Purification and molecular cloning of porcine intestinal glycerol-ester hydrolase--evidence for its identity with carboxylesterase

Eur J Biochem. 1998 Oct 1;257(1):142-8. doi: 10.1046/j.1432-1327.1998.2570142.x.

Abstract

A glycerol-ester hydrolase was purified to homogeneity from porcine intestinal mucosa using a partial delipidation method and an eight-step purification procedure. The isolation scheme used gave a 483-fold purification, resulting in a pure enzyme with a specific activity on tributyrin of 290 micromol x min(-1) x mg(-1). The molecular mass of the enzyme was estimated at 240 kDa, based on the results of size-exclusion chromatography, and at 60 kDa, as determined by SDS/PAGE analysis. The isoelectric focusing data obtained indicated that only one isoform with a pI of 5.1 was present. Complete identity was found to exist between the N-terminal sequence of the first 25 amino acid residues and that of a porcine liver carboxylesterase. A full-length cDNA coding for the enzyme was isolated from pig small intestine. We observed that the corresponding protein originally named intestinal glycerol-ester hydrolase definitely belongs to the carboxylesterase family. The deduced amino acid sequence consisted of 565 residues and showed 97% identity with that of porcine liver carboxylesterase and more than 50% identity with those of other carboxylesterases from different mammalian species.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Carbohydrates / analysis
  • Carboxylesterase
  • Carboxylic Ester Hydrolases / chemistry*
  • Cloning, Molecular
  • DNA, Complementary
  • Intestines / enzymology*
  • Molecular Sequence Data
  • Molecular Weight
  • Monoacylglycerol Lipases / chemistry
  • Monoacylglycerol Lipases / genetics
  • Monoacylglycerol Lipases / isolation & purification*
  • Sequence Homology, Amino Acid
  • Substrate Specificity
  • Swine

Substances

  • Carbohydrates
  • DNA, Complementary
  • Carboxylic Ester Hydrolases
  • Carboxylesterase
  • Monoacylglycerol Lipases

Associated data

  • GENBANK/AF064741