Microplate-Based Detection of Lytic Polysaccharide Monooxygenase Activity by Fluorescence-Labeling of Insoluble Oxidized Products.
Identifieur interne : 000174 ( Main/Curation ); précédent : 000173; suivant : 000175Microplate-Based Detection of Lytic Polysaccharide Monooxygenase Activity by Fluorescence-Labeling of Insoluble Oxidized Products.
Auteurs : Thu V. Vuong [Canada] ; Bing Liu [Suède] ; Mats Sandgren [Suède] ; Emma R. Master [Canada]Source :
- Biomacromolecules [ 1526-4602 ] ; 2017.
Descripteurs français
- KwdFr :
- Cellulose (composition chimique), Chitine (composition chimique), Fluorescence (MeSH), Microtechnologie (méthodes), Mixed function oxygenases (métabolisme), Oxydoréduction (MeSH), Phanerochaete (croissance et développement), Phanerochaete (enzymologie), Polyosides (composition chimique), Spectroscopie photoélectronique (MeSH), Spécificité du substrat (MeSH).
- MESH :
- composition chimique : Cellulose, Chitine, Polyosides.
- croissance et développement : Phanerochaete.
- enzymologie : Phanerochaete.
- métabolisme : Mixed function oxygenases.
- méthodes : Microtechnologie.
- Fluorescence, Oxydoréduction, Spectroscopie photoélectronique, Spécificité du substrat.
English descriptors
- KwdEn :
- Cellulose (chemistry), Chitin (chemistry), Fluorescence (MeSH), Microtechnology (methods), Mixed Function Oxygenases (metabolism), Oxidation-Reduction (MeSH), Phanerochaete (enzymology), Phanerochaete (growth & development), Photoelectron Spectroscopy (MeSH), Polysaccharides (chemistry), Substrate Specificity (MeSH).
- MESH :
- chemical , chemistry : Cellulose, Chitin, Polysaccharides.
- chemical , metabolism : Mixed Function Oxygenases.
- enzymology : Phanerochaete.
- growth & development : Phanerochaete.
- methods : Microtechnology.
- Fluorescence, Oxidation-Reduction, Photoelectron Spectroscopy, Substrate Specificity.
Abstract
Most existing methods for screening the activity of lytic polysaccharide mono-oxygenases (LPMOs) on polysaccharides are based on the detection of soluble oxidized sugars. This approach might underestimate the total performance of LPMOs since oxidation events that do not lead to oligosaccharide release are not detected. Using PcLPMO9D as a model enzyme, a microplate-based method has been developed to detect C1-oxidizing LPMO activity by covalently linking a water-soluble fluorophore to oxidized positions within the cellulose fiber. This fluorescence method was validated using X-ray photoelectron spectroscopy and then combined with high-performance anion-exchange chromatography to track total PcLPMO9D activity.
DOI: 10.1021/acs.biomac.6b01790
PubMed: 28125213
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pubmed:28125213Le document en format XML
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<term>Mixed Function Oxygenases (metabolism)</term>
<term>Oxidation-Reduction (MeSH)</term>
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<term>Phanerochaete (enzymologie)</term>
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<term>Spectroscopie photoélectronique (MeSH)</term>
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<front><div type="abstract" xml:lang="en">Most existing methods for screening the activity of lytic polysaccharide mono-oxygenases (LPMOs) on polysaccharides are based on the detection of soluble oxidized sugars. This approach might underestimate the total performance of LPMOs since oxidation events that do not lead to oligosaccharide release are not detected. Using PcLPMO9D as a model enzyme, a microplate-based method has been developed to detect C1-oxidizing LPMO activity by covalently linking a water-soluble fluorophore to oxidized positions within the cellulose fiber. This fluorescence method was validated using X-ray photoelectron spectroscopy and then combined with high-performance anion-exchange chromatography to track total PcLPMO9D activity.</div>
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