UDP-Glucose 4-Epimerase and β-1,4-Galactosyltransferase from the Oyster Magallana gigas as Valuable Biocatalysts for the Production of Galactosylated Products

Hui-Bo Song; Meng He; Zhi-Peng Cai; Kun Huang; Sabine L Flitsch; Li Liu; Josef Voglmeir

doi:10.3390/ijms19061600

UDP-Glucose 4-Epimerase and β-1,4-Galactosyltransferase from the Oyster Magallana gigas as Valuable Biocatalysts for the Production of Galactosylated Products

Int J Mol Sci. 2018 May 29;19(6):1600. doi: 10.3390/ijms19061600.

Authors

Hui-Bo Song^{1

2}, Meng He³, Zhi-Peng Cai⁴, Kun Huang⁵, Sabine L Flitsch⁶, Li Liu⁷, Josef Voglmeir⁸

Affiliations

¹ Glycomics and Glycan Bioengineering Research Center (GGBRC), College of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, China. 2013208001@njau.edu.cn.
² Department of Food Science, Zhejiang Pharmaceutical College, Ningbo 315100, China. 2013208001@njau.edu.cn.
³ Glycomics and Glycan Bioengineering Research Center (GGBRC), College of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, China. 2015108040@njau.edu.cn.
⁴ Glycomics and Glycan Bioengineering Research Center (GGBRC), College of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, China. caizhipeng@njau.edu.cn.
⁵ Manchester Institute of Biotechnology, University of Manchester, Manchester M1 7DN, UK. kun.huang-3@manchester.ac.uk.
⁶ Manchester Institute of Biotechnology, University of Manchester, Manchester M1 7DN, UK. sabine.flitsch@manchester.ac.uk.
⁷ Glycomics and Glycan Bioengineering Research Center (GGBRC), College of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, China. lichen.liu@njau.edu.cn.
⁸ Glycomics and Glycan Bioengineering Research Center (GGBRC), College of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, China. josef.voglmeir@njau.edu.cn.

Abstract

Uridine diphosphate galactose (UDP-galactose) is a valuable building block in the enzymatic synthesis of galactose-containing glycoconjugates. UDP-glucose 4-epimerase (UGE) is an enzyme which catalyzes the reversible conversion of abundantly available UDP-glucose to UDP-galactose. Herein, we described the cloning, expression, purification, and biochemical characterization of an unstudied UGE from the oyster Magallana gigas (MgUGE). Activity tests of recombinantly expressed MgUGE, using HPLC (high-performance liquid chromatography), mass spectrometry, and photometric assays, showed an optimal temperature of 16 °C, and reasonable thermal stability up to 37 °C. No metal ions were required for enzymatic activity. The simple nickel-affinity-purification procedure makes MgUGE a valuable biocatalyst for the synthesis of UDP-galactose from UDP-glucose. The biosynthetic potential of MgUGE was further exemplified in a coupled enzymatic reaction with an oyster-derived β-1,4-galactosyltransferase (MgGalT7), allowing the galactosylation of the model substrate para-nitrophenol xylose (pNP-xylose) using UDP-glucose as the starting material.

Keywords: Magallana gigas; UDP-galactose; UDP-glucose 4-epimerase; oyster metabolism.

MeSH terms

Animals
Galactosyltransferases / metabolism*
Glycoconjugates / biosynthesis*
Ostreidae / enzymology*
UDPglucose 4-Epimerase / metabolism*
Uridine Diphosphate Galactose / metabolism

Substances

Glycoconjugates
Uridine Diphosphate Galactose
Galactosyltransferases
UDPglucose 4-Epimerase