Decreasing the sialidase activity of multifunctional Pasteurella multocida α2-3-sialyltransferase 1 (PmST1) by site-directed mutagenesis

Go Sugiarto, Kam Lau, Yanhong Li, Zahra Khedri, Hai Yu, Diem Thuy Le, Xi Chen

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

Pasteurella multocida α2-3-sialyltransferase 1 (PmST1) is a multifunctional enzyme which has α2-6-sialyltransferase, α2-3-sialidase, and α2-3-trans-sialidase activities in addition to its major α2-3-sialyltransferase activity. The presence of the α2-3-sialidase activity of PmST1 complicates its application in enzymatic synthesis of α2-3-linked sialosides as the product formed can be hydrolyzed by the enzyme. Herein we show that the α2-3-sialidase activity of PmST1 can be significantly decreased by protein crystal structure-based site-directed mutagenesis. A PmST1 double mutant E271F/R313Y showed a significantly (6333-fold) decreased sialidase activity without affecting its α2-3-sialyltransferase activity. The double mutant E271F/R313Y, therefore, is a superior enzyme for enzymatic synthesis of α2-3-linked sialosides. This journal is

Original languageEnglish (US)
Pages (from-to)3021-3027
Number of pages7
JournalMolecular BioSystems
Volume7
Issue number11
DOIs
StatePublished - Nov 1 2011

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Sialyltransferases
Pasteurella multocida
Neuraminidase
Site-Directed Mutagenesis
Multifunctional Enzymes
Enzymes

ASJC Scopus subject areas

  • Biotechnology
  • Molecular Biology

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Decreasing the sialidase activity of multifunctional Pasteurella multocida α2-3-sialyltransferase 1 (PmST1) by site-directed mutagenesis. / Sugiarto, Go; Lau, Kam; Li, Yanhong; Khedri, Zahra; Yu, Hai; Le, Diem Thuy; Chen, Xi.

In: Molecular BioSystems, Vol. 7, No. 11, 01.11.2011, p. 3021-3027.

Research output: Contribution to journalArticle

Sugiarto, Go ; Lau, Kam ; Li, Yanhong ; Khedri, Zahra ; Yu, Hai ; Le, Diem Thuy ; Chen, Xi. / Decreasing the sialidase activity of multifunctional Pasteurella multocida α2-3-sialyltransferase 1 (PmST1) by site-directed mutagenesis. In: Molecular BioSystems. 2011 ; Vol. 7, No. 11. pp. 3021-3027.
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