FMN-dependent oligomerization of putative lactate oxidase from Pediococcus acidilactici

PLoS One. 2020 Feb 24;15(2):e0223870. doi: 10.1371/journal.pone.0223870. eCollection 2020.

Abstract

Lactate oxidases belong to a group of FMN-dependent enzymes and they catalyze a conversion of lactate to pyruvate with a release of hydrogen peroxide. Hydrogen peroxide is also utilized as a read out in biosensors to quantitate lactate levels in biological samples. Aerococcus viridans lactate oxidase is the best characterized lactate oxidase and our knowledge of lactate oxidases relies largely to studies conducted with that particular enzyme. Pediococcus acidilactici lactate oxidase is also commercially available for e.g. lactate measurements, but this enzyme has not been characterized in detail before. Here we report structural characterization of the recombinant enzyme and its co-factor dependent oligomerization. The crystal structures revealed two distinct conformations in the loop closing the active site, consistent with previous biochemical studies implicating the role of loop in catalysis. Despite the structural conservation of active site residues, we were not able to detect either oxidase or monooxygenase activity when L-lactate was used as a substrate. Pediococcus acidilactici lactate oxidase is therefore an example of a misannotation of an FMN-dependent enzyme, which catalyzes likely a so far unknown oxidation reaction.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Catalysis
  • Catalytic Domain
  • Crystallography, X-Ray
  • Flavin Mononucleotide / pharmacology*
  • Lactic Acid / metabolism
  • Mixed Function Oxygenases / metabolism*
  • Pediococcus acidilactici / enzymology*
  • Pediococcus acidilactici / metabolism
  • Protein Multimerization / drug effects*
  • Recombinant Proteins

Substances

  • Recombinant Proteins
  • Lactic Acid
  • Flavin Mononucleotide
  • Mixed Function Oxygenases
  • lactate 2-monooxygenase

Grants and funding

LL; grants 287063 and 294085; Academy of Finland; www.aka.fi PK and TK; grant LIIKUTPA; European Regional Development Fund; www.rakennerahastot.fi The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.