Characterization and structural analysis of a thermophilic GH11 xylanase from compost metatranscriptome

Yunlei Yi, Shenyuan Xu, Andrey Kovalevsky, Xia Zhang, Dongyang Liu, Qun Wan

Research output: Contribution to journalArticlepeer-review

14 Scopus citations

Abstract

Abstract: Xylanase is efficient for xylan degradation and widely applied in industries. We found a GH11 family xylanase (Xyn11A) with high thermostability and catalytic activity from compost metatranscriptome. This xylanase has the optimal reaction temperature at 80 °C with the activity of 2907.3 U/mg. The X-ray crystallographic structure shows a typical “right hand” architecture, which is the characteristics of the GH11 family enzymes. Comparing it with the mesophilic XYN II, a well-studied GH11 xylanase from Trichoderma reesei, Xyn11A is more compact with more H-bonds. Our mutagenic results show that the electrostatic interactions in the thumb and palm region of Xyn11A could result in its high thermostability and activity. Introducing a disulfide bond at the N-terminus further increased its optimal reaction temperature to 90 °C with augmented activity. Key points: • A hyperthermophilic xylanase with high activity was discovered using the metatranscriptomic method. • The mechanisms of thermophilicity and high activity were revealed using X-ray crystallography, mutagenesis, and molecular dynamics simulations. • The thermostability and activity were further improved by introducing a disulfide bond.

Original languageEnglish
Pages (from-to)7757-7767
Number of pages11
JournalApplied Microbiology and Biotechnology
Volume105
Issue number20
DOIs
StatePublished - Oct 2021

Funding

Research at ORNL’s Spallation Neutron Source was sponsored by the Scientific User Facilities Division, Office of Basic Energy Sciences, US Department of Energy. We thank the SSRF beamlines BL18U1 and BL19U1 for X-ray data collection. Q.W. was supported by the National Natural Science Foundation of China (Nos. 31670790 and 32071264) and the Fundamental Research Funds for the Central Universities (No. KYXK202009).

Keywords

  • Crystal structure
  • GH11 xylanase
  • MD simulations
  • Thermostability

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