Overproduction of Phenolic Compounds in <i>Pseudomonas putida</i> KT2440 Through Endogen Deregulation of the Shikimate Pathway
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 t...
Saved in:
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2025-03-01
|
| Series: | BioChem |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2673-6411/5/1/4 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850204001380859904 |
|---|---|
| author | William Merre Ricardo Andrade Cyril Perot Alexia Chandor-Proust Caroline Ranquet |
| author_facet | William Merre Ricardo Andrade Cyril Perot Alexia Chandor-Proust Caroline Ranquet |
| author_sort | William Merre |
| collection | DOAJ |
| description | 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 <i>Pseudomonas putida</i>. 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 <i>Pseudomonas putida</i> 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. |
| format | Article |
| id | doaj-art-9bf4a657bbca4dcba5f9e2884f7e6335 |
| institution | OA Journals |
| issn | 2673-6411 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | BioChem |
| spelling | doaj-art-9bf4a657bbca4dcba5f9e2884f7e63352025-08-20T02:11:22ZengMDPI AGBioChem2673-64112025-03-0151410.3390/biochem5010004Overproduction of Phenolic Compounds in <i>Pseudomonas putida</i> KT2440 Through Endogen Deregulation of the Shikimate PathwayWilliam Merre0Ricardo Andrade1Cyril Perot2Alexia Chandor-Proust3Caroline Ranquet4BGene Genetics, 7 Rue des Arts et Métiers, 38000 Grenoble, FranceBGene Genetics, 7 Rue des Arts et Métiers, 38000 Grenoble, FranceBGene Genetics, 7 Rue des Arts et Métiers, 38000 Grenoble, FranceBGene Genetics, 7 Rue des Arts et Métiers, 38000 Grenoble, FranceBGene Genetics, 7 Rue des Arts et Métiers, 38000 Grenoble, FranceMetabolic 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 <i>Pseudomonas putida</i>. 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 <i>Pseudomonas putida</i> 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.https://www.mdpi.com/2673-6411/5/1/4feedback resistanceDAHP synthase<i>Pseudomonas putida</i>enzyme engineeringshikimate pathwayp-coumaric acid |
| spellingShingle | William Merre Ricardo Andrade Cyril Perot Alexia Chandor-Proust Caroline Ranquet Overproduction of Phenolic Compounds in <i>Pseudomonas putida</i> KT2440 Through Endogen Deregulation of the Shikimate Pathway BioChem feedback resistance DAHP synthase <i>Pseudomonas putida</i> enzyme engineering shikimate pathway p-coumaric acid |
| title | Overproduction of Phenolic Compounds in <i>Pseudomonas putida</i> KT2440 Through Endogen Deregulation of the Shikimate Pathway |
| title_full | Overproduction of Phenolic Compounds in <i>Pseudomonas putida</i> KT2440 Through Endogen Deregulation of the Shikimate Pathway |
| title_fullStr | Overproduction of Phenolic Compounds in <i>Pseudomonas putida</i> KT2440 Through Endogen Deregulation of the Shikimate Pathway |
| title_full_unstemmed | Overproduction of Phenolic Compounds in <i>Pseudomonas putida</i> KT2440 Through Endogen Deregulation of the Shikimate Pathway |
| title_short | Overproduction of Phenolic Compounds in <i>Pseudomonas putida</i> KT2440 Through Endogen Deregulation of the Shikimate Pathway |
| title_sort | overproduction of phenolic compounds in i pseudomonas putida i kt2440 through endogen deregulation of the shikimate pathway |
| topic | feedback resistance DAHP synthase <i>Pseudomonas putida</i> enzyme engineering shikimate pathway p-coumaric acid |
| url | https://www.mdpi.com/2673-6411/5/1/4 |
| work_keys_str_mv | AT williammerre overproductionofphenoliccompoundsinipseudomonasputidaikt2440throughendogenderegulationoftheshikimatepathway AT ricardoandrade overproductionofphenoliccompoundsinipseudomonasputidaikt2440throughendogenderegulationoftheshikimatepathway AT cyrilperot overproductionofphenoliccompoundsinipseudomonasputidaikt2440throughendogenderegulationoftheshikimatepathway AT alexiachandorproust overproductionofphenoliccompoundsinipseudomonasputidaikt2440throughendogenderegulationoftheshikimatepathway AT carolineranquet overproductionofphenoliccompoundsinipseudomonasputidaikt2440throughendogenderegulationoftheshikimatepathway |