Cargando…
Combinatorial pathway balancing provides biosynthetic access to 2-fluoro-cis,cis-muconate in engineered Pseudomonas putida
The wealth of bio-based building blocks produced by engineered microorganisms seldom include halogen atoms. Muconate is a platform chemical with a number of industrial applications that could be broadened by introducing fluorine atoms to tune its physicochemical properties. The soil bacterium Pseudo...
Autores principales: | , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier B.V
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8711041/ https://www.ncbi.nlm.nih.gov/pubmed/34977847 http://dx.doi.org/10.1016/j.checat.2021.09.002 |
_version_ | 1784623295533940736 |
---|---|
author | Wirth, Nicolas T. Nikel, Pablo I. |
author_facet | Wirth, Nicolas T. Nikel, Pablo I. |
author_sort | Wirth, Nicolas T. |
collection | PubMed |
description | The wealth of bio-based building blocks produced by engineered microorganisms seldom include halogen atoms. Muconate is a platform chemical with a number of industrial applications that could be broadened by introducing fluorine atoms to tune its physicochemical properties. The soil bacterium Pseudomonas putida naturally assimilates benzoate via the ortho-cleavage pathway with cis,cis-muconate as intermediate. Here, we harnessed the native enzymatic machinery (encoded within the ben and cat gene clusters) to provide catalytic access to 2-fluoro-cis,cis-muconate (2-FMA) from fluorinated benzoates. The reactions in this pathway are highly imbalanced, leading to accumulation of toxic intermediates and limited substrate conversion. By disentangling regulatory patterns of ben and cat in response to fluorinated effectors, metabolic activities were adjusted to favor 2-FMA biosynthesis. After implementing this combinatorial approach, engineered P. putida converted 3-fluorobenzoate to 2-FMA at the maximum theoretical yield. Hence, this study illustrates how synthetic biology can expand the diversity of nature's biochemical catalysis. |
format | Online Article Text |
id | pubmed-8711041 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Elsevier B.V |
record_format | MEDLINE/PubMed |
spelling | pubmed-87110412021-12-30 Combinatorial pathway balancing provides biosynthetic access to 2-fluoro-cis,cis-muconate in engineered Pseudomonas putida Wirth, Nicolas T. Nikel, Pablo I. Chem Catal Article The wealth of bio-based building blocks produced by engineered microorganisms seldom include halogen atoms. Muconate is a platform chemical with a number of industrial applications that could be broadened by introducing fluorine atoms to tune its physicochemical properties. The soil bacterium Pseudomonas putida naturally assimilates benzoate via the ortho-cleavage pathway with cis,cis-muconate as intermediate. Here, we harnessed the native enzymatic machinery (encoded within the ben and cat gene clusters) to provide catalytic access to 2-fluoro-cis,cis-muconate (2-FMA) from fluorinated benzoates. The reactions in this pathway are highly imbalanced, leading to accumulation of toxic intermediates and limited substrate conversion. By disentangling regulatory patterns of ben and cat in response to fluorinated effectors, metabolic activities were adjusted to favor 2-FMA biosynthesis. After implementing this combinatorial approach, engineered P. putida converted 3-fluorobenzoate to 2-FMA at the maximum theoretical yield. Hence, this study illustrates how synthetic biology can expand the diversity of nature's biochemical catalysis. Elsevier B.V 2021-11-18 /pmc/articles/PMC8711041/ /pubmed/34977847 http://dx.doi.org/10.1016/j.checat.2021.09.002 Text en © 2021 The Author(s) https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Wirth, Nicolas T. Nikel, Pablo I. Combinatorial pathway balancing provides biosynthetic access to 2-fluoro-cis,cis-muconate in engineered Pseudomonas putida |
title | Combinatorial pathway balancing provides biosynthetic access to 2-fluoro-cis,cis-muconate in engineered Pseudomonas putida |
title_full | Combinatorial pathway balancing provides biosynthetic access to 2-fluoro-cis,cis-muconate in engineered Pseudomonas putida |
title_fullStr | Combinatorial pathway balancing provides biosynthetic access to 2-fluoro-cis,cis-muconate in engineered Pseudomonas putida |
title_full_unstemmed | Combinatorial pathway balancing provides biosynthetic access to 2-fluoro-cis,cis-muconate in engineered Pseudomonas putida |
title_short | Combinatorial pathway balancing provides biosynthetic access to 2-fluoro-cis,cis-muconate in engineered Pseudomonas putida |
title_sort | combinatorial pathway balancing provides biosynthetic access to 2-fluoro-cis,cis-muconate in engineered pseudomonas putida |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8711041/ https://www.ncbi.nlm.nih.gov/pubmed/34977847 http://dx.doi.org/10.1016/j.checat.2021.09.002 |
work_keys_str_mv | AT wirthnicolast combinatorialpathwaybalancingprovidesbiosyntheticaccessto2fluorociscismuconateinengineeredpseudomonasputida AT nikelpabloi combinatorialpathwaybalancingprovidesbiosyntheticaccessto2fluorociscismuconateinengineeredpseudomonasputida |