Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Crystal Structure and Functional Characterization of a Xylose Isomerase (PbXI) from the Psychrophilic Soil Microorganism, Paenibacillus sp.

  • Park, Sun-Ha (Unit of Polar Genomics, Korea Polar Research Institute) ;
  • Kwon, Sunghark (College of Pharmacy, Chung-Ang University) ;
  • Lee, Chang Woo (Unit of Polar Genomics, Korea Polar Research Institute) ;
  • Kim, Chang Min (College of Pharmacy, Chung-Ang University) ;
  • Jeong, Chang Sook (Unit of Polar Genomics, Korea Polar Research Institute) ;
  • Kim, Kyung-Jin (School of Life Sciences, KNU Creative BioResearch Group, Kyungpook National University) ;
  • Hong, Jong Wook (Department of Bionanotechnology, Graduate School, Hanyang University) ;
  • Kim, Hak Jun (Department of Chemistry, Pukyong National University) ;
  • Park, Hyun Ho (College of Pharmacy, Chung-Ang University) ;
  • Lee, Jun Hyuck (Unit of Polar Genomics, Korea Polar Research Institute)
  • Received : 2018.07.02
  • Accepted : 2018.09.15
  • Published : 2019.02.28
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Abstract

Xylose isomerase (XI; E.C. 5.3.1.5) catalyzes the isomerization of xylose to xylulose, which can be used to produce bioethanol through fermentation. Therefore, XI has recently gained attention as a key catalyst in the bioenergy industry. Here, we identified, purified, and characterized a XI (PbXI) from the psychrophilic soil microorganism, Paenibacillus sp. R4. Surprisingly, activity assay results showed that PbXI is not a cold-active enzyme, but displays optimal activity at $60^{\circ}C$. We solved the crystal structure of PbXI at $1.94-{\AA}$ resolution to investigate the origin of its thermostability. The PbXI structure shows a $({\beta}/{\alpha})_8$-barrel fold with tight tetrameric interactions and it has three divalent metal ions (CaI, CaII, and CaIII). Two metal ions (CaI and CaII) located in the active site are known to be involved in the enzymatic reaction. The third metal ion (CaIII), located near the ${\beta}4-{\alpha}6$ loop region, was newly identified and is thought to be important for the stability of PbXI. Compared with previously determined thermostable and mesophilic XI structures, the ${\beta}1-{\alpha}2$ loop structures near the substrate binding pocket of PbXI were remarkably different. Site-directed mutagenesis studies suggested that the flexible ${\beta}1-{\alpha}2$ loop region is essential for PbXI activity. Our findings provide valuable insights that can be applied in protein engineering to generate low-temperature purpose-specific XI enzymes.

Keywords

References

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