Characterization of a thermotolerant aryl-alcohol oxidase from Moesziomyces antarcticus oxidizing 5-hydroxymethyl-2-furancarboxylic acid
Abstract The development of enzymatic processes for the environmentally friendly production of 2,5-furandicarboxylic acid (FDCA), a renewable precursor for bioplastics, from 5-hydroxymethylfurfural (HMF) has gained increasing attention over the last years. Aryl-alcohol oxidases (AAOs) catalyze the o...
Published in: | Applied Microbiology and Biotechnology |
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Springer Science and Business Media LLC
2021
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Online Access: | http://dx.doi.org/10.1007/s00253-021-11557-8 https://link.springer.com/content/pdf/10.1007/s00253-021-11557-8.pdf https://link.springer.com/article/10.1007/s00253-021-11557-8/fulltext.html |
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crspringernat:10.1007/s00253-021-11557-8 2023-05-15T14:13:04+02:00 Characterization of a thermotolerant aryl-alcohol oxidase from Moesziomyces antarcticus oxidizing 5-hydroxymethyl-2-furancarboxylic acid Lappe, Alessa Jankowski, Nina Albrecht, Annemie Koschorreck, Katja ministerium für innovation, wissenschaft und forschung des landes nordrhein-westfalen Heinrich-Heine-Universität Düsseldorf 2021 http://dx.doi.org/10.1007/s00253-021-11557-8 https://link.springer.com/content/pdf/10.1007/s00253-021-11557-8.pdf https://link.springer.com/article/10.1007/s00253-021-11557-8/fulltext.html en eng Springer Science and Business Media LLC https://creativecommons.org/licenses/by/4.0 https://creativecommons.org/licenses/by/4.0 CC-BY Applied Microbiology and Biotechnology volume 105, issue 21-22, page 8313-8327 ISSN 0175-7598 1432-0614 Applied Microbiology and Biotechnology General Medicine Biotechnology journal-article 2021 crspringernat https://doi.org/10.1007/s00253-021-11557-8 2022-01-04T07:34:34Z Abstract The development of enzymatic processes for the environmentally friendly production of 2,5-furandicarboxylic acid (FDCA), a renewable precursor for bioplastics, from 5-hydroxymethylfurfural (HMF) has gained increasing attention over the last years. Aryl-alcohol oxidases (AAOs) catalyze the oxidation of HMF to 5-formyl-2-furancarboxylic acid (FFCA) through 2,5-diformylfuran (DFF) and have thus been applied in enzymatic reaction cascades for the production of FDCA. AAOs are flavoproteins that oxidize a broad range of benzylic and aliphatic allylic primary alcohols to the corresponding aldehydes, and in some cases further to acids, while reducing molecular oxygen to hydrogen peroxide. These promising biocatalysts can also be used for the synthesis of flavors, fragrances, and chemical building blocks, but their industrial applicability suffers from low production yield in natural and heterologous hosts. Here we report on heterologous expression of a new aryl-alcohol oxidase, Ma AAO, from Moesziomyces antarcticus at high yields in the methylotrophic yeast Pichia pastoris (recently reclassified as Komagataella phaffii ) . Fed-batch fermentation of recombinant P. pastoris yielded around 750 mg of active enzyme per liter of culture. Purified Ma AAO was highly stable at pH 2–9 and exhibited high thermal stability with almost 95% residual activity after 48 h at 57.5 °C. Ma AAO accepts a broad range of benzylic primary alcohols, aliphatic allylic alcohols, and furan derivatives like HMF as substrates and some oxidation products thereof like piperonal or perillaldehyde serve as building blocks for pharmaceuticals or show health-promoting effects. Besides this, Ma AAO oxidized 5-hydroxymethyl-2-furancarboxylic acid (HMFCA) to FFCA, which has not been shown for any other AAO so far. Combining Ma AAO with an unspecific peroxygenase oxidizing HMFCA to FFCA in one pot resulted in complete conversion of HMF to FDCA within 144 h. Ma AAO is thus a promising biocatalyst for the production of precursors for bioplastics and bioactive compounds. Key points • MaAAO from M. antarcticus was expressed in P. pastoris at 750 mg/l. • MaAAO oxidized 5-hydroxymethyl-2-furancarboxylic acid (HMFCA). • Complete conversion of HMF to 2,5-furandicarboxylic acid by combining MaAAO and UPO. Article in Journal/Newspaper Antarc* antarcticus Springer Nature (via Crossref) Applied Microbiology and Biotechnology 105 21-22 8313 8327 |
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Springer Nature (via Crossref) |
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English |
topic |
Applied Microbiology and Biotechnology General Medicine Biotechnology |
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Applied Microbiology and Biotechnology General Medicine Biotechnology Lappe, Alessa Jankowski, Nina Albrecht, Annemie Koschorreck, Katja Characterization of a thermotolerant aryl-alcohol oxidase from Moesziomyces antarcticus oxidizing 5-hydroxymethyl-2-furancarboxylic acid |
topic_facet |
Applied Microbiology and Biotechnology General Medicine Biotechnology |
description |
Abstract The development of enzymatic processes for the environmentally friendly production of 2,5-furandicarboxylic acid (FDCA), a renewable precursor for bioplastics, from 5-hydroxymethylfurfural (HMF) has gained increasing attention over the last years. Aryl-alcohol oxidases (AAOs) catalyze the oxidation of HMF to 5-formyl-2-furancarboxylic acid (FFCA) through 2,5-diformylfuran (DFF) and have thus been applied in enzymatic reaction cascades for the production of FDCA. AAOs are flavoproteins that oxidize a broad range of benzylic and aliphatic allylic primary alcohols to the corresponding aldehydes, and in some cases further to acids, while reducing molecular oxygen to hydrogen peroxide. These promising biocatalysts can also be used for the synthesis of flavors, fragrances, and chemical building blocks, but their industrial applicability suffers from low production yield in natural and heterologous hosts. Here we report on heterologous expression of a new aryl-alcohol oxidase, Ma AAO, from Moesziomyces antarcticus at high yields in the methylotrophic yeast Pichia pastoris (recently reclassified as Komagataella phaffii ) . Fed-batch fermentation of recombinant P. pastoris yielded around 750 mg of active enzyme per liter of culture. Purified Ma AAO was highly stable at pH 2–9 and exhibited high thermal stability with almost 95% residual activity after 48 h at 57.5 °C. Ma AAO accepts a broad range of benzylic primary alcohols, aliphatic allylic alcohols, and furan derivatives like HMF as substrates and some oxidation products thereof like piperonal or perillaldehyde serve as building blocks for pharmaceuticals or show health-promoting effects. Besides this, Ma AAO oxidized 5-hydroxymethyl-2-furancarboxylic acid (HMFCA) to FFCA, which has not been shown for any other AAO so far. Combining Ma AAO with an unspecific peroxygenase oxidizing HMFCA to FFCA in one pot resulted in complete conversion of HMF to FDCA within 144 h. Ma AAO is thus a promising biocatalyst for the production of precursors for bioplastics and bioactive compounds. Key points • MaAAO from M. antarcticus was expressed in P. pastoris at 750 mg/l. • MaAAO oxidized 5-hydroxymethyl-2-furancarboxylic acid (HMFCA). • Complete conversion of HMF to 2,5-furandicarboxylic acid by combining MaAAO and UPO. |
author2 |
ministerium für innovation, wissenschaft und forschung des landes nordrhein-westfalen Heinrich-Heine-Universität Düsseldorf |
format |
Article in Journal/Newspaper |
author |
Lappe, Alessa Jankowski, Nina Albrecht, Annemie Koschorreck, Katja |
author_facet |
Lappe, Alessa Jankowski, Nina Albrecht, Annemie Koschorreck, Katja |
author_sort |
Lappe, Alessa |
title |
Characterization of a thermotolerant aryl-alcohol oxidase from Moesziomyces antarcticus oxidizing 5-hydroxymethyl-2-furancarboxylic acid |
title_short |
Characterization of a thermotolerant aryl-alcohol oxidase from Moesziomyces antarcticus oxidizing 5-hydroxymethyl-2-furancarboxylic acid |
title_full |
Characterization of a thermotolerant aryl-alcohol oxidase from Moesziomyces antarcticus oxidizing 5-hydroxymethyl-2-furancarboxylic acid |
title_fullStr |
Characterization of a thermotolerant aryl-alcohol oxidase from Moesziomyces antarcticus oxidizing 5-hydroxymethyl-2-furancarboxylic acid |
title_full_unstemmed |
Characterization of a thermotolerant aryl-alcohol oxidase from Moesziomyces antarcticus oxidizing 5-hydroxymethyl-2-furancarboxylic acid |
title_sort |
characterization of a thermotolerant aryl-alcohol oxidase from moesziomyces antarcticus oxidizing 5-hydroxymethyl-2-furancarboxylic acid |
publisher |
Springer Science and Business Media LLC |
publishDate |
2021 |
url |
http://dx.doi.org/10.1007/s00253-021-11557-8 https://link.springer.com/content/pdf/10.1007/s00253-021-11557-8.pdf https://link.springer.com/article/10.1007/s00253-021-11557-8/fulltext.html |
genre |
Antarc* antarcticus |
genre_facet |
Antarc* antarcticus |
op_source |
Applied Microbiology and Biotechnology volume 105, issue 21-22, page 8313-8327 ISSN 0175-7598 1432-0614 |
op_rights |
https://creativecommons.org/licenses/by/4.0 https://creativecommons.org/licenses/by/4.0 |
op_rightsnorm |
CC-BY |
op_doi |
https://doi.org/10.1007/s00253-021-11557-8 |
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Applied Microbiology and Biotechnology |
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105 |
container_issue |
21-22 |
container_start_page |
8313 |
op_container_end_page |
8327 |
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1766285470938955776 |