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

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16 引文 斯高帕斯(Scopus)

摘要

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.

原文英語
文章編號215
期刊Biotechnology for Biofuels
11
發行號1
DOIs
出版狀態已發佈 - 八月 2 2018
對外發佈

ASJC Scopus subject areas

  • 生物技術
  • 應用微生物與生物技術
  • 可再生能源、永續發展與環境
  • 能源(全部)
  • 管理、監督、政策法律

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