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Chemostat enrichment and isolation of Hyphomicrobium EG

A dimethyl-sulphide oxidizing methylotroph and reevaluation of Thiobacillus MS1

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Abstract

A stable mixed bacterial culture was obtained by chemostat enrichment using dimethyl-sulphoxide as a carbon and energy source. This culture could not only rapidly oxidize dimethyl-sulphoxide but also dimethyl-sulphide. Enzyme determinations indicated that an important part of it consisted of methylotrophs, which assimilated carbon via the serine pathway. Indeed plate counts revealed the majority of the community to be a Hyphomicrobium species. This organism, designated Hyphomicrobium EG, is an obligate methylotroph which can only grow aerobically on several different C1-compounds. Its performance on dimethyl-sulphoxide was compared with that of the community and of another recently isolated strain, Hyphomicrobium S. The mixed culture, Hyphomicrobium EG and Hyphomicrobium S had a μmax of 0.08, 0.08 and 0.014 h-1 respectively. The KS for dimethyl-sulphoxide was the same for all three cultures (3–6 μM), whereas that for dimethyl-sulphide of Hyphomicrobium EG after growth on dimethyl-sulphoxide was 3-fold higher than that of the other two cultures (48 and 16 μM respectively). After growth on dimethyl-sulphide it improved to 3 μM. Dimethyl-sulphide respiration was maximal at a concentration of 100 μM; higher concentrations were inhibitory. One of the accompanying organisms, a pink methylotroph, was able to derive energy from the oxidation of thiosulphate. Available cultures of Thiobacillus MS1 that were reported to be able to utilize dimethyl-sulphide could no longer metabolize this compound.

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References

  • Andreae, M. O. (1980) Dimethyl-sulfoxide in marine and freshwaters. Limnol. Oceanogr. 25: 1054–1063

    Google Scholar 

  • Andreae, M. O. & Raemdonck, H. (1983) Dimethyl-sulphide in the surface ocean and the marine atmosphere: a global view. Science 221: 744–747

    Google Scholar 

  • Anness, B. J., Bamforth, C. W. & Wainwright, T. (1979) The measurement of dimethyl-sulphoxide in barley and malt and its reduction to dimethyl-sulphide by yeasts. J. Inst. Brew. 85: 346–349

    Google Scholar 

  • Anness, B. J. (1981) The determination of dimethyl-sulphoxide in aqueous solution. J. Sci. Food Agric. 32: 353–358

    Google Scholar 

  • Bamforth, C. W. (1980) Dimethyl-sulphoxide reductase of Saccharomyces spp. FEMS Microbiol. Lett. 7: 55–59

    Google Scholar 

  • Bamforth, C. W. & Anness, B. J. (1981) The role of dimethyl-sulphoxide reductase in the formation of dimethyl-sulphide during fermentations. J. Inst. Brew. 87: 30–34

    Google Scholar 

  • Banwart, W. L. & Bremner, J. M. (1976) Evolution of volatile sulfur compounds from soils treated with sulfur-containing organic materials. Soil Biol. Biochem. 8: 439–443

    Google Scholar 

  • Battat, E., Goldberg, I. & Mateles, R. I. (1974) Growth of Pseudomonas C on C1 compounds: continuous culture. Appl. Microbiol. 28: 906–911

    Google Scholar 

  • Beudeker, R. F., Cannon, G. C., Keunen, J. G. & Shively, J. M. (1980) Relations between D-ribulose-1,5-bisphosphate carboxylase, carboxysomes and CO2-fixing capacity in the obligate chemolithotroph Thiobacillus neapolitanus grown under different limitations in the chemostat. Arch. Microbiol. 124: 185–189

    Google Scholar 

  • Beudeker, R. F., Gottschal, J. C. & Kuenen, J. G. (1982) Reactivity versus flexibility in Thiobacilli. Antonie van Leeuwenhoek 48: 39–51

    Google Scholar 

  • Bont, J. A. M.de, Dijken, J. P.van & Harder, W. (1981) Dimethyl-sulphoxide as a carbon, sulphur and energy source for growth of Hyphomicrobium S. J. Gen. Microbiol. 127: 315–323

    Google Scholar 

  • Bremner, J. M. & Steele, C. G. (1978) Role of microorganisms in the atmospheric sulfur cycle. Adv. Microb. Ecol. 2: 155–201

    Google Scholar 

  • Cox, R. B. & Quayle, J. R. (1975) The autotrophic growth of Micrococcus denitrificans on methanol. Biochem. J. 150: 569–571

    Google Scholar 

  • Dijken, J. P.van, Harder, W., Beardsmore, A. J., & Quayle, J. R. (1978) Dihydroxyacetone: an intermediate in the assimilation of methanol by yeasts. FEMS Microbiol. Lett 4: 97–102

    Google Scholar 

  • Dodgson, K. S. (1961) Determination of inorganic sulphate in studies on the enzymic and nonenzymic hydrolysis of carbohydrate and other sulphate esters. Biochem. J. 78: 312–319

    Google Scholar 

  • Friedrich, C. G. & Mitrenga, G. (1981) Oxidation of thiosulfate by Paracoccus denitrificans and other hydrogen bacteria. FEMS Microbiol. Lett. 10: 209–212

    Google Scholar 

  • Ghisalba, O. & Küenzi, M. (1983) Biodegradation and utilization of monomethyl sulfate by specialized methylotrophs. Experientia 39: 1257–1263. GOA.J. (1953) A microbiuret method for protein determination; determination of total protein in cerebrospinal fluid. Scand. J. Clin. Lab. Invest. 5: 218

    Google Scholar 

  • Goldberg, I., Rock, J. S., Ben-Bassat, A. & Mateles, R. J. (1976) Bacterial yields on methanol, methylamine, formaldehyde, and formate. Biotechnol. Bioeng. 18: 1657–1668

    Google Scholar 

  • Gottschal, J. C. & Kuenen, J. G. (1980) Mixotrophic growth of Thiobacillus A2 on acetate and thiosulphate as growth limiting substrates in the chemostat. Arch. Microbiol. 126: 33–42

    Google Scholar 

  • Houle, M. J., Long, D. E. & Smette, D. (1970) A simplex optimized colorimetric method formaldehyde. Anal. Lett. 3: 401–409

    Google Scholar 

  • Imai, T. (1983) Odor removal from waste water from kraft pulp mill by activated sludge process. Chemical Abstracts 99: 27403q

  • Johnson, A. & Quayle, J. R. (1964) Microbial growth on C1-compounds. VI Oxidation of methanol, formaldehyde and formate by methanol grown Pseudomonas AM1. Biochem. J. 93: 281–290

    Google Scholar 

  • Kelly, D. P. & Wood, A. P. (1982) Autotrophic growth of Thiobacillus A2 on methanol. FEMS Microbiol. Lett. 15: 229–233

    Google Scholar 

  • Kushelev, Y. V., Libman, B. Y., Mazanko, A. F., Zemlyanskii, V. A. & Boikov, I. G. (1976) A few properties of solutions of dimethyl-sulfide, dimethyl-sulfoxide and water. Sov. Chem. Ind. 10: 775

    Google Scholar 

  • Large, P. J. & Quayle, J. R. (1963) Microbial growth on C1-compounds: Five enzyme activities in extracts of Pseudomonas AM1. Biochem. J. 87: 386–396

    Google Scholar 

  • Leach, J. M. & Chung, L. T. K. (1982) Gas concentrations and occupational health in kraft mills. Tappi J. 65, no. 11: 95–98

    Google Scholar 

  • Ljunggren, G. & Norberg, B. (1943) On the effect and toxicity of dimethyl-sulphide, dimethyl-disulphide and methyl-mercaptan. Acta Physiol. Scand. 5: 248–255

    Google Scholar 

  • Lorimer, G. H., Badger, M. R. & Andrews, T. J. (1977) D-ribulose-1,5-bisphosphate carboxylase-oxygenase. Anal. Biochem. 78: 66–75

    Google Scholar 

  • Lovelock, J. E., Maggs, R. J. & Rasmussen, R. A. (1972) Atmospheric dimethyl-sulphide and the natural sulphur cycle. Nature (London) 237: 452–453

    Google Scholar 

  • Mhatre, S. S., Chetty, K. G. & Pradhan, D. S. (1983) Uncoupling of oxidative phosphorylation in rat liver mitochondria following the administration of dimethyl-sulphoxide. Biochem. Biophys. Res. Commun. 110: 325–331

    Google Scholar 

  • Nguyen, B. C., Gaudry, A., Bonsang, B. & Lambert, G. (1978) Reevaluation of the role of dimethyl-sulphide in the sulphur budget. Nature (London) 275: 637–639

    Google Scholar 

  • Patel, R. N. & Hoare, D. S. (1971) Physiological studies of methane and methanol oxidizing bacteria: oxidation of C1-compounds by Methylococcus capsulatus. J. Bact. 107: 187–192

    Google Scholar 

  • Przyjazny, A., Janicki, W., Chrzanowski, W., & Staszewski, R. (1983) Headspace gas chromatographic determination of distribution coefficients of selected organosulphur compounds and their dependence on some parameters. J. Chromatogr. 280: 249–260

    Google Scholar 

  • Sivelä, S. & Sundman, V. (1975) Demonstration of Thiobacillus-type bacteria which utilize methyl sulphides. Arch. Microbiol. 103: 303–304

    Google Scholar 

  • Sivelä, S. (1980) Dimethyl-sulphide as a growth substrate for an obligately chemolithotrophic Thiobacillus. Commentationes physico-mathematicae dissertationes (Helsinki) 1: 1–69

    Google Scholar 

  • Sörbo, B. H. (1957) A colorimetric method for the determination of thiosulphate. Biochim. Biophys. Acta 23: 412–417

    Google Scholar 

  • Stirling, D. J. & Dalton, H. (1978) Purification and properties of a NAD(P)+-linked formaldehyde dehydrogenase from Methylococcus capsulatus (Bath). J. Gen. Microbiol. 107: 19–29

    Google Scholar 

  • Trüper, H. G. & Schlegel, H. G. (1964) Sulphur metabolism in Thiorhodaceae. 1: Quantitative measurements on growing cells of Chromatium okenii. Antonie van Leeuwenhoek 30: 225–238

    Google Scholar 

  • Vishniac, W. & Santer, M. (1957) The Thiobacilli. Bact. Rev. 21: 195–213

    Google Scholar 

  • Wood, P. M. (1981) The redox potential for dimethyl-sulphoxide reduction to dimethyl sulphide: Evaluation and biochemical implications. FEBS Lett. 124: 11–14

    Google Scholar 

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  1. Department of Microbiology and Enzymology, Delft University of Technology, Julianalaan 67a, 2628 BC, Delft, The Netherlands

    G. M. H. Suylen & J. G. Kuenen

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  1. G. M. H. Suylen
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  2. J. G. Kuenen
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Suylen, G.M.H., Kuenen, J.G. Chemostat enrichment and isolation of Hyphomicrobium EG. Antonie van Leeuwenhoek 52, 281–293 (1986). https://doi.org/10.1007/BF00428640

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