Applied Microbiology and Biotechnology

, Volume 27, Issue 5–6, pp 451–456 | Cite as

Purification and properties of an acetyl specific carboxylesterase from Nocardia mediterranei

  • Hans-Peter Schär
  • Daniel Gygax
  • Gerardo M. Ramos Tombo
  • Oreste Ghisalba
Biotechnology

Summary

An acetyl specific carboxylesterase has been purified from Nocardia mediterranei. The purified enzyme is homogeneous as shown by SDS polyacrylamide gel electrophoresis. The esterase has a molecular weight of 68,000 and is composed of two identical subunits. The enzyme exhibits optimal activity at pH 7.5 and at 35°C and is stable below 40°C. The enzyme activity is inhibited by several sulfhydryl reagents. The esterase hydrolyzes preferentially acetyl esters. Propionyl esters are cleaved very slowly whereas butyryl esters are no substrates at all. In addition, the esterase shows a pronounced regiospecificity. On the other hand the enantiospecificity is rather low as demonstrated by the hydrolysis of prochiral and racemic substrates.

Keywords

Enzyme Ester Hydrolysis Electrophoresis Polyacrylamide 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Aldridge WN (1953) Serum esterases. Biochem J 53:110–117Google Scholar
  2. Bergmann F, Segal R, Rimon S (1957) A new type of esterase in hog-kidney extract. Biochem J 67:481–486Google Scholar
  3. Bergmann F, Rimon S (1960) Fractionation of C-esterases from the hog's kidney extract. Biochem J 77:209–214Google Scholar
  4. 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 72:248–254Google Scholar
  5. Eubanks EF, Forney FW, Larson AD (1974) Purification and characterization of the nocardial acetylesterase involved in 2-butanone degradation. J Bacteriol 120:1133–1143Google Scholar
  6. Ghisalba O, Auden JA, Schupp T, Nüesch J (1984) The rifamycins: properties, biosynthesis and fermentation. In: Vandamme EJ (ed) Biotechnology of industrial antibiotics. Marcel Dekker Inc., New York and Basel, pp 281–327Google Scholar
  7. Higerd TB, Spizizen J (1973) Isolation of two acetyl esterases from extracts of Bacillus subtilis. J Bacteriol 114:1184–1192Google Scholar
  8. Jansen EF, Jang R, MacDonnell LR (1947) Citrus acetylesterase. Arch Biochem 15:415–431Google Scholar
  9. Kirsch K (1971) Carboxylic ester hydrolases. In: Boyer PD (ed), The enzymes, vol 5. Academic Press Inc., New York, pp 43–69Google Scholar
  10. Låas T, Fast-Johansson A (1979) Isoelectric focussing with PharmalyteTM in gel rods. In: Peeters H (ed) Protides of the biological fluids 27. Pergamon Press, Oxford, pp 693–697Google Scholar
  11. Lämmli UK, Favre M (1973) Maturation of the head of bacteriophage T4. J Mol Biol 80:575–599Google Scholar
  12. Oi S, Satomura Y (1967) Substrate specificity, mode of action, and of inhibition by organic acids of purified acetylesterase from Sclerotinia fungus. Agr Biol Chem 31:561–568Google Scholar
  13. Oi S (1974) Physical and chemical properties of crystalline acetylesterase from Sclerotinia fungus. Agr Biol Chem 38:809–816Google Scholar
  14. Oterholm A, Witter LD, Ordal ZJ (1972) Purification and properties of an acetyl ester hydrolase (acetylesterase) from Lactobacillus plantarum. J Dairy Sc 55:8–13Google Scholar
  15. Ramos Tombo GM, Schär HP, Fernandez I, Busquets X, Ghisalba O (1987) Enantioselective reactions in aqueous and in organic media using carboxyl esterase fractions obtained from crude porcine pancreas lipase preparations. Int. Symp. on Biocatalysis in Org. Media, Wageningen, The Netherlands, 1986, in: Biocatalysis in Organic Media, Elsevier, Amsterdam, pp 43–50Google Scholar
  16. Riefler JF, Higerd TB (1976) Characterization of intracellular esterase A from Bacillus subtilis. Biochem Biophys Acta 429:191–197Google Scholar
  17. Shum AC, Markovetz AJ (1974a) Purification and properties of undecyl esterase from Pseudomonas cepacia grown on 2-tridecanone. J Bacteriol 118:890–897Google Scholar
  18. Schär HP, Chemla P, Ghisalba O (1985) Methanol dehydrogenase from Hyphomicrobium MS 219. FEMS Microbiol Lett 26:117–122Google Scholar

Copyright information

© Springer-Verlag 1988

Authors and Affiliations

  • Hans-Peter Schär
    • 1
  • Daniel Gygax
    • 1
  • Gerardo M. Ramos Tombo
    • 1
  • Oreste Ghisalba
    • 1
  1. 1.Central Research Laboratories of Ciba-Geigy Ltd.BaselSwitzerland

Personalised recommendations