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Glycobiology Advance Access published online on September 20, 2007
Glycobiology, doi:10.1093/glycob/cwm096
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© The Author 2007. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oxfordjournals.org

The Chitin Catabolic Cascade in the Marine Bacterium Vibrio Cholerae: Characterization of a Unique Chitin Oligosaccharide Deacetylase

Xibing Li1, Lai Xi Wang1, Xuesong Wang2 and Saul Roseman1,*

1 Department of Biology, The Johns Hopkins University, MD 21218; Baltimore
2 Institute of Human Virology, University of Maryland Biotechnology Institute, Maryland 21201, Baltimore

* Address correspondence to: Dr. Saul Roseman, Department of Biology, The Johns Hopkins University, 3400 North Charles Street, Baltimore, MD 21218. E-mail: roseman{at}jhu.edu

Received on June 25, 2007; accepted on September 1, 2007

Chitin, one of the most abundant organic substances in nature, is consumed by marine bacteria, such as Vibrio cholerae, via a multitude of tightly regulated genes (Li and Roseman 2004). One such gene, cod, is reported here. It encodes a chitin oligosaccharide deacetylase (COD), when cells are induced by chitobiose, (GlcNH2)2, or crude crab shells. COD was molecularly cloned (COD-6His), overproduced, and purified to apparent homogeneity. COD is secreted at all stages of growth by induced V. cholerae. The gene sequence predicts a 26 N-terminal amino acid signal peptide not found in the isolated protein. COD is very active with chitin oligosaccharides, is virtually inactive with GlcNAc, and slightly active with colloidal ([3H]-N-acetyl)-chitin. The oligosaccharides are converted almost quantitatively to products lacking one acetyl group. The latter were characterized by mass spectrometry (ESI- MS), and treatment with nitrous acid. COD catalyzes the following reactions (n = 2–6): (GlcNAc)n GlcNAc-GlcNH2- (GlcNAc)n-2 + Ac–. That is, COD hydrolyzes the N-acetyl groups attached to the penultimate GlcNAc residue.

The gene bank sequence data show that cod is highly conserved in Vibrios and Photobacteria. One such gene encodes a deacetylase isolated from V. alginolytics (Ohishi et al. 1997; Ohishi et al. 2000), that is specific for (GlcNAc)2, but inactive with higher oligosaccharides.

The COD enzymatic products, GlcNAc-GlcNH2-(GlcNAc)n, closely resemble those obtained by hydrolysis of the chitooligosaccharides with Nod B: GlcNH2- (GlcNAc)3–4. The latter are key intermediates in the biosynthesis of Nod factors, critically important in communications between the symbiotic nitrogen fixing bacteria and plants. Conceivably, the COD products play equally important roles in cellular communications that remain to be defined.

Key words: Vibrios / chitin oligosaccharide deacetylase / extracellular / penultimate GlcNAc


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[Abstract] [Full Text] [PDF]


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