Improvement of GH10 family xylanase thermostability by introducing of an extra α-helix at the C-terminal

Biochem Biophys Res Commun. 2019 Jul 30;515(3):417-422. doi: 10.1016/j.bbrc.2019.05.163. Epub 2019 May 31.

Abstract

Xylanase is an important enzyme in industrial applications, which usually require the enzyme to maintain activity in high-temperature condition. In this study, a GH10 family xylanase XynAF0 from a thermophilic composting fungus, Aspergillus fumigatus Z5, was investigated to determine its thermostable mechanism. XynAF0 showed excellent thermostability, which could maintain 50% relative activity after incubation for 1 h at 70 °C. The homologous modeling structure of XynAF0 was constructed and an α-helix composed of poly-threonine has been found in the linker region between the catalytic domain and the carbohydrate-binding module domain. Both the molecular dynamics simulation and the biochemical experiments proved that the α-helix plays an important role in the thermostability of XynAF0. Introducing of this poly-threonine region to the C-terminus of another GH10 family xylanase improved its thermostability. Our results indicated that the poly-threonine α-helix at the C-terminus of the catalytic domain was important for improving the thermophilic of GH10 family xylanases, which provides a new strategy for the thermostability modification of xylanases.

Keywords: Enzyme engineering; GH10 family xylanase; Poly-threonine; Thermostability.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aspergillus fumigatus / enzymology*
  • Endo-1,4-beta Xylanases / chemistry*
  • Endo-1,4-beta Xylanases / metabolism*
  • Enzyme Stability
  • Models, Molecular
  • Protein Conformation, alpha-Helical
  • Recombinant Proteins / chemistry
  • Structure-Activity Relationship
  • Temperature*
  • Threonine / chemistry

Substances

  • Recombinant Proteins
  • Threonine
  • Endo-1,4-beta Xylanases