Fermentative Production of Bacterial Phenazines

Chincholkar, S. B.; Patil, Sandeep; Sarode, Prashant; Rane, M. R.

doi:10.1007/978-3-642-40573-0_5

Cited by 8 publications

(10 citation statements)

References 60 publications

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“…Therefore, it is necessary to enhance the yield of 2-OH-PHZ for its application in agriculture. Although it may be practical to synthesize phenazines by chemical methods, the yield is low [ 10 ], and toxic byproducts such as lead oxide, aniline, o-phenylenediamine, and azobenzoate are produced [ 11 ]. So the biocatalytic synthesis of 2-OH-PHZ using recombinant microorganisms provides an attractive alternative.…”

Section: Introductionmentioning

confidence: 99%

Genetic engineering of Pseudomonas chlororaphis GP72 for the enhanced production of 2-Hydroxyphenazine

et al. 2016

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BackgroundThe biocontrol strain Pseudomonas chlororaphis GP72 isolated from the green pepper rhizosphere synthesizes three antifungal phenazine compounds, 2-Hydroxyphenazine (2-OH-PHZ), 2-hydroxy-phenazine-1-carboxylic acid (2-OH-PCA) and phenazine-1-carboxylic acid (PCA). PCA has been a commercialized antifungal pesticide registered as “Shenqinmycin” in China since 2011. It is found that 2-OH-PHZ shows stronger fungistatic and bacteriostatic activity to some pathogens than PCA. 2-OH-PHZ could be developed as a potential antifungal pesticide. But the yield of 2-OH-PHZ generally is quite low, such as P. chlororaphis GP72, the production of 2-OH-PHZ by the wide-type strain is only 4.5 mg/L, it is necessary to enhance the yield of 2-OH-PHZ for its application in agriculture.ResultsDifferent strategies were used to improve the yield of 2-OH-PHZ: knocking out the negative regulatory genes, enhancing the shikimate pathway, deleting the competing pathways of 2-OH-PHZ synthesis based on chorismate, and improving the activity of PhzO which catalyzes the conversion of PCA to 2-OH-PHZ, although the last two strategies did not give us satisfactory results. In this study, four negative regulatory genes (pykF, rpeA, rsmE and lon) were firstly knocked out of the strain GP72 genome stepwise. The yield of 2-OH-PHZ improved more than 60 folds and increased from 4.5 to about 300 mg/L. Then six key genes (ppsA, tktA, phzC, aroB, aroD and aroE) selected from the gluconeogenesis, pentose phosphate and shikimate pathways which used to enhance the shikimate pathway were overexpressed to improve the production of 2-OH-PHZ. At last a genetically engineered strain that increased the 2-OH-PHZ production by 99-fold to 450.4 mg/L was obtained.ConclusionsThe 2-OH-PHZ production of P. chlororaphis GP72 was greatly improved through disruption of four negative regulatory genes and overexpression of six key genes, and it is shown that P. chlororaphis GP72 could be modified as a potential cell factory to produce 2-OH-PHZ and other phenazine biopesticides by genetic and metabolic engineering.Electronic supplementary materialThe online version of this article (doi:10.1186/s12934-016-0529-0) contains supplementary material, which is available to authorized users.

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Section: Introductionmentioning

confidence: 99%

Genetic engineering of Pseudomonas chlororaphis GP72 for the enhanced production of 2-Hydroxyphenazine

et al. 2016

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“…As a bacterial secondary metabolite, PCN has long been recognized for its broad-spectrum antibiotic activity in the biological control of a range of fungal phytopathogens. ,, Moreover, PCN has also been reported as a lead molecule in the development of antitumor drugs, antimicrobial coatings on silicone urethral catheters, and diamide derivatives for selective herbicides. ,, Chemical synthesis of PCN is technically feasible; however, the yield is low, and toxic byproducts such as aniline, azobenzoate, lead oxide, or o-phenylenediamine are produced. , Therefore, the biocatalytic synthesis of PCN using engineered microorganisms provides an attractive alternative.…”

Section: Discussionmentioning

confidence: 99%

“…8,38,39 Chemical synthesis of PCN is technically feasible; however, the yield is low, and toxic byproducts such as aniline, azobenzoate, lead oxide, or o-phenylenediamine are produced. 40,41 Therefore, the biocatalytic synthesis of PCN using engineered microorganisms provides an attractive alternative.…”

Section: ■ Discussionmentioning

confidence: 99%

Identification of a Strong Quorum Sensing- and Thermo-Regulated Promoter for the Biosynthesis of a New Metabolite Pesticide Phenazine-1-carboxamide in Pseudomonas strain PA1201

et al. 2020

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Phenazine-1-carboxamide (PCN) produced by multifarious Pseudomonas strains represents a promising candidate as a new metabolite pesticide due to its broad-spectrum antifungal activity and capacity to induce systemic resistance in plants. The rice rhizosphere Pseudomonas strain PA1201 contains two reiterated gene clusters, phz1 and phz2, for phenazine-1-carboxylic acid (PCA) biosynthesis; PCA is further converted into PCN by this strain using a functional phzH-encoding glutamine aminotransferase. However, PCN levels in PA1201 constitute approximately one-fifth of PCA levels and the optimal temperature for PCN synthesis is 28 °C. In this study, the phzH open reading frame (ORF) and promoter region were investigated and reannotated. phzH promoter P phzH was found to be a weak promoter, and PhzH levels were not sufficient to convert all of the native PCA into PCN. Following RNA Seq and promoter-lacZ fusion analyses, a strong quorum sensing (QS)-and thermo-regulated promoter P rhlI was identified and characterized. The activity of P phzH is approximately 1% of P rhlI in PA1201. After three rounds of promoter editing and swapping by P rhlI , a new PCNoverproducing strain UP46 was generated. The optimal fermentation temperature for PCN biosynthesis in UP46 was increased from 28 to 37 °C and the PCN fermentation titer increased 179.5-fold, reaching 14.1 g/L, the highest ever reported.

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“…The unique structure–activity relationship of phenazines plays a crucial role in drug design and potential applications. The redox-active tricyclic zwitterion has shown potential effects on various organs, including the respiratory, cardiovascular, urological, and central nervous systems through the formation of reactive oxygen species (ROS). , Previous studies have mainly focused on phenazines associated with antibacterial activities in vitro and respiratory tract infections. However, a comprehensive understanding of the effect of phenazines on the digestive system remains poorly described.…”

Section: Introductionmentioning

confidence: 99%

Pyocyanin Modulates Gastrointestinal Transformation and Microbiota

Peng

Zhao

et al. 2022

J. Agric. Food Chem.

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Phenazines are ubiquitously produced by Pseudomonas spp. in the environment and are widely used in agriculture and clinical therapies, making their accumulation through the food chain cause potential risks to human health. Here, we utilized pyocyanin (PYO) as a representative to study the effects of phenazines on digestive tracts. Pharmacokinetic analysis showed that PYO exhibited low systemic exposure, slow elimination, and low accumulation in both rat and pig models. PYO was subsequently found to induce intestinal microbiota dysbiosis, destroy the mucus layer and physical barrier, and even promote gut vascular barrier (GVB) impairment, consequently increasing the gut permeability. Additionally, integral and metabolomic analyses of the liver demonstrated that PYO induced liver inflammation and metabolic disorders. The metabolic analysis further confirmed that all of the metabolites of PYO retain the nitrogen-containing tricyclic structural skeleton of phenazines, which was the core bioactivity of phenazine compounds. These findings elucidated that PYO could be metabolized by animals. Meanwhile, high levels of PYO could induce intestinal barrier impairment and liver damage, suggesting that we should be alert to the accumulation of phenazines.

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Fermentative Production of Bacterial Phenazines

Cited by 8 publications

References 60 publications

Genetic engineering of Pseudomonas chlororaphis GP72 for the enhanced production of 2-Hydroxyphenazine

Genetic engineering of Pseudomonas chlororaphis GP72 for the enhanced production of 2-Hydroxyphenazine

Identification of a Strong Quorum Sensing- and Thermo-Regulated Promoter for the Biosynthesis of a New Metabolite Pesticide Phenazine-1-carboxamide in Pseudomonas strain PA1201

Pyocyanin Modulates Gastrointestinal Transformation and Microbiota

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