Abstract
From our previous studies on the mechanism of methane formation from acetate it was known that cell extracts of acetate-grown Methanosarcina barkeri (100 000 × g supernatant) catalyze the conversion of acetyl-CoA plus tetrahydromethanopterin (=H4MPT) to methyl-H4MPT, CoA, CO2 and presumably H2. We report here that these extracts, in the absence of H4MPT, mediated an isotope exchange between CO2 ([S]0.5 v=0.2% in the gas phase) and the carbonyl group of acetyl-CoA at almost the same specific rate as the above conversion (10 nmol · min−1 · mg protein−1). Both the exchange and the formation of methyl-H4MPT were inhibited by N2O, suggesting that a corrinoid could be the primary methyl group acceptor in the acetyl-CoA C-C-cleavage reaction. Both activities were dependent on the presence of H2 (E0′=−414 mV). Ti(III)citrate (E0′=−480 mV) was found to substitute for H2, indicating a reductive activation of the system. In the presence of Ti(III)citrate it was shown that the formation of CO2 from the carbonyl group of acetyl-CoA is associated with a 1:1 stoichiometric generation of H2. Free CO, a possible intermediate in CO2 and H2 formation, was not detected.
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Abbreviations
- AcCoA:
-
acetyl-CoA
- acetyl-P:
-
acetyl phosphate
- OH-B12 :
-
hydroxocobalamin
- H-S-CoM:
-
coenzyme M = 2-mercaptoethanesulfonate
- CH3-S-CoM:
-
methyl-coenzyme M = 2-(methylthio)ethanesulfonate
- H-S-HTP:
-
N-7-mercaptoheptanoylthreonine phosphate
- HTP-S-S-HTP:
-
disulfide of H-S-HTP
- CoM-S-S-HTP:
-
disulfide of H-S-CoM and H-S-HTP
- H4MPT:
-
tetrahydromethanopterin
- CH3-H4MPT:
-
N5-methyl-H4MPT
- DTT:
-
dithiothreitol
- MOPS:
-
morpholinopropane sulfonic acid
References
Banks RGS, Henderson RJ, Pratt JM (1967) Reactions of nitrous oxide with some transition-metal complexes. Chem Comm 246:387–388
Bhatnagar L, Krzycki JA, Zeikus JG (1987) Analysis of hydrogen metabolism in Methanosarcina barkeri: regulation of hydrogenase and role of CO-dehydrogenase in H2 production. FEMS Microbiol Lett 41:337–343
Börner G (1988) Isolierung von vier Coenzymen der Methanogenese aus Methanobacterium thermoautotrophicum Diploma thesis, Marburg
Boone DR, Menaja JAGF, Boone JE, Mah RA (1987) Effects of hydrogen pressure during growth and effects of pregrowth with hydrogen on acetate degradation by Methanosarcina species. Appl Environm Microbiol 53:83–87
Bott M, Thauer RK (1987) Proton-motive force-driven formation of CO from CO2 and H2 in methanogenic bacteria Eur J Biochem 168:407–412
Bott M, Thauer RK (1989a) Proton translocation coupled to the oxidation of carbon monoxide to CO2 and H2 in Methanosarcina barkeri. Eur J Biochem 179:469–472
Bott M, Thauer RK (1989b) The active species of “CO2” formed by carbon monoxide dehydrogenase from Peptostreptococcus productus. Z Naturforsch 440:392–396
Bott M, Eikmanns B, Thauer RK (1986) Coupling of carbon monoxide oxidation to CO2 and H2 with the phosphorylation of ADP in acetate-grown Methanosarcina barkeri. Eur J Biochem 159:393–398
Bryant MP, Boone DR (1987) Emended description of strain MST (DSM 800T), the type strain of Methanosarcina barkeri. Int J System Bacteriol 37:169–170
Dangel W, Schulz H, Dickert G, König H, Fuchs G (1987) Occurrence of corrinoid-containing membrane proteins in anaerobic bacteria. Arch Microbiol 148:52–56
Deppenmeier U, Blaut M, Jussofie A, Gottschalk G (1988) A methyl-CoM methylreductase system from methanogenic bacterium strain Gö1 not requiring ATP for activity. FEBS Lett 241:60–64
Eikmanns B, Thauer RK (1984) Catalysis of an isotopic exchange between CO2 and the carboxyl group of acetate by Methanosarcina barkeri grown on acetate. Arch Microbiol 138:365–370
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
Ellermann J, Hedderich R, Böcher R, Thauer RK (1988) The final step in methane formation. Investigations with highly purified methyl-CoM reductase (component C) from Methanobacterium thermoautotrophicum (strain Marburg). Eur J Biochem 172:669–677
Fauque G, Teixeira M, Moura I, Lespinat PA, Xavier AV, DerVartanian DV, Peck Jr HD, LeGall J, Moura JG (1984) Purification, characterization and redox properties of hydrogenase from Methanosarcina barkeri. Eur J Biochem 142:21–28
Fiebig K, Friedrich B (1989) Purification of the F420-reducing hydrogenase from Methanosarcina barkeri (strain Fusaro). Eur J Biochem 184:79–88
Fischer R, Thauer RK (1988) Methane formation from acetyl phosphate in cell extracts of Methanosarcina barkeri. Dependence of the reaction on coenzyme A. FEBS Lett 228:249–253
Fischer R, Thauer RK (1989) Methyltetrahydromethanopterin as an intermediate in methanogenesis from acetate in Methanosarcina barkeri. Arch Microbiol 151:459–465
Grahame DA, Stadtman TC (1987a) In vitro methane and methyl-coenzyme M formation from acetate: Evidence that acetyl-CoA is the required intermediate activated form of acetate. Biochem Biophys Res Commun 147:254–258
Grahame DA, Stadtman TC (1987b) Carbon monoxide dehydrogenase from Methanosarcina barkeri. J Biol Chem 262:3706–3712
Kandler O, Hippe H (1977) Lack of peptidoglycan in the cell walls of Methanosarcina barkeri. Arch Microbiol 113:57–60
Karrasch M, Bott M, Thauer RK (1989) Carbonic anhydrase activity in acetate grown Methanosarcina barkeri. Arch Microbiol 151:137–142
Kemner JM, Krzycki JA, Prince RC, Zeikus JG (1987) Spectroscopic and enzymatic evidence for membrane-bound electron transport carriers and hydrogenase and their relation to cytochrome b function in Methanosarcina barkeri. FEMS Microbiol Lett 48:267–272
Krone UE, Thauer RK, Hogenkamp HPC (1989) Reductive dehalogenation of chlorinated C1-hydrocarbons mediated by corrinoids. Biochemistry 28:4908–4914
Krzycki JA, Zeikus JG (1984) Acetate catabolism by Methanosarcina barkeri: hydrogen-dependent methane production from acetate by a soluble cell protein fraction. FEMS Microbiol Lett 25:27–32
Krzycki JA, Lehmann LJ, Zeikus JG (1985) Acetate catabolism by Methanosarcina barkeri: Evidence for involvement of carbon monoxide dehydrogenase, methyl coenzyme M, and methylreductase. J Bacteriol 163:1000–1006
Krzycki JA, Morgan JB, Conrad R, Zeikus JG (1987) Hydrogen metabolism during methanogenesis from acetate by Methanosarcina barkeri. FEMS Microbiol Lett 40:193–198
Krzycki JA, Mortenson LE, Prince RC (1989) Paramagnetic centers of carbon monoxide dehydrogenase from aceticlastic Methanosarcina barkeri. J Biol Chem 264:7217–7221
Laufer K, Eikmanns B, Frimmer U, Thauer RK (1987) Methanogenesis from acetate by Methanosarcina barkeri: Catalysis of acetate formation from methyl iodide, CO2 and H2 by the enzyme system involved. Z Naturforsch 42c:360–372
Lovley DR, Ferry JG (1985) Production and consumption of H2 during growth of Methanosarcina spp. on acetate. Appl Environm Microbiol 49:247–249
Peinemann S, Müller V, Blaut M, Gottschalk G (1988) Bioenergetics of methanogenesis from acetate by Methanosarcina barkeri. J Bacteriol 170:1369–1372
Perski HJ, Schönheit P, Thauer RK (1982) Sodium dependence of methane formation in methanogenic bacteria. FEBS Lett 143:323–326
Pezacka E, Wood HG (1986) The autotrophic pathway of acetogenic bacteria. J Biol Chem 261:1609–1615
Pezacka E, Wood HG (1988) Acetyl-CoA pathway of autotrophic growth. J Biol Chem 263:16000–16006
Ragsdale SW, Lindahl PA, Münck E (1987) Mössbauer, EPR, and optical studies of the corrinoid/iron-sulfur protein involved in the synthesis of acetyl coenzyme A by Clostridium pasteurianum. J Biol Chem 262:14289–14297
Raybuck SA, Bastian NR, Orme-Johnson WH, Walsh CT (1988) Kinetic characterization of the carbon monoxide-acetyl-CoA (carbonyl group) exchange activity of the acetyl-CoA synthesizing CO dehydrogenase from Clostridium thermoaceticum. Biochemistry 27:7698–7702
Schönheit P, Moll J, Thauer RK (1980) Growth parameters (K s, μ max , Y s) of Methanobacterium thermoautotrophicum. Arch Microbiol 127:59–65
Schulz H, Fuchs G (1986) Cobamide-containing membrane protein complex in Methanobacterium. FEBS Lett 198:279–282
Schulz H, Albracht SPJ, Coremans JMCC, Fuchs G (1988) Purification and some properties of the corrinoid-containing membrane protein from Methanobacterium thermoautotrophicum. Eur J Biochem 171:589–597
Shanmugasundaram T, Kumar GK, Wood HG (1988) Involvement of tryptophan residues at the coenzyme A binding site of carbon monoxide dehydrogenase from Clostridium thermoaceticum. Biochemistry 27:6499–6503
Spormann AM, Thauer RK (1989) Anaerobic acetate oxidation to CO2 by Desulfotomaculum acetoxidans: isotopic exchange between CO and the carbonyl group of acetyl-CoA and topology of enzymes involved. Arch Microbiol 152:189–195
Terlesky KC, Ferry JG (1988a) Ferredoxin requirement for electron transport from the carbon monoxide dehydrogenase complex to a membrane-bound hydrogenase in acetate-grown Methanosarcina thermophila. J Biol Chem 263:4075–4079
Terlesky KC, Ferry JG (1988b) Purification and characterization of a ferredoxin from acetate-grown Methanosarcina thermophila. J Biol Chem 263:4080–4082
Terlesky KC, Nelson MJK, Ferry JG (1986) Isolation of an enzyme complex with carbon monoxide dehydrogenase activity containing corrinoid and nickel from acetate-grown Methanosarcina thermophila. J Bacteriol 168:1053–1058
Terlesky KC, Barber MJ, Aceti DJ, Ferry JG (1987) EPR properties of the Ni-Fe-C center in an enzyme complex with carbon monoxide dehydrogenase activity from acetate-grown Methanosarcina thermophila. J Biol Chem 262:15392–15395
Thauer RK, Möller-Zinkhan D, Spormann AM (1989) Biochemistry of acetate catabolism in anaerobic chemotrophic bacteria. Annu Rev Microbiol 43:43–67
Wijngaard WMH van de, Drift C van der, Vogels GD (1988) Involvement of a corrinoid enzyme in methanogenesis from acetate in Methanosarcina barkeri. FEMS Microbiol Lett 52:165–172
Wood HG (1985) Then and now. Ann Rev Biochem 54:1–41
Wood HG, Ragsdale SW, Pezacka E (1986a) The acetyl-CoA pathway: a newly discovered pathway of autotrophic growth. Trends in Biochemical Sciences 11:14–18
Wood HG, Ragsdale SW, Pezacka E (1986b) The acetyl-CoA pathway of autotrophic growth. FEMS Microbiol Rev 39:345–362
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Fischer, R., Thauer, R.K. Methanogenesis from acetate in cell extracts of Methanosarcina barkeri: Isotope exchange between CO2 and the carbonyl group of acetyl-CoA, and the role of H2 . Arch. Microbiol. 153, 156–162 (1990). https://doi.org/10.1007/BF00247814
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DOI: https://doi.org/10.1007/BF00247814