2019 ; Vol.29-2: 244~255
|Author||Sun-Ha Park, Sunghark Kwon, Chang Woo Lee, Chang Min Kim, Chang Sook Jeong, Kyung-Jin Kim, Jong Wook Hong, Hak Jun Kim, Hyun Ho Park, Jun Hyuck Lee|
|Place of duty||Unit of Polar Genomics, Korea Polar Research Institute, Republic of Korea|
|Title||Crystal Structure and Functional Characterization of a Xylose Isomerase (PbXI) from the Psychrophilic Soil Microorganism, Paenibacillus sp.|
J. Microbiol. Biotechnol.2019 ;
|Abstract||Xylose isomerase (XI; E.C. 18.104.22.168) 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°C. We solved the crystal structure of PbXI at 1.94-Å resolution to
investigate the origin of its thermostability. The PbXI structure shows a (β/α)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 β4-α6 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 β1-α2 loop structures near the substrate
binding pocket of PbXI were remarkably different. Site-directed mutagenesis studies
suggested that the flexible β1-α2 loop region is essential for PbXI activity. Our findings
provide valuable insights that can be applied in protein engineering to generate lowtemperature
purpose-specific XI enzymes.|
|Key_word||Cold-active protein, crystal structure, paenibacillus species, xylose isomerase, X-ray crystallography|
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