Cloning, expression and characterization of a UDP-galactose 4-epimerase from Escherichia coli

Xi Chen, Przemyslaw Kowal, Sarah Hamad, Hongni Fan, Peng George Wang

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

The gene galE encoding UDP-galactose 4-epimerase was cloned into E. coli BL21(DE3) from the chromosomal DNA of E. coli strain K-12. High expression of the soluble recombinant epimerase was achieved in the cell lysate. In order to evaluate the use of this epimerase in enzymatic synthesis of important α- Gal epitopes (oligosaccharides with a terminal Galα1,3Gal sequence), a new radioactivity assay (α1,3-galactosyltransferase coupled assay) was established to characterize its activity in producing UDP-galactose from UDP- glucose. Approximately 2700 units (100 mg) enzyme with a specific activity of 27 U mg-1 protein could be obtained from one liter of bacterial culture. The epimerase was active in a wide pH range with an optimum at pH 7.0. This expression system established a viable route to the enzymatic production of α-Gal oligosaccharides to support xenotransplantation research.

Original languageEnglish (US)
Pages (from-to)1131-1135
Number of pages5
JournalBiotechnology Letters
Volume21
Issue number12
DOIs
StatePublished - 1999
Externally publishedYes

Fingerprint

UDPglucose 4-Epimerase
Racemases and Epimerases
Oligosaccharides
Cloning
Escherichia coli
Organism Cloning
Assays
Epitopes
Gene encoding
Radioactivity
Uridine Diphosphate Galactose
Galactosyltransferases
Uridine Diphosphate Glucose
Glucose
Heterologous Transplantation
DNA
Enzymes
Proteins
Research
Genes

Keywords

  • α 1,3-galactosyltransferase
  • Cloning
  • E. coli
  • Expression
  • UDP-galactose 4- epimerase

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology
  • Microbiology

Cite this

Cloning, expression and characterization of a UDP-galactose 4-epimerase from Escherichia coli. / Chen, Xi; Kowal, Przemyslaw; Hamad, Sarah; Fan, Hongni; Wang, Peng George.

In: Biotechnology Letters, Vol. 21, No. 12, 1999, p. 1131-1135.

Research output: Contribution to journalArticle

Chen, Xi ; Kowal, Przemyslaw ; Hamad, Sarah ; Fan, Hongni ; Wang, Peng George. / Cloning, expression and characterization of a UDP-galactose 4-epimerase from Escherichia coli. In: Biotechnology Letters. 1999 ; Vol. 21, No. 12. pp. 1131-1135.
@article{42a22b99231c4c5aa5960671db9ec132,
title = "Cloning, expression and characterization of a UDP-galactose 4-epimerase from Escherichia coli",
abstract = "The gene galE encoding UDP-galactose 4-epimerase was cloned into E. coli BL21(DE3) from the chromosomal DNA of E. coli strain K-12. High expression of the soluble recombinant epimerase was achieved in the cell lysate. In order to evaluate the use of this epimerase in enzymatic synthesis of important α- Gal epitopes (oligosaccharides with a terminal Galα1,3Gal sequence), a new radioactivity assay (α1,3-galactosyltransferase coupled assay) was established to characterize its activity in producing UDP-galactose from UDP- glucose. Approximately 2700 units (100 mg) enzyme with a specific activity of 27 U mg-1 protein could be obtained from one liter of bacterial culture. The epimerase was active in a wide pH range with an optimum at pH 7.0. This expression system established a viable route to the enzymatic production of α-Gal oligosaccharides to support xenotransplantation research.",
keywords = "α 1,3-galactosyltransferase, Cloning, E. coli, Expression, UDP-galactose 4- epimerase",
author = "Xi Chen and Przemyslaw Kowal and Sarah Hamad and Hongni Fan and Wang, {Peng George}",
year = "1999",
doi = "10.1023/A:1005678225031",
language = "English (US)",
volume = "21",
pages = "1131--1135",
journal = "Biotechnology Letters",
issn = "0141-5492",
publisher = "Springer Netherlands",
number = "12",

}

TY - JOUR

T1 - Cloning, expression and characterization of a UDP-galactose 4-epimerase from Escherichia coli

AU - Chen, Xi

AU - Kowal, Przemyslaw

AU - Hamad, Sarah

AU - Fan, Hongni

AU - Wang, Peng George

PY - 1999

Y1 - 1999

N2 - The gene galE encoding UDP-galactose 4-epimerase was cloned into E. coli BL21(DE3) from the chromosomal DNA of E. coli strain K-12. High expression of the soluble recombinant epimerase was achieved in the cell lysate. In order to evaluate the use of this epimerase in enzymatic synthesis of important α- Gal epitopes (oligosaccharides with a terminal Galα1,3Gal sequence), a new radioactivity assay (α1,3-galactosyltransferase coupled assay) was established to characterize its activity in producing UDP-galactose from UDP- glucose. Approximately 2700 units (100 mg) enzyme with a specific activity of 27 U mg-1 protein could be obtained from one liter of bacterial culture. The epimerase was active in a wide pH range with an optimum at pH 7.0. This expression system established a viable route to the enzymatic production of α-Gal oligosaccharides to support xenotransplantation research.

AB - The gene galE encoding UDP-galactose 4-epimerase was cloned into E. coli BL21(DE3) from the chromosomal DNA of E. coli strain K-12. High expression of the soluble recombinant epimerase was achieved in the cell lysate. In order to evaluate the use of this epimerase in enzymatic synthesis of important α- Gal epitopes (oligosaccharides with a terminal Galα1,3Gal sequence), a new radioactivity assay (α1,3-galactosyltransferase coupled assay) was established to characterize its activity in producing UDP-galactose from UDP- glucose. Approximately 2700 units (100 mg) enzyme with a specific activity of 27 U mg-1 protein could be obtained from one liter of bacterial culture. The epimerase was active in a wide pH range with an optimum at pH 7.0. This expression system established a viable route to the enzymatic production of α-Gal oligosaccharides to support xenotransplantation research.

KW - α 1,3-galactosyltransferase

KW - Cloning

KW - E. coli

KW - Expression

KW - UDP-galactose 4- epimerase

UR - http://www.scopus.com/inward/record.url?scp=0033392291&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033392291&partnerID=8YFLogxK

U2 - 10.1023/A:1005678225031

DO - 10.1023/A:1005678225031

M3 - Article

AN - SCOPUS:0033392291

VL - 21

SP - 1131

EP - 1135

JO - Biotechnology Letters

JF - Biotechnology Letters

SN - 0141-5492

IS - 12

ER -