Skip to main content
SpringerLink
Log in

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

  • Original Papers
  • Published:
Archives of Microbiology Aims and scope Submit manuscript

Abstract

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

Abbreviations

DTE:

dithioerythritol

TES:

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

References

  • AndreesenJR, GottschalkG, SchlegelHG (1970) Clostridium formicoaceticum nov spec. Isolation, description and distinction from C. aceticum and C. thermoaceticum. Arch Mikrobiol 72: 154–174

    Google Scholar 

  • BarnardGF, AkhtarM (1979) Mechanistic and stereochemical studies on the glycine reductase of Clostridium sticklandii. Eur J Biochem 99:593–603

    Google Scholar 

  • 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 25:563–567

    Google Scholar 

  • BradfordMM (1976) A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein dye-binding. Anal Biochem 72:248–254

    Google Scholar 

  • BreznakJA, SwitzerJM, SeitzHJ (1988) Sporomusa termitida sp nov, an H2/CO2-utilizing acetogen isolated from termites. Arch Microbiol 150:282–288

    Google Scholar 

  • CarruthersA, OldfieldJET, TeagueHJ (1960) The removal of interfering ions in the detection of betaine in sugar-beet juices and plant material. Analyst 85:272–275

    Google Scholar 

  • 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 73:2659–2663

    Google Scholar 

  • ConeJE, Martin del RioR, StadtmanTC (1977) Clostridial glycine reductase complex. Purification and characterization of the selenoprotein component. J Biol Chem 252:5337–5344

    Google Scholar 

  • CsonkaLN (1989) Physiological and genetic responses of bacteria to osmotic stress. Microbiol Rev 53:121–147

    Google Scholar 

  • DornM, AndreesenJR, GottschalkG (1978) Fermentation of fumarate and L-malate by Clostridium formicoaceticum. J Bacteriol 133:26–32

    Google Scholar 

  • EnerothP, LindstedtG (1965) Thin-layer chromatography of betaines and other compounds related to carnitine. Anal Biochem 10:479–485

    Google Scholar 

  • FinkelsteinJD, MartinJJ, HarrisJ (1988) Methionine metabolism in mammals. The methionine-sparing effect of cystine. J Biol Chem 263:11750–11754

    Google Scholar 

  • FochtRL, SchmidtFH (1956) Colorimetric determination of betaine in glutamate process and liquor. J Agric Food Chem 4:579–585

    Google Scholar 

  • FreudenbergW, AndreesenJR (1989) Purification and partial characterization of the glycine decarboxylase multienzyme complex from Eubacterium acidaminophilum. J Bacteriol 171:2209–2215

    Google Scholar 

  • FreudenbergW, DietrichsD, LebertzH, AndreesenJR (1989a) Isolation of an atypically small lipoamide dehydrogenase involved in the glycine decarboxylase complex from Eubacterium acidaminophilum. J Bacteriol 171:1346–1354

    Google Scholar 

  • 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 25–28

    Google Scholar 

  • 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 152:182–188

    Google Scholar 

  • GalinskiEA, TrüperHG (1982) Betaine, a compatible solute in the extremely halophilic phototrophic bacterium Ectothiorhodospira halochloris. FEMS Microbiol Lett 13:357–360

    Google Scholar 

  • Gauglitz U (1988) Anaerober mikrobieller Abbau von Kreatin, Kreatinin and N-Methylhydantoin. PhD thesis, Univ Göttingen

  • GenthnerBRS, DavisCL, BryantBP (1981) Features of rumen and sewage sludge strains of Eubacterium limosum, a methanol- and H2−CO2-utilizing species. Appl Environ Microbiol 42:12–19

    Google Scholar 

  • GoodwinJF, StampwalaS (1973) Spectrophotometric quantification of glycine in serum and urine. Clin Chem 19:1010–1015

    Google Scholar 

  • GorhamJ (1984) Separation of plant betaines and their sulphur analogues by cation-exchange high-performance liquid chromatography. J Chromatogr 287:345–351

    Google Scholar 

  • GreenbergDM (1961) Biosynthesis of amino acids and related compounds. In: GreenbergDM (ed) Metabolic pathways. Academic Press, New York, pp 173–236

    Google Scholar 

  • 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 93:81–113

    Google Scholar 

  • HeijthuijsenJHFG, HansenTA (1989) Betaine fermentation and oxidation by marine Desulfuromonas strains. Appl Environ Microbiol 55:965–969

    Google Scholar 

  • IkutaS, MatuuraK, ImamuraS, MisakiH, HoriutiY (1977) Oxidative pathways of choline to betaine in the soluble fraction prepared from Arthrobacter globiformis. J Biochem 82:157–163

    Google Scholar 

  • ImhoffJF (1986) Osmoregulation and compatible solutes in eubacteria. FEMS Microbiol Rev 39:57–66

    Google Scholar 

  • 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 71:235–244

    Google Scholar 

  • LaemmliUK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Google Scholar 

  • LangE, LangH (1972) Spezifische Farbreaktion zum direkten Nachweis der Ameisensäure. Z Anal Chem 260:8–10

    Google Scholar 

  • 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 24:201–210

    Google Scholar 

  • 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 12:19–25

    Google Scholar 

  • LovittRW, KellDD, MorrisJG (1986) Proline reduction by Clostridium sporogenes is coupled to vectorial proton ejection. FEMS Microbiol Lett 36:269–273

    Google Scholar 

  • MargolisJ, KenrickKG (1967) Polyacrylamide gel electrophoresis across a molecular sieve gradient. Nature 214:1334–1336

    Google Scholar 

  • 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 139:388–396

    Google Scholar 

  • MöllerB, HippeH, GottschalkG (1986) Degradation of various amine compounds by mesophilic clostridia. Arch Microbiol 145:85–90

    Google Scholar 

  • MüllerE, FahlbuschK, WaltherR, GottschalkG (1981) Formation of N,N-dimethylglycine, acetic acid and butyric acid from betaine by Eubacterium limosum. Appl Environ Microbiol 42:439–445

    Google Scholar 

  • NakajimaM, ShirokaneY, MizusawaK (1980) A new amidino-hydrolase, methylguanidine amidinohydrolase from Alcaligenes sp N-42. FEBS Lett 110:43–46

    Google Scholar 

  • Naumann E (1983) Methanbildung aus Betain über Trimethylamin als Zwischenprodukt. PhD thesis, Univ Göttingen

  • 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 45:474–483

    Google Scholar 

  • Rieth M (1987) Untersuchungen zur selenabhängigen Glycin-Reduktase aus Eubacterium acidaminophilum. PhD thesis, Univ Göttingen

  • SetoB (1980) The Stickland reaction. In: KnowlesCJ (ed) Diversity of bacterial respiratory chains, vol II. CRC Press, Boca Raton, pp 49–64

    Google Scholar 

  • ShimizuS, KimJM, ShinmenY, YamadaH (1986) Evaluation of two alternative metabolic pathways for creatinine degradation in microorganisms. Arch Microbiol 145:322–328

    Google Scholar 

  • SliwskowskiMX, StadtmanTC (1987) Purification and immunological studies of selenoprotein A of the clostridial glycine reductase complex. J Biol Chem 262:4899–4904

    Google Scholar 

  • StadtmanTC (1970) Glycine reductase systems (Clostridium). Meth Enzymol 17A:959–966

    Google Scholar 

  • 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 1–47

    Google Scholar 

  • ThauerRK, JungermannK, DeckerK (1977) Energy conservation in chemotrophic anaerobic bacteria. Bacteriol Rev 41:100–180

    Google Scholar 

  • VanEykHG, VermaatHJ, Leijnse-YbemaHJ, LeijnseB (1968) The conversion of creatinine by creatininase of bacterial origin. Enzymologia 34:198–202

    Google Scholar 

  • WiddelF (1988) Microbiology and ecology of sulfate- and sulfur-reducing bacteria. In: ZehnderAJB (ed) Biology of anaerobic microorganisms. John Wiley & Sons, New York, pp 469–585

    Google Scholar 

  • YanceyPH, ClarkME, HandSC, BowlusRD, SomeroGN (1982) Living with water stress: Evolution of osmolyte systems. Science 217:1214–1222

    Google Scholar 

  • 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 150:254–266

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institut für Mikrobiologie der Universität Göttingen, Grisebachstrasse 8, D-3400, Göttingen, Federal Republic of Germany

    K. Hormann & J. R. Andreesen

Authors
  1. K. Hormann
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. J. R. Andreesen
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hormann, K., Andreesen, J.R. Reductive cleavage of sarcosine and betaine by Eubacterium acidaminophilum via enzyme systems different from glycine reductase. Arch. Microbiol. 153, 50–59 (1989). https://doi.org/10.1007/BF00277541

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00277541

Key words

Search

Navigation