Structure and function of a broad-specificity chitin deacetylase from Aspergillus nidulans FGSC A4

Zhanliang Liu, Laurie M. Gay, Tina R. Tuveng, Jane W. Agger, Bjørge Westereng, Geir Mathiesen, Svein J. Horn, Gustav Vaaje-Kolstad, Daan M. F. van Aalten, Vincent G. H. Eijsink (Lead / Corresponding author)

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Abstract

Enzymatic conversion of chitin, a β-1,4 linked polymer of N-acetylglucosamine, is of major interest in areas varying from the biorefining of chitin-rich waste streams to understanding how medically relevant fungi remodel their chitin-containing cell walls. Although numerous chitinolytic enzymes have been studied in detail, relatively little is known about enzymes capable of deacetylating chitin. We describe the structural and functional characterization of a 237 residue deacetylase (AnCDA) from Aspergillus nidulans FGSC A4. AnCDA acts on chito-oligomers, crystalline chitin, chitosan, and acetylxylan, but not on peptidoglycan. The K m and k cat of AnCDA for the first deacetylation of penta-N-acetyl-chitopentaose are 72 µM and 1.4 s(-1), respectively. Combining mass spectrometry and analyses of acetate release, it was shown that AnCDA catalyses mono-deacetylation of (GlcNAc)2 and full deacetylation of (GlcNAc)3-6 in a non-processive manner. Deacetylation of the reducing end sugar was much slower than deacetylation of the other sugars in chito-oligomers. These enzymatic characteristics are discussed in the light of the crystal structure of AnCDA, providing insight into how the chitin deacetylase may interact with its substrates. Interestingly, AnCDA activity on crystalline chitin was enhanced by a lytic polysaccharide monooxygenase that increases substrate accessibility by oxidative cleavage of the chitin chains.

Original languageEnglish
Article number1746
Pages (from-to)1-12
Number of pages12
JournalScientific Reports
Volume7
DOIs
Publication statusPublished - 11 May 2017

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chitin deacetylase
Aspergillus nidulans
Chitin
Acetylglucosamine
Peptidoglycan
Chitosan
Enzymes
Mixed Function Oxygenases
Cell Wall
Polysaccharides
Mass Spectrometry
Polymers
Acetates
Cats
Fungi

Keywords

  • Journal article
  • Enzymes
  • Polysaccharides
  • X-ray crystallography

Cite this

Liu, Z., Gay, L. M., Tuveng, T. R., Agger, J. W., Westereng, B., Mathiesen, G., ... Eijsink, V. G. H. (2017). Structure and function of a broad-specificity chitin deacetylase from Aspergillus nidulans FGSC A4. Scientific Reports, 7, 1-12. [1746]. https://doi.org/10.1038/s41598-017-02043-1
Liu, Zhanliang ; Gay, Laurie M. ; Tuveng, Tina R. ; Agger, Jane W. ; Westereng, Bjørge ; Mathiesen, Geir ; Horn, Svein J. ; Vaaje-Kolstad, Gustav ; van Aalten, Daan M. F. ; Eijsink, Vincent G. H. / Structure and function of a broad-specificity chitin deacetylase from Aspergillus nidulans FGSC A4. In: Scientific Reports. 2017 ; Vol. 7. pp. 1-12.
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abstract = "Enzymatic conversion of chitin, a β-1,4 linked polymer of N-acetylglucosamine, is of major interest in areas varying from the biorefining of chitin-rich waste streams to understanding how medically relevant fungi remodel their chitin-containing cell walls. Although numerous chitinolytic enzymes have been studied in detail, relatively little is known about enzymes capable of deacetylating chitin. We describe the structural and functional characterization of a 237 residue deacetylase (AnCDA) from Aspergillus nidulans FGSC A4. AnCDA acts on chito-oligomers, crystalline chitin, chitosan, and acetylxylan, but not on peptidoglycan. The K m and k cat of AnCDA for the first deacetylation of penta-N-acetyl-chitopentaose are 72 µM and 1.4 s(-1), respectively. Combining mass spectrometry and analyses of acetate release, it was shown that AnCDA catalyses mono-deacetylation of (GlcNAc)2 and full deacetylation of (GlcNAc)3-6 in a non-processive manner. Deacetylation of the reducing end sugar was much slower than deacetylation of the other sugars in chito-oligomers. These enzymatic characteristics are discussed in the light of the crystal structure of AnCDA, providing insight into how the chitin deacetylase may interact with its substrates. Interestingly, AnCDA activity on crystalline chitin was enhanced by a lytic polysaccharide monooxygenase that increases substrate accessibility by oxidative cleavage of the chitin chains.",
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note = "This work was supported by grants from the Norwegian Research Council (grant numbers 164653, 197388 and 214138). Daan van Aalten is supported by an MRC Programme Grant M004139.",
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Liu, Z, Gay, LM, Tuveng, TR, Agger, JW, Westereng, B, Mathiesen, G, Horn, SJ, Vaaje-Kolstad, G, van Aalten, DMF & Eijsink, VGH 2017, 'Structure and function of a broad-specificity chitin deacetylase from Aspergillus nidulans FGSC A4', Scientific Reports, vol. 7, 1746, pp. 1-12. https://doi.org/10.1038/s41598-017-02043-1

Structure and function of a broad-specificity chitin deacetylase from Aspergillus nidulans FGSC A4. / Liu, Zhanliang; Gay, Laurie M.; Tuveng, Tina R.; Agger, Jane W.; Westereng, Bjørge; Mathiesen, Geir; Horn, Svein J.; Vaaje-Kolstad, Gustav; van Aalten, Daan M. F.; Eijsink, Vincent G. H. (Lead / Corresponding author).

In: Scientific Reports, Vol. 7, 1746, 11.05.2017, p. 1-12.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Structure and function of a broad-specificity chitin deacetylase from Aspergillus nidulans FGSC A4

AU - Liu, Zhanliang

AU - Gay, Laurie M.

AU - Tuveng, Tina R.

AU - Agger, Jane W.

AU - Westereng, Bjørge

AU - Mathiesen, Geir

AU - Horn, Svein J.

AU - Vaaje-Kolstad, Gustav

AU - van Aalten, Daan M. F.

AU - Eijsink, Vincent G. H.

N1 - This work was supported by grants from the Norwegian Research Council (grant numbers 164653, 197388 and 214138). Daan van Aalten is supported by an MRC Programme Grant M004139.

PY - 2017/5/11

Y1 - 2017/5/11

N2 - Enzymatic conversion of chitin, a β-1,4 linked polymer of N-acetylglucosamine, is of major interest in areas varying from the biorefining of chitin-rich waste streams to understanding how medically relevant fungi remodel their chitin-containing cell walls. Although numerous chitinolytic enzymes have been studied in detail, relatively little is known about enzymes capable of deacetylating chitin. We describe the structural and functional characterization of a 237 residue deacetylase (AnCDA) from Aspergillus nidulans FGSC A4. AnCDA acts on chito-oligomers, crystalline chitin, chitosan, and acetylxylan, but not on peptidoglycan. The K m and k cat of AnCDA for the first deacetylation of penta-N-acetyl-chitopentaose are 72 µM and 1.4 s(-1), respectively. Combining mass spectrometry and analyses of acetate release, it was shown that AnCDA catalyses mono-deacetylation of (GlcNAc)2 and full deacetylation of (GlcNAc)3-6 in a non-processive manner. Deacetylation of the reducing end sugar was much slower than deacetylation of the other sugars in chito-oligomers. These enzymatic characteristics are discussed in the light of the crystal structure of AnCDA, providing insight into how the chitin deacetylase may interact with its substrates. Interestingly, AnCDA activity on crystalline chitin was enhanced by a lytic polysaccharide monooxygenase that increases substrate accessibility by oxidative cleavage of the chitin chains.

AB - Enzymatic conversion of chitin, a β-1,4 linked polymer of N-acetylglucosamine, is of major interest in areas varying from the biorefining of chitin-rich waste streams to understanding how medically relevant fungi remodel their chitin-containing cell walls. Although numerous chitinolytic enzymes have been studied in detail, relatively little is known about enzymes capable of deacetylating chitin. We describe the structural and functional characterization of a 237 residue deacetylase (AnCDA) from Aspergillus nidulans FGSC A4. AnCDA acts on chito-oligomers, crystalline chitin, chitosan, and acetylxylan, but not on peptidoglycan. The K m and k cat of AnCDA for the first deacetylation of penta-N-acetyl-chitopentaose are 72 µM and 1.4 s(-1), respectively. Combining mass spectrometry and analyses of acetate release, it was shown that AnCDA catalyses mono-deacetylation of (GlcNAc)2 and full deacetylation of (GlcNAc)3-6 in a non-processive manner. Deacetylation of the reducing end sugar was much slower than deacetylation of the other sugars in chito-oligomers. These enzymatic characteristics are discussed in the light of the crystal structure of AnCDA, providing insight into how the chitin deacetylase may interact with its substrates. Interestingly, AnCDA activity on crystalline chitin was enhanced by a lytic polysaccharide monooxygenase that increases substrate accessibility by oxidative cleavage of the chitin chains.

KW - Journal article

KW - Enzymes

KW - Polysaccharides

KW - X-ray crystallography

U2 - 10.1038/s41598-017-02043-1

DO - 10.1038/s41598-017-02043-1

M3 - Article

VL - 7

SP - 1

EP - 12

JO - Scientific Reports

JF - Scientific Reports

SN - 2045-2322

M1 - 1746

ER -