A fast, sensitive and fluorescent LPMO activity assay

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A fast, sensitive and fluorescent LPMO activity assay. / Ipsen, Johan Ø.; Johansen, Katja S.; Brander, Søren.

I: Frontiers in Microbiology, Bind 14, 1128470, 2023.

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt

Harvard

Ipsen, JØ, Johansen, KS & Brander, S 2023, 'A fast, sensitive and fluorescent LPMO activity assay', Frontiers in Microbiology, bind 14, 1128470. https://doi.org/10.3389/fmicb.2023.1128470

APA

Ipsen, J. Ø., Johansen, K. S., & Brander, S. (2023). A fast, sensitive and fluorescent LPMO activity assay. Frontiers in Microbiology, 14, [1128470]. https://doi.org/10.3389/fmicb.2023.1128470

Vancouver

Ipsen JØ, Johansen KS, Brander S. A fast, sensitive and fluorescent LPMO activity assay. Frontiers in Microbiology. 2023;14. 1128470. https://doi.org/10.3389/fmicb.2023.1128470

Author

Ipsen, Johan Ø. ; Johansen, Katja S. ; Brander, Søren. / A fast, sensitive and fluorescent LPMO activity assay. I: Frontiers in Microbiology. 2023 ; Bind 14.

Bibtex

@article{b404ba30fcf34528b12354e4f2b36278,
title = "A fast, sensitive and fluorescent LPMO activity assay",
abstract = "Lytic polysaccharide monooxygenases (LPMOs) are industrially relevant enzymes that utilize a copper co-factor and an oxygen species to break down recalcitrant polysaccharides. These enzymes are secreted by microorganisms and are used in lignocellulosic refineries. As such, they are interesting from both the ecological/biological and industrial perspectives. Here we describe the development of a new fluorescence-based kinetic LPMO activity assay. The assay is based on the enzymatic production of fluorescein from its reduced counterpart. The assay can detect as little as 1 nM LPMO with optimized assay conditions. Furthermore, the reduced fluorescein substrate can also be used to identify peroxidase activity as seen by the formation of fluorescein by horseradish peroxidase. The assay was shown to work well at relatively low H2O2 and dehydroascorbate concentrations. The applicability of the assay was demonstrated.",
keywords = "copper enzyme, enzyme assay, fluorescin, LPMO, peroxide, redox activity",
author = "Ipsen, {Johan {\O}.} and Johansen, {Katja S.} and S{\o}ren Brander",
note = "Publisher Copyright: Copyright {\textcopyright} 2023 Ipsen, Johansen and Brander.",
year = "2023",
doi = "10.3389/fmicb.2023.1128470",
language = "English",
volume = "14",
journal = "Frontiers in Microbiology",
issn = "1664-302X",
publisher = "Frontiers Media S.A.",

}

RIS

TY - JOUR

T1 - A fast, sensitive and fluorescent LPMO activity assay

AU - Ipsen, Johan Ø.

AU - Johansen, Katja S.

AU - Brander, Søren

N1 - Publisher Copyright: Copyright © 2023 Ipsen, Johansen and Brander.

PY - 2023

Y1 - 2023

N2 - Lytic polysaccharide monooxygenases (LPMOs) are industrially relevant enzymes that utilize a copper co-factor and an oxygen species to break down recalcitrant polysaccharides. These enzymes are secreted by microorganisms and are used in lignocellulosic refineries. As such, they are interesting from both the ecological/biological and industrial perspectives. Here we describe the development of a new fluorescence-based kinetic LPMO activity assay. The assay is based on the enzymatic production of fluorescein from its reduced counterpart. The assay can detect as little as 1 nM LPMO with optimized assay conditions. Furthermore, the reduced fluorescein substrate can also be used to identify peroxidase activity as seen by the formation of fluorescein by horseradish peroxidase. The assay was shown to work well at relatively low H2O2 and dehydroascorbate concentrations. The applicability of the assay was demonstrated.

AB - Lytic polysaccharide monooxygenases (LPMOs) are industrially relevant enzymes that utilize a copper co-factor and an oxygen species to break down recalcitrant polysaccharides. These enzymes are secreted by microorganisms and are used in lignocellulosic refineries. As such, they are interesting from both the ecological/biological and industrial perspectives. Here we describe the development of a new fluorescence-based kinetic LPMO activity assay. The assay is based on the enzymatic production of fluorescein from its reduced counterpart. The assay can detect as little as 1 nM LPMO with optimized assay conditions. Furthermore, the reduced fluorescein substrate can also be used to identify peroxidase activity as seen by the formation of fluorescein by horseradish peroxidase. The assay was shown to work well at relatively low H2O2 and dehydroascorbate concentrations. The applicability of the assay was demonstrated.

KW - copper enzyme

KW - enzyme assay

KW - fluorescin

KW - LPMO

KW - peroxide

KW - redox activity

U2 - 10.3389/fmicb.2023.1128470

DO - 10.3389/fmicb.2023.1128470

M3 - Journal article

C2 - 36998406

AN - SCOPUS:85151089645

VL - 14

JO - Frontiers in Microbiology

JF - Frontiers in Microbiology

SN - 1664-302X

M1 - 1128470

ER -

ID: 344364462