Abstract
An enzymatic complex from Rhodotorula was characterized and it was indicated that it possessed thiosulfate-oxidizing activity, forming tetrathionate as well as sulfite oxidase activity. Both activities coupled with ferricyanide and native cytochrome c but no with mammalian cytochrome c. Activities of these enzymes were inhibited by thiol inhibitors. Chelating agents did not affect thiosulfate oxidizing activity and only moderately inhibited sulfite oxidase. Both activities disappeared after treatment with proteolytic enzymes or sodium deoxycholate which indicates an essential role played not only by protein but also by phospholipids in the enzymatic activity of the complex. Thiosulfate oxidizing enzyme had a K m for thiosulfate of 0.16 mM with ferricyanide as electron acceptor and of 14 μM with native cytochrome c and of 0.34 mM for ferricyanide. Optimum pH for this activity was 7.8. Other properties of this enzyme were similar to those of thiobacilli and heterotrophic bacteria. The activity of sulfite oxidase was inhibited by 50% with 10 μM AMP. The K m values of this enzyme were 1 mM with ferricyanide as electron acceptor and 60 μM with native cytochrome c for sulfite and 0.42 mM for ferricyanide. The enzyme did not show a specific optimum pH value with ferricyanide as electron acceptor. However, with native cytochrome c optimum pH was 7.8 for its activity. In many properties the sulfite oxidase from Rhodotorula was similar to the enzyme from Thiobacillus ferrooxidans, T. concretivorus, T. thioparus and T. novellus.
Similar content being viewed by others
Abbreviations
- CSH:
-
reduced glutathion
- APS:
-
reductase, adenosine-S′-phosphosulfate reductase
- pHMB:
-
p-hydroxymercuribenzoate
- NEM:
-
N-ethylmalcimide
- TCA:
-
trichloroacetic acid
- PPO:
-
2,5-diphenyloxazole
- POPOP:
-
2,2-p-phenylen-bis 5-phenyloxazol
References
Aminuddin M, Nicholas DJD (1974) An AMP independent sulfite oxidase from Thiobacillus denitrificans. Purification and properties. J Gen Microbiol 83:103–113
Barrett J (1969) The incorporation of 2-14C glycine and δ-amino [4-14C] laevulinic acid into the prosthetic groups of cytochrome oxidase and other cytochromes in yeast adapting to oxygen. Biochim Biophys Acta 177:442–455
Basford RE, Tisdale HD, Glenn JL, Green DE (1957) Studies on the terminal electron transport system. VII. Further studies on the succinic dehydrogenase complex. Biochim Biophys Acta 24:107–115
Bowen TJ, Butler PJ, Chappola FC (1965) Some properties of the rhodanese system from Thiobacillus denitrificans. Biochem J 97:651–657
Burton CP, Akagi JM (1971) Observations on the rhodanese activity of Desulfotomaculum nigrificans. J Bacteriol 107:375–376
Charles AM, Suzuki I (1966) Purification and properties of sulfite: cytochrome c oxidoreductase from Thiobacillus novellus. Biochim Biophys Acta 128:522–534
Hall MR, Berk R (1972) Thiosulfate oxidase from an Alcaligenes grown on mercaptosuccinate. Can J Microbiol 18:235–245
Johnson JL, Cohen HJ, Rajagopolan KV (1974) Molecular basis of the biological function of molybdenum: molybdenum-free sulfite oxidase from livers of tungsten treated rats. J Biol Chem 249:5046–5055
King TE, Morris RO (1967) Determination of acid-labile sulfide and sulfhydryl groups. In: Colowick SP, Kaplan NO (eds) Methods in enzymology, vol 10. Academic Press, New York London, pp 634–637
Kurek E (1979) Oxidation of inorganic sulfur compounds by yeast. Acta Microbiol Pol 28:169–172
Kurek EJ (1983) An enzymatic complex active in sulfite and thiosulfate oxidation by Rhodotorula sp. Arch Microbiol 134:143–147
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275
Lu WP, Kelly DP (1984) Properties and role of sulphite: cytochrome c oxidoreductase purified from Thiobacillus versutus (A2). J Gen Microbiol 130:1683–1692
Lyric RM, Suzuki I (1970a) Enzymes involved in the metabolism of thiosulphate by Thiobacillus thipparus. I. Survey of enzymes and properties of sulfite: cytochrome c oxidoreductase. Can J Biochem 48:334–343
Lyric RM, Suzuki I (1970b) Enzymes involved in the metabolism of thiosulfate by Thiobacillus thioparus. II. Properties of adenosine-5′-phosphosulfate reductase. Can J Biochem 48:344–354
Lyric RM, Suzuki I (1970c) Enzymes involved in the metabolism of thiosulfate by Thiobacillus thiparus. III. Properties of thiosulfate-oxidizing enzyme and proposed pathway of thiosulfate oxidation. Can J Biochem 48:355–363
Moriarty DJW (1969) Inorganic sulfur oxidation in Thiobacillus concretivorus. Ph. D. Thesis University of Adelaide
Morrison M, Connelly JC, Petrix J, Stotz E (1960) Properties and structural consideration of hemin a. J Biol Chem 245:1202–1205
Oh JK, Suzuki I (1977a) Isolation and characterization of a membrane-associated thiosulfate-oxidizing system of Thiobacillus novellus. J Gen Microbiol 99:397–412
Oh JK, Suzuki I (1977b) Resolution of a membrane-associated thiosulfate-oxidizing complex of Thiobacillus novellus. J Gen Microbiol 99:413–423
Schook LB, Berk RS (1978) Nutritional studies with Pseudomonas aeruginosa grown on inorganic sulfur sources. J Bacteriol 133:1377–1382
Schook LB, Berk RS (1979) Partial purification and characterization of thiosulfate oxidase from Pseudomonas aeruginosa. J Bacteriol 140:306–308
Silver M, Lundgren DG (1968) The thiosulfate-oxidizing enzyme of Ferrobacillus ferrooxidans (Thiobacillus ferrooxidans). Can J Biochem 46:1215–1220
Suzuki I (1974) Mechanisms of inorganic oxidation and energy coupling. Ann Rev Microbiol 28:85–101
Suzuki I, Silver M (1966) The initial product and properties of the sulfur-oxidizing enzyme of thiobacilli. Biochim Biophys Acta 122:22–23
Toghrol F, Southerland WM (1983) Purification of Thiobacillus novellus sulfite oxidase. J Biol Chem 258:6762–6766
Trudinger PA (1961) Thlosulfate oxidation and cytochromes in Thiobacillus X2. Thiosulfate oxidizing enzyme. Biochem J 78:680–686
Trudinger PA (1964) The effect of thiosulfate and oxygen concentration on tetrathionate oxidation by Thiobacillus X and T. thioparus. Biochem J 90:640–646
Trudinger PA (1967) Metabolism of thiosulfate and tetrathionate by heterotrophic bacteria from soil. J Bacteriol 93:550–559
Tuttle JH, Schwartz JH, Whited GM (1983) Some properties of thiosulfate-oxidizing enzyme from marine heterotroph 16B. Appl Environ Microbiol 46:438–445
Vestal JR, Lundgren DG (1971) Sullite oxidase of Thiobacillus ferrooxidans (Ferrobacillus ferrooxidans). Can J Biochem 49:1125–1130
Yamanaka T, Yoshioka T, Kimura K (1981) Purification of sulfitecytochrome c reductase of Thiobacillus novellus and reconstitution of its sulfite oxidase system with the purified constituents. Plant Cell Physiol 22:613–622
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Kurek, E.J. Properties of an enzymatic complex active in sulfite and thiosulfate oxidation by Rhodotorula sp. . Arch. Microbiol. 143, 277–282 (1985). https://doi.org/10.1007/BF00411250
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00411250