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Applied Microbiology and Biotechnology

, Volume 94, Issue 4, pp 977–985 | Cite as

PmST3 from Pasteurella multocida encoded by Pm1174 gene is a monofunctional α2–3-sialyltransferase

  • Vireak Thon
  • Yanhong Li
  • Hai Yu
  • Kam Lau
  • Xi ChenEmail author
Biotechnologically relevant enzymes and proteins

Abstract

Pasteurella multocida (Pm) strain Pm70 has three putative sialyltransferase genes including Pm0188, Pm0508, and Pm1174. A Pm0188 gene homolog in Pm strain P-1059 encodes a multifunctional α2–3-sialyltransferase, PmST1, that prefers oligosaccharide acceptors. A Pm0508 gene homolog in the same strain encodes a monofunctional sialyltransferase PmST2 that prefers glycolipid acceptors. Here, we report that the third sialyltransferase from Pm (PmST3) encoded by gene Pm1174 in strain Pm70 is a monofunctional α2–3-sialyltransferase that can use both oligosaccharides and glycolipids as efficient acceptors. Despite the existence of both Pm0188 and Pm0508 gene homologs encoding PmST1 and PmST2, respectively, in Pm strain P-1059, a Pm1174 gene homolog appears to be absent from Pm strains P-1059 and P-934. PmST3 was successfully obtained by cloning and expression using a synthetic gene of Pm1174 with codons optimized for Escherichia coli expression system as the DNA template for polymer chain reactions. Truncation of 35 amino acid residues from the carboxyl terminus was shown to improve the expression of a soluble and active enzyme in E. coli as a C-His6-tagged fusion protein. This sialidase-free monofunctional α2–3-sialyltransferase is a useful tool for synthesizing sialylated oligosaccharides and glycolipids.

Keywords

Carbohydrate synthesis Glycosyltransferase Pasteurella multocida Pasteurella multocida sialyltransferase 3 Sialic acid Sialyltransferase 

Notes

Acknowledgments

This work was support by NSF grant CHE1012511, NIH grant R01HD065122, the Camille Dreyfus Teacher-Scholarship, and the UC-Davis Chancellor's Fellowship.

Supplementary material

253_2011_3676_MOESM1_ESM.docx (212 kb)
ESM 1 (DOCX 211 kb)

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Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Vireak Thon
    • 1
  • Yanhong Li
    • 1
  • Hai Yu
    • 1
  • Kam Lau
    • 1
  • Xi Chen
    • 1
    Email author
  1. 1.Department of ChemistryUniversity of CaliforniaDavisUSA

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