Purification and biochemical characterization of a turkey preduodenal esterase

Comp Biochem Physiol B Biochem Mol Biol. 2006 Nov-Dec;145(3-4):358-64. doi: 10.1016/j.cbpb.2006.08.011. Epub 2006 Sep 5.

Abstract

A preduodenal esterase was purified to homogeneity from turkey (Meleagris gallopavo) pharyngial tissue. Pure turkey pregastric esterase (TPrE) was obtained after anion exchange chromatography (DEAE-cellulose), Sephacryl S-200 gel filtration, anion exchange chromatography (Mono-Q sepharose) and affinity chromatography (Blue-Gel Affi Gel). The pure enzyme has an apparent molecular mass of 50 kDa, as determined by SDS/PAGE analysis. The optimum pH and temperature for enzyme activity using tributyrin as substrate were 8.5 and 48 degrees C, respectively. Under these conditions, the specific activity measured was 650 U/mg. No significant lipolytic activity was found when was tested on triolein or liprocil as substrates or with monolayer dicaprin with TPrE. In contrast, TPrE displayed a maximal activities of 800, 680 and 520 U/mg with vinyl acetate, vinyl propionate and vinyl butyrate, respectively. This enzyme retained 75% of its maximal activity when incubated for 30 min at pH 4 and 50 degrees C, but it was completely inactivated after an incubation for 10 min at 60 degrees C. The TPrE N-terminal amino acid sequence showed similarities to the N-terminal sequence of a thioesterase from mallard duck, but no similarity with known preduodenal lipases was found.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Bile Acids and Salts / pharmacology
  • Chromatography, Affinity
  • Chromatography, Gel
  • Chromatography, Ion Exchange
  • Crop, Avian / enzymology*
  • Enzyme Stability
  • Esterases / chemistry
  • Esterases / isolation & purification*
  • Esterases / metabolism*
  • Hydrogen-Ion Concentration
  • Molecular Sequence Data
  • Temperature
  • Turkeys / metabolism*

Substances

  • Bile Acids and Salts
  • Esterases