Characterization and Engineering of Pseudomonas chlororaphis LX24 with High Production of 2-Hydroxyphenazine

Abstract

The take-all disease of wheat is one of the most serious diseases in the field of food security in the world. There is no effective biological pesticide to prevent the take-all disease of wheat. 2-Hydroxyphenazine (2-OH-PHZ) was reported to possess a better inhibitory effect on the take-all disease of wheat than phenazine-1-carboxylic acid, which was registered as “Shenqinmycin” in China in 2011. The aim of this study was to construct a 2-OH-PHZ high-producing strain by strain screening, genome sequencing, genetic engineering, and fermentation optimization. First, the metabolites of the previously screened new phenazine-producing Pseudomonas sp. strain were identified, and the taxonomic status of the new Pseudomonas sp. strain was confirmed through 16S rRNA and matrix-assisted laser desorption ionization–time-of-flight mass spectrometry (MALDI-TOF MS). Then, the new Pseudomonas sp. strain was named Pseudomonas chlororaphis subsp. aurantiaca LX24, which is a new subspecies of P. chlororaphis that can synthesize 2-OH-PHZ. Next, the draft genome of strain LX24 was determined, and clusters of orthologous group (COG) analysis, KEGG analysis, and gene ontology (GO) analysis of strain LX24 were performed. Furthermore, the production of 2-OH-PHZ increased to 351.7 from 158.6 mg/L by deletion of the phenazine synthesis negative regulatory genes rpeA and rsmE in strain LX24. Finally, the 2-OH-PHZ production of strain LX24 reached 677.1 mg/L after fermentation optimization, which is the highest production through microbial fermentation reported to date. This work provides a reference for the efficient production of other pesticides and antibiotics.

Metal complexes with valuable biomolecules produced by Pseudomonas aeruginosa: a review of the coordination properties of pyocyanin, pyochelin and pyoverdines

“…3,4 Due to the broad-spectrum antimicrobial activity against various bacterial and fungal pathogens, several phenazines, such as phenazine-1-carboxylic acid and phenazine 5-oxide, have been approved as pesticides in agricultural crops. 5,6 Phenazines have low molecular weight and zwitterionic properties, allowing them to easily penetrate cell membranes and cross through the blood–brain barrier, resulting in cell damage. 7 Moreover, the redox-active tricyclic zwitterion forms reactive oxygen species (ROS), a process which can influence the cardiovascular, respiratory, urological, and central nervous systems.…”

Enhanced Biosynthesis of Phenazine-1,6-dicarboxylic Acid in Streptomyces coelicolor

“…27 The production of PCA and its derivatives has been widely studied. The highest production of PCA, PCN, 2hydroxyphenazine (2-OH-PHZ), and 1-hydroxyphenazine (1-OH-PHZ) reached 9.9 g/L, 27 14.1 g/L, 31 677.1 mg/L, 29 and 3.6 g/L, 32 respectively. Meanwhile, many efforts have been made to elevate PDC production.…”

Pyocyanin Modulates Gastrointestinal Transformation and Microbiota

“…5 The electron-shuttling properties of phenazines make them widely used in the clinic and ecological environment. Besides their usage as antimicrobial agents, phenazines such as phenazine-1-carboxylic acid (PCA) and phenazine 5-oxide have also been approved as pesticides in agricultural crops 6 because they possess a broad-spectrum antibiotic activity against diverse fungal, bacterial, and oomycete plant pathogens. 7 Additionally, PYO can be exploited as food colorants in the food and beverage industries for their brilliant color.…”