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

Scientific Reports
Zhanliang LiuVincent G H Eijsink

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. Int...Continue Reading

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Citations

Feb 2, 2018·International Journal of Molecular Sciences·Hugo AragundeAntoni Planas
Nov 7, 2017·PloS One·Tina Rise TuvengVincent G H Eijsink
Apr 11, 2019·Polymers·Laia Grifoll-RomeroAntoni Planas
Oct 16, 2019·Frontiers in Bioengineering and Biotechnology·Michal Benedykt KaczmarekMaurycy Daroch
Feb 6, 2020·Marine Drugs·Nathanael D ArnoldThomas B Brück
Mar 3, 2020·Frontiers in Cellular and Infection Microbiology·Isabelle MouynaJean-Paul Latgé
Feb 5, 2021·Environmental Microbiology·Mathias ChoquerNathalie Poussereau
Feb 15, 2020·International Journal of Biological Macromolecules·Yang BaiSheng Yuan
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Oct 9, 2021·Annual Review of Microbiology·Devanshi KhokhaniJean-Michel Ané
Nov 3, 2021·Acta Crystallographica. Section F, Structural Biology Communications·Kohei SasamotoTsutomu Nakamura

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Datasets Mentioned

BETA
AF93062
AAT68493
9

Methods Mentioned

BETA
acetylation
affinity column chromatography
Protein Assay
size exclusion chromatography

Software Mentioned

CCP4
Scala
Chromeleon
Refmac5
PyMod
Mosflm
PyMol
Molrep
Coot

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