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Production of Antibacterial Questiomycin A in Metabolically Engineered Pseudomonas chlororaphis HT66
Journal of Agricultural and Food Chemistry ( IF 5.7 ) Pub Date : 2022-06-15 , DOI: 10.1021/acs.jafc.2c03216 Shuqi Guo 1, 2 , Hongbo Hu 2, 3 , Wei Wang 2 , Muhammad Bilal 4 , Xuehong Zhang 2
Journal of Agricultural and Food Chemistry ( IF 5.7 ) Pub Date : 2022-06-15 , DOI: 10.1021/acs.jafc.2c03216 Shuqi Guo 1, 2 , Hongbo Hu 2, 3 , Wei Wang 2 , Muhammad Bilal 4 , Xuehong Zhang 2
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Pseudomonas chlororaphis has been demonstrated as a valuable source of antimicrobial metabolites for plant disease biocontrol and biopesticide development. Although phenazine-1-carboxylic acid (PCA) secreted by P. chlororaphis has been commercialized as an antifungal biopesticide, it shows poor antibacterial activity. Questiomycin A, with versatile antibacterial activities, is mainly discovered in some well-known phenazine-producing strains but not in Pseudomonas. Its low titer hinders practical applications. In this work, a metabolite was first identified as Questiomycin A in P. chlororaphis-derived strain HT66ΔphzBΔNat. Subsequently, Questiomycin A has been elucidated to share the same biosynthesis process with PCA by gene deletion and in vitro assays. Through rational metabolic engineering, heterologous phenoxazinone synthase introduction, and medium optimization, the titer reached 589.78 mg/L in P. chlororaphis, the highest production reported to date. This work contributes to a better understanding of Questiomycin A biosynthesis and demonstrates a promising approach to developing a new antibacterial biopesticide in Pseudomonas.
中文翻译：
在代谢工程绿刺假单胞菌 HT66 中生产抗菌奎丝霉素 A
Pseudomonas chlororaphis已被证明是植物病害生物防治和生物农药开发的重要抗菌代谢物来源。虽然P. chlororaphis分泌的吩嗪-1-羧酸 (PCA)已作为抗真菌生物农药商业化，但其抗菌活性较差。具有多种抗菌活性的Questiomycin A主要存在于一些著名的吩嗪生产菌株中，而假单胞菌中则没有。其低滴度阻碍了实际应用。在这项工作中，首先在P. chlororaphis衍生菌株 HT66Δ phzB Δ Nat中鉴定出一种代谢物为 Questiomycin A. 随后，通过基因缺失和体外测定，已阐明 Questiomycin A 与 PCA 共享相同的生物合成过程。通过合理的代谢工程、异源吩恶嗪酮合酶的引入和培养基的优化，绿刺假单胞菌的滴度达到589.78 mg/L ，是迄今为止报道的最高产量。这项工作有助于更好地了解 Questiomycin A 的生物合成，并展示了一种在假单胞菌中开发新型抗菌生物农药的有前景的方法。
更新日期：2022-06-15
中文翻译：
在代谢工程绿刺假单胞菌 HT66 中生产抗菌奎丝霉素 A
Pseudomonas chlororaphis已被证明是植物病害生物防治和生物农药开发的重要抗菌代谢物来源。虽然P. chlororaphis分泌的吩嗪-1-羧酸 (PCA)已作为抗真菌生物农药商业化，但其抗菌活性较差。具有多种抗菌活性的Questiomycin A主要存在于一些著名的吩嗪生产菌株中，而假单胞菌中则没有。其低滴度阻碍了实际应用。在这项工作中，首先在P. chlororaphis衍生菌株 HT66Δ phzB Δ Nat中鉴定出一种代谢物为 Questiomycin A. 随后，通过基因缺失和体外测定，已阐明 Questiomycin A 与 PCA 共享相同的生物合成过程。通过合理的代谢工程、异源吩恶嗪酮合酶的引入和培养基的优化，绿刺假单胞菌的滴度达到589.78 mg/L ，是迄今为止报道的最高产量。这项工作有助于更好地了解 Questiomycin A 的生物合成，并展示了一种在假单胞菌中开发新型抗菌生物农药的有前景的方法。
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