Glycoside hydrolase DisH from Desulfovibrio vulgaris degrades the N-acetylgalactosamine component of diverse biofilms

Lei Zhu, Venkata G. Poosarla, Sooyeon Song, Thammajun Leungsakul Wood, Daniel S. Miller, Bei Yin, Thomas Keith Wood

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Biofilms of sulfate-reducing bacteria (SRB) produce H2S, which contributes to corrosion. Although bacterial cells in biofilms are cemented together, they often dissolve their own biofilm to allow the cells to disperse. Using Desulfovibrio vulgaris as a model SRB, we sought polysaccharide-degrading enzymes that disperse its biofilm. Using a whole-genome approach, we identified eight enzymes as putative extracellular glycoside hydrolases including DisH (DVU2239, dispersal hexosaminidase), an enzyme that we demonstrated here, by utilizing various p-nitrooligosaccharide substrates, to be an N-acetyl-β-D-hexosaminidase. For N-acetyl-β-D-galactosamine (GalNAc), Vmax was 3.6 µmol of p-nitrophenyl/min (mg protein)−1 and Km was 0.8 mM; the specific activity for N-acetyl β-D-glucosamine (GlcNAc) was 7.8 µmol of p-nitrophenyl/min (mg protein)−1. Since GalNAc is one of the three exopolysaccharide matrix components of D. vulgaris, purified DisH was found to disperse 63 ± 2% biofilm as well as inhibit biofilm formation up to 47 ± 4%. The temperature and pH optima are 60°C and pH 6, respectively; DisH is also inhibited by copper and is secreted. In addition, since polymers of GalNAc and GlcNAc are found in the matrix of diverse bacteria, DisH dispersed biofilms of Pseudomonas aeruginosa, Escherichia coli and Bacillus subtilis. Therefore, DisH has the potential to inhibit and disperse a wide-range of biofilms.

Original languageEnglish (US)
Pages (from-to)2026-2037
Number of pages12
JournalEnvironmental microbiology
Volume20
Issue number6
DOIs
StatePublished - Jun 1 2018

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Desulfovibrio vulgaris
beta-N-acetylhexosaminidase
Acetylgalactosamine
Hexosaminidases
Glycoside Hydrolases
Biofilms
hydrolases
biofilm
glycosides
sulfate-reducing bacteria
sulfate-reducing bacterium
enzyme
Bacteria
Sulfates
Enzymes
enzymes
galactosamine
matrix
protein
Corrosion

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Ecology, Evolution, Behavior and Systematics

Cite this

@article{605b432be9794ace8f6f295f9929db56,
title = "Glycoside hydrolase DisH from Desulfovibrio vulgaris degrades the N-acetylgalactosamine component of diverse biofilms",
abstract = "Biofilms of sulfate-reducing bacteria (SRB) produce H2S, which contributes to corrosion. Although bacterial cells in biofilms are cemented together, they often dissolve their own biofilm to allow the cells to disperse. Using Desulfovibrio vulgaris as a model SRB, we sought polysaccharide-degrading enzymes that disperse its biofilm. Using a whole-genome approach, we identified eight enzymes as putative extracellular glycoside hydrolases including DisH (DVU2239, dispersal hexosaminidase), an enzyme that we demonstrated here, by utilizing various p-nitrooligosaccharide substrates, to be an N-acetyl-β-D-hexosaminidase. For N-acetyl-β-D-galactosamine (GalNAc), Vmax was 3.6 µmol of p-nitrophenyl/min (mg protein)−1 and Km was 0.8 mM; the specific activity for N-acetyl β-D-glucosamine (GlcNAc) was 7.8 µmol of p-nitrophenyl/min (mg protein)−1. Since GalNAc is one of the three exopolysaccharide matrix components of D. vulgaris, purified DisH was found to disperse 63 ± 2{\%} biofilm as well as inhibit biofilm formation up to 47 ± 4{\%}. The temperature and pH optima are 60°C and pH 6, respectively; DisH is also inhibited by copper and is secreted. In addition, since polymers of GalNAc and GlcNAc are found in the matrix of diverse bacteria, DisH dispersed biofilms of Pseudomonas aeruginosa, Escherichia coli and Bacillus subtilis. Therefore, DisH has the potential to inhibit and disperse a wide-range of biofilms.",
author = "Lei Zhu and Poosarla, {Venkata G.} and Sooyeon Song and Wood, {Thammajun Leungsakul} and Miller, {Daniel S.} and Bei Yin and Wood, {Thomas Keith}",
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Glycoside hydrolase DisH from Desulfovibrio vulgaris degrades the N-acetylgalactosamine component of diverse biofilms. / Zhu, Lei; Poosarla, Venkata G.; Song, Sooyeon; Wood, Thammajun Leungsakul; Miller, Daniel S.; Yin, Bei; Wood, Thomas Keith.

In: Environmental microbiology, Vol. 20, No. 6, 01.06.2018, p. 2026-2037.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Glycoside hydrolase DisH from Desulfovibrio vulgaris degrades the N-acetylgalactosamine component of diverse biofilms

AU - Zhu, Lei

AU - Poosarla, Venkata G.

AU - Song, Sooyeon

AU - Wood, Thammajun Leungsakul

AU - Miller, Daniel S.

AU - Yin, Bei

AU - Wood, Thomas Keith

PY - 2018/6/1

Y1 - 2018/6/1

N2 - Biofilms of sulfate-reducing bacteria (SRB) produce H2S, which contributes to corrosion. Although bacterial cells in biofilms are cemented together, they often dissolve their own biofilm to allow the cells to disperse. Using Desulfovibrio vulgaris as a model SRB, we sought polysaccharide-degrading enzymes that disperse its biofilm. Using a whole-genome approach, we identified eight enzymes as putative extracellular glycoside hydrolases including DisH (DVU2239, dispersal hexosaminidase), an enzyme that we demonstrated here, by utilizing various p-nitrooligosaccharide substrates, to be an N-acetyl-β-D-hexosaminidase. For N-acetyl-β-D-galactosamine (GalNAc), Vmax was 3.6 µmol of p-nitrophenyl/min (mg protein)−1 and Km was 0.8 mM; the specific activity for N-acetyl β-D-glucosamine (GlcNAc) was 7.8 µmol of p-nitrophenyl/min (mg protein)−1. Since GalNAc is one of the three exopolysaccharide matrix components of D. vulgaris, purified DisH was found to disperse 63 ± 2% biofilm as well as inhibit biofilm formation up to 47 ± 4%. The temperature and pH optima are 60°C and pH 6, respectively; DisH is also inhibited by copper and is secreted. In addition, since polymers of GalNAc and GlcNAc are found in the matrix of diverse bacteria, DisH dispersed biofilms of Pseudomonas aeruginosa, Escherichia coli and Bacillus subtilis. Therefore, DisH has the potential to inhibit and disperse a wide-range of biofilms.

AB - Biofilms of sulfate-reducing bacteria (SRB) produce H2S, which contributes to corrosion. Although bacterial cells in biofilms are cemented together, they often dissolve their own biofilm to allow the cells to disperse. Using Desulfovibrio vulgaris as a model SRB, we sought polysaccharide-degrading enzymes that disperse its biofilm. Using a whole-genome approach, we identified eight enzymes as putative extracellular glycoside hydrolases including DisH (DVU2239, dispersal hexosaminidase), an enzyme that we demonstrated here, by utilizing various p-nitrooligosaccharide substrates, to be an N-acetyl-β-D-hexosaminidase. For N-acetyl-β-D-galactosamine (GalNAc), Vmax was 3.6 µmol of p-nitrophenyl/min (mg protein)−1 and Km was 0.8 mM; the specific activity for N-acetyl β-D-glucosamine (GlcNAc) was 7.8 µmol of p-nitrophenyl/min (mg protein)−1. Since GalNAc is one of the three exopolysaccharide matrix components of D. vulgaris, purified DisH was found to disperse 63 ± 2% biofilm as well as inhibit biofilm formation up to 47 ± 4%. The temperature and pH optima are 60°C and pH 6, respectively; DisH is also inhibited by copper and is secreted. In addition, since polymers of GalNAc and GlcNAc are found in the matrix of diverse bacteria, DisH dispersed biofilms of Pseudomonas aeruginosa, Escherichia coli and Bacillus subtilis. Therefore, DisH has the potential to inhibit and disperse a wide-range of biofilms.

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