Characterisation of a 1,4-β-fucoside hydrolase degrading colanic acid

René Verhoef, Gerrit Beldman, Henk A. Schols, Matti Siika-aho, Marjaana Rättö, Johanna Buchert, Alphons G.J. Voragen (Corresponding Author)

Research output: Contribution to journalArticleScientificpeer-review

13 Citations (Scopus)

Abstract

A novel colanic acid-degrading enzyme was isolated from a mixed culture filtrate obtained by enrichment culturing of a compost sample using colanic acid as carbon source. The enzyme was partially purified resulting in a 17-fold increase in specific activity. Further purification by Native PAGE revealed that the enzyme is part of a high-molecular weight multi protein complex of at least six individual proteins. The enzyme showed a temperature optimum at 50 °C while after 5 h at 50 °C and pH 7 still 70% of the total activity was left. The pH optimum was found to be pH 7. Analysis of the degradation products showed that the enzyme is a novel 1,4-β-fucoside hydrolase that liberates repeating units of colanic acid with varying degrees of acetylation. Km and Vmax of the enzyme were determined against the native substrate as well as its de-O-acetylated and depyruvated forms. Compared to the native substrate the affinity of the enzyme for the modified substrates was much lower. However, for the de-O-acetylated sample a dramatic increase in catalytic efficiency was observed. The native form of the substrate showed substrate inhibition at high concentrations, probably due to the formation of nonproductive substrate complexes. Removal of the acetyl groups probably prevents this effect resulting in a higher catalytic efficiency.
Original languageEnglish
Pages (from-to)1780 - 1788
Number of pages9
JournalCarbohydrate Research
Volume340
Issue number11
DOIs
Publication statusPublished - 2005
MoE publication typeA1 Journal article-refereed

Keywords

  • Colanic acid
  • Exopolysaccharide (EPS)
  • Enzymes
  • Fucoside hydrolase
  • Paper industry
  • Biofouling

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    Verhoef, R., Beldman, G., Schols, H. A., Siika-aho, M., Rättö, M., Buchert, J., & Voragen, A. G. J. (2005). Characterisation of a 1,4-β-fucoside hydrolase degrading colanic acid. Carbohydrate Research, 340(11), 1780 - 1788. https://doi.org/10.1016/j.carres.2005.06.003