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对绿针假单胞菌ATCC 9446进行代谢重建,以了解其作为吩嗪-1-甲酰胺生产菌株的代谢潜力。

Metabolic reconstruction of Pseudomonas chlororaphis ATCC 9446 to understand its metabolic potential as a phenazine-1-carboxamide-producing strain.

作者信息

Moreno-Avitia Fabián, Utrilla José, Bolívar Francisco, Nogales Juan, Escalante Adelfo

机构信息

Departamento de Ingeniería Celular y Biocatálisis, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca, Morelos, México.

Programa de Biología de Sistemas y Biología Sintética, Centro de Ciencias Genómicas, Universidad Nacional Autónoma de México, Cuernavaca, Morelos, México.

出版信息

Appl Microbiol Biotechnol. 2020 Dec;104(23):10119-10132. doi: 10.1007/s00253-020-10913-4. Epub 2020 Sep 28.

Abstract

Pseudomonas chlororaphis is a plant-associated bacterium with reported antagonistic activity against different organisms and plant growth-promoting properties. P. chlororaphis possesses exciting biotechnological features shared with another Pseudomonas with a nonpathogenic phenotype. Part of the antagonistic role of P. chlororaphis is due to its production of a wide variety of phenazines. To expand the knowledge of the metabolic traits of this organism, we constructed the first experimentally validated genome-scale model of P. chlororaphis ATCC 9446, containing 1267 genes and 2289 reactions, and analyzed strategies to maximize its potential for the production of phenazine-1-carboxamide (PCN). The resulting model also describes the capability of P. chlororaphis to carry out the denitrification process and its ability to consume sucrose (Scr), trehalose, mannose, and galactose as carbon sources. Additionally, metabolic network analysis suggested fatty acids as the best carbon source for PCN production. Moreover, the optimization of PCN production was performed with glucose and glycerol. The optimal PCN production phenotype requires an increased carbon flux in TCA and glutamine synthesis. Our simulations highlight the intrinsic HO flux associated with PCN production, which may generate cellular stress in an overproducing strain. These results suggest that an improved antioxidative strategy could lead to optimal performance of phenazine-producing strains of P. chlororaphis. KEY POINTS : • This is the first publication of a metabolic model for a strain of P. chlororaphis. • Genome-scale model is worthy tool to increase the knowledge of a non model organism. • Fluxes simulations indicate a possible effect of HO on phenazines production. • P. chlororaphis can be a suitable model for a wide variety of compounds.

摘要

绿针假单胞菌是一种与植物相关的细菌,据报道对不同生物体具有拮抗活性,并具有促进植物生长的特性。绿针假单胞菌具有令人感兴趣的生物技术特性,与另一种具有非致病表型的假单胞菌相同。绿针假单胞菌的部分拮抗作用归因于其产生的多种吩嗪。为了扩展对该生物体代谢特征的认识,我们构建了首个经过实验验证的绿针假单胞菌ATCC 9446基因组规模模型,该模型包含1267个基因和2289个反应,并分析了最大化其吩嗪 - 1 - 甲酰胺(PCN)生产潜力的策略。所得模型还描述了绿针假单胞菌进行反硝化过程的能力以及其消耗蔗糖(Scr)、海藻糖、甘露糖和半乳糖作为碳源的能力。此外,代谢网络分析表明脂肪酸是PCN生产的最佳碳源。此外,还使用葡萄糖和甘油对PCN生产进行了优化。最佳的PCN生产表型需要三羧酸循环(TCA)和谷氨酰胺合成中碳通量增加。我们的模拟突出了与PCN生产相关的内在HO通量,这可能在过量生产菌株中产生细胞应激。这些结果表明,改进的抗氧化策略可能导致绿针假单胞菌吩嗪生产菌株的最佳性能。要点:• 这是关于绿针假单胞菌菌株代谢模型的首次发表。• 基因组规模模型是增加对非模式生物认识的有价值工具。• 通量模拟表明HO对吩嗪生产可能有影响。• 绿针假单胞菌可以是多种化合物的合适模型。

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