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In planta production of the nylon precursor beta-ketoadipate

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Title: In planta production of the nylon precursor beta-ketoadipate
Authors: Sami Kazaz, Jaya Tripathi, Yang Tian, Halbay Turumtay, Dylan Chin, İrem Pamukçu, Monikaben Nimavat, Emine Akyuz Turumtay, Edward E.K. Baidoo, Corinne D. Scown, Aymerick Eudes
Source: Journal of Biotechnology, vol 404
Publisher Information: Elsevier BV, 2025.
Publication Year: 2025
Subject Terms: 3001 Agricultural biotechnology (for-2020), Theory and Criticism (for-2020), 7 Affordable and Clean Energy (sdg), 30 Agricultural, Adipates (mesh), Lignin, Veterinary and Food Sciences (for-2020), theory and criticism (for-2020), 3206 Medical Biotechnology (for-2020), 3601 Art History, Arabidopsis (mesh), 32 Biomedical and Clinical Sciences (for-2020), Plant engineering, Techno-economic analysis, Metabolic Engineering (mesh), 3206 Medical biotechnology (for-2020), Shikimic Acid (mesh), Shikimate pathway, Nicotiana (mesh), Biotechnology (science-metrix), 3001 Agricultural Biotechnology (for-2020), Plants, Genetically Modified (mesh), Beta-ketoadipate, 06 Biological Sciences (for), 09 Engineering (for), 36 Creative Arts and Writing (for-2020), Bioproduct, 10 Technology (for), 3601 Art history
Description: Beta-ketoadipate (βKA) is an intermediate of the βKA pathway involved in the degradation of aromatic compounds in several bacteria and fungi. Beta-ketoadipate also represents a promising chemical for the manufacturing of performance-advantaged nylons. We established a strategy for the in planta synthesis of βKA via manipulation of the shikimate pathway and the expression of bacterial enzymes from the βKA pathway. Using Nicotiana benthamiana as a transient expression system, we demonstrated the efficient conversion of protocatechuate (PCA) to βKA when plastid-targeted bacterial-derived PCA 3,4-dioxygenase (PcaHG) and 3-carboxy-cis,cis-muconate cycloisomerase (PcaB) were co-expressed with 3-deoxy-D-arabinoheptulosonate 7-phosphate synthase (AroG) and 3-dehydroshikimate dehydratase (QsuB). This metabolic pathway was reconstituted in Arabidopsis by introducing a construct (pAtβKA) with stacked pcaG, pcaH, and pcaB genes into a PCA-overproducing genetic background that expresses AroG and QsuB (referred as QsuB-2). The resulting QsuB-2 x pAtβKA stable lines displayed βKA titers as high as 0.25 % on a dry weight basis in stems, along with a drastic reduction in lignin content and improvement of biomass saccharification efficiency compared to wild-type controls, and without any significant reduction in biomass yields. Using biomass sorghum as a potential crop for large-scale βKA production, techno-economic analysis indicated that βKA accumulated at titers of 0.25 % and 4 % on a dry weight basis could be competitively priced in the range of $2.04-34.49/kg and $0.47-2.12/kg, respectively, depending on the selling price of the residual biomass recovered after βKA extraction. This study lays the foundation for a more environmentally-friendly synthesis of βKA using plants as production hosts.
Document Type: Article
File Description: application/pdf
Language: English
ISSN: 0168-1656
DOI: 10.1016/j.jbiotec.2025.04.008
Access URL: https://pubmed.ncbi.nlm.nih.gov/40228630
https://escholarship.org/content/qt6g61625x/qt6g61625x.pdf
https://escholarship.org/uc/item/6g61625x
Rights: CC BY
Accession Number: edsair.doi.dedup.....15c5286072c99e97031b6603098a4888
Database: OpenAIRE
Description
Abstract:Beta-ketoadipate (βKA) is an intermediate of the βKA pathway involved in the degradation of aromatic compounds in several bacteria and fungi. Beta-ketoadipate also represents a promising chemical for the manufacturing of performance-advantaged nylons. We established a strategy for the in planta synthesis of βKA via manipulation of the shikimate pathway and the expression of bacterial enzymes from the βKA pathway. Using Nicotiana benthamiana as a transient expression system, we demonstrated the efficient conversion of protocatechuate (PCA) to βKA when plastid-targeted bacterial-derived PCA 3,4-dioxygenase (PcaHG) and 3-carboxy-cis,cis-muconate cycloisomerase (PcaB) were co-expressed with 3-deoxy-D-arabinoheptulosonate 7-phosphate synthase (AroG) and 3-dehydroshikimate dehydratase (QsuB). This metabolic pathway was reconstituted in Arabidopsis by introducing a construct (pAtβKA) with stacked pcaG, pcaH, and pcaB genes into a PCA-overproducing genetic background that expresses AroG and QsuB (referred as QsuB-2). The resulting QsuB-2 x pAtβKA stable lines displayed βKA titers as high as 0.25 % on a dry weight basis in stems, along with a drastic reduction in lignin content and improvement of biomass saccharification efficiency compared to wild-type controls, and without any significant reduction in biomass yields. Using biomass sorghum as a potential crop for large-scale βKA production, techno-economic analysis indicated that βKA accumulated at titers of 0.25 % and 4 % on a dry weight basis could be competitively priced in the range of $2.04-34.49/kg and $0.47-2.12/kg, respectively, depending on the selling price of the residual biomass recovered after βKA extraction. This study lays the foundation for a more environmentally-friendly synthesis of βKA using plants as production hosts.
ISSN:01681656
DOI:10.1016/j.jbiotec.2025.04.008