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In vivo characterization of the activities of novel cyclodipeptide oxidases: new tools for increasing chemical diversity of bioproduced 2,5-diketopiperazines in Escherichia coli.
Le Chevalier, Fabien; Correia, Isabelle; Matheron, Lucrèce; Babin, Morgan; Moutiez, Mireille; Canu, Nicolas; Gondry, Muriel; Lequin, Olivier; Belin, Pascal.
Afiliação
  • Le Chevalier F; Université Paris-Saclay, CEA, CNRS, Institute for Integrative Biology of the Cell (I2BC), 91198, Gif-sur-Yvette, France.
  • Correia I; Laboratoire des Biomolécules (LBM), Sorbonne Université, Ecole Normale Supérieure, PSL University, CNRS, 75005, Paris, France.
  • Matheron L; Sorbonne Université, Institut de Biologie Paris Seine (IBPS), FRE3631, 75005, Paris, France.
  • Babin M; Université Paris-Saclay, CEA, CNRS, Institute for Integrative Biology of the Cell (I2BC), 91198, Gif-sur-Yvette, France.
  • Moutiez M; Université Paris-Saclay, CEA, CNRS, Institute for Integrative Biology of the Cell (I2BC), 91198, Gif-sur-Yvette, France.
  • Canu N; Université Paris-Saclay, CEA, CNRS, Institute for Integrative Biology of the Cell (I2BC), 91198, Gif-sur-Yvette, France.
  • Gondry M; Université Paris-Saclay, CEA, CNRS, Institute for Integrative Biology of the Cell (I2BC), 91198, Gif-sur-Yvette, France.
  • Lequin O; Laboratoire des Biomolécules (LBM), Sorbonne Université, Ecole Normale Supérieure, PSL University, CNRS, 75005, Paris, France.
  • Belin P; Université Paris-Saclay, CEA, CNRS, Institute for Integrative Biology of the Cell (I2BC), 91198, Gif-sur-Yvette, France. pascal.belin@cea.fr.
Microb Cell Fact ; 19(1): 178, 2020 Sep 07.
Article em En | MEDLINE | ID: mdl-32894164
BACKGROUND: Cyclodipeptide oxidases (CDOs) are enzymes involved in the biosynthesis of 2,5-diketopiperazines, a class of naturally occurring compounds with a large range of pharmaceutical activities. CDOs belong to cyclodipeptide synthase (CDPS)-dependent pathways, in which they play an early role in the chemical diversification of cyclodipeptides by introducing Cα-Cß dehydrogenations. Although the activities of more than 100 CDPSs have been determined, the activities of only a few CDOs have been characterized. Furthermore, the assessment of the CDO activities on chemically-synthesized cyclodipeptides has shown these enzymes to be relatively promiscuous, making them interesting tools for cyclodipeptide chemical diversification. The purpose of this study is to provide the first completely microbial toolkit for the efficient bioproduction of a variety of dehydrogenated 2,5-diketopiperazines. RESULTS: We mined genomes for CDOs encoded in biosynthetic gene clusters of CDPS-dependent pathways and selected several for characterization. We co-expressed each with their associated CDPS in the pathway using Escherichia coli as a chassis and showed that the cyclodipeptides and the dehydrogenated derivatives were produced in the culture supernatants. We determined the biological activities of the six novel CDOs by solving the chemical structures of the biologically produced dehydrogenated cyclodipeptides. Then, we assessed the six novel CDOs plus two previously characterized CDOs in combinatorial engineering experiments in E. coli. We co-expressed each of the eight CDOs with each of 18 CDPSs selected for the diversity of cyclodipeptides they synthesize. We detected more than 50 dehydrogenated cyclodipeptides and determined the best CDPS/CDO combinations to optimize the production of 23. CONCLUSIONS: Our study establishes the usefulness of CDPS and CDO for the bioproduction of dehydrogenated cyclodipeptides. It constitutes the first step toward the bioproduction of more complex and diverse 2,5-diketopiperazines.
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Oxirredutases / Peptídeo Sintases / Biotecnologia / Escherichia coli / Dicetopiperazinas Idioma: En Ano de publicação: 2020 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Oxirredutases / Peptídeo Sintases / Biotecnologia / Escherichia coli / Dicetopiperazinas Idioma: En Ano de publicação: 2020 Tipo de documento: Article