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Isolation of cobamides from Methanothrix soehngenii: 5-methylbenzimidazole as the α-ligand of the predominant cobamide

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Abstract

Methanothrix soehngenii was found to contain five different cobamides when grown on vitamin B12 supplemented as well as vitamin B12 free media. In both cases, it was shown by HPLC-chromatography and UV/VIS spectroscopy, that α-5-methylbenzimidazolyl-β-cyanocobamide was the predominant cobamide, accounting for 27% and 23%, respectively, of the total corrinoid content. Vitamin B12 and α-5-hydroxybenzimidazolyl-β-cyanocobamide could also be detected in both cell batches in varying amounts. Cells grown on vitamin B12 free medium contained significantly more baseless cobamides, indicating biosynthesis of cobamides.

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Abbreviations

(5-MeBza)CNCba:

α-5-methylbenzimidazolyl-β-cyanocobamide

(5-HOBza)CNCBa:

α-5-hydroxybenzimidazolyl-β-cyanocobamide (factor III)

(5-MeOBza)CNCba:

α-5-methoxybenzimidazolyl-β-cyanocobamide (factor IIIm)

(Bza)CNCba:

α-benzimidazolyl-β-cyanocobamide

References

  • Bernhauer K, Müller O, Wagner F (1964) Fortschritte auf dem Vitamin B12-Gebiet. Adv Enzymol 26:233–281

    Google Scholar 

  • Dellweg H, Becher E, Bernhauer K (1956) Biosynthesen in der Cobalamin-Reihe. I. Mitteilung. Cobalamin-Analoga der Benzimidazol-Reihe. Biochem 2:422–449

    Google Scholar 

  • Eikmanns B, Thauer RK (1985) Evidence for the involvement and role of a corrinoid enzyme in methane formation from acetate in Methanosarcina barkeri. Arch Microbiol 142:175–179

    Google Scholar 

  • Frenkel EP, Kitchens RL, Prough R (1979) High-performance liquid chromatographic seperation of cobalamins. J Chromatogr 174:393–400

    Google Scholar 

  • Friedrich W (1975) Vitamin B12 und verwandte Corrinoide In: Ammon R, Dirscherl W (eds) Fermente-Hormone-Vitamine, vol III/2. Thieme, Stuttgart

    Google Scholar 

  • Friedrich W, Bernhauer K (1958a) Über die 5- und 6-Methylbenzimidazol-cobalamin-Analoga. Chem Ber 91:1665–1670

    Google Scholar 

  • Friedrich W, Bernhauer K (1958b) Die Isolierung der Benzimidazol-und 5-Methyl-benzimidazol-cobalamin-Analoga aus Faulschlamm. Chem Ber 91:2061–2065

    Google Scholar 

  • Golding BT (1982) Mechanisms of action of the B12 coenzyme: theory and models. In: Dolphin D (ed) B12, vol 1. John Wiley and Sons, New York, pp 543–582

    Google Scholar 

  • Grate JH, Schrauzer GN (1979) Sterically induced, spontaneous dealkylation of secondary alkyl-cobalamins due to axial base coordination and conformational changes of the corrin ligand. J Am Chem Soc 101:4601–4611

    Google Scholar 

  • Höllriegel V, Scherrer P, Renz P (1983) Isolation and characterisation of the Co-methyl and the Co-aquo derivative of 5-hydroxybenzimidazolylcobamide (factor III) from Methanosarcina barkeri grown on methanol. FEBS Lett 151:156–158

    Google Scholar 

  • Holder U, Schmidt DE, Stupperich E, Fuchs G (1985) Autotropic synthesis of activated acetic acid from two CO2 in Methanobacterium thermoautotrophicum. III. Evidence for a common one-carbon percursor pool and the role of corrinoids. Arch Microbiol 141:229–238

    Google Scholar 

  • Hu SI, Pezacka E, Wood HG (1984) Acetate synthesis from carbon monoxide by Clostridium thermoautotrophicum. J Biol Chem 259:8892–8897

    Google Scholar 

  • Huennekens FM, Vitols KS, Fujii K, Jacobsen DW (1982) Biosynthesis of cobalamin coenzymes. In: Dolphin D (ed) B12, vol 1. John Wiley and Sons, New York, pp 145–167

    Google Scholar 

  • Huser BA, Wuhrmann K, Zehnder AJB (1982) Methanothrix soehngenii gen. nov. sp. nov., a new acetotrophic non-hydrogenoxidizing methane bacterium. Arch Microbiol 132:1–9

    Google Scholar 

  • Irion E, Ljungdahl L (1965) Isolation of factor IIIm coenzyme and cobyric acid coenzyme plus other B12 factors from Clostridium thermoaceticum. Biochem 12:2780–2790

    Google Scholar 

  • Kenealy WR, Zeikus JG (1982) One carbon metabolism in methanogens: Evidence for synthesis of a two carbon cellular intermediate and unification of catabolism and anabolism in Methanosarcina barkeri. J Bacteriol 151:932–941

    Google Scholar 

  • Kohler HPE, Zehnder AJB (1984) Carbon monoxide dehydrogenase and acetate thiokinase in Methanothrix soehngenii FEMS Microbiol Lett 21:287–292

    Google Scholar 

  • Kräutler B (1984) Acetyl-cobalamin from photoinduced carbonylation of methyl-cobalamin. Helv Chim Acta 67:1053–1059

    Google Scholar 

  • Kräutler B (1985) Zur biologischen Funktion der Vitamin B12-Derivate: Beziehungen, Struktur, Reaktivität. Habilitationsschrift, ETH Zürich

  • Kräutler B (1987) Thermodynamic trans-effects of the nucleotide base in the B12 coenzymes. Helv Chim Acta 70:1268–1278

    Google Scholar 

  • Kräutler B, Moll J, Thauer RK (1987) The corrinoid from Methanobacterium thermoautotrophicum (Marburg strain) Spectroscopic structure analysis and identification as Coβ-cyano-5′-hydroxybenzimidazolyl-cobamide (factor III). Eur J Biochem 162:275–278

    Google Scholar 

  • Lezius AG, Barker HA (1965) Corrinoid compounds of Methanobacillus omelianskii. I. Fractionations of the corrinoid compounds and identification of factor III and factor III coenzyme. Biochem 4:510–518

    Google Scholar 

  • Ljungdahl L, Wood HG (1982) Acetate biosynthesis. In: Dolphin D (ed) B12, vol 2. John Wiley and Sons, New York, pp 165–202

    Google Scholar 

  • Perlman D, Barret JM (1959) Biosynthesis of cobalamins by cell suspensions of Propionibacteria and Streptomycetes. J Bacteriol 78:171–174

    Google Scholar 

  • Pol A, Van der Drift C, Vogels GD (1982) Corrinoids from Methanosarcina barkeri: Structure of the α-ligand. Biochem Biophys Res Commun 108:731–737

    Google Scholar 

  • Pol A, Gage RA, Neis JM, Reijenen JWM, Van der Drift C, Vogels GD (1984) Corrinoids from Methanosarcina barkeri. The β-ligands. Biochim Biophys Acta 797:83–93

    Google Scholar 

  • Scherer P, Sahm H (1981) Effect of trace elements and vitamins on the growth of Methanosarcina barkeri. Acta Biotech 1:57–65

    Google Scholar 

  • Scherer P, Höllriegel V, Krug C, Bockel M, Renz P (1984) On the biosynthesis of 5-hydroxybenzimidazolyl cobamide (Vitamin B12 — factor III) in Methanosarcina barkeri. Arch Microbiol 138:354–359

    Google Scholar 

  • Shapiro S (1982) Do corrinoids function in the methanogenic dissimilation of methanol by Methanosarcina barkeri? Can J Microbiol 28:629–635

    Google Scholar 

  • Stadtman TC, Blaylock BA (1966) Role of B12 compounds in methane formation. Fed Proc 25:1657–1661

    Google Scholar 

  • Stupperich E, Kräutler B (1988) Pseudo vitamin B12 or 5-hydroxybenzimidazolyl-cobamide are the corrinoids found in methanogenic bacteria. Arch Microbiol 149:268–271

    Google Scholar 

  • Stupperich E, Steiner I, Rühlmann M (1986) Isolation and analysis of bacterial cobamides by high-performance liquid chromatography. Anal Biochem 155:365–370

    Google Scholar 

  • Taylor RT (1982) B12-dependent methionine biosynthesis. In: Dolphin D (ed) B12, vol 2. John Wiley and Sons, New York, pp 307–355

    Google Scholar 

  • Thauer RK (1985) Nickelenzyme im Stoffwechsel von methanogenen Bakterien. Biol Chem Hoppe-Seyler 366:103–112

    Google Scholar 

  • Van der Meijden P, Te Brörmmelstroet BW, Poirot CM, Van der Drift C, Vogels GD (1984) Purification and properties of methanol: 5-hydroxybenzimidazolylcobamide methyltransferase from Methanosarcina barkeri. J Bacteriol 160:629–635

    Google Scholar 

  • Whitmann WB, Wolfe RS (1984) Purification and analysis of cobamides of Methanobacterium bryyantii by high-performance liquid chromatography. Anal Biochem 137:261–265

    Google Scholar 

  • Wood JM (1982) Mechanisms for B12-dependent methyl transfer. In: Dolphin D (ed) B12, vol 2. John Wiley and Sons, New york, pp 151–164

    Google Scholar 

  • Wood JM, Moura I, Moura JJG, Santos MH, Xavier AV, LeGall J, Scandellari M (1982) Role of Vitamin B12 In methyl transfer for methane biosynthesis by Methanosarcina barkeri. Science 216:303–305

    Google Scholar 

  • Zagalak B (1982) Vitamin B12 als biologisch aktive Modellsubstanz. Naturwissenschaften 69:63–74

    Google Scholar 

  • Zehnder AJB, Huser BA, Brock TD, Wuhrmann K (1980) Characterisation of an acetate-decarboxylating, non-hydrogen oxidizing methane bacterium. Arch Microbiol 124:1–11

    Google Scholar 

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Kohler, HP.E. Isolation of cobamides from Methanothrix soehngenii: 5-methylbenzimidazole as the α-ligand of the predominant cobamide. Arch. Microbiol. 150, 219–223 (1988). https://doi.org/10.1007/BF00407783

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