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...

Descripción completa

Detalles Bibliográficos
Autores principales: Peng, Huasong, Zhang, Pingyuan, Bilal, Muhammad, Wang, Wei, Hu, Hongbo, Zhang, Xuehong
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