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Purification and molecular cloning of porcine intestinal glycerol-ester hydrolase--evidence for its identity with carboxylesterase.
- Source :
-
European journal of biochemistry [Eur J Biochem] 1998 Oct 01; Vol. 257 (1), pp. 142-8. - Publication Year :
- 1998
-
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.
- Subjects :
- Amino Acid Sequence
Animals
Base Sequence
Carbohydrates analysis
Carboxylesterase
Cloning, Molecular
DNA, Complementary
Molecular Sequence Data
Molecular Weight
Monoacylglycerol Lipases chemistry
Monoacylglycerol Lipases genetics
Sequence Homology, Amino Acid
Substrate Specificity
Swine
Carboxylic Ester Hydrolases chemistry
Intestines enzymology
Monoacylglycerol Lipases isolation & purification
Subjects
Details
- Language :
- English
- ISSN :
- 0014-2956
- Volume :
- 257
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- European journal of biochemistry
- Publication Type :
- Academic Journal
- Accession number :
- 9799112
- Full Text :
- https://doi.org/10.1046/j.1432-1327.1998.2570142.x