A colorimetric assay to rapidly determine the activities of lytic polysaccharide monooxygenases

Damao Wang, Jing Li, Ann C.Y. Wong, Finn L. Aachmann, Yves S.Y. Hsieh

Research output: Contribution to journalArticlepeer-review

11 Citations (Scopus)

Abstract

Background: Lytic polysaccharide monooxygenase (LPMOs) are enzymes that catalyze the breakdown of polysaccharides in biomass and have excellent potential for biorefinery applications. However, their activities are relatively low, and methods to measure these activities are costly, tedious or often reflect only an apparent activity to the polysaccharide substrates. Here, we describe a new method we have developed that is simple to use to determine the activities of type-1 (C1-oxidizing) LPMOs. The method is based on quantifying the ionic binding of cations to carboxyl groups formed by the action of type-1 LPMOs on polysaccharides. It allows comparisons to be made of activities under different conditions. Results: Based on the colorimetric detection and quantification of the pyrocatechol violet (PV)-Ni2+ complex, we have developed an assay to reliably detect and quantify carboxylate moieties introduced by type-1 LPMOs. Conditions were optimized for determining the activities of specific LPMOs. Comparisons were made of the activities against cellulose and chitin of a novel AA10 LPMO and a recently reported family AA11 LPMO. Activities of both LPMOs were boosted by hydrogen peroxide in the 1st hour of the reaction, with a 16-fold increase for the family AA11 LPMO, and up to a 34-fold increase for the family AA10 LPMO. Conclusions: We developed a versatile colorimetric cation-based assay to determine the activities of type-1 LPMOs. The assay is quick, low cost and could be adapted for use in industrial biorefineries.

Original languageEnglish
Article number215
JournalBiotechnology for Biofuels
Volume11
Issue number1
DOIs
Publication statusPublished - Aug 2 2018
Externally publishedYes

Keywords

  • Biomass deconstruction
  • Enzyme assay
  • Lytic polysaccharide monooxygenase

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology
  • Renewable Energy, Sustainability and the Environment
  • Energy(all)
  • Management, Monitoring, Policy and Law

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