Structural analysis of β-glucosidase mutants derived from a hyperthermophilic tetrameric structure

Makoto Nakabayashi, Misumi Kataoka, Yumiko Mishima, Yuka Maeno, Kazuhiko Ishikawa

Research output: Contribution to journalArticlepeer-review

5 Citations (Scopus)

Abstract

β-Glucosidase from Pyrococcus furiosus (BGLPf) is a hyperthermophilic tetrameric enzyme which can degrade cellooligosaccharides to glucose under hyperthermophilic conditions and thus holds promise for the saccharification of lignocellulosic biomass at high temperature. Prior to the production of large amounts of this enzyme, detailed information regarding the oligomeric structure of the enzyme is required. Several crystals of BGLPf have been prepared over the past ten years, but its crystal structure had not been solved until recently. In 2011, the first crystal structure of BGLPf was solved and a model was constructed at somewhat low resolution (2.35 Å). In order to obtain more detailed structural data on BGLPf, the relationship between its tetrameric structure and the quality of the crystal was re-examined. A dimeric form of BGLPf was constructed and its crystal structure was solved at a resolution of 1.70 Å using protein-engineering methods. Furthermore, using the high-resolution crystal structural data for the dimeric form, a monomeric form of BGLPf was constructed which retained the intrinsic activity of the tetrameric form. The thermostability of BGLPf is affected by its oligomeric structure. Here, the biophysical and biochemical properties of engineered dimeric and monomeric BGLPfs are reported, which are promising prototype models to apply to the saccharification reaction. Furthermore, details regarding the oligomeric structures of BGLPf and the reasons why the mutations yielded improved crystal structures are discussed.

Original languageEnglish
Pages (from-to)877-888
Number of pages12
JournalActa Crystallographica Section D: Biological Crystallography
Volume70
Issue number3
DOIs
Publication statusPublished - Mar 1 2014

Keywords

  • Biomass
  • Crystal engineering
  • Intermolecular interactions
  • Protein engineering
  • Thermostable enzymes

ASJC Scopus subject areas

  • Structural Biology

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