Identification, molecular cloning and expression of an α-N-acetylgalactosaminidase gene from Clostridium perfringens

Michael J. Calcutt, Hsin Yeh Hsieh, Linda F. Chapman, Daniel S. Smith

Research output: Contribution to journalArticle

17 Scopus citations

Abstract

The Clostridium perfringens gene encoding the previously characterized α-N-acetylgalactosaminidase (αNAG) was identified by protein microsequencing and database searching. The αNAG protein, designated AagA, was found to be encoded by a hypothetical gene of unknown function in the recently completed genome sequence of C. perfringens strain 13. The deduced translation product of 629 amino acid residues possessed a region of limited homology to several hypothetical open reading frames, an enterotoxin of unknown function and several known or predicted α-galactosidases, but did not exhibit homology to any of the multiple sequenced eukaryotic αNAG proteins. The C. perfringens aagA gene, encoding AagA, was cloned in an Escherichia coli T7 expression system, resulting in recombinants exhibiting high-level expression of the expected αNAG activity. To our knowledge, this is the first report of the cloning and expression of a bacterial αNAG-encoding gene and represents an important step in the development of recombinant αNAG as a tool in the enzymatic conversion of blood group antigens.

Original languageEnglish (US)
Pages (from-to)77-80
Number of pages4
JournalFEMS Microbiology Letters
Volume214
Issue number1
DOIs
StatePublished - Aug 27 2002

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Keywords

  • Blood type A epitope
  • Enzymatic conversion
  • Glycosidase

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology
  • Genetics

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