Directed evolution of biosynthetic pathways. Recruitment of cysteine thioethers for constructing the cell wall of Escherichia coli.

C Richaud,D Mengin-Lecreulx,S Pochet,E J Johnson,G N Cohen,P Marlière

doi:10.1016/s0021-9258(19)74186-6

C Richaud, D Mengin-Lecreulx + Show 4 more

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https://doi.org/10.1016/s0021-9258(19)74186-6

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Abstract

We report that expansion of thioether biosynthesis in Escherichia coli generates sulfur-containing amino acids that can replace meso-diaminopimelate, the essential amino acid used for cross-linking the cell wall. This was accomplished by jointly overexpressing the metB gene coding for L-cystathionine gamma-synthase and disrupting the metC gene, whose product, L-cystathionine beta-lyase, is responsible for the destruction of L-cystathionine and other L-cysteine thioethers. As a result, meso-lanthionine and L-allo-cystathionine were produced endogenously and incorporated in the peptidoglycan, thereby enabling E. coli strains auxotrophic for diaminopimelate to grow in its absence. Thus, current techniques of metabolic engineering can be applied to evolving the chemical constitution of living cells beyond its present state.

Highlights

Chimie Organique (CNRS URA 487), the 11 Unite de Biochimie Cellulaire (CNRS URA 1129),Znstitut Pasteur, 25, rue du Docteur Roux, 75015 Paris, France, the SLaboratoire de Biochimie MoEculaire et Cellulaire (CNRS URA 1131), Bhtiment 432, Universite Paris-Sud, 91405 Orsay, France, and the VDepartment of Microbiology and Immunology, Tulane University School of Medicine, New Orleans,LOuisiaG 70112
We report that expansion of thioether biosynthesis in Escherichia coli generatessulfur-containing amino acids that can replace meso-diaminopimelate, the essential amino acid used for cross-linking the cell wall
Current techniques of metabolic engineering can be applied to evolving the chemical constitution of living cells beyond its present state

Summary

Introduction

Chimie Organique (CNRS URA 487), the 11 Unite de Biochimie Cellulaire (CNRS URA 1129),Znstitut Pasteur, 25, rue du Docteur Roux, 75015 Paris, France, the SLaboratoire de Biochimie MoEculaire et Cellulaire (CNRS URA 1131), Bhtiment 432, Universite Paris-Sud, 91405 Orsay, France, and the VDepartment of Microbiology and Immunology, Tulane University School of Medicine, New Orleans,LOuisiaG 70112. We report that expansion of thioether biosynthesis in Escherichia coli generatessulfur-containing amino acids that can replace meso-diaminopimelate, the essential amino acid used for cross-linking the cell wall This was accomplished by jointly overexpressing the metB gene coding for L-cystathionine y-synthase and disrupting the metC gene, whose product, L-cystathionine &lyase, is responsible for the destruction of Lcystathionine and other L-cysteine thioethers. Bridging the gap between the two parts, we have constructed strains of Escherichia coli whose lack of meso-diaminopimelate in their cell wallis compensated by abnormal side products of the methionine biosynthesis pathway. Thioethers, can be formed readily and remain stable in water [11].In addition, the sulfur atom of thioethers (-S-) makes versatile connections within organic molecules, with geometric parameters close to those of the methylene group (-CH2-) [12]

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