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...
Autores principales: | , , , , , , |
---|---|
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 |