Understanding xylose isomerase from burkholderia cenocepacia: Insights into structure and functionality for ethanol production

Igor P.V. Vieira, Gabrielle T. Cordeiro, Diego E.B. Gomes, Rafael D. Melani, Leonardo F. Vilela, Gilberto B. Domont, Rafael D. Mesquita, Elis C.A. Eleutherio, Bianca C. Neves*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

4 Scopus citations

Abstract

The inability of the yeast Saccharomyces cerevisiae to produce ethanol from xylose has hampered the biofuel production from lignocellulosic biomass. However, prior studies reveal that functional expression of xylose isomerase (XI) from Burkholderia cenocepacia (XylABc) in S. cerevisiae has remarkably improved xylose consumption and ethanol productivity. Yet, little is known about kinetic and structural properties of this enzyme. Hereby, a purified recombi-nant XylA was assayed in vitro, showing optimal enzyme activity at 37 °C and pH 7.2. The Km of XylA for d-xylose was at least threefold lower than the Km results for any XI published to date (e.g. XylA from Piromyces sp.). In addition, oligomerization behavior as a tetramer was observed for XylA in solution. Functional and structural comparative analyses amongst three microbial XIs were further performed as theoretical models, showing that xylose orientation at the active site was highly conserved among the XIs. Mg2+ ions anchor the sugar and guide its pyranoside oxygen towards a histidine residue present at the active site, allowing an acid–base reaction, linearizing xylose. Electrostatic surface analyses showed that small variations in the net charge distribution and dipole moment could directly affect the way the substrate interacts with the protein, thus altering its kinetic properties. Accordingly, in silico modeling suggested the tetramer may be the major functional form. These analyses and the resulting model promote a better understanding of this protein family and pave the way to further protein engineering and application of XylA in the ethanol industry.

Original languageEnglish (US)
Article number73
Pages (from-to)1-12
Number of pages12
JournalAMB Express
Volume9
Issue number1
DOIs
StatePublished - 2019
Externally publishedYes

Keywords

  • Burkholderia cenocepacia
  • Enzyme kinetics
  • Ethanol
  • XylA
  • Xylose fermentation
  • Xylose isomerase

ASJC Scopus subject areas

  • Biophysics
  • Applied Microbiology and Biotechnology

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