Archives of Microbiology

, Volume 153, Issue 1, pp 50–59 | Cite as

Reductive cleavage of sarcosine and betaine by Eubacterium acidaminophilum via enzyme systems different from glycine reductase

  • K. Hormann
  • J. R. Andreesen
Original Papers


The obligate anaerobe Eubacterium acidaminophilum metabolized the glycine derivatives sarcosine (N-monomethyl glycine) and betaine (N-trimethyl glycine) only by reduction in a reaction analogous to glycine reductase. Using formate as electron donor, sarcosine and betaine were stoichiometrically reduced to acetate and methylamine or trimethylamine, respectively. The N-methyl groups of the cosubstrates or of the amines produced were not transformed to CO2 or acetate. Under optimum conditions (formate/acceptor ratio of 1 to 1.2, 34°C, pH 7.3) the doubling times were 4.2 h on formate/sarcosine and 3.6 h on formate/betaine. The molar growth yields were 8.15 and 8.5 g dry cell mass per mol sarcosine and betaine, respectively. The assays for sarcosine reductase and betaine reductase were optimized in cell extracts; NADPH was preferred as physiological electron donor compared to NADH, dithioerythritol was used as artificial donor; no requirements for AMP and ADP could be detected. Growth experiments mostly revealed diauxic substrate utilization pattern using different combinations of glycine, sarcosine, and betaine (plus formate) and inocula from different precultures. Glycine was always utilized first, what coincided with the presence of glycine reductase activity under all growth conditions except for serine as substrate. Sarcosine reductase and betaine reductase were only induced when E. acidaminophilum was grown on sarcosine and betaine, respectively. Creatine was metabolized via sarcosine. [75Se]-selenite labeling revealed about the same pattern of predominant labeled proteins in glycine-, sarcosine-, and betaine-grown cells.

Key words

Eubacterium acidaminophilum Diauxic growth Growth yields Formate metabolism Glycine reductase Sarcosine reductase Betaine reductase Creatine metabolism Selenium incorporation 





N-Tris (hydroxymethyl) methyl-2-amino-ethane sulfonic acid


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  1. AndreesenJR, GottschalkG, SchlegelHG (1970) Clostridium formicoaceticum nov spec. Isolation, description and distinction from C. aceticum and C. thermoaceticum. Arch Mikrobiol 72: 154–174Google Scholar
  2. BarnardGF, AkhtarM (1979) Mechanistic and stereochemical studies on the glycine reductase of Clostridium sticklandii. Eur J Biochem 99:593–603Google Scholar
  3. BlundenG, GordonSM, McLeanWFH, GuiryMD (1982) The distribution and possible taxonomic significance of quarternary ammonium and other Dragendorff-positive compounds in some genera of marine algae. Bot Mar 25:563–567Google Scholar
  4. BradfordMM (1976) A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein dye-binding. Anal Biochem 72:248–254Google Scholar
  5. BreznakJA, SwitzerJM, SeitzHJ (1988) Sporomusa termitida sp nov, an H2/CO2-utilizing acetogen isolated from termites. Arch Microbiol 150:282–288Google Scholar
  6. CarruthersA, OldfieldJET, TeagueHJ (1960) The removal of interfering ions in the detection of betaine in sugar-beet juices and plant material. Analyst 85:272–275Google Scholar
  7. ConeJE, Martin del RioR, DavisJN, StadtmanTC (1976) Chemical characterization of the selenoprotein component of clostridial glycine reductase: Identification of selenocysteine as the organoselenium moiety. Proc Natl Acad Sci USA 73:2659–2663Google Scholar
  8. ConeJE, Martin del RioR, StadtmanTC (1977) Clostridial glycine reductase complex. Purification and characterization of the selenoprotein component. J Biol Chem 252:5337–5344Google Scholar
  9. CsonkaLN (1989) Physiological and genetic responses of bacteria to osmotic stress. Microbiol Rev 53:121–147Google Scholar
  10. DornM, AndreesenJR, GottschalkG (1978) Fermentation of fumarate and L-malate by Clostridium formicoaceticum. J Bacteriol 133:26–32Google Scholar
  11. EnerothP, LindstedtG (1965) Thin-layer chromatography of betaines and other compounds related to carnitine. Anal Biochem 10:479–485Google Scholar
  12. FinkelsteinJD, MartinJJ, HarrisJ (1988) Methionine metabolism in mammals. The methionine-sparing effect of cystine. J Biol Chem 263:11750–11754Google Scholar
  13. FochtRL, SchmidtFH (1956) Colorimetric determination of betaine in glutamate process and liquor. J Agric Food Chem 4:579–585Google Scholar
  14. FreudenbergW, AndreesenJR (1989) Purification and partial characterization of the glycine decarboxylase multienzyme complex from Eubacterium acidaminophilum. J Bacteriol 171:2209–2215Google Scholar
  15. FreudenbergW, DietrichsD, LebertzH, AndreesenJR (1989a) Isolation of an atypically small lipoamide dehydrogenase involved in the glycine decarboxylase complex from Eubacterium acidaminophilum. J Bacteriol 171:1346–1354Google Scholar
  16. FreudenbergW, HormannK, RiethM, AndreesenJR (1989b) Involvement of a selenoprotein in glycine, sarcosine, and betaine reduction by Eubacterium acidaminophilum. In: WendelA (ed) Selenium in biology and medicine. Springer, Heidelberg, pp 25–28Google Scholar
  17. FreudenbergW, MayerF, AndreesenJR (1989c) Immunocytochemical localization of proteins P1, P2, P3 of glycine decarboxylase, and of the selenoprotein PA of glycine reductase, all involved in anaerobic glycine metabolism of Eubacterium acidaminophilum. Arch Microbiol 152:182–188Google Scholar
  18. GalinskiEA, TrüperHG (1982) Betaine, a compatible solute in the extremely halophilic phototrophic bacterium Ectothiorhodospira halochloris. FEMS Microbiol Lett 13:357–360Google Scholar
  19. Gauglitz U (1988) Anaerober mikrobieller Abbau von Kreatin, Kreatinin and N-Methylhydantoin. PhD thesis, Univ GöttingenGoogle Scholar
  20. GenthnerBRS, DavisCL, BryantBP (1981) Features of rumen and sewage sludge strains of Eubacterium limosum, a methanol- and H2−CO2-utilizing species. Appl Environ Microbiol 42:12–19Google Scholar
  21. GoodwinJF, StampwalaS (1973) Spectrophotometric quantification of glycine in serum and urine. Clin Chem 19:1010–1015Google Scholar
  22. GorhamJ (1984) Separation of plant betaines and their sulphur analogues by cation-exchange high-performance liquid chromatography. J Chromatogr 287:345–351Google Scholar
  23. GreenbergDM (1961) Biosynthesis of amino acids and related compounds. In: GreenbergDM (ed) Metabolic pathways. Academic Press, New York, pp 173–236Google Scholar
  24. HeigenerH (1935) Verwertung von Aminosäuren als gemeinsame C- und N-Quelle durch bekannte Bodenbakterien nebst botanischer Beschreibung neu isolierter Betain- und Valin-Abbauer. Zentralbl Bakt Abt II 93:81–113Google Scholar
  25. HeijthuijsenJHFG, HansenTA (1989) Betaine fermentation and oxidation by marine Desulfuromonas strains. Appl Environ Microbiol 55:965–969Google Scholar
  26. IkutaS, MatuuraK, ImamuraS, MisakiH, HoriutiY (1977) Oxidative pathways of choline to betaine in the soluble fraction prepared from Arthrobacter globiformis. J Biochem 82:157–163Google Scholar
  27. ImhoffJF (1986) Osmoregulation and compatible solutes in eubacteria. FEMS Microbiol Rev 39:57–66Google Scholar
  28. KortsteeGJJ (1970) The aerobic decomposition of choline by micro-organisms. I. The ability of aerobic organisms, particularly coryneform bacteria, to utilize choline as the sole carbon and nitrogen source. Arch Mikrobiol 71:235–244Google Scholar
  29. LaemmliUK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685Google Scholar
  30. LangE, LangH (1972) Spezifische Farbreaktion zum direkten Nachweis der Ameisensäure. Z Anal Chem 260:8–10Google Scholar
  31. LarherF, JolivetY, BriensU, GoasU (1982) Osmoregulation in higher plants halophytes: organic nitrogen accumulation in glycine, betaine, and proline during the growth of Aster tripolinum and Sueda macrocarpa under saline conditions. Plant Sci Lett 24:201–210Google Scholar
  32. LeveringPR, BinnemaDJ, VanDijkenJP, HarderW (1981) Enzymatic evidence for a simultaneous operation of two one-carbon assimilation pathways during growth of Arthrobacter P1 on choline. FEMS Microbiol Lett 12:19–25Google Scholar
  33. LovittRW, KellDD, MorrisJG (1986) Proline reduction by Clostridium sporogenes is coupled to vectorial proton ejection. FEMS Microbiol Lett 36:269–273Google Scholar
  34. MargolisJ, KenrickKG (1967) Polyacrylamide gel electrophoresis across a molecular sieve gradient. Nature 214:1334–1336Google Scholar
  35. MöllerB, OssmerR, HowardBH, GottschalkG, HippeH (1984) Sporomusa, a new genus of Gram-negative anaerobic bacteria including Sporomusa sphaeroides spec. nov. and Sporomusa ovata spec. nov. Arch Microbiol 139:388–396Google Scholar
  36. MöllerB, HippeH, GottschalkG (1986) Degradation of various amine compounds by mesophilic clostridia. Arch Microbiol 145:85–90Google Scholar
  37. MüllerE, FahlbuschK, WaltherR, GottschalkG (1981) Formation of N,N-dimethylglycine, acetic acid and butyric acid from betaine by Eubacterium limosum. Appl Environ Microbiol 42:439–445Google Scholar
  38. NakajimaM, ShirokaneY, MizusawaK (1980) A new amidino-hydrolase, methylguanidine amidinohydrolase from Alcaligenes sp N-42. FEBS Lett 110:43–46Google Scholar
  39. Naumann E (1983) Methanbildung aus Betain über Trimethylamin als Zwischenprodukt. PhD thesis, Univ GöttingenGoogle Scholar
  40. NaumannE, HippeH, GottschalkG (1983) Betaine: a new oxidant in the Stickland reaction and methanogenesis from betaine and L-alanine by a Clostridium sporogenes-Methanosarcina barkeri coculture. Appl Environ Microbiol 45:474–483Google Scholar
  41. Rieth M (1987) Untersuchungen zur selenabhängigen Glycin-Reduktase aus Eubacterium acidaminophilum. PhD thesis, Univ GöttingenGoogle Scholar
  42. SetoB (1980) The Stickland reaction. In: KnowlesCJ (ed) Diversity of bacterial respiratory chains, vol II. CRC Press, Boca Raton, pp 49–64Google Scholar
  43. ShimizuS, KimJM, ShinmenY, YamadaH (1986) Evaluation of two alternative metabolic pathways for creatinine degradation in microorganisms. Arch Microbiol 145:322–328Google Scholar
  44. SliwskowskiMX, StadtmanTC (1987) Purification and immunological studies of selenoprotein A of the clostridial glycine reductase complex. J Biol Chem 262:4899–4904Google Scholar
  45. StadtmanTC (1970) Glycine reductase systems (Clostridium). Meth Enzymol 17A:959–966Google Scholar
  46. StouthamerAH (1979) The search for correlation between theoretical and experimental growth yields. In: QuayleJR (ed) Microbial biochemistry, Intern Rev Biochem, vol 21. University Park Press, Baltimore, pp 1–47Google Scholar
  47. ThauerRK, JungermannK, DeckerK (1977) Energy conservation in chemotrophic anaerobic bacteria. Bacteriol Rev 41:100–180Google Scholar
  48. VanEykHG, VermaatHJ, Leijnse-YbemaHJ, LeijnseB (1968) The conversion of creatinine by creatininase of bacterial origin. Enzymologia 34:198–202Google Scholar
  49. WiddelF (1988) Microbiology and ecology of sulfate- and sulfur-reducing bacteria. In: ZehnderAJB (ed) Biology of anaerobic microorganisms. John Wiley & Sons, New York, pp 469–585Google Scholar
  50. YanceyPH, ClarkME, HandSC, BowlusRD, SomeroGN (1982) Living with water stress: Evolution of osmolyte systems. Science 217:1214–1222Google Scholar
  51. ZindelU, FreudenbergW, RiethM, AndreesenJR, SchnellJ, WiddelF (1988) Eubacterium acidaminophilum sp. nov., a versatile amino acid-degrading anaerobe producing or utilizing H2 or formate. Arch Microbiol 150:254–266Google Scholar

Copyright information

© Springer-Verlag 1989

Authors and Affiliations

  • K. Hormann
    • 1
  • J. R. Andreesen
    • 1
  1. 1.Institut für Mikrobiologie der Universität GöttingenGöttingenFederal Republic of Germany

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