An Engineered Synthetic Pathway for Discovering Nonnatural Nonribosomal Peptides in <italic toggle="yes">Escherichia coli</italic>

ABSTRACT Peptides that are synthesized independently of the ribosome in plants, fungi, and bacteria can have clinically relevant anticancer, antihemochromatosis, and antiviral activities, among many other. Despite their natural origin, discovering new natural products is challenging, and there is a...

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Autores principales:	Sara Cleto, Timothy K. Lu
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2017
Materias:	nonribosomal peptides pathway engineering genome engineering heterologous gene expression mutasynthesis polyamines Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/860c57c092144fd5817584415a0e1329
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spelling	oai:doaj.org-article:860c57c092144fd5817584415a0e13292021-11-15T15:51:50ZAn Engineered Synthetic Pathway for Discovering Nonnatural Nonribosomal Peptides in <italic toggle="yes">Escherichia coli</italic>10.1128/mBio.01474-172150-7511https://doaj.org/article/860c57c092144fd5817584415a0e13292017-11-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01474-17https://doaj.org/toc/2150-7511ABSTRACT Peptides that are synthesized independently of the ribosome in plants, fungi, and bacteria can have clinically relevant anticancer, antihemochromatosis, and antiviral activities, among many other. Despite their natural origin, discovering new natural products is challenging, and there is a need to expand the chemical diversity that is accessible. In this work, we created a novel, compressed synthetic pathway for the heterologous expression and diversification of nonribosomal peptides (NRPs) based on homologs of siderophore pathways from Escherichia coli and Vibrio cholerae. To enhance the likelihood of successful molecule production, we established a selective pressure via the iron-chelating properties of siderophores. By supplementing cells containing our synthetic pathway with different precursors that are incorporated into the pathway independently of NRP enzymes, we generated over 20 predesigned, novel, and structurally diverse NRPs. This engineering approach, where phylogenetically related genes from different organisms are integrated and supplemented with novel precursors, should enable heterologous expression and molecular diversification of NRPs. IMPORTANCE Nonribosomal peptides (NRPs) constitute a source of bioactive molecules with potential therapeutic applications. However, discovering novel NRPs by rational engineering of biosynthetic pathways remains challenging. Here, we show that a synthetic compressed pathway in which we replaced biosynthetic genes with their ancestral homologs and orthologs enabled successful heterologous NRP expression. Polyamines added exogenously were incorporated into nascent NRPs, and molecular production was pressured by growing the host under conditions that make such NRPs beneficial for survival. This multilayered approach resulted in the assembly of over 20 distinct and novel molecules. We envision this strategy being used to enable the production of NRPs from heterologous pathways.Sara CletoTimothy K. LuAmerican Society for Microbiologyarticlenonribosomal peptidespathway engineeringgenome engineeringheterologous gene expressionmutasynthesispolyaminesMicrobiologyQR1-502ENmBio, Vol 8, Iss 5 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	nonribosomal peptides pathway engineering genome engineering heterologous gene expression mutasynthesis polyamines Microbiology QR1-502
spellingShingle	nonribosomal peptides pathway engineering genome engineering heterologous gene expression mutasynthesis polyamines Microbiology QR1-502 Sara Cleto Timothy K. Lu An Engineered Synthetic Pathway for Discovering Nonnatural Nonribosomal Peptides in <italic toggle="yes">Escherichia coli</italic>
description	ABSTRACT Peptides that are synthesized independently of the ribosome in plants, fungi, and bacteria can have clinically relevant anticancer, antihemochromatosis, and antiviral activities, among many other. Despite their natural origin, discovering new natural products is challenging, and there is a need to expand the chemical diversity that is accessible. In this work, we created a novel, compressed synthetic pathway for the heterologous expression and diversification of nonribosomal peptides (NRPs) based on homologs of siderophore pathways from Escherichia coli and Vibrio cholerae. To enhance the likelihood of successful molecule production, we established a selective pressure via the iron-chelating properties of siderophores. By supplementing cells containing our synthetic pathway with different precursors that are incorporated into the pathway independently of NRP enzymes, we generated over 20 predesigned, novel, and structurally diverse NRPs. This engineering approach, where phylogenetically related genes from different organisms are integrated and supplemented with novel precursors, should enable heterologous expression and molecular diversification of NRPs. IMPORTANCE Nonribosomal peptides (NRPs) constitute a source of bioactive molecules with potential therapeutic applications. However, discovering novel NRPs by rational engineering of biosynthetic pathways remains challenging. Here, we show that a synthetic compressed pathway in which we replaced biosynthetic genes with their ancestral homologs and orthologs enabled successful heterologous NRP expression. Polyamines added exogenously were incorporated into nascent NRPs, and molecular production was pressured by growing the host under conditions that make such NRPs beneficial for survival. This multilayered approach resulted in the assembly of over 20 distinct and novel molecules. We envision this strategy being used to enable the production of NRPs from heterologous pathways.
format	article
author	Sara Cleto Timothy K. Lu
author_facet	Sara Cleto Timothy K. Lu
author_sort	Sara Cleto
title	An Engineered Synthetic Pathway for Discovering Nonnatural Nonribosomal Peptides in <italic toggle="yes">Escherichia coli</italic>
title_short	An Engineered Synthetic Pathway for Discovering Nonnatural Nonribosomal Peptides in <italic toggle="yes">Escherichia coli</italic>
title_full	An Engineered Synthetic Pathway for Discovering Nonnatural Nonribosomal Peptides in <italic toggle="yes">Escherichia coli</italic>
title_fullStr	An Engineered Synthetic Pathway for Discovering Nonnatural Nonribosomal Peptides in <italic toggle="yes">Escherichia coli</italic>
title_full_unstemmed	An Engineered Synthetic Pathway for Discovering Nonnatural Nonribosomal Peptides in <italic toggle="yes">Escherichia coli</italic>
title_sort	engineered synthetic pathway for discovering nonnatural nonribosomal peptides in <italic toggle="yes">escherichia coli</italic>
publisher	American Society for Microbiology
publishDate	2017
url	https://doaj.org/article/860c57c092144fd5817584415a0e1329
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An Engineered Synthetic Pathway for Discovering Nonnatural Nonribosomal Peptides in <italic toggle="yes">Escherichia coli</italic>

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