedione and progesterone in the sediment of a river receiving paper mill effluent. Toxicol Sci 73:53–59
Lee CY, Liu WH (1992) Production of androsta-1,4-diene-3,17-dione from cholesterol using immobilized growing cells of Mycobacterium sp. NRRL B-3683 adsorbed on solid carriers. Appl Microbiol Biotechnol 36:598–603
Leppik RA, Park RJ, Smith MG (1982) Aerobic catabolism of bile acids. Appl Environ Microbiol 44:771–776
van Niel CB, Allen MB (1952) A note on Pseudomonas stutzeri. J Bacteriol 64:413–422
Nikaido H (2003) Molecular basis of bacterial outer membrane permeability revisited. Microbiol Mol Biol Rev 67:593–656
Owen RW, Bilton RF (1983) The degradation of cholic acid by Pseudomonas sp. N.C.I.B. 10590 under anaerobic conditions. Biochem J 216:641–654
Palleroni NJ, Doudoroff M, Stanier RY, Solanes RE, Mandel M (1970) Taxonomy of the aerobic pseudomonads: the properties of the Pseudomonas stutzeri group. J Gen Microbiol 60:215–231
Park RJ (1984) Phenolic 9,10-secosteroids as products of the catabolism of bile acids by a Pseudomonas sp. Steroids 44:175–193
Park RJ, Dunn NW, Ide JA (1986) A catecholic 9,10-seco steroid as a product of aerobic catabolism of cholic acid by a Pseudomonas sp. Steroids 48:439–450
Perez C, Falero A, Llanes N, Hung BR, Herve ME, Palmero A, Marti E (2003) Resistance to androstanes as an approach for androstandienedione yield enhancement in industrial mycobacteria. J Ind Microbiol Biotechnol 30:623–626
Plesiat P, Nikaido H (1992) Outer membranes of gram-negative bacteria are permeable to steroid probes. Mol Microbiol 6:1323–1333
Poole K (2004) Efflux-mediated multiresistance in Gram-negative bacteria. Clin Microbiol Infect 10:12–26
Schmitt-Wagner D, Friedrich MW, Wagner B, Brune A (2003) Phylogenetic diversity, abundance, and axial distribution of bacteria in the intestinal tract of two soil-feeding termites (Cubitermes spp.). Appl Environ Microbiol 69:6007–6017
Sih CJ, Lee SS, Tsong YY, Wang KC (1966) Mechanisms of steroid oxidation by microorganisms. 8. 3,4-Dihydroxy-9,10-secoandrosta-1,3,5(10)-triene-9,17-dione, an intermediate in the microbiological degradation of ring A of androst-4-ene-3,17-dione. J Biol Chem 241:540–550
Smith MG, Park RJ (1984) Effect of restricted aeration on catabolism of cholic acid by two Pseudomonas species. Appl Environ Microbiol 48:108–113
Strijewski A (1982) The steroid-9 alpha-hydroxylation system from Nocardia species. Eur J Biochem 128:125–135
Tai HH, Sih CJ (1970) 3,4-Dihydroxy-9,10-secoandrosta-1,3,5(10)-triene-9,17-dione 4,5-dioxygenase from Nocardia restrictus. I. Isolation of the enzyme and study of its physical and chemical properties. J Biol Chem 245:5062–5071
Tenneson ME, Owen RW, Mason AN (1977) The anaerobic side-chain cleavage of bile acids by Escherichia coli isolated from human faeces. Biochem Soc Trans 5:1758–1760
Tenneson ME, Baty JD, Bilton RF, Mason AN (1979) The degradation of cholic acid by Pseudomonas sp. N.C.I.B. 10590. Biochem J 184:613–618
Thanassi DG, Cheng LW, Nikaido H (1997) Active efflux of bile salts by Escherichia coli. J Bacteriol 179:2512–2518
Widdel F, Pfennig N (1981) Studies on dissimilatory sulfate-reducing bacteria that decompose fatty acids. I. Isolation of new sulfate-reducing bacteria enriched with acetate from saline environments. Description of Desulfobacter postgatei gen. nov., sp. nov. Arch Microbiol 129:395–400
Widdel F, Kohring GW, Mayer F (1983) Studies on dissimilatory sulfate-reducing bacteria that decompose fatty acids. III. Characterization of the filamentous gliding Desulfonema limicola gen. nov. sp. nov., and Desulfonema magnum sp. nov. Arch Microbiol 134:286–294
Zietz E, Spiteller G (1974) Zur Lokalisierung funktioneller Gruppen mit Hilfe der Massenspektrometrie-XI. Tetrahedron 30:585–596
Acknowledgements
The authors like to thank Antje Karst and René Schoenenberger for excellent technical assistance and Normen Szesni, Dirk Schmitt-Wagner, Melanie Hempel, and Steffi Schneider for help with some experiments. Janosch Klebensberger is acknowledged for valuable discussions. Financial support to B.P. by the Deutsche Forschungsgemeinschaft (project PH71/2-1) is acknowledged.
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Philipp, B., Erdbrink, H., Suter, M.J.F. et al. Degradation of and sensitivity to cholate in Pseudomonas sp. strain Chol1. Arch Microbiol 185, 192–201 (2006). https://doi.org/10.1007/s00203-006-0085-9
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DOI: https://doi.org/10.1007/s00203-006-0085-9