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Conversion of methionine biosynthesis in Escherichia coli from trans- to direct-sulfurylation enhances extracellular methionine levels

Methionine is an essential amino acid in mammals and a precursor for vital metabolites required for the survival of all organisms. Consequently, its inclusion is required in diverse applications, such as food, feed, and pharmaceuticals. Although amino acids and other metabolites are commonly produce...

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Autores principales: Gruzdev, Nadya, Hacham, Yael, Haviv, Hadar, Stern, Inbar, Gabay, Matan, Bloch, Itai, Amir, Rachel, Gal, Maayan, Yadid, Itamar
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10416483/
https://www.ncbi.nlm.nih.gov/pubmed/37568230
http://dx.doi.org/10.1186/s12934-023-02150-x
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author Gruzdev, Nadya
Hacham, Yael
Haviv, Hadar
Stern, Inbar
Gabay, Matan
Bloch, Itai
Amir, Rachel
Gal, Maayan
Yadid, Itamar
author_facet Gruzdev, Nadya
Hacham, Yael
Haviv, Hadar
Stern, Inbar
Gabay, Matan
Bloch, Itai
Amir, Rachel
Gal, Maayan
Yadid, Itamar
author_sort Gruzdev, Nadya
collection PubMed
description Methionine is an essential amino acid in mammals and a precursor for vital metabolites required for the survival of all organisms. Consequently, its inclusion is required in diverse applications, such as food, feed, and pharmaceuticals. Although amino acids and other metabolites are commonly produced through microbial fermentation, high-yield biosynthesis of L-methionine remains a significant challenge due to the strict cellular regulation of the biosynthesis pathway. As a result, methionine is produced primarily synthetically, resulting in a racemic mixture of D,L-methionine. This study explores methionine bio-production in E. coli by replacing its native trans-sulfurylation pathway with the more common direct-sulfurylation pathway used by other bacteria. To this end, we generated a methionine auxotroph E. coli strain (MG1655) by simultaneously deleting metA and metB genes and complementing them with metX and metY from different bacteria. Complementation of the genetically modified E. coli with metX/metY from Cyclobacterium marinum or Deinococcus geothermalis, together with the deletion of the global repressor metJ and overexpression of the transporter yjeH, resulted in a substantial increase of up to 126 and 160-fold methionine relative to the wild-type strain, respectively, and accumulation of up to 700 mg/L using minimal MOPS medium and 2 ml culture. Our findings provide a method to study methionine biosynthesis and a chassis for enhancing L-methionine production by fermentation. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12934-023-02150-x.
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spelling pubmed-104164832023-08-12 Conversion of methionine biosynthesis in Escherichia coli from trans- to direct-sulfurylation enhances extracellular methionine levels Gruzdev, Nadya Hacham, Yael Haviv, Hadar Stern, Inbar Gabay, Matan Bloch, Itai Amir, Rachel Gal, Maayan Yadid, Itamar Microb Cell Fact Research Methionine is an essential amino acid in mammals and a precursor for vital metabolites required for the survival of all organisms. Consequently, its inclusion is required in diverse applications, such as food, feed, and pharmaceuticals. Although amino acids and other metabolites are commonly produced through microbial fermentation, high-yield biosynthesis of L-methionine remains a significant challenge due to the strict cellular regulation of the biosynthesis pathway. As a result, methionine is produced primarily synthetically, resulting in a racemic mixture of D,L-methionine. This study explores methionine bio-production in E. coli by replacing its native trans-sulfurylation pathway with the more common direct-sulfurylation pathway used by other bacteria. To this end, we generated a methionine auxotroph E. coli strain (MG1655) by simultaneously deleting metA and metB genes and complementing them with metX and metY from different bacteria. Complementation of the genetically modified E. coli with metX/metY from Cyclobacterium marinum or Deinococcus geothermalis, together with the deletion of the global repressor metJ and overexpression of the transporter yjeH, resulted in a substantial increase of up to 126 and 160-fold methionine relative to the wild-type strain, respectively, and accumulation of up to 700 mg/L using minimal MOPS medium and 2 ml culture. Our findings provide a method to study methionine biosynthesis and a chassis for enhancing L-methionine production by fermentation. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12934-023-02150-x. BioMed Central 2023-08-11 /pmc/articles/PMC10416483/ /pubmed/37568230 http://dx.doi.org/10.1186/s12934-023-02150-x Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Gruzdev, Nadya
Hacham, Yael
Haviv, Hadar
Stern, Inbar
Gabay, Matan
Bloch, Itai
Amir, Rachel
Gal, Maayan
Yadid, Itamar
Conversion of methionine biosynthesis in Escherichia coli from trans- to direct-sulfurylation enhances extracellular methionine levels
title Conversion of methionine biosynthesis in Escherichia coli from trans- to direct-sulfurylation enhances extracellular methionine levels
title_full Conversion of methionine biosynthesis in Escherichia coli from trans- to direct-sulfurylation enhances extracellular methionine levels
title_fullStr Conversion of methionine biosynthesis in Escherichia coli from trans- to direct-sulfurylation enhances extracellular methionine levels
title_full_unstemmed Conversion of methionine biosynthesis in Escherichia coli from trans- to direct-sulfurylation enhances extracellular methionine levels
title_short Conversion of methionine biosynthesis in Escherichia coli from trans- to direct-sulfurylation enhances extracellular methionine levels
title_sort conversion of methionine biosynthesis in escherichia coli from trans- to direct-sulfurylation enhances extracellular methionine levels
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10416483/
https://www.ncbi.nlm.nih.gov/pubmed/37568230
http://dx.doi.org/10.1186/s12934-023-02150-x
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