Abstract
Cell-free extracts of Syntrophomonas wolfei subsp. wolfei grown with crotonate in pure culture or in coculture with Methanospirillum hungatei contained crotonyl-coenzyme A (CoA): acetate CoA-transferase activity. This activity was not detected in cell-free extracts from the butyrate-grown coculture which suggests that the long lag times observed before S. wolfei grew with crotonate were initially due to the inability to activate crotonate. Cell-free extracts of S. wolfei grown in pure culture contained high specific activities of hydrogenase and very low levels of formate dehydrogenase. The low levels suggest a biosynthethic rather than a catabolic role for the latter enzyme when S. wolfei is grown in pure culture. CO dehydrogenase activity was not detected. S. wolfei can form butyrate using a CoA transferase activity, but not by a phosphotransbutyrylase or enoate reductase activity. A c-type cytochrome was detected in S. wolfei grown in pure culture or in coculture indicating the presence of an electron transport system. This is a characteristic which separates S. wolfei from other known crotonate-using bacteria.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Amos DA, McInerney MJ (1990) Growth of Synthrophomonas wolfei on short chain unsaturated fatty acids. Arch Microbiol 154: 31–36
Bader J, Günther H, Schleicher E, Simon H, Pohl S, Mannheim W (1980) Utilization of (E)-2-butenoate (crotonate) by Clostridium kluyveri and some other Clostridium species. Arch Microbiol 125: 159–165
Blach WE, Wolfe RS (1976) New approach to cultivation of methanogenic bacteria: 2-mercaptoethanesulfonic acid (HS-CoM)-dependent growth of Methanobacterium ruminantium in a pressurized atmosphere. Appl Environ Microbiol 32: 781–791
Barker HA, Jeng J-M, Neff N, Robertson JM, Tam FK, Hosaka S (1978) Butyryl CoA: acetoacetate CoA-transferase from a lysine-fermenting Clostridium. J Biol Chem 253: 1219–1225
Barker HA, Stadtman ER, Kornberg A (1955) Coenzyme A transphorase from Clostridium kluyveri. Methods Enzymol 1: 599–602
Beaty PS, McInerney MJ (1989) Effects of organic acid anions on the growth and metabolism of Syntrophomonas wolfei in pure culture and in defined consortia. Appl Environ Microbiol 55: 977–983
Beaty PS, McInerney MJ (1987) Growth of Syntrophomonas wolfei in pure culture on crotonate. Arch Microbiol 147: 389–393
Beaty PS, Wofford NQ, McInerney MJ (1987) Separation of Syntrophomonas wolfei from Methanospirillum hungatei in syntrophic cocultures by using Percoll gradients. Appl Environ Microbiol 53: 1183–1185
Boone DR, Bryant MP (1980) Propionate-degrading bacterium Syntrophobacter wolinii gen. nov., sp. nov., from methanogenic ecosystem. Appl Environ Microbiol 40: 626–632
Boone DR, Johnson RL, Liu Y (1989) Diffusion of interspecies electron carriers H2 and formate in methanogenic ecosystems and its implication in the measurement of K m for H2 or formate uptake. Appl Environ Microbiol 55: 1735–1741
Bryant MP, Wolin EA, Wolin MJ, Wolfe RS (1967) Methanobacillus omelianskii, a symbiotic association of two species of bacteria. Arch Mikrobiol 59: 20–31
Clark JE, Ragsdale SW, Ljungdahl LG, Wiegel J (1982) Levels of enzymes involved in the synthesis of acetate from CO2 in Clostridium thermoautotrophicum. J Bacteriol 151: 507–509
Decker K, Jungermann K, Thauer RK (1970) Energy production in anaerobic organisms. Angew Chem Int Ed Engl 9: 138–158
Falk JE (1964) Porphyrins and metalloporphyrins. Elsevier, Amsterdam, p 241
Gustafson WG, Feinberg BA, McFarland JT (1986) Energetics of β-oxidation. Reduction potentials of general fatty acyl-CoA dehydrogenase, electron transfer flavoprotein, and fatty acyl-CoA substrates. J Biol Chem 261: 7733–7741
Hartmanis MGN, Gatenbeck S (1984) Intermediary metabolism in Clostridium acetobutylicum: levels of enzymes involved in the formation of acetate and butyrate. Appl Environ Microbiol 47: 1277–1283
Jones CW, Redfearn ER (1967) Electron transport in Azotobacter vinelandii. Biochim Biophys Acta 113: 467–481
Krumholz LR, Bryant MP (1986) Eubacterium oxidoreducens sp. nov. requiring H2 or formate to degrade gallate, pyrogallol, phloroglucinol and guercetin. Arch Microbiol 144: 8–14
Ljungdahl LG (1986) The autotrophic pathway of acetate synthesis in acetogenic bacteria. Ann Rey Microbiol 40: 415–450
McInerney MJ, Bryant MP (1981) Basic principles of bioconversion in anaerobic digestion and methanogenesis. In: Sofer S, Zaborsky OR (eds) Biomass conversion processes for energy and fuels. Plenum, New York, pp 277–296
McInerney MJ, Bryant MP, Hespell RB, Costerton JW (1981) Syntrophomonas wolfei gen. nov., sp. nov., an anaerobic, syntrophic, fatty acid-oxidizing bacterium. Appl Environ Microbiol 41: 1029–1039
McInerney MJ, Bryant MP, Pfenning N (1979) Anaerobic bacterium that degrades fatty acids in syntrophic association with methanogens. Arch Microbiol 122: 129–135
Mountfort DO, Bryant MP (1982) Isolation and characterization of an anaerobic, syntrophic benzoate-degrading bacterium from sewage sludge. Arch Microbiol 133: 249–256
Schulman M, Valentino D (1976) Kinetic and catalytic properties of coenzyme A transferase from Petostreptococcus elsdenii. J Bacteriol 128: 372–381
Smith PK, Krohn RI, Hermanson GT, Mallia AK, Gartner FH, Provenzano MD, Fujimoto EK, Goeke NM, Olson BJ, Klenk DC (1985) Measurement of protein using bicinchoninic acid. Anal Biochem 150: 76–86
Sperry JE, Wilkins TD (1976) Cytochrome spectrum of an obligate anaerobe, Eubacterium lentum. J Bacteriol 125: 905–909
Sramek SJ, Frerman FE (1975) Purification and properties of E. coli coenzyme A-transferase. Arch Biochem Biophys 171: 14–26
Stadtman ER (1953) The coenzyme A transferase system in Clostridium kluyveri. J Biol Chem 203: 501–512
Stieb M, Schink B (1984) A new 3-hydroxybutyrate fermenting anaerobe, Ilyobacter polytropus, gen. nov., sp. nov., possessing various fermentation pathways. Arch Microbiol 140: 139–146
Thauer RK, Jungermann K, Decker K (1977) Energy conservation in chemotrophic anaerobic bacteria. Bacteriol Rev 47: 100–180
Thauer RK, Jungermann K, Henninger H, Wenning J, Decker K (1968) The energy metabolism of Clostridium kluyveri. Eur J Biochem 4: 173–180
Thiele JH; Zeikus JG (1988) Control of interspecies electron flow during anaerobic digestion: role of floc formation in syntrophic methanogenesis. Appl Environ Microbiol 54: 20–29
Tischer W, Bader J, Simon H (1979) Purification and some properties of a hitherto-unknown enzyme reducing the carbon-carbon double bond of α,β-unsaturated carboxylate anions. Eur J Biochem 97: 103–112
Twarog R, Wolfe RS (1963) Role of butyryl phosphate in the energy metabolism of Clostridium tetanomorphum. J Bacteriol 86: 112–117
Wofford NQ, Beaty PS, McInerney MJ (1986) Preparation of cell-free extracts and the enzymes involved in fatty acid metabolism in Syntrophomonas wolfei. J Bacteriol 167: 179–185
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
McInerney, M.J., Wofford, N.Q. Enzymes involved in crotonate metabolism in Syntrophomonas wolfei . Arch. Microbiol. 158, 344–349 (1992). https://doi.org/10.1007/BF00245363
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00245363