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

Vireak Thon, Yanhong Li, Hai Yu, Kam Lau, Xi Chen

Research output: Contribution to journalArticle

14 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)977-985
Number of pages9
JournalApplied Microbiology and Biotechnology
Volume94
Issue number4
DOIs
StatePublished - May 2012

Fingerprint

Sialyltransferases
Pasteurella multocida
Glycolipids
Oligosaccharides
Genes
Escherichia coli
Synthetic Genes
Neuraminidase
Codon
Organism Cloning
Polymers
Amino Acids
DNA
Enzymes

Keywords

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

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology

Cite this

PmST3 from Pasteurella multocida encoded by Pm1174 gene is a monofunctional α2-3-sialyltransferase. / Thon, Vireak; Li, Yanhong; Yu, Hai; Lau, Kam; Chen, Xi.

In: Applied Microbiology and Biotechnology, Vol. 94, No. 4, 05.2012, p. 977-985.

Research output: Contribution to journalArticle

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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.",
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T1 - PmST3 from Pasteurella multocida encoded by Pm1174 gene is a monofunctional α2-3-sialyltransferase

AU - Thon, Vireak

AU - Li, Yanhong

AU - Yu, Hai

AU - Lau, Kam

AU - Chen, Xi

PY - 2012/5

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N2 - 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.

AB - 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.

KW - Carbohydrate synthesis

KW - Glycosyltransferase

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KW - Pasteurella multocida sialyltransferase 3

KW - Sialic acid

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