Cargando…
Biosynthesis and metabolic engineering of 1-hydroxyphenazine in Pseudomonas chlororaphis H18
BACKGROUND: 1-Hydroxyphenazine (1-OH-PHZ) is a phenazine microbial metabolite with broad-spectrum antibacterial activities against a lot of plant pathogens. However, its use is hampered by the low yield all along. Metabolic engineering of microorganisms is an increasingly powerful method for the pro...
Autores principales: | , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8717658/ https://www.ncbi.nlm.nih.gov/pubmed/34965873 http://dx.doi.org/10.1186/s12934-021-01731-y |
_version_ | 1784624578633400320 |
---|---|
author | Wan, Yupeng Liu, Hongchen Xian, Mo Huang, Wei |
author_facet | Wan, Yupeng Liu, Hongchen Xian, Mo Huang, Wei |
author_sort | Wan, Yupeng |
collection | PubMed |
description | BACKGROUND: 1-Hydroxyphenazine (1-OH-PHZ) is a phenazine microbial metabolite with broad-spectrum antibacterial activities against a lot of plant pathogens. However, its use is hampered by the low yield all along. Metabolic engineering of microorganisms is an increasingly powerful method for the production of valuable organisms at high levels. Pseudomonas chlororaphis is recognized as a safe and effective plant rhizosphere growth-promoting bacterium, and faster growth rate using glycerol or glucose as a renewable carbon source. Therefore, Pseudomonas chlororaphis is particularly suitable as the chassis cell for the modification and engineering of phenazines. RESULTS: In this study, enzyme PhzS (monooxygenase) was heterologously expressed in a phenazine-1-carboxylic acid (PCA) generating strain Pseudomonas chlororaphis H18, and 1-hydroxyphenazine was isolated, characterized in the genetically modified strain. Next, the yield of 1-hydroxyphenazine was systematically engineered by the strategies including (1) semi-rational design remodeling of crucial protein PhzS, (2) blocking intermediate PCA consumption branch pathway, (3) enhancing the precursor pool, (4) engineering regulatory genes, etc. Finally, the titer of 1-hydroxyphenazine reached 3.6 g/L in 5 L fermenter in 54 h. CONCLUSIONS: The 1-OH-PHZ production of Pseudomonas chlororaphis H18 was greatly improved through systematically engineering strategies, which is the highest, reported to date. This work provides a promising platform for 1-hydroxyphenazine engineering and production. GRAPHICAL ABSTRACT: [Image: see text] SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12934-021-01731-y. |
format | Online Article Text |
id | pubmed-8717658 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-87176582022-01-05 Biosynthesis and metabolic engineering of 1-hydroxyphenazine in Pseudomonas chlororaphis H18 Wan, Yupeng Liu, Hongchen Xian, Mo Huang, Wei Microb Cell Fact Research BACKGROUND: 1-Hydroxyphenazine (1-OH-PHZ) is a phenazine microbial metabolite with broad-spectrum antibacterial activities against a lot of plant pathogens. However, its use is hampered by the low yield all along. Metabolic engineering of microorganisms is an increasingly powerful method for the production of valuable organisms at high levels. Pseudomonas chlororaphis is recognized as a safe and effective plant rhizosphere growth-promoting bacterium, and faster growth rate using glycerol or glucose as a renewable carbon source. Therefore, Pseudomonas chlororaphis is particularly suitable as the chassis cell for the modification and engineering of phenazines. RESULTS: In this study, enzyme PhzS (monooxygenase) was heterologously expressed in a phenazine-1-carboxylic acid (PCA) generating strain Pseudomonas chlororaphis H18, and 1-hydroxyphenazine was isolated, characterized in the genetically modified strain. Next, the yield of 1-hydroxyphenazine was systematically engineered by the strategies including (1) semi-rational design remodeling of crucial protein PhzS, (2) blocking intermediate PCA consumption branch pathway, (3) enhancing the precursor pool, (4) engineering regulatory genes, etc. Finally, the titer of 1-hydroxyphenazine reached 3.6 g/L in 5 L fermenter in 54 h. CONCLUSIONS: The 1-OH-PHZ production of Pseudomonas chlororaphis H18 was greatly improved through systematically engineering strategies, which is the highest, reported to date. This work provides a promising platform for 1-hydroxyphenazine engineering and production. GRAPHICAL ABSTRACT: [Image: see text] SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12934-021-01731-y. BioMed Central 2021-12-30 /pmc/articles/PMC8717658/ /pubmed/34965873 http://dx.doi.org/10.1186/s12934-021-01731-y Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Research Wan, Yupeng Liu, Hongchen Xian, Mo Huang, Wei Biosynthesis and metabolic engineering of 1-hydroxyphenazine in Pseudomonas chlororaphis H18 |
title | Biosynthesis and metabolic engineering of 1-hydroxyphenazine in Pseudomonas chlororaphis H18 |
title_full | Biosynthesis and metabolic engineering of 1-hydroxyphenazine in Pseudomonas chlororaphis H18 |
title_fullStr | Biosynthesis and metabolic engineering of 1-hydroxyphenazine in Pseudomonas chlororaphis H18 |
title_full_unstemmed | Biosynthesis and metabolic engineering of 1-hydroxyphenazine in Pseudomonas chlororaphis H18 |
title_short | Biosynthesis and metabolic engineering of 1-hydroxyphenazine in Pseudomonas chlororaphis H18 |
title_sort | biosynthesis and metabolic engineering of 1-hydroxyphenazine in pseudomonas chlororaphis h18 |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8717658/ https://www.ncbi.nlm.nih.gov/pubmed/34965873 http://dx.doi.org/10.1186/s12934-021-01731-y |
work_keys_str_mv | AT wanyupeng biosynthesisandmetabolicengineeringof1hydroxyphenazineinpseudomonaschlororaphish18 AT liuhongchen biosynthesisandmetabolicengineeringof1hydroxyphenazineinpseudomonaschlororaphish18 AT xianmo biosynthesisandmetabolicengineeringof1hydroxyphenazineinpseudomonaschlororaphish18 AT huangwei biosynthesisandmetabolicengineeringof1hydroxyphenazineinpseudomonaschlororaphish18 |