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Comparative Analysis of Catabolic and Anabolic Dehydroshikimate Dehydratases for 3,4-DHBA Production in Escherichia coli

The production of 3,4-dihydroxybenzoic acid (3,4-DHBA or protocatechuate) is a relevant task owing to 3,4-DHBA’s pharmaceutical properties and its use as a precursor for subsequent synthesis of high value-added chemicals. The microbial production of 3,4-DHBA using dehydroshikimate dehydratase (DSD)...

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Autores principales: Shmonova, Ekaterina A., Savrasova, Ekaterina A., Fedorova, Elizaveta N., Doroshenko, Vera G.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9324987/
https://www.ncbi.nlm.nih.gov/pubmed/35889076
http://dx.doi.org/10.3390/microorganisms10071357
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author Shmonova, Ekaterina A.
Savrasova, Ekaterina A.
Fedorova, Elizaveta N.
Doroshenko, Vera G.
author_facet Shmonova, Ekaterina A.
Savrasova, Ekaterina A.
Fedorova, Elizaveta N.
Doroshenko, Vera G.
author_sort Shmonova, Ekaterina A.
collection PubMed
description The production of 3,4-dihydroxybenzoic acid (3,4-DHBA or protocatechuate) is a relevant task owing to 3,4-DHBA’s pharmaceutical properties and its use as a precursor for subsequent synthesis of high value-added chemicals. The microbial production of 3,4-DHBA using dehydroshikimate dehydratase (DSD) (EC: 4.2.1.118) has been demonstrated previously. DSDs from soil-dwelling organisms (where DSD is involved in quinate/shikimate degradation) and from Bacillus spp. (synthesizing the 3,4-DHBA-containing siderophore) were compared in terms of the kinetic properties and their ability to produce 3,4-DHBA. Catabolic DSDs from Corynebacterium glutamicum (QsuB) and Neurospora crassa (Qa-4) had higher K(m) (1 and 0.6 mM, respectively) and k(cat) (61 and 220 s(−1), respectively) than biosynthetic AsbF from Bacillus thuringiensis (K(m)~0.04 mM, k(cat)~1 s(−1)). Product inhibition was found to be a crucial factor when choosing DSD for strain development. AsbF was more inhibited by 3,4-DHBA (IC(50)~0.08 mM), and Escherichia coli MG1655 ΔaroE P(lacUV5)-asbF(attφ80) strain provided only 0.2 g/L 3,4-DHBA in test-tube fermentation. Isogenic strains MG1655 ΔaroE P(lacUV5)-qsuB(attφ80) and MG1655 ΔaroE P(lacUV5)-qa-4(attφ80) expressing QsuB and Qa-4 with IC(50) ~0.35 mM and ~0.64 mM, respectively, accumulated 2.7 g/L 3,4-DHBA under the same conditions.
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spelling pubmed-93249872022-07-27 Comparative Analysis of Catabolic and Anabolic Dehydroshikimate Dehydratases for 3,4-DHBA Production in Escherichia coli Shmonova, Ekaterina A. Savrasova, Ekaterina A. Fedorova, Elizaveta N. Doroshenko, Vera G. Microorganisms Article The production of 3,4-dihydroxybenzoic acid (3,4-DHBA or protocatechuate) is a relevant task owing to 3,4-DHBA’s pharmaceutical properties and its use as a precursor for subsequent synthesis of high value-added chemicals. The microbial production of 3,4-DHBA using dehydroshikimate dehydratase (DSD) (EC: 4.2.1.118) has been demonstrated previously. DSDs from soil-dwelling organisms (where DSD is involved in quinate/shikimate degradation) and from Bacillus spp. (synthesizing the 3,4-DHBA-containing siderophore) were compared in terms of the kinetic properties and their ability to produce 3,4-DHBA. Catabolic DSDs from Corynebacterium glutamicum (QsuB) and Neurospora crassa (Qa-4) had higher K(m) (1 and 0.6 mM, respectively) and k(cat) (61 and 220 s(−1), respectively) than biosynthetic AsbF from Bacillus thuringiensis (K(m)~0.04 mM, k(cat)~1 s(−1)). Product inhibition was found to be a crucial factor when choosing DSD for strain development. AsbF was more inhibited by 3,4-DHBA (IC(50)~0.08 mM), and Escherichia coli MG1655 ΔaroE P(lacUV5)-asbF(attφ80) strain provided only 0.2 g/L 3,4-DHBA in test-tube fermentation. Isogenic strains MG1655 ΔaroE P(lacUV5)-qsuB(attφ80) and MG1655 ΔaroE P(lacUV5)-qa-4(attφ80) expressing QsuB and Qa-4 with IC(50) ~0.35 mM and ~0.64 mM, respectively, accumulated 2.7 g/L 3,4-DHBA under the same conditions. MDPI 2022-07-05 /pmc/articles/PMC9324987/ /pubmed/35889076 http://dx.doi.org/10.3390/microorganisms10071357 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Shmonova, Ekaterina A.
Savrasova, Ekaterina A.
Fedorova, Elizaveta N.
Doroshenko, Vera G.
Comparative Analysis of Catabolic and Anabolic Dehydroshikimate Dehydratases for 3,4-DHBA Production in Escherichia coli
title Comparative Analysis of Catabolic and Anabolic Dehydroshikimate Dehydratases for 3,4-DHBA Production in Escherichia coli
title_full Comparative Analysis of Catabolic and Anabolic Dehydroshikimate Dehydratases for 3,4-DHBA Production in Escherichia coli
title_fullStr Comparative Analysis of Catabolic and Anabolic Dehydroshikimate Dehydratases for 3,4-DHBA Production in Escherichia coli
title_full_unstemmed Comparative Analysis of Catabolic and Anabolic Dehydroshikimate Dehydratases for 3,4-DHBA Production in Escherichia coli
title_short Comparative Analysis of Catabolic and Anabolic Dehydroshikimate Dehydratases for 3,4-DHBA Production in Escherichia coli
title_sort comparative analysis of catabolic and anabolic dehydroshikimate dehydratases for 3,4-dhba production in escherichia coli
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9324987/
https://www.ncbi.nlm.nih.gov/pubmed/35889076
http://dx.doi.org/10.3390/microorganisms10071357
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