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J Bacteriol. Dec 1986; 168(3): 1053–1058.
PMCID: PMC213601

Isolation of an enzyme complex with carbon monoxide dehydrogenase activity containing corrinoid and nickel from acetate-grown Methanosarcina thermophila.

Abstract

Fast protein liquid chromatography of cell extract from methanol- or acetate-grown Methanosarcina thermophila resolved two peaks of CO dehydrogenase activity. The activity of one of the CO dehydrogenases was sixfold greater in acetate-grown compared with methanol-grown cells. This CO dehydrogenase was purified to apparent homogeneity (70 mumol of methyl viologen reduced per min per mg of protein) and made up greater than 10% of the cellular protein of acetate-grown cells. The native enzyme (Mr 250,000) formed aggregates with an Mr of approximately 1,000,000. The enzyme contained five subunits (Mrs 89,000, 71,000, 60,000, 58,000, and 19,000), suggesting a multifunctional enzyme complex. Nickel, iron, cobalt, zinc, inorganic sulfide, and a corrinoid were present in the complex. The UV-visible spectrum suggested the presence of iron-sulfur centers. The electron paramagnetic resonance spectrum contained g values of 2.073, 2.049, and 2.028; these features were broadened in enzyme that was purified from cells grown in the presence of medium enriched with 61Ni, indicating the involvement of this metal in the spectrum. The pattern of potassium cyanide inhibition indicated that cyanide binds at or near the CO binding site. The properties of the enzyme imply an involvement in the dissimilation of acetate to methane, possibly by cleavage of acetate or activated acetate.

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Selected References

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  • Baresi L. Methanogenic cleavage of acetate by lysates of Methanosarcina barkeri. J Bacteriol. 1984 Oct;160(1):365–370. [PMC free article] [PubMed]
  • Beinert H. Semi-micro methods for analysis of labile sulfide and of labile sulfide plus sulfane sulfur in unusually stable iron-sulfur proteins. Anal Biochem. 1983 Jun;131(2):373–378. [PubMed]
  • Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. [PubMed]
  • Hu SI, Pezacka E, Wood HG. Acetate synthesis from carbon monoxide by Clostridium thermoaceticum. Purification of the corrinoid protein. J Biol Chem. 1984 Jul 25;259(14):8892–8897. [PubMed]
  • Krzycki JA, Zeikus JG. Characterization and purification of carbon monoxide dehydrogenase from Methanosarcina barkeri. J Bacteriol. 1984 Apr;158(1):231–237. [PMC free article] [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Lovley DR, White RH, Ferry JG. Identification of methyl coenzyme M as an intermediate in methanogenesis from acetate in Methanosarcina spp. J Bacteriol. 1984 Nov;160(2):521–525. [PMC free article] [PubMed]
  • Margolis J, Kenrick KG. Polyacrylamide gel electrophoresis in a continuous molecular sieve gradient. Anal Biochem. 1968 Oct 24;25(1):347–362. [PubMed]
  • Nelson MJ, Ferry JG. Carbon monoxide-dependent methyl coenzyme M methylreductase in acetotrophic Methosarcina spp. J Bacteriol. 1984 Nov;160(2):526–532. [PMC free article] [PubMed]
  • Pol A, van der Drift C, Vogels GD. Corrinoids from Methanosarcina barkeri: structure of the alpha-ligand. Biochem Biophys Res Commun. 1982 Sep 30;108(2):731–737. [PubMed]
  • Ragsdale SW, Clark JE, Ljungdahl LG, Lundie LL, Drake HL. Properties of purified carbon monoxide dehydrogenase from Clostridium thermoaceticum, a nickel, iron-sulfur protein. J Biol Chem. 1983 Feb 25;258(4):2364–2369. [PubMed]
  • Ragsdale SW, Ljungdahl LG, DerVartanian DV. Isolation of carbon monoxide dehydrogenase from Acetobacterium woodii and comparison of its properties with those of the Clostridium thermoaceticum enzyme. J Bacteriol. 1983 Sep;155(3):1224–1237. [PMC free article] [PubMed]
  • Ragsdale SW, Wood HG. Acetate biosynthesis by acetogenic bacteria. Evidence that carbon monoxide dehydrogenase is the condensing enzyme that catalyzes the final steps of the synthesis. J Biol Chem. 1985 Apr 10;260(7):3970–3977. [PubMed]
  • Ragsdale SW, Wood HG, Antholine WE. Evidence that an iron-nickel-carbon complex is formed by reaction of CO with the CO dehydrogenase from Clostridium thermoaceticum. Proc Natl Acad Sci U S A. 1985 Oct;82(20):6811–6814. [PMC free article] [PubMed]
  • Schauer NL, Ferry JG. Properties of formate dehydrogenase in Methanobacterium formicicum. J Bacteriol. 1982 Apr;150(1):1–7. [PMC free article] [PubMed]
  • Smith MR, Lequerica JL, Hart MR. Inhibition of methanogenesis and carbon metabolism in Methanosarcina sp. by cyanide. J Bacteriol. 1985 Apr;162(1):67–71. [PMC free article] [PubMed]
  • van der Meijden P, Heythuysen HJ, Pouwels A, Houwen F, van der Drift C, Vogels GD. Methyltransferases involved in methanol conversion by Methanosarcina barkeri. Arch Microbiol. 1983 Jun;134(3):238–242. [PubMed]
  • WOLIN EA, WOLIN MJ, WOLFE RS. FORMATION OF METHANE BY BACTERIAL EXTRACTS. J Biol Chem. 1963 Aug;238:2882–2886. [PubMed]
  • Zinder SH, Elias AF. Growth substrate effects on acetate and methanol catabolism in Methanosarcina sp. strain TM-1. J Bacteriol. 1985 Jul;163(1):317–323. [PMC free article] [PubMed]

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