Cargando…

Droplet-Microfluidic-Based Promoter Engineering and Expression Fine-Tuning for Improved Erythromycin Production in Saccharopolyspora erythraea NRRL 23338

Erythromycin is a clinically important drug produced by the rare actinomycete Saccharopolyspora erythraea. In the wide-type erythromycin producer S. erythraea NRRL 23338, there is a lack of systematical method for promoter engineering as well as a well-characterized promoter panel for comprehensive...

Descripción completa

Detalles Bibliográficos
Autores principales: Yun, Kaiyue, Zhang, Yue, Li, Shixin, Wang, Yan, Tu, Ran, Liu, Hao, Wang, Meng
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9013967/
https://www.ncbi.nlm.nih.gov/pubmed/35445005
http://dx.doi.org/10.3389/fbioe.2022.864977
_version_ 1784688112961585152
author Yun, Kaiyue
Zhang, Yue
Li, Shixin
Wang, Yan
Tu, Ran
Liu, Hao
Wang, Meng
author_facet Yun, Kaiyue
Zhang, Yue
Li, Shixin
Wang, Yan
Tu, Ran
Liu, Hao
Wang, Meng
author_sort Yun, Kaiyue
collection PubMed
description Erythromycin is a clinically important drug produced by the rare actinomycete Saccharopolyspora erythraea. In the wide-type erythromycin producer S. erythraea NRRL 23338, there is a lack of systematical method for promoter engineering as well as a well-characterized promoter panel for comprehensive metabolic engineering. Here we demonstrated a systematical promoter acquiring process including promoter characterization, engineering and high-throughput screening by the droplet-microfluidic based platform in S. erythraea NRRL 23338, and rapidly obtained a panel of promoters with 21.5-fold strength variation for expression fine-tuning in the native host. By comparative qRT-PCR of S. erythraea NRRL 23338 and a high-producing strain S0, potential limiting enzymes were identified and overexpressed individually using two screened synthetic promoters. As a result, erythromycin production in the native host was improved by as high as 137.24 folds by combinational gene overexpression. This work enriches the accessible regulatory elements in the important erythromycin-producing strain S. erythraea NRRL 23338, and also provides a rapid and systematic research paradigm of promoter engineering and expression fine-tuning in the similar filamentous actinomycete hosts.
format Online
Article
Text
id pubmed-9013967
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-90139672022-04-19 Droplet-Microfluidic-Based Promoter Engineering and Expression Fine-Tuning for Improved Erythromycin Production in Saccharopolyspora erythraea NRRL 23338 Yun, Kaiyue Zhang, Yue Li, Shixin Wang, Yan Tu, Ran Liu, Hao Wang, Meng Front Bioeng Biotechnol Bioengineering and Biotechnology Erythromycin is a clinically important drug produced by the rare actinomycete Saccharopolyspora erythraea. In the wide-type erythromycin producer S. erythraea NRRL 23338, there is a lack of systematical method for promoter engineering as well as a well-characterized promoter panel for comprehensive metabolic engineering. Here we demonstrated a systematical promoter acquiring process including promoter characterization, engineering and high-throughput screening by the droplet-microfluidic based platform in S. erythraea NRRL 23338, and rapidly obtained a panel of promoters with 21.5-fold strength variation for expression fine-tuning in the native host. By comparative qRT-PCR of S. erythraea NRRL 23338 and a high-producing strain S0, potential limiting enzymes were identified and overexpressed individually using two screened synthetic promoters. As a result, erythromycin production in the native host was improved by as high as 137.24 folds by combinational gene overexpression. This work enriches the accessible regulatory elements in the important erythromycin-producing strain S. erythraea NRRL 23338, and also provides a rapid and systematic research paradigm of promoter engineering and expression fine-tuning in the similar filamentous actinomycete hosts. Frontiers Media S.A. 2022-04-04 /pmc/articles/PMC9013967/ /pubmed/35445005 http://dx.doi.org/10.3389/fbioe.2022.864977 Text en Copyright © 2022 Yun, Zhang, Li, Wang, Tu, Liu and Wang. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Bioengineering and Biotechnology
Yun, Kaiyue
Zhang, Yue
Li, Shixin
Wang, Yan
Tu, Ran
Liu, Hao
Wang, Meng
Droplet-Microfluidic-Based Promoter Engineering and Expression Fine-Tuning for Improved Erythromycin Production in Saccharopolyspora erythraea NRRL 23338
title Droplet-Microfluidic-Based Promoter Engineering and Expression Fine-Tuning for Improved Erythromycin Production in Saccharopolyspora erythraea NRRL 23338
title_full Droplet-Microfluidic-Based Promoter Engineering and Expression Fine-Tuning for Improved Erythromycin Production in Saccharopolyspora erythraea NRRL 23338
title_fullStr Droplet-Microfluidic-Based Promoter Engineering and Expression Fine-Tuning for Improved Erythromycin Production in Saccharopolyspora erythraea NRRL 23338
title_full_unstemmed Droplet-Microfluidic-Based Promoter Engineering and Expression Fine-Tuning for Improved Erythromycin Production in Saccharopolyspora erythraea NRRL 23338
title_short Droplet-Microfluidic-Based Promoter Engineering and Expression Fine-Tuning for Improved Erythromycin Production in Saccharopolyspora erythraea NRRL 23338
title_sort droplet-microfluidic-based promoter engineering and expression fine-tuning for improved erythromycin production in saccharopolyspora erythraea nrrl 23338
topic Bioengineering and Biotechnology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9013967/
https://www.ncbi.nlm.nih.gov/pubmed/35445005
http://dx.doi.org/10.3389/fbioe.2022.864977
work_keys_str_mv AT yunkaiyue dropletmicrofluidicbasedpromoterengineeringandexpressionfinetuningforimprovederythromycinproductioninsaccharopolysporaerythraeanrrl23338
AT zhangyue dropletmicrofluidicbasedpromoterengineeringandexpressionfinetuningforimprovederythromycinproductioninsaccharopolysporaerythraeanrrl23338
AT lishixin dropletmicrofluidicbasedpromoterengineeringandexpressionfinetuningforimprovederythromycinproductioninsaccharopolysporaerythraeanrrl23338
AT wangyan dropletmicrofluidicbasedpromoterengineeringandexpressionfinetuningforimprovederythromycinproductioninsaccharopolysporaerythraeanrrl23338
AT turan dropletmicrofluidicbasedpromoterengineeringandexpressionfinetuningforimprovederythromycinproductioninsaccharopolysporaerythraeanrrl23338
AT liuhao dropletmicrofluidicbasedpromoterengineeringandexpressionfinetuningforimprovederythromycinproductioninsaccharopolysporaerythraeanrrl23338
AT wangmeng dropletmicrofluidicbasedpromoterengineeringandexpressionfinetuningforimprovederythromycinproductioninsaccharopolysporaerythraeanrrl23338