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• W3104304150 endingPage "2402" @default.
• W3104304150 startingPage "2385" @default.
• W3104304150 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." @default.
• W3104304150 created "2020-11-23" @default.
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• W3104304150 date "2020-11-10" @default.
• W3104304150 modified "2023-10-18" @default.
• W3104304150 title "Channelling carbon flux through the <i>meta</i> ‐cleavage route for improved poly(3‐hydroxyalkanoate) production from benzoate and lignin‐based aromatics in <i>Pseudomonas putida</i> H" @default.
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• W3104304150 doi "https://doi.org/10.1111/1751-7915.13705" @default.
• W3104304150 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/8601166" @default.
• W3104304150 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/33171015" @default.
• W3104304150 hasPublicationYear "2020" @default.
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