Abstract
The capacity of mesophilic sulfate-reducing bacteria to grow lithoautotrophically with H2, sulfate and CO2 was investigated with enrichment cultures and isolated species. (a) Enrichments in liquid mineral media with H2, sulfate and CO2 consistently yielded mixed cultures of nonautotrophic, acetate-requiring Desulfovibrio species and autotrophic, acetate-producing Acetobacterium species (cell ratio approx. 20:1). (b) By direct dilution of mud samples in agar, various non-sporing sulfate reducers were isolated in pure cultures that did grow autotrophically. Two oval cell types (strains HRM2, HRM4) and one curved cell type (strain HRM6) from marine sediment were studied in detail. The strains grew in mineral medium supplemented only with vitamins (biotin, p-aminobenzoate, nicotinate). Carbon autotrophy was evident (i) from comparative growth experiments with non-autotrophic, acetate-requiring species, (ii) from high cell densities ruling out a cell synthesis from organic impurities in the mineral media, and (iii) by demonstrating that 96–99% of the cell carbon was derived from 14C-labelled CO2. Autotrophic growth occurred with a doubling time of 16–20 h at 24–28°C. Formate, fatty acids up to palmitate, ethanol, lactate, succinate, fumarate, malate and other organic acids were also used and completely oxidized. The three strains possessed cytochromes of the b-and c-type, but no desulfoviridin. Strain HRM2 is described as a new species of a new genus, Desulfobacterium autotrophicum. (c) The capacity for autotrophic growth was also tested with sulfate-reducing bacteria that originally had been isolated on organic substrates. The incompletely oxidizing, non-sporing types such as Desulfovibrio and Desulfobulbus species and Desulfomonas pigra were confirmed to be obligate heterotrophs that required acetate for growth with H2 and sulfate. In contrast, several of the completely oxidizing sulfate reducers were facultative autotrophs, such as Desulfosarcina variabilis, Desulfonema limicola, Desulfococcus niacini, and the newly isolated Desulfobacterium vacuolatum and Desulfobacter hydrogenophilus. The only incompletely oxidizing sulfate reducer that could grow autotrophically was the sporing Desulfotomaculum orientis, which obtained 96% of its cell carbon from 14C-labelled CO2. Desulfovibrio baarsii and Desulfococcus multivorans may also be regarded as types of facultative autotrophs; they could not oxidize H2, but grew on sulfate with formate as the only organic substrate.
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References
Alvarez M, Barton LL (1977) Evidence for the presence of phosphoriboisomerase and ribulose-1,5-diphosphate carboxylase in extracts of Desulfovibrio vulgaris. J Bacteriol 131:133–135
Bache R, Pfennig, N (1981) Selective isolation of Acetobacterium woodii on methoxylated aromatic acids and determination of growth yields. Arch Microbiol 130:225–261
Badziong W, Thauer RK (1978) Growth yields and growth rates of Desulfovibrio vulgaris (Marburg) growing on hydrogen plus sulfate and hydrogen plus thiosulfate as the sole energy sources. Arch Microbiol 117:209–214
Badziong W, Thauer RK, Zeikus JG (1978) Isolation and characterization of Desulfovibrio growing on hydrogen plus sulfate as the sole energy source. Arch Microbiol 116:41–49
Badziong W, Ditter B, Thauer RK (1979) Acetate and carbon dioxide assimilation by Desulfovibrio vulgaris (Marburg), growing on hydrogen and sulfate as sole energy source. Arch Microbiol 123:301–305
Bak F, Widdel F (1986) Anaerobic degradation of indolic compounds by sulfate-reducing enrichment cultures, and description of Desulfobacterium indolicum gen. nov., sp. nov. Arch Microbiol 146:170–176
Balch WE, Fox GE, Magrum LJ, Woese CR, Wolfe RS (1979) Methanogens: reevaluation of a unique biological group. Microbiol Rev 43:260–296
Beijerinck WM (1895) Über Spirillum desulfuricans als Ursache von Sulfatreduction. Centralbl Bakteriol II. Abt 1:1–9, 49–59, 104–114
Brandis A, Thauer RK (1981) Growth of Desulfovibrio species on hydrogen and sulphate as sole energy source. J Gen Microbiol 126:249–252
Brandis-Heep A, Gebhardt NA, Thauer RK, Widdel F, Pfennig N (1983) Anaerobic acetate oxidation to CO2 by Desulfobacter postgatei. 1. Demonstration of all enzymes required for the operation of the citric acid cycle. Arch Microbiol 136:222–229
Bryant MP (1972) Commentary on the, Hungate technique for culture of anaerobic bacteria. Am J Clin Nutr 25:1324–1328
Butlin KR, Adams ME (1947) Autotrophic growth of sulphatereducing bacteria. Nature 160:145–155
Collins MD, Widdel F (1986) Respiratory quinones of sulphatereducing and sulphur-reducing bacteria: a systematic investigation. Syst Appl Microbiol 8:8–18
Cord-Ruwisch R (1985) A quick method for the determination of dissolved and precipitated sulfides in cultures of sulfate-reducing bacteria. J Microbiol Meth 4:33–36
Cypionka, H, Pfennig N (1986) Growth yields of Desulfotomaculum orientis with hydrogen in chemostat culture. Arch Microbiol 143:396–399
Fowler VJ, Widdel F, Pfennig, N, Woese CR, Stackebrandt E (1986) Phylogenetic relationships of sulfate- and sulfur-reducing eubacteria. Syst Appl Microbiol 8:32–41
Fuchs G (1986) CO2 fixation in acetogenic bacteria: variations on a theme. FEMS Microbiol Rev 39:181–213
Fuchs G, Thauer RK, Ziegler H, Stichler W (1979) Carbon isotope fractionation by Methanobacterium thermoautotrophicum. Arch Microbiol 120:135–139
Gebhardt NA, Linder D, Thauer RK (1983) Anaerobic acetate oxidation to CO2 by Desulfobacter postgatei. 2. Evidence from 14C-labelling studies for the operation of the citric acid cycle. Arch Microbiol 136:230–233
Gregersen T (1978) Rapid method for distinction of Gram-negative from Gram-positive bacteria. Eur J Appl Microbiol Biotechnol 5:123–127
Imhoff-Stuckle D, Pfennig N (1983) Isolation and characterization of a nicotinic acid-degrading sulfate-reducing bacterium, Desulfococcus niacini sp. nov. Arch Microbiol 136:194–198
Jansen K, Thauer RK, Widdel F, Fuchs G (1984) Carbon assimilation pathways in sulfate-reducing bacteria. Formate, carbon dioxide, carbon monoxide, and acetate assimilation by Desulfovibrio baarsii. Arch Microbiol 138:257–262
Jansen K, Fuchs G, Thauer RK (1985) Autotrophic CO2 fixation by Desulfovibrio baarsii: demonstration of enzyme activities characteristic for the acetyl-CoA pathway. FEMS Microbiol Lett 28:311–315
Jørgensen BB (1978) A comparison of methods for the quantification of bacterial sulfate reduction in coastal marine sediments. III. Estimation from chemical and bacteriological field data. Geomicrobiol J 1:49–64
Klemps R, Cypionka H, Widdel F, Pfennig N (1985) Growth with hydrogen, and further physiological characteristics of Desulfotomaculum species. Arch Microbiol 143:203–208
Laanbroek HJ, Pfennig N (1981) Oxidation of short-chain fatty acids by sulfate-reducing bacteria in freshwater and in marine sediments. Arch Microbiol 128:330–335
Lijungdahl LG (1986) The autotrophic pathway, of acetate synthesis in acetogenic bacteria. Ann Rev Microbiol 40:415–450
Marmur J, Doty P (1962) Determination of the base composition of deoxyribonucleic acid from its thermal denaturation temperature. J Mol Biol 5:109–118
Mechalas BJ, Rittenberg SC (1960) Energy coupling in Desulfovibrio desulfuricans. J Bacteriol 80:501–507
Pfennig N, Widdel F (1981) Ecology and physiology of some anaerobic bacteria from the microbial sulfur cycle. In: Bothe H, Trebst A (eds) Biology of inorganic nitrogen and sulfur. Springer, Berlin Heidelberg New York, pp 169–177
Pfennig N, Widdel F, Trüper HG (1981) The dissimilatory sulfatereducing bacteria. In: Starr MP, Stolp H, Trüper HG, Balows A, Schlegel HG (eds) The prokaryotes, vol 1. Springer, Berlin Heidelberg New York, pp 926–940
Postgate JR (1959) A diagnostic reaction of Desulphovibrio desulphuricans. Nature 183:481–482
Postgate JR (1960) On the autotrophy of Desulfovibrio desulphuricans. Z Allg Mikrobiol 1:53–56
Schauder R, Eikmanns B, Thauer RK, Widdel F, Fuchs G (1986) Acetate oxidation to CO2 in anaerobic bacteria via a novel pathway not involving reactions of the citric acid cycle. Arch Microbiol 145:162–172
Schauder R, Widdel F, Fuchs G (1987) Carbon assimilation pathways in sulfate-reducing bacteria. II. Enzymes of a reductive citric acid cycle in the autotrophic Desulfobacter hydrogenophilus. Arch Microbiol 148:218–225
Schink B, Pfennig N (1982) Propionigenium modestum gen. nov. sp. nov., a new strictly anaerobic, nonsporing bacterium growing on succinate. Arch Microbiol 133:209–216
Simon H, Floss HG (1967) Bestimmung der Isotopenverteilung in markierten Verbindungen. Springer, Berlin Heidelberg New York
Sisler FD, ZoBell CE (1951) Hydrogen utilization by some marine sulfate-reducing bacteria. J Bacteriol 62:117–127
Sorokin YuI (1966a) Sources of energy and carbon for biosynthesis in sulfate-reducing bacteria. Microbiology (translated from Mikrobiologiya) 35:643–647
Sorokin YuI (1966b) Investigation of the structural metabolism of sulfate-reducing bacteria with C14. Microbiology (translated from Mikrobiologiva) 35:806–814
Sorokin YuI (1966c) Role of carbon dioxide and acetate in the biosynthesis by sulfate reducing bacteria. Nature 210:551–552
Stephenson M, Strickland LH (1931) Hydrogenase. II. The reduction of sulphate to sulphide by molecular hydrogen. Biochem J 25:215–220
Tschech A, Pfennig N (1984) Growth yield increase linked to caffeate reduction in Acetobacterium woodii. Arch Microbiol 137:163–167
Watson GR, Williams JP (1970) Rapid method for wet combustion and scintillation counting of 14C-labeled organic materials. Anal Biochem 33:356–365
Widdel F (1980) Anaerober Abbau von Fettsäuren und Benzoesäure durch neu isolierte Arten Sulfat-reduzierender Bakterien. Doctoral thesis, Univ Göttingen
Widdel F (1987a) New types of acetate-oxidizing, sulfate-reducing Desulfobacter species, D. hydrogenophilus sp. nov., D. latus sp. nov., and D. curvatus sp. nov. Arch Microbiol 148:286–291
Widdel F (1987b) Microbiology and ecology, of sulfate- and sulfurreducing bacteria. In: Zehnder AJB (ed) Environmental microbiology of anaerobic bacteria, chapter 10. John Wiley, & Sons, New York (in press)
Widdel F, Pfennig N (1981) Studies on dissimilatory sulfate-reducing bacteria that decompose fatty acids. I. Isolation of new sulfate-reducing bacteria enriched with acetate from saline environments. Description of Desulfobacter postgatei gen. nov., sp. nov. Arch Microbiol 129:395–400
Widdel F, Pfennig N (1982) Studies on dissimilatory sulfate-reducing bacteria that decompose fatty acids. II. Incomplete oxidation of propionate by Desulfobulbus propionicus gen. nov., sp. nov. Arch Microbiol 131:360–365
Widdel F, Pfennig N (1984) Dissimilatory sulfate- or sulfur-reducing bacteria. In: Krieg NR, Holt JG (eds) Bergey's manual of systematic bacteriology, vol 1. Williams & Wilkins, Baltimore London, pp 663–379
Widdel F, Kohring G-W, Mayer F (1983) Studies on dissimilatory sulfate-reducing bacteria that decompose fatty acids. III. Characterization of the filamentous gliding Desulfonema limicola gen. nov. sp. nov. and Desulfonema magnum sp. nov. Arch Microbiol 134:286–294
Wight KM, Starkey RL (1945) Utilization of hydrogen by sulfatereducing bacteria and its significance in anaerobic corrosion. J Bacteriol 50:238
Wood HG, Ragsdale SW, Pezacka E (1986) The acety-CoA pathway of autotrophic growth. FEMS Microbiol Rev 30:345–362
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Brysch, K., Schneider, C., Fuchs, G. et al. Lithoautotrophic growth of sulfate-reducing bacteria, and description of Desulfobacterium autotrophicum gen. nov., sp. nov.. Arch. Microbiol. 148, 264–274 (1987). https://doi.org/10.1007/BF00456703
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DOI: https://doi.org/10.1007/BF00456703