Archives of Microbiology

, Volume 150, Issue 5, pp 465–470 | Cite as

The carbon monoxide- and oxygen-reacting haemoproteins of Streptomyces clavuligerus: cytochrome aa3 is the predominant terminal oxidase of the respiratory chain

  • Robert I. Scott
  • Robert K. Poole
Original Papers

Abstract

The nature of the carbon monoxide- and oxygen-reacting haemoproteins in the respiratory chain of the filamentous antibiotic-producing bacterium Streptomyces clavuligerus has been investigated. CO-difference (i.e. CO+ reduced minus reduced) spectra of intact cells showed the presence of cytochrome aa3, a CO binding b-type cytochrome, and a pigment resembling cytochrome d. In addition, cells that were approaching the end of the growth phase showed the presence of cytochrome P450: this pigment was undetectable in cells harvested early in the growth cycle. High speed centrifugation of cell-free extracts prepared from cells broken by sonication showed that cytochrome aa3 was tightly membrane-bound and that cytochrome P450 was soluble. Inhibition of oxygen uptake rates of cells by cyanide indicated that one component, which showed 50% inhibition at 2–4 mM CN, was acting as major terminal oxidase: this was observed in cells harvested from all stages of growth. Photodissociation (i. e. photolysed, CO reduced minus CO reduced) spectra at-118°C, in the absence of oxygen, showed cytochrome aa3 to be the sole photolysable CO-reacting haemoprotein. At higher temperature (-87°C), in the presence of oxygen, cytochrome aa3 formed a complex with oxygen that could not be photolysed by similar intensities of light. By raising the temperature to-43°C, the oxidation of c-type cytochromes was observed. It is concluded that cytochrome aa3 is the predominant terminal oxidase in S. clavuligerus and that the other CO reacting haemoproteins, of unknown function, are unlikely to be oxidases.

Key words

Cytochrome aa3 Cytochrome P450 Respiratory chain Streptomyces clavuligerus Oxygen uptake CO-reacting haemoproteins Oxygen metabolism 

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References

  1. Appleby CA (1978) Purification of Rhizobium cytochromes P450. Meth Enzymol 52:157–166Google Scholar
  2. Birk Y, Silver WS, Heim AH (1957) A b type cytochrome from Streptomyces fradiae. Biochim Biophys Acta 25:227–228Google Scholar
  3. Castor LN, Chance B (1955) Photochemical action spectra of carbon monoxide inhibited respiration. J Biol Chem 217:453–465Google Scholar
  4. Castor LN, Chance B (1959) Photochemical determinations of the oxidases of bacteria. J Biol Chem 234:1587–1592Google Scholar
  5. Chance B, Saronio C, Leigh JS (1975) Functional intermediates in the reaction of membrane bound cytochrome oxidase with oxygen. J Biol Chem 250:9226–9237Google Scholar
  6. Corcoran JW (1981) Biochemical mechanisms in the biosynthesis of the erythromycins. In: Corcoran JW (ed) Antibiotics. IV. Biosynthesis. Springer, Berlin Heidelberg New York, pp 132–174Google Scholar
  7. Edwards C, Ball AS (1987) Respiratory chain composition and activity in some thermotolerant streptomycetes. FEMS Microbiol Lett 40:61–66Google Scholar
  8. Heim AH, Silver WS, Birk Y (1957) Cytochrome composition of some strains of Streptomyces. Nature 180:608–609Google Scholar
  9. Ingledew WJ, Poole RK (1984) The respiratory chains of Escherichia coli. Microbiol Rev 48:222–271Google Scholar
  10. Inoue Y (1958) The metabolism of Streptomyces griseus. IV. The terminal pathway of the respiration of Streptomyces griseus. J Antibiotics Ser A 11:109–115Google Scholar
  11. Inoue Y (1973) The metabolism of Streptomyces griseus. XI. Further observation on the electron transfer system of the Streptomyces griseus respiratory particle. Bot Mag Tokyo 86:121–132Google Scholar
  12. Inoue Y (1976) Presence of cytochrome d in respiratory particles of Streptomyces griseus and its function in the terminal oxidation system. Bot Mag Tokyo 89:183–190Google Scholar
  13. Jones CW, Poole RK (1985) The analysis of cytochromes. In: Gottschalk G (ed) Methods in microbiology, vol 18. Academic Press, London Orlando, pp 285–328Google Scholar
  14. Jurtshuk P, Mueller TJ, Acord WC (1975) Bacterial terminal oxidases. CRC Crit Rev Microbiol 3:399–468Google Scholar
  15. Katagiri M, Ganguli BM, Gunsalus IC (1968) A soluble cytochrome P450 functional in methylene hydroxylation. J Biol Chem 243:3543–3546Google Scholar
  16. Lloyd D, Scott RI, Edwards SW, Chance B (1982) Cytochrome a 620 in Tetrahymena pyriformis. Reactions with carbon monoxide and oxygen at subzero temperatures and photochemical action spectra. Biochem J 206:367–372Google Scholar
  17. Markwell MAK, Haas SM, Bieber LL, Tolbert NE (1978) A modification of the Lowry procedure to simplify protein determination in membrane and lipoprotein samples. Anal Biochem 87:206–210Google Scholar
  18. Petersen JA (1971) Camphor binding by Pseudomonas putida cytochrome P450. Arch Biochem Biophys 144:678–693Google Scholar
  19. Poole RK (1983) Bacterial cytochrome oxidases. A structurally and functionally diverse group of electron transfer proteins. Biochim Biophys Acta 726:205–243Google Scholar
  20. Poole RK, Ingledew WJ (1987) Pathways of electrons to oxygen. In: Neidhardt FC, Ingraham JK, Brooks Low K, Magasonik B, Schaechter M, Umbarger HE (eds) Escherichia coli and Salmonella typhimurium. Cellular and molecular biology, vol 1. American Society for Microbiology. Washington, pp 170–200Google Scholar
  21. Poole RK, Scott RI, Baines BS, Salmon I, Lloyd D (1982a) Identification of cytochromes o and a 3 as functional terminal oxidases in the thermophilic bacterium PS3. FEBS Lett 150:281–285Google Scholar
  22. Poole RK, Sivaram A, Salmon I, Chance B (1982b) Photolysis at very low temperatures of CO-liganded cytochrome oxidase (cytochrome d) in oxygen limited Escherichia coli. FEBS Lett 141:237–241Google Scholar
  23. Scott RI, Lloyd D (1983) Photochemical action spectra indicate that cytochrome aa 3 is the predominant haemoprotein terminal oxidase in Acanthamoeba castellanii. Biochem J 210:721–725Google Scholar
  24. Scott RI, Edwards SW, Chance B, Lloyd D (1983) Terminal oxidase of Crithidia fasciculata. Reactions with carbon monoxide and oxygen at subzero temperatures and photochemical action spectra. J Gen Microbiol 129:1983–1989Google Scholar
  25. Scott RI, Sladen S, Maidment M, Rashid T, Pratsis C, Perry D (1988) The effects of oxygen on β-lactam biosynthesis by alginate entrapped Streptomyces clavuligerus. J Chem Tech Biotech 41:145–154Google Scholar
  26. Stevenson PM, Ruettinger RT, Fulco AJ (1983) Cytochrome P450 revealed: The effect of respiratory cytochromes on the spectrum of bacterial cytochrome P450. Biochem Biophys Res Comm 112:927–934Google Scholar
  27. Williams HD, Poole RK (1987) The cytochromes of Acetobacter pasteurianus NCIB 6428. Evidence of a role for a cytochrome a 1 like haemoprotein in electron transfer to cytochrome oxidase d. J Gen Microbiol 133:2461–2472Google Scholar
  28. Wood PM (1984) Bacterial proteins with CO binding b or c-type haem. Functions and absorption spectroscopy. Biochim Biophys Acta 768:293–317Google Scholar

Copyright information

© Springer-Verlag 1988

Authors and Affiliations

  • Robert I. Scott
    • 1
  • Robert K. Poole
    • 2
  1. 1.School of BiotechnologyPolytechnic of Central LondonLondonUK
  2. 2.Department of MicrobiologyKings College LondonLondonUK

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