Palafox-Félix Michel, Huerta-Ocampo José Ángel, Hernández-Ortíz Magdalena, Encarnación-Guevara Sergio, Vázquez-Moreno Luz, Guzmán-Partida Ana María, Cabrera Rosina
Centro de Investigación en Alimentación y Desarrollo, A.C., Unidad Regional Hidalgo, Pachuca Ciudad del Conocimiento y la Cultura, Blvd. Santa Catarina, C.P. 42163 San Agustin Tlaxiaca, Hidalgo, Mexico.
Conacyt-Centro de Investigación en Alimentación y Desarrollo, A.C., Carretera Gustavo Enrique Astiazarán Rosas, No. 46 Colonia La Victoria, C.P. 83304 Hermosillo, Sonora, Mexico.
J Proteomics. 2022 Feb 20;253:104461. doi: 10.1016/j.jprot.2021.104461. Epub 2021 Dec 16.
Amycolatopsis sp. BX17 is an actinobacterium isolated from milpa soils, which antagonizes the phytopathogenic fungus Fusarium graminearum. Metabolites secreted by the actinobacterium cultured in glucose-free medium inhibited 100% of the mycelial growth of F. graminearum RH1, while the inhibition rate was 65% in medium supplemented with 20 g/L glucose. With the aim of studying how the metabolism of strain BX17 is modulated by glucose as the main carbon source, media with 0 and 20 g/L glucose were selected to analyze the intracellular proteins by quantitative label-free proteomic analysis. Data are available via ProteomeXchange with identifier PXD028644. Proteins identified in bacteria cultured in medium without glucose were involved in glutamate metabolism, the Krebs cycle and the shikimate pathway, suggesting that amino acids are metabolized to synthesize antifungal compounds. In glucose-containing medium, carbon flux was directed mainly toward the synthesis of energy and cell growth. This study shows the metabolic versatility of Amycolatopsis BX17, and strengthens its potential use in designing biotechnological strategies for phytopathogen control. SIGNIFICANCE: Amycolatopsis BX17 is a bacterium isolated from milpa agroecosystems that antagonizes the phytopathogenic fungus Fusarium graminearum. Currently, there is scarce information about the metabolism involved in the biosynthesis of antifungal agents by this genus. We used a label-free proteomic approach to identify the differences in metabolic routes for antifungal biosynthesis in Amycolatopsis BX17 grown in media with 0 and 20 g/L glucose. Taken together the results suggest that the BX17 strain could be synthesizing the antifungal metabolite(s) from the Shikimate pathway through the synthesis and degradation of the amino acid tyrosine, which is a known precursor of glycopeptides with antibiotic and antifungal activity. While the lower antifungal activity of the metabolites secreted by Amycolatopsis BX17 when grown in a medium with glucose as the main carbon source, may be correlated with a lower synthesis of antifungal compounds, due to the directing of carbon flux toward metabolic pathways involved with energy synthesis and cell growth. Likewise, it is possible that the bacteria synthesize other compounds with biological activity, such as glycopeptides with antibiotic activity. These findings are relevant because they represent the first stage to understand the metabolic regulation involved in the biosynthesis of antifungal metabolites by the genus Amycolatopsis. Finally, improving our understanding of the metabolic regulation involved in the biosynthesis of antifungal metabolites is essential to design of strategies in agricultural biotechnology for phytopathogen control.
嗜甲基杆菌属菌株BX17是一种从玉米地土壤中分离出的放线菌,它能拮抗植物病原真菌禾谷镰刀菌。在无葡萄糖培养基中培养该放线菌所分泌的代谢产物可100%抑制禾谷镰刀菌RH1的菌丝生长,而在添加了20 g/L葡萄糖的培养基中,抑制率为65%。为了研究作为主要碳源的葡萄糖如何调节菌株BX17的代谢,选择了含0和20 g/L葡萄糖的培养基,通过无标记定量蛋白质组学分析来分析细胞内蛋白质。数据可通过ProteomeXchange获得,标识符为PXD028644。在无葡萄糖培养基中培养的细菌中鉴定出的蛋白质参与谷氨酸代谢、三羧酸循环和莽草酸途径,这表明氨基酸被代谢以合成抗真菌化合物。在含葡萄糖的培养基中,碳流主要导向能量合成和细胞生长。本研究展示了嗜甲基杆菌属BX17的代谢多样性,并增强了其在设计控制植物病原体的生物技术策略中的潜在用途。意义:嗜甲基杆菌属BX17是一种从玉米农业生态系统中分离出的细菌,可拮抗植物病原真菌禾谷镰刀菌。目前关于该属在抗真菌剂生物合成中涉及的代谢的信息很少。我们使用无标记蛋白质组学方法来鉴定嗜甲基杆菌属BX17在含0和20 g/L葡萄糖的培养基中生长时抗真菌生物合成代谢途径的差异。综合结果表明,BX17菌株可能通过氨基酸酪氨酸的合成和降解,从莽草酸途径合成抗真菌代谢物,酪氨酸是具有抗生素和抗真菌活性的糖肽的已知前体。虽然嗜甲基杆菌属BX17在以葡萄糖为主要碳源的培养基中生长时分泌的代谢产物的抗真菌活性较低,可能与抗真菌化合物的合成减少有关,因为碳流导向了与能量合成和细胞生长有关的代谢途径。同样,细菌也可能合成其他具有生物活性的化合物,如具有抗生素活性的糖肽。这些发现很重要,因为它们代表了了解嗜甲基杆菌属抗真菌代谢物生物合成中涉及的代谢调节的第一步。最后,增进我们对抗真菌代谢物生物合成中涉及的代谢调节的理解对于设计农业生物技术中控制植物病原体的策略至关重要。