Cargando…
Enhanced biosynthesis of phenazine-1-carboxamide by engineered Pseudomonas chlororaphis HT66
BACKGROUND: Phenazine-1-carboxamide (PCN), a phenazine derivative, is strongly antagonistic to fungal phytopathogens. The high PCN biocontrol activity fascinated researcher’s attention in isolating and identifying novel bacterial strains combined with engineering strategies to target PCN as a lead m...
Autores principales: | , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2018
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6060551/ https://www.ncbi.nlm.nih.gov/pubmed/30045743 http://dx.doi.org/10.1186/s12934-018-0962-3 |
_version_ | 1783342056092991488 |
---|---|
author | Peng, Huasong Zhang, Pingyuan Bilal, Muhammad Wang, Wei Hu, Hongbo Zhang, Xuehong |
author_facet | Peng, Huasong Zhang, Pingyuan Bilal, Muhammad Wang, Wei Hu, Hongbo Zhang, Xuehong |
author_sort | Peng, Huasong |
collection | PubMed |
description | BACKGROUND: Phenazine-1-carboxamide (PCN), a phenazine derivative, is strongly antagonistic to fungal phytopathogens. The high PCN biocontrol activity fascinated researcher’s attention in isolating and identifying novel bacterial strains combined with engineering strategies to target PCN as a lead molecule. The chemical route for phenazines biosynthesis employs toxic chemicals and display low productivities, require harsh reaction conditions, and generate toxic by-products. Phenazine biosynthesis using some natural phenazine-producers represent remarkable advantages of non-toxicity and possibly high yield in environmentally-friendlier settings. RESULTS: A biocontrol bacterium with antagonistic activity towards fungal plant pathogens, designated as strain HT66, was isolated from the rice rhizosphere. The strain HT66 was identified as Pseudomonas chlororaphis based on the colony morphology, gas chromatography of cellular fatty acids and 16S rDNA sequence analysis. The secondary metabolite produced by HT66 strain was purified and identified as PCN through mass spectrometry, and (1)H, (13)C nuclear magnetic resonance spectrum. The yield of PCN by wild-type strain HT66 was 424.87 mg/L at 24 h. The inactivation of psrA and rpeA increased PCN production by 1.66- and 3.06-fold, respectively, which suggests that psrA and rpeA are PCN biosynthesis repressors. qRT-PCR analysis showed that the expression of phzI, phzR, and phzE was markedly increased in the psrA and rpeA double mutant than in psrA or rpeA mutant. However, the transcription level of rpeA and rpeB in strain HT66ΔpsrA increased by 3.52- and 11.58-folds, respectively. The reduced psrA expression in HT66ΔrpeA strain evidenced a complex regulation mechanism for PCN production in HT66. CONCLUSION: In conclusion, the results evidence that P. chlororaphis HT66 could be modified as a potential cell factory for industrial-scale biosynthesis of PCN and other phenazine derivatives by metabolic engineering strategies. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12934-018-0962-3) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-6060551 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-60605512018-07-31 Enhanced biosynthesis of phenazine-1-carboxamide by engineered Pseudomonas chlororaphis HT66 Peng, Huasong Zhang, Pingyuan Bilal, Muhammad Wang, Wei Hu, Hongbo Zhang, Xuehong Microb Cell Fact Research BACKGROUND: Phenazine-1-carboxamide (PCN), a phenazine derivative, is strongly antagonistic to fungal phytopathogens. The high PCN biocontrol activity fascinated researcher’s attention in isolating and identifying novel bacterial strains combined with engineering strategies to target PCN as a lead molecule. The chemical route for phenazines biosynthesis employs toxic chemicals and display low productivities, require harsh reaction conditions, and generate toxic by-products. Phenazine biosynthesis using some natural phenazine-producers represent remarkable advantages of non-toxicity and possibly high yield in environmentally-friendlier settings. RESULTS: A biocontrol bacterium with antagonistic activity towards fungal plant pathogens, designated as strain HT66, was isolated from the rice rhizosphere. The strain HT66 was identified as Pseudomonas chlororaphis based on the colony morphology, gas chromatography of cellular fatty acids and 16S rDNA sequence analysis. The secondary metabolite produced by HT66 strain was purified and identified as PCN through mass spectrometry, and (1)H, (13)C nuclear magnetic resonance spectrum. The yield of PCN by wild-type strain HT66 was 424.87 mg/L at 24 h. The inactivation of psrA and rpeA increased PCN production by 1.66- and 3.06-fold, respectively, which suggests that psrA and rpeA are PCN biosynthesis repressors. qRT-PCR analysis showed that the expression of phzI, phzR, and phzE was markedly increased in the psrA and rpeA double mutant than in psrA or rpeA mutant. However, the transcription level of rpeA and rpeB in strain HT66ΔpsrA increased by 3.52- and 11.58-folds, respectively. The reduced psrA expression in HT66ΔrpeA strain evidenced a complex regulation mechanism for PCN production in HT66. CONCLUSION: In conclusion, the results evidence that P. chlororaphis HT66 could be modified as a potential cell factory for industrial-scale biosynthesis of PCN and other phenazine derivatives by metabolic engineering strategies. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12934-018-0962-3) contains supplementary material, which is available to authorized users. BioMed Central 2018-07-25 /pmc/articles/PMC6060551/ /pubmed/30045743 http://dx.doi.org/10.1186/s12934-018-0962-3 Text en © The Author(s) 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Peng, Huasong Zhang, Pingyuan Bilal, Muhammad Wang, Wei Hu, Hongbo Zhang, Xuehong Enhanced biosynthesis of phenazine-1-carboxamide by engineered Pseudomonas chlororaphis HT66 |
title | Enhanced biosynthesis of phenazine-1-carboxamide by engineered Pseudomonas chlororaphis HT66 |
title_full | Enhanced biosynthesis of phenazine-1-carboxamide by engineered Pseudomonas chlororaphis HT66 |
title_fullStr | Enhanced biosynthesis of phenazine-1-carboxamide by engineered Pseudomonas chlororaphis HT66 |
title_full_unstemmed | Enhanced biosynthesis of phenazine-1-carboxamide by engineered Pseudomonas chlororaphis HT66 |
title_short | Enhanced biosynthesis of phenazine-1-carboxamide by engineered Pseudomonas chlororaphis HT66 |
title_sort | enhanced biosynthesis of phenazine-1-carboxamide by engineered pseudomonas chlororaphis ht66 |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6060551/ https://www.ncbi.nlm.nih.gov/pubmed/30045743 http://dx.doi.org/10.1186/s12934-018-0962-3 |
work_keys_str_mv | AT penghuasong enhancedbiosynthesisofphenazine1carboxamidebyengineeredpseudomonaschlororaphisht66 AT zhangpingyuan enhancedbiosynthesisofphenazine1carboxamidebyengineeredpseudomonaschlororaphisht66 AT bilalmuhammad enhancedbiosynthesisofphenazine1carboxamidebyengineeredpseudomonaschlororaphisht66 AT wangwei enhancedbiosynthesisofphenazine1carboxamidebyengineeredpseudomonaschlororaphisht66 AT huhongbo enhancedbiosynthesisofphenazine1carboxamidebyengineeredpseudomonaschlororaphisht66 AT zhangxuehong enhancedbiosynthesisofphenazine1carboxamidebyengineeredpseudomonaschlororaphisht66 |