Tannerella forsythia, a Gram-negative member of the Bacteroidetes has evolved to harvest and utilize sialic acid. The most common sialic acid in humans is a mono-N-acetylated version termed Neu5Ac (5-N-acetyl-neuraminic acid). Many bacteria are known to access sialic acid using sialidase enzymes. However, in humans a high proportion of sialic acid contains a second acetyl group attached via an O-group, i.e. chiefly O-acetylated Neu5,9Ac2 or Neu5,4Ac2. This diacetylated sialic acid is not cleaved efficiently by many sialidases and in order to access diacetylated sialic acid, some organisms produce sialate-O-acetylesterases that catalyse the removal of the second acetyl group. In the present study, we performed bioinformatic and biochemical characterization of a putative sialate-O-acetylesterase from T. forsythia (NanS), which contains two putative SGNH-hydrolase domains related to sialate-O-acetylesterases from a range of organisms. Purification of recombinant NanS revealed an esterase that has activity against Neu5,9Ac2 and its glycolyl form Neu5Gc,9Ac. Importantly, the enzyme did not remove acetyl groups positioned at the 4-O position (Neu5,4Ac2). In addition NanS can act upon complex N-glycans released from a glycoprotein [erythropoietin (EPO)], bovine submaxillary mucin and oral epithelial cell-bound glycans. When incubated with its cognate sialidase, NanS increased sialic acid release from mucin and oral epithelial cell surfaces, implying that this esterase improves sialic acid harvesting for this pathogen and potentially other members of the oral microbiome. In summary, we have characterized a novel sialate-O-acetylesterase that contributes to the sialobiology of this important human pathogen and has potential applications in the analysis of sialic acid diacetylation of biologics in the pharmaceutical industry.
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December 2015
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Research Article|
November 13 2015
Characterization of a sialate-O-acetylesterase (NanS) from the oral pathogen Tannerella forsythia that enhances sialic acid release by NanH, its cognate sialidase
Chatchawal Phansopa
;
Chatchawal Phansopa
*Integrated BioSciences, School of Clinical Dentistry, University of Sheffield, Sheffield, S10 2TA, U.K.
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Radoslaw P. Kozak
;
Radoslaw P. Kozak
†Ludger Ltd, Culham Science Centre, Oxfordshire OX14 3EB, U.K.
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Li Phing Liew
;
Li Phing Liew
†Ludger Ltd, Culham Science Centre, Oxfordshire OX14 3EB, U.K.
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Andrew M. Frey
;
Andrew M. Frey
*Integrated BioSciences, School of Clinical Dentistry, University of Sheffield, Sheffield, S10 2TA, U.K.
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Thomas Farmilo
;
Thomas Farmilo
*Integrated BioSciences, School of Clinical Dentistry, University of Sheffield, Sheffield, S10 2TA, U.K.
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Jennifer L. Parker
;
Jennifer L. Parker
*Integrated BioSciences, School of Clinical Dentistry, University of Sheffield, Sheffield, S10 2TA, U.K.
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David J. Kelly
;
David J. Kelly
‡Department of Molecular Biology and Biotechnology, University of Sheffield, Sheffield S10 2TN, U.K.
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Robert J. Emery
;
Robert J. Emery
†Ludger Ltd, Culham Science Centre, Oxfordshire OX14 3EB, U.K.
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Rebecca I. Thomson
;
Rebecca I. Thomson
§Reading School of Pharmacy, University of Reading, Whiteknights, Reading RG6 6AP, U.K.
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Louise Royle
;
Louise Royle
†Ludger Ltd, Culham Science Centre, Oxfordshire OX14 3EB, U.K.
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Richard A. Gardner
;
Richard A. Gardner
†Ludger Ltd, Culham Science Centre, Oxfordshire OX14 3EB, U.K.
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Daniel I.R. Spencer
;
Daniel I.R. Spencer
†Ludger Ltd, Culham Science Centre, Oxfordshire OX14 3EB, U.K.
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Graham P. Stafford
*Integrated BioSciences, School of Clinical Dentistry, University of Sheffield, Sheffield, S10 2TA, U.K.
2To whom correspondence should be addressed (email g.stafford@sheffield.ac.uk).
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Biochem J (2015) 472 (2): 157–167.
Article history
Received:
March 30 2015
Revision Received:
September 03 2015
Accepted:
September 16 2015
Accepted Manuscript online:
September 16 2015
Citation
Chatchawal Phansopa, Radoslaw P. Kozak, Li Phing Liew, Andrew M. Frey, Thomas Farmilo, Jennifer L. Parker, David J. Kelly, Robert J. Emery, Rebecca I. Thomson, Louise Royle, Richard A. Gardner, Daniel I.R. Spencer, Graham P. Stafford; Characterization of a sialate-O-acetylesterase (NanS) from the oral pathogen Tannerella forsythia that enhances sialic acid release by NanH, its cognate sialidase. Biochem J 1 December 2015; 472 (2): 157–167. doi: https://doi.org/10.1042/BJ20150388
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