Cress and potato soluble epoxide hydrolases: Purification, biochemical characterization, and comparison to mammalian enzymes

Christophe Morisseau, Jeffrey K. Beetham, Franck Pinot, Stéphane Debernard, John W. Newman, Bruce D. Hammock

Research output: Contribution to journalArticle

58 Citations (Scopus)

Abstract

Affinity chromatographic methods were developed for the one-step purification to homogeneity of recombinant soluble epoxide hydrolases (sEHs) from cress and potato. The enzymes are monomeric, with masses of 36 and 39 kDa and pI values of 4.5 and 5.0, respectively. In spite of a large difference in sequence, the two plant enzymes have properties of inhibition and substrate selectivity which differ only slightly from mammalian sEHs. Whereas mammalian sEHs are highly selective for trans- versus cis-substituted stilbene oxide and 1,3-diphenylpropene oxide (DPPO), plant sEHs exhibit far greater selectivity for trans- versus cis-stilbene oxide, but little to no selectivity for DPPO isomers. The isolation of a covalently linked plant sEH- substrate complex indicated that the plant and mammalian sEHs have a similar mechanism of action. We hypothesize an in vivo role for plant sEH in cutin biosynthesis, based on relatively high plant sEH activity on epoxystearate to form a cutin precursor, 9,10-dihydroxystearate. Plant sEHs display a high thermal stability relative to mammalian sEHs. This stability and their high enantioselectivity for a single substrate suggest that their potential as biocatalysts for the preparation of enantiopure epoxides should be evaluated. (C) 2000 Academic Press.

Original languageEnglish (US)
Pages (from-to)321-332
Number of pages12
JournalArchives of Biochemistry and Biophysics
Volume378
Issue number2
DOIs
StatePublished - Jun 15 2000

Fingerprint

Brassicaceae
Epoxide Hydrolases
Solanum tuberosum
Purification
Enzymes
Substrates
Enantioselectivity
Epoxy Compounds
Biosynthesis
Isomers
Oxides
Thermodynamic stability
Hot Temperature

Keywords

  • Affinity purification
  • Cutin biosynthesis
  • Epoxide hydrolase
  • Epoxy fatty acid
  • Mechanism

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Cress and potato soluble epoxide hydrolases : Purification, biochemical characterization, and comparison to mammalian enzymes. / Morisseau, Christophe; Beetham, Jeffrey K.; Pinot, Franck; Debernard, Stéphane; Newman, John W.; Hammock, Bruce D.

In: Archives of Biochemistry and Biophysics, Vol. 378, No. 2, 15.06.2000, p. 321-332.

Research output: Contribution to journalArticle

Morisseau, Christophe ; Beetham, Jeffrey K. ; Pinot, Franck ; Debernard, Stéphane ; Newman, John W. ; Hammock, Bruce D. / Cress and potato soluble epoxide hydrolases : Purification, biochemical characterization, and comparison to mammalian enzymes. In: Archives of Biochemistry and Biophysics. 2000 ; Vol. 378, No. 2. pp. 321-332.
@article{0bddf240dc1943ec968cdc27a1ad7914,
title = "Cress and potato soluble epoxide hydrolases: Purification, biochemical characterization, and comparison to mammalian enzymes",
abstract = "Affinity chromatographic methods were developed for the one-step purification to homogeneity of recombinant soluble epoxide hydrolases (sEHs) from cress and potato. The enzymes are monomeric, with masses of 36 and 39 kDa and pI values of 4.5 and 5.0, respectively. In spite of a large difference in sequence, the two plant enzymes have properties of inhibition and substrate selectivity which differ only slightly from mammalian sEHs. Whereas mammalian sEHs are highly selective for trans- versus cis-substituted stilbene oxide and 1,3-diphenylpropene oxide (DPPO), plant sEHs exhibit far greater selectivity for trans- versus cis-stilbene oxide, but little to no selectivity for DPPO isomers. The isolation of a covalently linked plant sEH- substrate complex indicated that the plant and mammalian sEHs have a similar mechanism of action. We hypothesize an in vivo role for plant sEH in cutin biosynthesis, based on relatively high plant sEH activity on epoxystearate to form a cutin precursor, 9,10-dihydroxystearate. Plant sEHs display a high thermal stability relative to mammalian sEHs. This stability and their high enantioselectivity for a single substrate suggest that their potential as biocatalysts for the preparation of enantiopure epoxides should be evaluated. (C) 2000 Academic Press.",
keywords = "Affinity purification, Cutin biosynthesis, Epoxide hydrolase, Epoxy fatty acid, Mechanism",
author = "Christophe Morisseau and Beetham, {Jeffrey K.} and Franck Pinot and St{\'e}phane Debernard and Newman, {John W.} and Hammock, {Bruce D.}",
year = "2000",
month = "6",
day = "15",
doi = "10.1006/abbi.2000.1810",
language = "English (US)",
volume = "378",
pages = "321--332",
journal = "Archives of Biochemistry and Biophysics",
issn = "0003-9861",
publisher = "Academic Press Inc.",
number = "2",

}

TY - JOUR

T1 - Cress and potato soluble epoxide hydrolases

T2 - Purification, biochemical characterization, and comparison to mammalian enzymes

AU - Morisseau, Christophe

AU - Beetham, Jeffrey K.

AU - Pinot, Franck

AU - Debernard, Stéphane

AU - Newman, John W.

AU - Hammock, Bruce D.

PY - 2000/6/15

Y1 - 2000/6/15

N2 - Affinity chromatographic methods were developed for the one-step purification to homogeneity of recombinant soluble epoxide hydrolases (sEHs) from cress and potato. The enzymes are monomeric, with masses of 36 and 39 kDa and pI values of 4.5 and 5.0, respectively. In spite of a large difference in sequence, the two plant enzymes have properties of inhibition and substrate selectivity which differ only slightly from mammalian sEHs. Whereas mammalian sEHs are highly selective for trans- versus cis-substituted stilbene oxide and 1,3-diphenylpropene oxide (DPPO), plant sEHs exhibit far greater selectivity for trans- versus cis-stilbene oxide, but little to no selectivity for DPPO isomers. The isolation of a covalently linked plant sEH- substrate complex indicated that the plant and mammalian sEHs have a similar mechanism of action. We hypothesize an in vivo role for plant sEH in cutin biosynthesis, based on relatively high plant sEH activity on epoxystearate to form a cutin precursor, 9,10-dihydroxystearate. Plant sEHs display a high thermal stability relative to mammalian sEHs. This stability and their high enantioselectivity for a single substrate suggest that their potential as biocatalysts for the preparation of enantiopure epoxides should be evaluated. (C) 2000 Academic Press.

AB - Affinity chromatographic methods were developed for the one-step purification to homogeneity of recombinant soluble epoxide hydrolases (sEHs) from cress and potato. The enzymes are monomeric, with masses of 36 and 39 kDa and pI values of 4.5 and 5.0, respectively. In spite of a large difference in sequence, the two plant enzymes have properties of inhibition and substrate selectivity which differ only slightly from mammalian sEHs. Whereas mammalian sEHs are highly selective for trans- versus cis-substituted stilbene oxide and 1,3-diphenylpropene oxide (DPPO), plant sEHs exhibit far greater selectivity for trans- versus cis-stilbene oxide, but little to no selectivity for DPPO isomers. The isolation of a covalently linked plant sEH- substrate complex indicated that the plant and mammalian sEHs have a similar mechanism of action. We hypothesize an in vivo role for plant sEH in cutin biosynthesis, based on relatively high plant sEH activity on epoxystearate to form a cutin precursor, 9,10-dihydroxystearate. Plant sEHs display a high thermal stability relative to mammalian sEHs. This stability and their high enantioselectivity for a single substrate suggest that their potential as biocatalysts for the preparation of enantiopure epoxides should be evaluated. (C) 2000 Academic Press.

KW - Affinity purification

KW - Cutin biosynthesis

KW - Epoxide hydrolase

KW - Epoxy fatty acid

KW - Mechanism

UR - http://www.scopus.com/inward/record.url?scp=0034660209&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034660209&partnerID=8YFLogxK

U2 - 10.1006/abbi.2000.1810

DO - 10.1006/abbi.2000.1810

M3 - Article

C2 - 10860549

AN - SCOPUS:0034660209

VL - 378

SP - 321

EP - 332

JO - Archives of Biochemistry and Biophysics

JF - Archives of Biochemistry and Biophysics

SN - 0003-9861

IS - 2

ER -