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      Lignin conversion to β-ketoadipic acid by Pseudomonas putida via metabolic engineering and bioprocess development

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          Abstract

          Bioconversion of a heterogeneous mixture of lignin-related aromatic compounds (LRCs) to a single product via microbial biocatalysts is a promising approach to valorize lignin. Here, Pseudomonas putida KT2440 was engineered to convert mixed p-coumaroyl– and coniferyl-type LRCs to β-ketoadipic acid, a precursor for performance-advantaged polymers. Expression of enzymes mediating aromatic O-demethylation, hydroxylation, and ring-opening steps was tuned, and a global regulator was deleted. β-ketoadipate titers of 44.5 and 25 grams per liter and productivities of 1.15 and 0.66 grams per liter per hour were achieved from model LRCs and corn stover-derived LRCs, respectively, the latter representing an overall yield of 0.10 grams per gram corn stover-derived lignin. Technoeconomic analysis of the bioprocess and downstream processing predicted a β-ketoadipate minimum selling price of $2.01 per kilogram, which is cost competitive with fossil carbon-derived adipic acid ($1.10 to 1.80 per kilogram). Overall, this work achieved bioproduction metrics with economic relevance for conversion of lignin-derived streams into a performance-advantaged bioproduct.

          Abstract

          An engineered bacterium efficiently converts plant biomass-derived aromatic compounds to a performance-advantaged bioproduct.

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          Most cited references76

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          The ecoinvent database version 3 (part I): overview and methodology

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            Lignin valorization: improving lignin processing in the biorefinery.

            Research and development activities directed toward commercial production of cellulosic ethanol have created the opportunity to dramatically increase the transformation of lignin to value-added products. Here, we highlight recent advances in this lignin valorization effort. Discovery of genetic variants in native populations of bioenergy crops and direct manipulation of biosynthesis pathways have produced lignin feedstocks with favorable properties for recovery and downstream conversion. Advances in analytical chemistry and computational modeling detail the structure of the modified lignin and direct bioengineering strategies for future targeted properties. Refinement of biomass pretreatment technologies has further facilitated lignin recovery, and this coupled with genetic engineering will enable new uses for this biopolymer, including low-cost carbon fibers, engineered plastics and thermoplastic elastomers, polymeric foams, fungible fuels, and commodity chemicals.
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              The catalytic valorization of lignin for the production of renewable chemicals.

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                Author and article information

                Contributors
                Role: ConceptualizationRole: Formal analysisRole: Funding acquisitionRole: InvestigationRole: MethodologyRole: Project administrationRole: ValidationRole: VisualizationRole: Writing - original draftRole: Writing - review & editing
                Role: InvestigationRole: MethodologyRole: ValidationRole: Writing - original draftRole: Writing - review & editing
                Role: InvestigationRole: MethodologyRole: ResourcesRole: ValidationRole: VisualizationRole: Writing - review & editing
                Role: Formal analysisRole: MethodologyRole: VisualizationRole: Writing - original draft
                Role: Data curationRole: Formal analysisRole: InvestigationRole: MethodologyRole: ResourcesRole: SupervisionRole: ValidationRole: Writing - review & editing
                Role: Formal analysisRole: MethodologyRole: ResourcesRole: Visualization
                Role: InvestigationRole: MethodologyRole: Resources
                Role: InvestigationRole: MethodologyRole: Resources
                Role: InvestigationRole: ValidationRole: Visualization
                Role: InvestigationRole: ValidationRole: Writing - review & editing
                Role: ConceptualizationRole: MethodologyRole: Writing - review & editing
                Role: Project administrationRole: SupervisionRole: Writing - original draftRole: Writing - review & editing
                Role: ConceptualizationRole: Funding acquisitionRole: MethodologyRole: Project administrationRole: SupervisionRole: Writing - original draftRole: Writing - review & editing
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: Funding acquisitionRole: MethodologyRole: Project administrationRole: SupervisionRole: ValidationRole: Writing - original draftRole: Writing - review & editing
                Journal
                Sci Adv
                Sci Adv
                sciadv
                advances
                Science Advances
                American Association for the Advancement of Science
                2375-2548
                September 2023
                06 September 2023
                : 9
                : 36
                : eadj0053
                Affiliations
                [ 1 ]Renewable Resources and Enabling Sciences Center, National Renewable Energy Laboratory, Golden, CO, USA.
                [ 2 ]Department of Chemical and Biological Engineering, University of Wisconsin-Madison, Madison, WI, USA.
                [ 3 ]Catalytic Carbon Transformation and Scale-Up Center, National Renewable Energy Laboratory, Golden, CO, USA.
                Author notes
                [* ]Corresponding author. Email: davinia.salvachua@ 123456nrel.gov (D.S.); gregg.beckham@ 123456nrel.gov (G.T.B.)
                [†]

                These authors contributed equally to this work.

                [‡]

                Present address: School of Food Science and Biotechnology, Kyungpook National University, Daegu, South Korea.

                [§]

                Present address: CMC Drug Product Development, Novo Nordisk A/S, 2760 Måløv, Denmark.

                Author information
                https://orcid.org/0000-0001-7147-2863
                https://orcid.org/0000-0001-7910-8206
                https://orcid.org/0000-0002-1794-2921
                https://orcid.org/0000-0003-3438-253X
                https://orcid.org/0000-0002-7791-6389
                https://orcid.org/0000-0002-9110-2410
                https://orcid.org/0000-0002-7350-4862
                https://orcid.org/0000-0002-5114-742X
                https://orcid.org/0000-0001-6958-0136
                https://orcid.org/0000-0002-2979-4751
                https://orcid.org/0000-0002-9232-9959
                https://orcid.org/0000-0002-3480-212X
                https://orcid.org/0000-0003-0799-061X
                Article
                adj0053
                10.1126/sciadv.adj0053
                10482344
                37672573
                d31b1eed-2721-4087-b119-f02bddf7a551
                Copyright © 2023 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).

                This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial license, which permits use, distribution, and reproduction in any medium, so long as the resultant use is not for commercial advantage and provided the original work is properly cited.

                History
                : 31 May 2023
                : 04 August 2023
                Funding
                Funded by: DOE Bioenergy Technologies Office;
                Award ID: DE-AC36-08GO28308
                Funded by: DOE Office of Science Graduate Student Research;
                Award ID: DE-SC0014664
                Categories
                Research Article
                Physical and Materials Sciences
                SciAdv r-articles
                Microbiology
                Applied Sciences and Engineering
                Applied Sciences and Engineering
                Custom metadata
                Eunice Diego

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