Enhanced Protocatechuic Acid Production From Glucose Using Pseudomonas putida 3-Dehydroshikimate Dehydratase Expressed in a Phenylalanine-Overproducing Mutant of Escherichia coli
(2021) In Frontiers in Bioengineering and Biotechnology 9.- Abstract
Protocatechuic acid (PCA) is a strong antioxidant and is also a potential platform for polymer building blocks like vanillic acid, vanillin, muconic acid, and adipic acid. This report presents a study on PCA production from glucose via the shikimate pathway precursor 3-dehydroshikimate by heterologous expression of a gene encoding 3-dehydroshikimate dehydratase in Escherichia coli. The phenylalanine overproducing E. coli strain, engineered to relieve the allosteric inhibition of 3-deoxy-7-phosphoheptulonate synthase by the aromatic amino acids, was shown to give a higher yield of PCA than the unmodified strain under aerobic conditions. Highest PCA yield of 18 mol% per mol glucose and concentration of 4.2 g/L was obtained... (More)
Protocatechuic acid (PCA) is a strong antioxidant and is also a potential platform for polymer building blocks like vanillic acid, vanillin, muconic acid, and adipic acid. This report presents a study on PCA production from glucose via the shikimate pathway precursor 3-dehydroshikimate by heterologous expression of a gene encoding 3-dehydroshikimate dehydratase in Escherichia coli. The phenylalanine overproducing E. coli strain, engineered to relieve the allosteric inhibition of 3-deoxy-7-phosphoheptulonate synthase by the aromatic amino acids, was shown to give a higher yield of PCA than the unmodified strain under aerobic conditions. Highest PCA yield of 18 mol% per mol glucose and concentration of 4.2 g/L was obtained at a productivity of 0.079 g/L/h during cultivation in fed-batch mode using a feed of glucose and ammonium salt. Acetate was formed as a major side-product indicating a shift to catabolic metabolism as a result of feedback inhibition of the enzymes including 3-dehydroshikimate dehydratase by PCA when reaching a critical concentration. Indirect measurement of proton motive force by flow cytometry revealed no membrane damage of the cells by PCA, which was thus ruled out as a cause for affecting PCA formation.
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- author
- Örn, Oliver Englund LU ; Sacchetto, Stefano LU ; van Niel, Ed W. J. LU and Hatti-Kaul, Rajni LU
- organization
- publishing date
- 2021-06-24
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- 3-dehydroshikimate dehydratase, allosteric inhibition, aromatic building block, protocatechuic acid, proton motive force, shikimate pathway
- in
- Frontiers in Bioengineering and Biotechnology
- volume
- 9
- article number
- 695704
- pages
- 10 pages
- publisher
- Frontiers Media S. A.
- external identifiers
-
- scopus:85109676700
- pmid:34249890
- ISSN
- 2296-4185
- DOI
- 10.3389/fbioe.2021.695704
- project
- STEPS – Sustainable Plastics and Transition Pathways, Phase 2
- language
- English
- LU publication?
- yes
- id
- 0d4f71ca-b774-480e-8f29-e584385554a4
- date added to LUP
- 2021-09-08 10:12:00
- date last changed
- 2024-06-15 15:55:36
@article{0d4f71ca-b774-480e-8f29-e584385554a4,
abstract = {{<p>Protocatechuic acid (PCA) is a strong antioxidant and is also a potential platform for polymer building blocks like vanillic acid, vanillin, muconic acid, and adipic acid. This report presents a study on PCA production from glucose via the shikimate pathway precursor 3-dehydroshikimate by heterologous expression of a gene encoding 3-dehydroshikimate dehydratase in <i>Escherichia coli</i>. The phenylalanine overproducing <i>E. coli</i> strain, engineered to relieve the allosteric inhibition of 3-deoxy-7-phosphoheptulonate synthase by the aromatic amino acids, was shown to give a higher yield of PCA than the unmodified strain under aerobic conditions. Highest PCA yield of 18 mol% per mol glucose and concentration of 4.2 g/L was obtained at a productivity of 0.079 g/L/h during cultivation in fed-batch mode using a feed of glucose and ammonium salt. Acetate was formed as a major side-product indicating a shift to catabolic metabolism as a result of feedback inhibition of the enzymes including 3-dehydroshikimate dehydratase by PCA when reaching a critical concentration. Indirect measurement of proton motive force by flow cytometry revealed no membrane damage of the cells by PCA, which was thus ruled out as a cause for affecting PCA formation.</p>}},
author = {{Örn, Oliver Englund and Sacchetto, Stefano and van Niel, Ed W. J. and Hatti-Kaul, Rajni}},
issn = {{2296-4185}},
keywords = {{3-dehydroshikimate dehydratase; allosteric inhibition; aromatic building block; protocatechuic acid; proton motive force; shikimate pathway}},
language = {{eng}},
month = {{06}},
publisher = {{Frontiers Media S. A.}},
series = {{Frontiers in Bioengineering and Biotechnology}},
title = {{Enhanced Protocatechuic Acid Production From Glucose Using <i>Pseudomonas putida</i> 3-Dehydroshikimate Dehydratase Expressed in a Phenylalanine-Overproducing Mutant of <i>Escherichia coli</i>}},
url = {{http://dx.doi.org/10.3389/fbioe.2021.695704}},
doi = {{10.3389/fbioe.2021.695704}},
volume = {{9}},
year = {{2021}},
}