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Overproduction of Phenolic Compounds in Pseudomonas putida KT2440 Through Endogen Deregulation of the Shikimate Pathway

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Názov: Overproduction of Phenolic Compounds in Pseudomonas putida KT2440 Through Endogen Deregulation of the Shikimate Pathway
Autori: William Merre, Ricardo Andrade, Cyril Perot, Alexia Chandor-Proust, Caroline Ranquet
Zdroj: BioChem ; Volume 5 ; Issue 1 ; Pages: 4
Informácie o vydavateľovi: Multidisciplinary Digital Publishing Institute
Rok vydania: 2025
Zbierka: MDPI Open Access Publishing
Predmety: feedback resistance, DAHP synthase, Pseudomonas putida, enzyme engineering, shikimate pathway, p-coumaric acid
Geografické téma: agris
Popis: Metabolic engineering of the shikimate pathway offers a promising strategy for enhancing the production of aromatic compounds in microbial hosts. However, feedback inhibition of key enzymes, such as the 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase (DAHP synthase), often limits the yield of target products. In this study, we focused on the DAHP synthase (AroF-I) from Pseudomonas putida. Through computational modeling and experimental validation, we identified specific amino-acid residues responsible for tyrosine-mediated feedback inhibition. By targeted mutagenesis, we engineered DAHP synthase variants that exhibit reduced sensitivity to feedback inhibition. The introduction of these engineered enzymes into a metabolically engineered Pseudomonas putida strain resulted in significantly increased production of p-coumaric acid. Our findings provide valuable insights into the regulation of the shikimate pathway and demonstrate the potential of protein engineering to improve microbial production of aromatic compounds.
Druh dokumentu: text
Popis súboru: application/pdf
Jazyk: English
Relation: https://dx.doi.org/10.3390/biochem5010004
DOI: 10.3390/biochem5010004
Dostupnosť: https://doi.org/10.3390/biochem5010004
Rights: https://creativecommons.org/licenses/by/4.0/
Prístupové číslo: edsbas.D5242A64
Databáza: BASE
Popis
Abstrakt:Metabolic engineering of the shikimate pathway offers a promising strategy for enhancing the production of aromatic compounds in microbial hosts. However, feedback inhibition of key enzymes, such as the 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase (DAHP synthase), often limits the yield of target products. In this study, we focused on the DAHP synthase (AroF-I) from Pseudomonas putida. Through computational modeling and experimental validation, we identified specific amino-acid residues responsible for tyrosine-mediated feedback inhibition. By targeted mutagenesis, we engineered DAHP synthase variants that exhibit reduced sensitivity to feedback inhibition. The introduction of these engineered enzymes into a metabolically engineered Pseudomonas putida strain resulted in significantly increased production of p-coumaric acid. Our findings provide valuable insights into the regulation of the shikimate pathway and demonstrate the potential of protein engineering to improve microbial production of aromatic compounds.
DOI:10.3390/biochem5010004