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 language | English (US) |
|---|---|
| Pages (from-to) | 1131-1135 |
| Number of pages | 5 |
| Journal | Biotechnology Letters |
| Volume | 21 |
| Issue number | 12 |
| DOIs | |
| State | Published - 1999 |
| Externally published | Yes |
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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 journal › Article
}
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 -