Crystal structure and substrate specificity modification of acetyl xylan esterase from Aspergillus luchuensis

Dai Komiya, Akane Hori, Takuya Ishida, Kiyohiko Igarashi, Masahiro Samejima, Takuya Koseki, Shinya Fushinobu

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6 Citations (Scopus)

Abstract

Acetyl xylan esterase (AXE) catalyzes the hydrolysis of the acetyl bonds present in plant cell wall polysaccharides. Here, we determined the crystal structure of AXE from Aspergillus luchuensis (AlAXEA), providing the three-dimensional structure of an enzyme in the Esterase_phb family. AlAXEA shares its core α/β-hydrolase fold structure with esterases in other families, but it has an extended central β-sheet at both its ends and an extra loop. Structural comparison with a ferulic acid esterase (FAE) from Aspergillus niger indicated that AlAXEA has a conserved catalytic machinery: a catalytic triad (Ser119, His259, and Asp202) and an oxyanion hole (Cys40 and Ser120). Near the catalytic triad of AlAXEA, two aromatic residues (Tyr39 and Trp160) form small pockets at both sides. Homology models of fungal FAEs in the same Esterase_phb family have wide pockets at the corresponding sites because they have residues with smaller side chains (Pro, Ser, and Gly). Mutants with sitedirected mutations at Tyr39 showed a substrate specificity similar to that of the wild-type enzyme, whereas those with mutations at Trp160 acquired an expanded substrate specificity. Interestingly, the Trp160 mutants acquired weak but significant type B-like FAE activity. Moreover, the engineered enzymes exhibited ferulic acid-releasing activity from wheat arabinoxylan.

Original languageEnglish
Article numbere01251-17
JournalApplied and Environmental Microbiology
Volume83
Issue number20
Early online date2017
DOIs
Publication statusPublished - 2017
MoE publication typeA1 Journal article-refereed

Keywords

  • aspergillus luchuensis
  • carboxylesterase
  • catalytic triad
  • enzyme structure
  • ESTHER database
  • X-ray crystallography
  • a/ß-hydrolase fold

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    Komiya, D., Hori, A., Ishida, T., Igarashi, K., Samejima, M., Koseki, T., & Fushinobu, S. (2017). Crystal structure and substrate specificity modification of acetyl xylan esterase from Aspergillus luchuensis. Applied and Environmental Microbiology, 83(20), [e01251-17]. https://doi.org/10.1128/AEM.01251-17