dc.creator | Borrero de Acuña, José Manuel | es |
dc.creator | Gutiérrez Urrutia, Izabook | es |
dc.creator | Hidalgo Dumont, Cristian | es |
dc.creator | Aravena Carrasco, Carla | es |
dc.creator | Orellana Saez, Matias | es |
dc.creator | Palominos González, Nestor | es |
dc.date.accessioned | 2022-10-03T16:04:51Z | |
dc.date.available | 2022-10-03T16:04:51Z | |
dc.date.issued | 2020-11-10 | |
dc.identifier.citation | Borrero de Acuña, J.M., Gutiérrez Urrutia, I., Hidalgo Dumont, C., Aravena Carrasco, C., Orellana Saez, M. y Palominos González, N. (2020). Channelling carbon flux through the meta-cleavage route for improved poly(3-hydroxyalkanoate) production from benzoate and lignin-based aromatics in Pseudomonas putida H. Microbial Biotechnology, 14 (6), 2385-2402. https://doi.org/10.1111/1751-7915.13705. | |
dc.identifier.issn | 1751-7915;1751-7907 | es |
dc.identifier.uri | https://hdl.handle.net/11441/137579 | |
dc.description.abstract | Lignin-based aromatics are attractive raw materials to derive medium-chain length poly(3-hydroxyalkanoates) (mcl-PHAs), biodegradable polymers of commercial value. So far, this conversion has exclusively used the ortho-cleavage route of Pseudomonas putida KT2440, which results in the secretion of toxic intermediates and limited performance. Pseudomonas putida H exhibits the ortho- and the meta-cleavage pathways where the latter appears promising because it stoichiometrically yields higher levels of acetyl-CoA. Here, we created a double-mutant H-ΔcatAΔA2 that utilizes the meta route exclusively and synthesized 30% more PHA on benzoate than the parental strain but suffered from catechol accumulation. The single deletion of the catA2 gene in the H strain provoked a slight attenuation on the enzymatic capacity of the ortho route (25%) and activation of the meta route by nearly 8-fold, producing twice as much mcl-PHAs compared to the wild type. Inline, the mutant H-ΔcatA2 showed a 2-fold increase in the intracellular malonyl-CoA abundance – the main precursor for mcl-PHAs synthesis. As inferred from flux simulation and enzyme activity assays, the superior performance of H-ΔcatA2 benefited from reduced flux through the TCA cycle and malic enzyme and diminished by-product formation. In a benzoate-based fed-batch, P. putida H-ΔcatA2 achieved a PHA titre of 6.1 g l–1 and a volumetric productivity of 1.8 g l–1 day–1. Using Kraft lignin hydrolysate as feedstock, the engineered strain formed 1.4 g l- 1 PHA. The balancing of carbon flux between the parallel catechol-degrading routes emerges as an important strategy to prevent intermediate accumulation and elevate mcl-PHA production in P. putida H and, as shown here, sets the next level to derive this sustainable biopolymer from lignin hydrolysates and aromatics. | es |
dc.format | application/pdf | es |
dc.format.extent | 18 p. | es |
dc.language.iso | eng | es |
dc.publisher | Wiley Open Access | es |
dc.relation.ispartof | Microbial Biotechnology, 14 (6), 2385-2402. | |
dc.rights | Attribution-NonCommercial-NoDerivatives 4.0 Internacional | * |
dc.rights.uri | http://creativecommons.org/licenses/by-nc-nd/4.0/ | * |
dc.title | Channelling carbon flux through the meta-cleavage route for improved poly(3-hydroxyalkanoate) production from benzoate and lignin-based aromatics in Pseudomonas putida H | es |
dc.type | info:eu-repo/semantics/article | es |
dcterms.identifier | https://ror.org/03yxnpp24 | |
dc.type.version | info:eu-repo/semantics/publishedVersion | es |
dc.rights.accessRights | info:eu-repo/semantics/openAccess | es |
dc.contributor.affiliation | Universidad de Sevilla. Departamento de Microbiología | es |
dc.relation.publisherversion | https://sfamjournals.onlinelibrary.wiley.com/doi/10.1111/1751-7915.13705 | es |
dc.identifier.doi | 10.1111/1751-7915.13705 | es |
dc.journaltitle | Microbial Biotechnology | es |
dc.publication.volumen | 14 | es |
dc.publication.issue | 6 | es |
dc.publication.initialPage | 2385 | es |
dc.publication.endPage | 2402 | es |