Skip to main content
Log in

Crotonobetaine reductase fromEscherichia coli — a new inducible enzyme of anaerobic metabolization of L(-)-carnitine

  • Research Articles
  • Published:
Antonie van Leeuwenhoek Aims and scope Submit manuscript

Abstract

Crotonobetaine reductase fromEscherichia coli 044 K74 is an inducible enzyme detectable only in cells grown anaerobically in the presence of L(-)-carnitine or crotonobetaine as inducers. Enzyme activity was not detected in cells cultivated in the presence of inducer plus glucose, nitrate, γ-butyrobetaine or oxygen, respectively. Fumarate caused an additional stimulation of growth and an increased expression of crotonobetaine reductase. The reaction product, γ-butyrobetaine, was identified by autoradiography. Crotonobetaine reductase is localized in the cytoplasm, and has been characterized with respect to pH (pH 7.8) and temperature optimum (40–45 °C). The K m value for crotonobetaine was determined to be 1.1×10−2M. γ-Butyrobetaine,D(+)-carnitine and choline are inhibitors of crotonobetaine reduction. For γ-butyrobetaine (K i =3×10−5M) a competitive inhibition type was determined. Various properties suggest that crotonobetaine reductase is different from other reductases of anaerobic respiration.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Bergmeyer HU (ed) (1970) Methods of Enzymic Analysis, pp 1714–1715, Akademie Verlag, Berlin

    Google Scholar 

  • Bilous PT & Weiner JH (1985) Dimethyl sulfoxide reductase activity by anaerobically grownEscherichia coli HB 101. J. Bacteriol. 162: 1151–1155

    Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72: 248–254

    Google Scholar 

  • Dickie P & Weiner JH (1979) Purification and characterization of membrane-bound fumarate reductase from anaerobically grownEscherichia coli. Can. J. Biochem. 57: 813–821

    Google Scholar 

  • Haddock BA & Jones CW (1977) Bacterial respiration. Bacteriol. Rev. 48: 47–99

    Google Scholar 

  • Ishimoto MS & Shimokawa O (1978) Reduction of trimethylamine N-oxide byEscherichia coli as anaerobic respiration. Z. Allg. Mikrobiol. 18: 173–181

    Google Scholar 

  • Jung K, Jung H & Kleber H-P (1987) Regulation of L-carnitine metabolism inEscherichia coli. J. Basic. Microbiol. 27: 131–137

    Google Scholar 

  • Jung H, Jung K & Kleber H-P (1989) Purification and properties of carnitine dehydratase fromEscherichia coli — a new enzyme of carnitine metabolization. Biochim. Biophys. Acta. 1003: 270–276

    Google Scholar 

  • Jung H, Jung K & Kleber H-P (1990a) L-Carnitine metabolization and osmotic stress response inEscherichia coli. J. Basic. Microbiol. 30: 409–413

    Google Scholar 

  • Jung H, Jung K & Kleber H-P (1990b) L-Carnitine uptake byEscherichia coli. J. Basic. Microbiol. 30: 507–514

    Google Scholar 

  • Jung H, Jung K & Kleber H-P (1993) Synthesis of L-carnitine by microorganisms and isolated enzymes. Adv. Biochem. Engin. Biotechnol. 50: 21–44

    Google Scholar 

  • Kaback HR (1971) Bacterial membranes. Meth. Enzymol. 22: 99–120

    Google Scholar 

  • Kleber H-P, Seim H, Aurich H & Strack E (1977) Verwertung von Trimethylammoniumverbindungen durchAcinetobacter calcoaceticus. Arch. Microbiol. 112: 201–206

    Google Scholar 

  • Konings WN & Boonstra J (1977) Anaerobic electron transfer and active transport in bacteria. Curr. Top. Membr. Transp. 9: 177–231

    Google Scholar 

  • Kröger A (1978) Fumarate as terminal acceptor of phosphorylative electron transport. Biochim. Biophys. Acta 505: 129–145

    Google Scholar 

  • Myers CR & Myers JM (1992) Fumarate reductase is a soluble enzyme in anaerobically grownShewanella putrefaciens MR-I. FEMS Microbiol. Lett. 98: 13–20

    Google Scholar 

  • Roth S (1992) Anreicherung und Charakterisierung der Crotonobetainreduktase ausE. coli. Diplomarbeit, Univ. Leipzig

  • Seim H, Ezold R, Kleber H-P & Strack E (1980) Stoffwechsel des L-Carnitins bei Enterobakterien. Z. Allg. Mikrobiol. 20: 591–594

    Google Scholar 

  • Seim H, Jung H, Löster H & Kleber H-P (1985) Wachstumsstimulation und Stoffwechsel des Carnitins beiEscherichia coli. Wiss. Z. Karl-Marx-Univ. Leipzig, Math-Nat. R. 34: 287–292

    Google Scholar 

  • Seim H, Löster H, Claus R, Kleber H-P & Strack E (1982a) Stimulation of the anaerobic growth ofSalmonella typhimurium by reduction of L-carnitine, carnitine derivatives and structure-related trimethylammonium compounds. Arch. Microbiol. 132: 91–95

    Google Scholar 

  • Seim H, Löster H & Kleber H-P (1982b) Reduktiver Stoffwechsel des L-Carnitins und strukturverwandter Trimethylammoniumverbindungen inEscherichia coli. Acta Biol. Med. Germ. 41: 1009–1018

    Google Scholar 

  • Shimokawa O & Ishimoto M (1979) Purification and some properties of inducible tertiary amine N-oxide reductase fromEscherichia coli. J. Biochem. 86: 1709–1717

    Google Scholar 

  • Spencer ME & Guest JR (1973) Isolation and properties of fumarate reductase mutants ofEscherichia coli. J. Bacteriol. 114: 563–570

    Google Scholar 

  • Violet M, Medani C-L & Giordano G (1985) Trimethylamine N-oxide (TMAO) reductases fromEscherichia coli K-12. FEMS Microbiol. Lett. 27: 85–91

    Google Scholar 

  • Weiner JH, Rothery RA, Sambasivarao D & Trieber CA (1992) Molecular analysis of dimethylsulfoxide reductase: a complex iron-sulfur molybdoenzyme ofEscherichia coli. Biochim. Biophys. Acta 1102: 1–18

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Roth, S., Jung, K., Jung, H. et al. Crotonobetaine reductase fromEscherichia coli — a new inducible enzyme of anaerobic metabolization of L(-)-carnitine. Antonie van Leeuwenhoek 65, 63–69 (1994). https://doi.org/10.1007/BF00878280

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00878280

Key words

Navigation