A New Phenolic Acid Decarboxylase from the Brown-Rot Fungus <i>Neolentinus lepideus</i> Natively Decarboxylates Biosourced Sinapic Acid into Canolol, a Bioactive Phenolic Compound
Rapeseed meal (RSM) is a cheap, abundant and renewable feedstock, whose biorefinery is a current challenge for the sustainability of the oilseed sector. RSM is rich in sinapic acid (SA), a <i>p</i>-hydroxycinnamic acid that can be decarboxylated into canolol (2,6-dimethoxy-4-vinylphenol)...
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| Format: | Article |
| Language: | English |
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MDPI AG
2024-02-01
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| Series: | Bioengineering |
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| Online Access: | https://www.mdpi.com/2306-5354/11/2/181 |
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| author | Elise Odinot Alexandra Bisotto-Mignot Toinou Frezouls Bastien Bissaro David Navarro Eric Record Frédéric Cadoret Annick Doan Didier Chevret Frédéric Fine Anne Lomascolo |
| author_facet | Elise Odinot Alexandra Bisotto-Mignot Toinou Frezouls Bastien Bissaro David Navarro Eric Record Frédéric Cadoret Annick Doan Didier Chevret Frédéric Fine Anne Lomascolo |
| author_sort | Elise Odinot |
| collection | DOAJ |
| description | Rapeseed meal (RSM) is a cheap, abundant and renewable feedstock, whose biorefinery is a current challenge for the sustainability of the oilseed sector. RSM is rich in sinapic acid (SA), a <i>p</i>-hydroxycinnamic acid that can be decarboxylated into canolol (2,6-dimethoxy-4-vinylphenol), a valuable bioactive compound. Microbial phenolic acid decarboxylases (PADs), mainly described for the non-oxidative decarboxylation of ferulic and <i>p</i>-coumaric acids, remain very poorly documented to date, for SA decarboxylation. The species <i>Neolentinus lepideus</i> has previously been shown to biotransform SA into canolol in vivo, but the enzyme responsible for bioconversion of the acid has never been characterized. In this study, we purified and characterized a new PAD from the canolol-overproducing strain <i>N. lepideus</i> BRFM15. Proteomic analysis highlighted a sole PAD-type protein sequence in the intracellular proteome of the strain. The native enzyme (<i>Nle</i>PAD) displayed an unusual outstanding activity for decarboxylating SA (V<sub>max</sub> of 600 U.mg<sup>−1</sup>, k<sub>cat</sub> of 6.3 s<sup>−1</sup> and k<sub>cat</sub>/K<sub>M</sub> of 1.6 s<sup>−1</sup>.mM<sup>−1</sup>). We showed that <i>Nle</i>PAD (a homodimer of 2 × 22 kDa) is fully active in a pH range of 5.5–7.5 and a temperature range of 30–55 °C, with optima of pH 6–6.5 and 37–45 °C, and is highly stable at 4 °C and pH 6–8. Relative ratios of specific activities on ferulic, sinapic, <i>p</i>-coumaric and caffeic acids, respectively, were 100:24.9:13.4:3.9. The enzyme demonstrated in vitro effectiveness as a biocatalyst for the synthesis of canolol in aqueous medium from commercial SA, with a molar yield of 92%. Then, we developed processes to biotransform naturally-occurring SA from RSM into canolol by combining the complementary potentialities of an <i>Aspergillus niger</i> feruloyl esterase type-A, which is able to release free SA from the raw meal by hydrolyzing its conjugated forms, and <i>Nle</i>PAD, in aqueous medium and mild conditions. <i>Nle</i>PAD decarboxylation of biobased SA led to an overall yield of 1.6–3.8 mg canolol per gram of initial meal. Besides being the first characterization of a fungal PAD able to decarboxylate SA, this report shows that <i>Nle</i>PAD is very promising as new biotechnological tool to generate biobased vinylphenols of industrial interest (especially canolol) as valuable platform chemicals for health, nutrition, cosmetics and green chemistry. |
| first_indexed | 2024-03-07T22:41:35Z |
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| institution | Directory Open Access Journal |
| issn | 2306-5354 |
| language | English |
| last_indexed | 2024-03-07T22:41:35Z |
| publishDate | 2024-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Bioengineering |
| spelling | doaj.art-4b222f03baff45729dd5e774d4fbae532024-02-23T15:08:02ZengMDPI AGBioengineering2306-53542024-02-0111218110.3390/bioengineering11020181A New Phenolic Acid Decarboxylase from the Brown-Rot Fungus <i>Neolentinus lepideus</i> Natively Decarboxylates Biosourced Sinapic Acid into Canolol, a Bioactive Phenolic CompoundElise Odinot0Alexandra Bisotto-Mignot1Toinou Frezouls2Bastien Bissaro3David Navarro4Eric Record5Frédéric Cadoret6Annick Doan7Didier Chevret8Frédéric Fine9Anne Lomascolo10OléoInnov, 19 Rue du Musée, F-13001 Marseille, FranceINRAE, Aix-Marseille Université, UMR1163 BBF Fungal Biodiversity and Biotechnology, 163 Avenue de Luminy, F-13009 Marseille, FranceINRAE, Aix-Marseille Université, UMR1163 BBF Fungal Biodiversity and Biotechnology, 163 Avenue de Luminy, F-13009 Marseille, FranceINRAE, Aix-Marseille Université, UMR1163 BBF Fungal Biodiversity and Biotechnology, 163 Avenue de Luminy, F-13009 Marseille, FranceINRAE, Aix-Marseille Université, UMR1163 BBF Fungal Biodiversity and Biotechnology, 163 Avenue de Luminy, F-13009 Marseille, FranceINRAE, Aix-Marseille Université, UMR1163 BBF Fungal Biodiversity and Biotechnology, 163 Avenue de Luminy, F-13009 Marseille, FranceINRAE, Aix-Marseille Université, UMR1163 BBF Fungal Biodiversity and Biotechnology, 163 Avenue de Luminy, F-13009 Marseille, FranceINRAE, Aix-Marseille Université, UMR1163 BBF Fungal Biodiversity and Biotechnology, 163 Avenue de Luminy, F-13009 Marseille, FranceINRAE, UMR1319 MICALIS Institute, PAPPSO, Domaine de Vilvert, F-78350 Jouy-en-Josas, FranceTERRES INOVIA, Parc Industriel, 11 Rue Monge, F-33600 Pessac, FranceINRAE, Aix-Marseille Université, UMR1163 BBF Fungal Biodiversity and Biotechnology, 163 Avenue de Luminy, F-13009 Marseille, FranceRapeseed meal (RSM) is a cheap, abundant and renewable feedstock, whose biorefinery is a current challenge for the sustainability of the oilseed sector. RSM is rich in sinapic acid (SA), a <i>p</i>-hydroxycinnamic acid that can be decarboxylated into canolol (2,6-dimethoxy-4-vinylphenol), a valuable bioactive compound. Microbial phenolic acid decarboxylases (PADs), mainly described for the non-oxidative decarboxylation of ferulic and <i>p</i>-coumaric acids, remain very poorly documented to date, for SA decarboxylation. The species <i>Neolentinus lepideus</i> has previously been shown to biotransform SA into canolol in vivo, but the enzyme responsible for bioconversion of the acid has never been characterized. In this study, we purified and characterized a new PAD from the canolol-overproducing strain <i>N. lepideus</i> BRFM15. Proteomic analysis highlighted a sole PAD-type protein sequence in the intracellular proteome of the strain. The native enzyme (<i>Nle</i>PAD) displayed an unusual outstanding activity for decarboxylating SA (V<sub>max</sub> of 600 U.mg<sup>−1</sup>, k<sub>cat</sub> of 6.3 s<sup>−1</sup> and k<sub>cat</sub>/K<sub>M</sub> of 1.6 s<sup>−1</sup>.mM<sup>−1</sup>). We showed that <i>Nle</i>PAD (a homodimer of 2 × 22 kDa) is fully active in a pH range of 5.5–7.5 and a temperature range of 30–55 °C, with optima of pH 6–6.5 and 37–45 °C, and is highly stable at 4 °C and pH 6–8. Relative ratios of specific activities on ferulic, sinapic, <i>p</i>-coumaric and caffeic acids, respectively, were 100:24.9:13.4:3.9. The enzyme demonstrated in vitro effectiveness as a biocatalyst for the synthesis of canolol in aqueous medium from commercial SA, with a molar yield of 92%. Then, we developed processes to biotransform naturally-occurring SA from RSM into canolol by combining the complementary potentialities of an <i>Aspergillus niger</i> feruloyl esterase type-A, which is able to release free SA from the raw meal by hydrolyzing its conjugated forms, and <i>Nle</i>PAD, in aqueous medium and mild conditions. <i>Nle</i>PAD decarboxylation of biobased SA led to an overall yield of 1.6–3.8 mg canolol per gram of initial meal. Besides being the first characterization of a fungal PAD able to decarboxylate SA, this report shows that <i>Nle</i>PAD is very promising as new biotechnological tool to generate biobased vinylphenols of industrial interest (especially canolol) as valuable platform chemicals for health, nutrition, cosmetics and green chemistry.https://www.mdpi.com/2306-5354/11/2/181biorefinery processcanololferulic acid<i>Neolentinus lepideus</i>phenolic acid decarboxylaserapeseed meal |
| spellingShingle | Elise Odinot Alexandra Bisotto-Mignot Toinou Frezouls Bastien Bissaro David Navarro Eric Record Frédéric Cadoret Annick Doan Didier Chevret Frédéric Fine Anne Lomascolo A New Phenolic Acid Decarboxylase from the Brown-Rot Fungus <i>Neolentinus lepideus</i> Natively Decarboxylates Biosourced Sinapic Acid into Canolol, a Bioactive Phenolic Compound Bioengineering biorefinery process canolol ferulic acid <i>Neolentinus lepideus</i> phenolic acid decarboxylase rapeseed meal |
| title | A New Phenolic Acid Decarboxylase from the Brown-Rot Fungus <i>Neolentinus lepideus</i> Natively Decarboxylates Biosourced Sinapic Acid into Canolol, a Bioactive Phenolic Compound |
| title_full | A New Phenolic Acid Decarboxylase from the Brown-Rot Fungus <i>Neolentinus lepideus</i> Natively Decarboxylates Biosourced Sinapic Acid into Canolol, a Bioactive Phenolic Compound |
| title_fullStr | A New Phenolic Acid Decarboxylase from the Brown-Rot Fungus <i>Neolentinus lepideus</i> Natively Decarboxylates Biosourced Sinapic Acid into Canolol, a Bioactive Phenolic Compound |
| title_full_unstemmed | A New Phenolic Acid Decarboxylase from the Brown-Rot Fungus <i>Neolentinus lepideus</i> Natively Decarboxylates Biosourced Sinapic Acid into Canolol, a Bioactive Phenolic Compound |
| title_short | A New Phenolic Acid Decarboxylase from the Brown-Rot Fungus <i>Neolentinus lepideus</i> Natively Decarboxylates Biosourced Sinapic Acid into Canolol, a Bioactive Phenolic Compound |
| title_sort | new phenolic acid decarboxylase from the brown rot fungus i neolentinus lepideus i natively decarboxylates biosourced sinapic acid into canolol a bioactive phenolic compound |
| topic | biorefinery process canolol ferulic acid <i>Neolentinus lepideus</i> phenolic acid decarboxylase rapeseed meal |
| url | https://www.mdpi.com/2306-5354/11/2/181 |
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