1887

Abstract

(previously named ) is a Gram-negative, anaerobic, fusiform bacterium that is a primary or secondary aetiological agent in periodontal disease in humans. expresses several putative virulence factors, including a sialidase; however, there has been no molecular genetic characterization of this enzyme. A sialidase clone (pHI-1) was screened from a total of 455 recombinant clones of a genomic DNA library using the 2′- (4-methylumbelliferyl)-α---acetylneuraminic acid (MUNeuAc) filter-paper spot assay. The sialidase gene ORF () consists of a 1395 bp coding sequence and encodes a protein with 465 amino acids with an overall molecular mass of 52 kDa. The sialidase does not have sequence similarity to any other bacterial sialidase. The entire sialidase ORF was expressed in . Furthermore, the sialidase was purified from the type strain of and from a recombinant clone, pHI-1 : 1, and was analysed using a non-denaturing gel, revealing that the enzyme preparations were respectively separated as two major bands and as a single band. Southern blot hybridization analysis revealed similar patterns of -hybridizing bands among clinical isolates of from periodontitis patients. This is the first study on the cloning and expression of a sialidase gene and the purification of the SiaHI enzyme from ATCC 43037 and recombinant .

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2003-12-01
2019-11-15
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