Abstract
Wolinella succinogenes can grow at the expense of sulphur reduction by formate. The enzymes involved in the catalysis of this catabolic reaction have been investigated. From the results the following conclusions are drawn: 1. The enzyme isolated as a sulphide dehydrogenase from the cytoplasmic membrane of W. succinogenes is the functional sulphur reductase that operates in the electron transport from formate to sulphur. 2. The enzyme (Mr 200,000) consists essentially of one type of subunit with the Mr 85,000 and contains equal amounts of free iron and sulphide (120 μmol/g protein), but no heme. It represents the first functional sulphur reductase ever isolated. 3. The electron transport chain catalyzing sulphur reduction by formate consists merely of formate dehydrogenase and sulphur reductase. A lipophilic quinone which mediates the transfer of electrons between enzymes in other chains, is apparently not involved. This is the first known example of a phosphorylative electron transport chain that operates without a quinone. 4. The same formate dehydrogenase appears to operate in the electron transport both with sulphur and with fumarate as the terminal electron acceptor in W. succinogenes.
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Abbreviations
- DMN:
-
2,3-Dimethyl-1,4-naphthoquinone
- DTT:
-
dithiothreitol
- MK:
-
menaquinone (vitamin K2)
- PMSF:
-
phenylmethane sulfonylfluoride
- Tricine:
-
N-[2-hydroxy-1,1-bis(hydroxymethyl)ethyl]-glycine
- Tea:
-
triethanolamine
- Hepes:
-
4-(2-hydroxyethyl)-1-piperazineethane sulfonate
References
Bergmeyer HU (1970) Methoden der enzymatischen Analyse. Verlag Chemie, Weinheim, pp 1596–1600
Bode C, Goebell H, Stähler E (1968) Zur Eliminierung von Trübungsfehlern bei der Eiweißbestimmung mit der Biuretmethode. Z Klin Chem Klin Biochem 6:419–422
Bokranz M, Mörschel E, Kröger A (1985) Structural and ATP hydrolyzing properties of the ATP synthase isolated from Wolinella succinogenes. Biochim Biophys Acta 810:84–93
Brumby PE, Massey V (1967) Determination of non heme iron, total iron, and copper. Meth Enzymol 10:463–474
Chen PS, Toribara TY, Warner H (1956) Microdetermination of phosphorus. Anal Chem 28:1756–1758
Collins MD, Fernandez F (1984) Menaquinone-6 and thermoplasmaquinone-6 in Wolinella succinogenes. FEMS Microbiol Lett 22:273–276
Fauque G, Herve D, Le Gall J (1979) Structure-function relationship in hemoproteins: The role of cytochrome c3 in the reduction of colloidal sulfur by sulfate-reducing bacteria. Arch Microbiol 121:261–264
Fauque GD, Barton LL, Le Gall J (1980) Oxidative phosphorylation linked to the dissimilatory reduction of elemental sulphur by Desulfovibrio. Ciba Foundation Symposium 72:71–86
Fliermans CB, Brock TD (1973) Assay of elemental sulfur in soil. Soil Science 115:120–122
Francis RT, Becker RR (1984) Specific indication of hemoproteins in polyacrylamide gels using a double-staining process. Anal Biochem 136:509–514
Gebhardt NA, Thauer RK, Linder D, Kaulfers PM, Pfennig N (1985) Mechanism of acetate oxidation to CO2 with elemental sulfur in Desulfuromonas acedoxidans. Arch Microbiol 141: 392–398
Holloway PW (1973) A simple procedure for removal of Triton X-100 from protein samples. Anal Biochem 53:304–308
King TE, Morris RO (1967) Determination of acid-labile sulfide and sulfhydryl groups. Meth Enzymol 10:634–641
Kröger A, Innerhofer A (1976a) The function of menaquinone, covalently bound FAD and iron-sulfur protein in the electron transport from formate to fumarate of Vibrio succinogenes. Eur J Biochem 69:487–495
Kröger A, Innerhofer A (1976b) The function of the b cytochromes in the electron transport from formate to fumarate of Vibrio succinogenes. Eur J Biochem 69:497–506
Kröger A, Unden G (1985) The function of menaquinone in bacterial electron transport. In: Lenaz G (ed) Coenzyme Q. John Wiley, New York, pp 285–300
Kröger A, Winkler E, Innerhofer A, Hackenberg H, Schägger H (1979) The formate dehydrogenase involved in electron transport from formate to fumarate in Vibrio succinogenes. Eur J Biochem 94:465–475
Lenaz G, Fato R (1986) Is ubiquinone diffusion rate-limiting for electron transfer? J Bioenerg Biomembr 18:369–401
Macy JM, Schröder I, Thauer RK, Kröger A (1986) Growth of Wolinella succinogenes on H-S plus fumarate and on formate plus sulfur as energy sources. Arch Microbiol 144:147–150
Martin RG, Ames BN (1961) A method for determining the sedimentation behavior of enzymes: Application to protein mixtures. J Biol Chem 236:1372–1379
Neuhoff V, Philipp K, Zimmer HG, Mesecke S (1979) A simple, versatile, sensitive and volume-independent method for quantitative protein determination which is independent of other external influences. Z Physiol Chem 360:1657–1670
Paulsen J, Kröger A, Thauer RK (1986) ATP-driven succinate oxidation in the catabolism of Desulfuromonas acetoxidans. Arch Microbiol 144:78–83
Pfennig N, Biebl H (1976) Desulfuromonas acetoxidans gen. nov. and sp. nov., a new anaerobic, sulfur-reducing, acetate-oxidizing bacterium. Arch Microbiol 110:3–12
Schröder I, Roberton AM, Bokranz M, Unden G, Böcher R, Kröger A (1985) The membraneous nitrite reductase involved in the electron transport of Wolinella succinogenes. Arch Microbiol 140:380–386
Tanford C, Nozaki Y, Reynolds JA, Makino S (1974) Molecular characterization of proteins in detergent solution. Biochemistry 13:2369–2376
Unden G, Hackenberg H, Kröger A (1980) Isolation and functional aspects of the fumarate reductase involved in the phosphorylative electron transport of Vibrio succinogenes. Biochim Biophys Acta 591:275–288
Unden G, Kröger A (1981) The function of the subunits of the fumarate reductase of Vibrio succinogenes. Eur J Biochem 120:577–584
Unden G, Kröger A (1982) Reconstitution in liposomes of the electron-transport chain catalyzing fumarate reduction by formate. Biochim Biophys Acta 682:258–263
Unden G, Kröger A (1983) Low potential cytochrome b as essential electron transport component of menaquinone reduction by formate in Vibrio succinogenes. Biochim Biophys Acta 725:325–331
Unden G, Kröger A (1986) Reconstitution of a functional electron transport chain from isolated enzymes in liposomes. Meth Enzymol 126:387–399
Unden G, Böcher R, Knecht A, Kröger A (1982) Hydrogenase from Vibrio succinogenes, a nickel protein. FEBS Lett 145:230–234
Unden G, Mörschel E, Bokranz M, Kröger A (1983) Structural properties of the proteoliposomes catalyzing electron transport from formate to fumarate. Biochim Biophys Acta 725:41–48
Widdel F (1988) Microbiology and ecology of sulfate- and sulfurreducing bacteria. In: Zehnder AJB (ed) Environmental microbiology of anaerobes. John Wiley, New York (in press)
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Dedicated to Professor F. Schneider (Philipps-Universität Marburg) on the occasion of his 60th birthday
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Schröder, I., Kröger, A. & Macy, J.M. Isolation of the sulphur reductase and reconstitution of the sulphur respiration of Wolinella succinogenes . Arch. Microbiol. 149, 572–579 (1988). https://doi.org/10.1007/BF00446763
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DOI: https://doi.org/10.1007/BF00446763