Channelling carbon flux through the meta-cleavage route for improved poly(3-hydroxyalkanoate) production from benzoate and lignin-based aromatics in Pseudomonas putida H

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dc.contributor.authorBorrero-de Acuna, J.
dc.contributor.authorGutierrez-Urrutia, I.
dc.contributor.authorHidalgo-Dumont, C.
dc.contributor.authorAravena-Carrasco, C.
dc.contributor.authorOrellana-Saez, M.
dc.contributor.authorPalominos-Gonzalez, N.
dc.contributor.authorDuuren, J.
dc.contributor.authorWagner, V.
dc.contributor.authorGlaser, L.
dc.contributor.authorBecker, J.
dc.contributor.authorKohlstedt, M.
dc.contributor.authorZacconi, F.
dc.date.accessioned2021-07-26T16:45:51Z
dc.date.available2021-07-26T16:45:51Z
dc.date.issued2020
dc.descriptionIndexación: Scopus.es
dc.description.abstractLignin-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.description.urihttps://sfamjournals.onlinelibrary.wiley.com/doi/10.1111/1751-7915.13705
dc.identifier.citationMicrobial Biotechnology, 2020es
dc.identifier.doiDOI: 10.1111/1751-7915.13705
dc.identifier.issn17517907
dc.identifier.urihttp://repositorio.unab.cl/xmlui/handle/ria/19505
dc.language.isoenes
dc.publisherWiley Open Access [University Publisher] Society for Applied Microbiology [Associate Organisation]es
dc.subjectPolyhydroxyalkanoateses
dc.subjectCupriavidus Necatores
dc.subjectBurkholderia Saccharies
dc.titleChannelling carbon flux through the meta-cleavage route for improved poly(3-hydroxyalkanoate) production from benzoate and lignin-based aromatics in Pseudomonas putida Hes
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