Purification and renaturation of membrane neuraminidase from Haemophilus parasuis

Carol A. Lichtensteiger, Eric R. Vimr

Research output: Contribution to journalArticlepeer-review

Abstract

Haemophilus parasuis, which causes polyserositis, polysynovitis, meningitis, septicemia, and pneumonia in pigs, has emerged as an increasing problem in modern swine production systems. Co-factors for and the pathogenesis of H. parasuis disease are not defined. One of the potential virulence factors of H. parasuis is its neuraminidase (sialidase). While purifying the H. parasuis neuraminidase from the membrane fraction, we developed a protocol to renature enzymatic activity after enzyme preparations were resolved electrophorectically in denaturing polyacrylamide gels. The H. parasuis neuraminidase co-resolved with recombinant neuraminidase of Vibrio cholera; thus its apparent molecular mass is 82kilodalton (kDa). The H. parasuis neuraminidase was associated with the membrane fraction and the purification protocol removed over 99% of the H. parasuis cell protein while retaining over 90% of the neuraminidase activity. Purified protein will provide another avenue to clone the neuraminidase gene that has been refractory to cloning and the protocol will be a means to purify recombinant protein.

Original languageEnglish (US)
Pages (from-to)79-87
Number of pages9
JournalVeterinary Microbiology
Volume93
Issue number1
DOIs
StatePublished - May 2 2003

Keywords

  • Haemophilus parasuis
  • Neuraminidase
  • Pig-bacteria
  • Protein purification
  • Protein renaturization
  • Sialidase

ASJC Scopus subject areas

  • Microbiology
  • veterinary(all)

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