Abstract
Thiobacillus tepidarius, isolated from the hot springs at Bath, Avon, UK, grew optimally at 43–45°C and pH 6.0–7.5 on thiosulphate or tetrathionate. In batch culture, thiosulphate was oxidized stoichiometrically to tetrathionate, with a rise in pH. The tetrathionate was then oxidized to sulphate, supporting growth and producing a fall in pH to a minimum of ph 4.8. The organism contained high levels of thiosulphate-oxidizing enzyme, rhodanese and ribulose bisphosphate carboxylase. It was obligately chemolithotrophic and autotrophic. In chemostat culture, T. tepidarius grew autotrophically with the following sole energy-substrates: sulphide, thiosulphate, trithionate, tetrathionate, hexathionate or heptathionate. Thiocyanate, dithionate and sulphite were not used as sole substrates, although sulphite enhanced growth yields in the presence of thiosulphate. Maximum specific growth rate on tetrathionate was 0.44 h-1. ‘True growth yields’ (Y max) and maintenance coefficients (m) were calculated for sulphide, thiosulphate, trithionate and tetrathionate and observed yields at a single fixed dilution rate compared with those on hexathionate and heptathionate. Mean values for Y max, determined from measurements of absorbance, dry wt, total organic carbon and cell protein, were similar for sulphide, thiosulphate and trithionate (10.9 g dry wt/mol substrate) as expected from their equivalent oxygen consumption for oxidation. Y max for tetrathionate (20.5) and the relative Y o values (as g dry wt/g atom oxygen consumed) for thiosulphate and all four polythionates indicated that substrate level phosphorylation did not contribute significantly to energy conservation. These Y max values were 40–70% higher than any of those previously reported for obligately aerobic thiobacilli. Mean values for m were 6.7 mmol substrate oxidized/g dry wt·h for sulphide, thiosulphate and trithionate, and 2.6 for tetrathionate.
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Abbreviations
- PIPES:
-
Piperazine-N,N′-bis(ethane sulphonic acid)
References
Auerbach F, Koppel I (1927) Darstellung der Polythionate und Polythionsäuren. In: Abegg R, Auerbach F, Koppel I (eds) Handbuch der anorganischen Chemie, vol 4. S. Hirzel, Leipzig, p 554
Eccleston M, Kelly DP (1978) Oxidation kinetics and chemostat growth kinetics of Thiobacillus ferrooxidans on tetrathionate and thiosulfate. J Bacteriol 134:718–727
Fieschko J, Humphrey AE (1984) Statistical analysis in the estimation of maintenance and true growth yield coefficients. Biotechnol Bioeng 26:394–396
Goehring M, Feldmann U (1948) Neue Verfahren zur Darstellung von Kaliumpentathionat und von Kaliumhexathionat. Z Anorg Chem 257:223–226
Hempfling WP, Vishniac W (1967) Yield coefficients of Thiobacillus neapolitanus in continuous culture. J Bacteriol 93:874–878
Justin PM, Kelly DP (1978) Growth kinetics of Thiobacillus denitrificans in anaerobic and aerobic chemostat culture. J Gen Microbiol 107:123–130
Kelly DP (1978) Bioenergetics of chemolithotrophic bacteria. In: Bull AT, Meadow PM (eds) Companion to microbiology. Longman, London, pp 363–386
Kelly DP (1982) Biochemistry of the chemolithotrophic oxidation of inorganic sulphur. Phil Trans Roy Soc Lond B 298:499–528
Kelly DP (1985) Physiology of the thiobacilli: elucidating the sulphur oxidation pathway. Microbiol Sci 2:105–109
Kelly DP, Harrison AP (1985) Thiobacillus. In: Holt JG (ed) Bergey's manual of determinative bacteriology, 9th ed. Williams and Wilkins, Baltimore (in press)
Kelly DP, Chambers LA, Trudinger PA (1969) Cyanolysis and spectrophotometric estimation of trithionate in mixture with thiosulphate and tetrathionate. Anal Chem 41:898–901
Koh T, Iwasaki I (1965) The determination of microamounts of polythionates. II. A photometric method for the determination of hexathionate by means of its cyanolysis. Bull Chem Soc Japan 38:2135–2138
Koh T, Iwasaki I (1966a) The determination of microamounts of polythionates. III. A photometric method for the determination of tetrathionate by means of its cyanolysis. Bull Chem Soc Japan 39:352–356
Koh T, Iwasaki I (1966b) The determination of microamounts of polythionates. V. A photometric method for the determination of polythionates when two species of them, tetra-, penta- and hexathionate are present together. Bull Chem Soc Japan 39:703–708
Koh T, Taniguchi K (1973) Spectrophotometric determination of total amounts of polythionates (tetra-, penta-and hexathionate) in mixtures with thiosulfate and sulfite. Anal Chem 45:2018–2022
Kolthoff IM, Belcher P (1957) Volumetric analysis, vol III. Interscience, New York, p 184
Kuenen JG (1979) Growth yields and “maintenance energy requirement” in Thiobacillus species under energy limitation. Arch Microbiol 122:183–188
Lu WP, Kelly DP (1983) Thiosulfate oxidation, electron transport and phosphorylation in cell-free systems from Thiobacillus A2. J Gen Microbiol 129:1661–1671
Lu WP, Kelly DP (1984) Oxidation of inorganic sulfur compounds by thiobacilli. In: Crawford RL, Hanson RS (eds) Microbial growth on C1 compounds. American Society for Microbiology, Washington, pp 34–41
Lu WP, Swoboda BEP, Kelly DP (1985) Properties of the thiosulfate-oxidizing multi-enzyme system from Thiobacillus versutus. Biochim Biophys Acta 828:116–122
Pachmayr F (1960) Vorkommen und Bestimmung von Schwefelverbindungen in Mineralwasser. Diss Univ, München
Peck HD (1960) Adenosine 5′-phosphosulfate as an intermediate in the oxidation of thiosulfate by Thiobacillus thioparus. Proc Natl Acad Sci USA 46:1053–1057
Pirt SJ (1965) The maintenance energy of bacteria in growing cultures. Proc R Soc London B 163:224–231
Pirt SJ (1975) Principles of microbe and cell cultivation. Blackwell Scientific Publications, Oxford
Roy AP, Trudinger PA (1970) The biochemistry of inorganic compounds of sulphur. Cambridge University Press, p 48
Smith AL, Kelly DP, Wood AP (1980) Metabolism of Thiobacillus A2 under autotrophic, mixotrophic and heterotrophic conditions in the chemostat. J Gen Microbiol 121:127–138
Stamm H, Seipold O, Goehring M (1941) Zur Kenntnis der Polythionsäuren und ihrer Bildung. 4. Die Reaktionen zwischen Polythionsäuren und schwefliger Säure bzw. Thioschwefelsäure. Z Anorg Allg Chem 247:277–306
Tempest DW, Neijssel OM (1984) The status of Y ATP and maintenance energy as biologically interpretable phenomena. Annu Rev Microbiol 38:459–486
Timmer-ten Hoor A (1981) Cell yield and bioenergetics of Thiomicrospira denitrificans compared with Thiobacillus denitrificans. Antonie van Leeuwenhoek J Microbiol Serol 47:231–243
Tuovinen OH, Kelly DP (1973) Studies on the growth of Thiobacillus ferrooxidans. Arch Mikrobiol 88:285–298
Vishniac W (1953) The metabolism of Thiobacillus thioparus. I. The oxidation of thiosulfate. J Bacteriol 64:363–373
Weitz E, Achterberg F (1928) Über höhere Polythionsäuren. I. Die Hexathionsäure. Ber Dt Chem Ges 61:399–408
Willstätter R (1903) Über die Einwirkung von Hydrogenperoxyd auf Natriumthiosulfat. Ber Dt Chem Ges 36:1831–1833
Wood AP, Kelly DP (1981) Mixotrophic growth of Thiobacillus A2 in chemostat culture on formate and glucose. J Gen Microbiol 125:55–62
Wood AP, Kelly DP (1983) Autotrophic, mixotrophic and heterotrophic growth with denitrification by Thiobacillus A2 under anaerobic conditions. FEMS Microbiol Lett 16:363–370
Wood AP, Kelly DP (1984) Growth and sugar metabolism of a thermoacidophilic iron-oxidizing mixotrophic bacterium. J Gen Microbiol 130:1337–1349
Wood AP, Kelly DP (1985) Physiological characteristics of a new thermophilic, obligately chemolithotrophic Thiobacillus species, Thiobacillus tepidarius sp. nov. Int J Syst Bacteriol 35:91–94
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Wood, A.P., Kelly, D.P. Chemolithotrophic metabolism of the newly-isolated moderately thermophilic, obligately autotrophic Thiobacillus tepidarius . Arch. Microbiol. 144, 71–77 (1986). https://doi.org/10.1007/BF00454959
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DOI: https://doi.org/10.1007/BF00454959