Identification of redox activators for continuous reactivation of glyoxal oxidase from Trametes versicolor in a two-enzyme reaction cascade
Abstract Glyoxal oxidases, belonging to the group of copper radical oxidases (CROs), oxidize aldehydes to carboxylic acids, while reducing O2 to H2O2. Their activity on furan derivatives like 5-hydroxymethylfurfural (HMF) makes these enzymes promising biocatalysts for the environmentally friendly sy...
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ftdoajarticles:oai:doaj.org/article:0374f97d3b8b4d47902bbca0acdbd09e 2024-09-15T17:44:25+00:00 Identification of redox activators for continuous reactivation of glyoxal oxidase from Trametes versicolor in a two-enzyme reaction cascade Saadet Alpdağtaş Nina Jankowski Vlada B. Urlacher Katja Koschorreck 2024-03-01T00:00:00Z https://doi.org/10.1038/s41598-024-56429-z https://doaj.org/article/0374f97d3b8b4d47902bbca0acdbd09e EN eng Nature Portfolio https://doi.org/10.1038/s41598-024-56429-z https://doaj.org/toc/2045-2322 doi:10.1038/s41598-024-56429-z 2045-2322 https://doaj.org/article/0374f97d3b8b4d47902bbca0acdbd09e Scientific Reports, Vol 14, Iss 1, Pp 1-9 (2024) Glyoxal oxidase Copper radical oxidase Redox activators FDCA (2,5-furandicarboxylic acid) Medicine R Science Q article 2024 ftdoajarticles https://doi.org/10.1038/s41598-024-56429-z 2024-08-05T17:49:48Z Abstract Glyoxal oxidases, belonging to the group of copper radical oxidases (CROs), oxidize aldehydes to carboxylic acids, while reducing O2 to H2O2. Their activity on furan derivatives like 5-hydroxymethylfurfural (HMF) makes these enzymes promising biocatalysts for the environmentally friendly synthesis of the bioplastics precursor 2,5-furandicarboxylic acid (FDCA). However, glyoxal oxidases suffer from inactivation, which requires the identification of suitable redox activators for efficient substrate conversion. Furthermore, only a few glyoxal oxidases have been expressed and characterized so far. Here, we report on a new glyoxal oxidase from Trametes versicolor (TvGLOX) that was expressed at high levels in Pichia pastoris (reclassified as Komagataella phaffii). TvGLOX was found to catalyze the oxidation of aldehyde groups in glyoxylic acid, methyl glyoxal, HMF, 2,5-diformylfuran (DFF) and 5-formyl-2-furancarboxylic acid (FFCA), but barely accepted alcohol groups as in 5-hydroxymethyl-2-furancarboxylic acid (HMFCA), preventing formation of FDCA from HMF. Various redox activators were tested for TvGLOX reactivation during catalyzed reactions. Among them, a combination of horseradish peroxidase and its substrate 2,2′-azino-di-(3-ethylbenzthiazoline sulfonic acid) (ABTS) most efficiently reactivated TvGLOX. Through continuous reactivation of TvGLOX in a two-enzyme system employing a recombinant Moesziomyces antarcticus aryl-alcohol oxidase (MaAAO) almost complete conversion of 8 mM HMF to FDCA was achieved within 24 h. Article in Journal/Newspaper Antarc* antarcticus Directory of Open Access Journals: DOAJ Articles Scientific Reports 14 1 |
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op_collection_id |
ftdoajarticles |
language |
English |
topic |
Glyoxal oxidase Copper radical oxidase Redox activators FDCA (2,5-furandicarboxylic acid) Medicine R Science Q |
spellingShingle |
Glyoxal oxidase Copper radical oxidase Redox activators FDCA (2,5-furandicarboxylic acid) Medicine R Science Q Saadet Alpdağtaş Nina Jankowski Vlada B. Urlacher Katja Koschorreck Identification of redox activators for continuous reactivation of glyoxal oxidase from Trametes versicolor in a two-enzyme reaction cascade |
topic_facet |
Glyoxal oxidase Copper radical oxidase Redox activators FDCA (2,5-furandicarboxylic acid) Medicine R Science Q |
description |
Abstract Glyoxal oxidases, belonging to the group of copper radical oxidases (CROs), oxidize aldehydes to carboxylic acids, while reducing O2 to H2O2. Their activity on furan derivatives like 5-hydroxymethylfurfural (HMF) makes these enzymes promising biocatalysts for the environmentally friendly synthesis of the bioplastics precursor 2,5-furandicarboxylic acid (FDCA). However, glyoxal oxidases suffer from inactivation, which requires the identification of suitable redox activators for efficient substrate conversion. Furthermore, only a few glyoxal oxidases have been expressed and characterized so far. Here, we report on a new glyoxal oxidase from Trametes versicolor (TvGLOX) that was expressed at high levels in Pichia pastoris (reclassified as Komagataella phaffii). TvGLOX was found to catalyze the oxidation of aldehyde groups in glyoxylic acid, methyl glyoxal, HMF, 2,5-diformylfuran (DFF) and 5-formyl-2-furancarboxylic acid (FFCA), but barely accepted alcohol groups as in 5-hydroxymethyl-2-furancarboxylic acid (HMFCA), preventing formation of FDCA from HMF. Various redox activators were tested for TvGLOX reactivation during catalyzed reactions. Among them, a combination of horseradish peroxidase and its substrate 2,2′-azino-di-(3-ethylbenzthiazoline sulfonic acid) (ABTS) most efficiently reactivated TvGLOX. Through continuous reactivation of TvGLOX in a two-enzyme system employing a recombinant Moesziomyces antarcticus aryl-alcohol oxidase (MaAAO) almost complete conversion of 8 mM HMF to FDCA was achieved within 24 h. |
format |
Article in Journal/Newspaper |
author |
Saadet Alpdağtaş Nina Jankowski Vlada B. Urlacher Katja Koschorreck |
author_facet |
Saadet Alpdağtaş Nina Jankowski Vlada B. Urlacher Katja Koschorreck |
author_sort |
Saadet Alpdağtaş |
title |
Identification of redox activators for continuous reactivation of glyoxal oxidase from Trametes versicolor in a two-enzyme reaction cascade |
title_short |
Identification of redox activators for continuous reactivation of glyoxal oxidase from Trametes versicolor in a two-enzyme reaction cascade |
title_full |
Identification of redox activators for continuous reactivation of glyoxal oxidase from Trametes versicolor in a two-enzyme reaction cascade |
title_fullStr |
Identification of redox activators for continuous reactivation of glyoxal oxidase from Trametes versicolor in a two-enzyme reaction cascade |
title_full_unstemmed |
Identification of redox activators for continuous reactivation of glyoxal oxidase from Trametes versicolor in a two-enzyme reaction cascade |
title_sort |
identification of redox activators for continuous reactivation of glyoxal oxidase from trametes versicolor in a two-enzyme reaction cascade |
publisher |
Nature Portfolio |
publishDate |
2024 |
url |
https://doi.org/10.1038/s41598-024-56429-z https://doaj.org/article/0374f97d3b8b4d47902bbca0acdbd09e |
genre |
Antarc* antarcticus |
genre_facet |
Antarc* antarcticus |
op_source |
Scientific Reports, Vol 14, Iss 1, Pp 1-9 (2024) |
op_relation |
https://doi.org/10.1038/s41598-024-56429-z https://doaj.org/toc/2045-2322 doi:10.1038/s41598-024-56429-z 2045-2322 https://doaj.org/article/0374f97d3b8b4d47902bbca0acdbd09e |
op_doi |
https://doi.org/10.1038/s41598-024-56429-z |
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Scientific Reports |
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14 |
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1 |
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1810491974643351552 |