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首页> 外文期刊>Applied Microbiology and Biotechnology >Isolation, identification, and accumulation of 2-acetamidophenol in liquid cultures of the wheat take-all biocontrol agent Pseudomonas fluorescens 2-79
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Isolation, identification, and accumulation of 2-acetamidophenol in liquid cultures of the wheat take-all biocontrol agent Pseudomonas fluorescens 2-79

机译：小麦全杀生物控制剂荧光假单胞菌2-79液体培养物中2-乙酰氨基酚的分离，鉴定和积累

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摘要

Pseudomonas fluorescens strain 2-79 (NRRL B-15132) is a classic biological control agent known to produce phenazine-1-carboxylic acid (PCA) as its primary means of suppressing take-all disease of wheat. In addition to PCA, an unknown metabolite was discovered in a liquid culture used to produce the biocontrol agent. The objective of the current study was to isolate, identify, and evaluate the accumulation of this compound in production cultures. Upon centrifugal fractionation of a production culture, thin-layer chromatography (TLC) analyses of extracts of the cells and cell-free supernatant indicated the compound to be primarily in the supernatant. Purified compound was obtained by extraction of culture supernatant, followed by flash chromatography of the extract and preparative TLC. The H-1 and C-13 nuclear magnetic resonance and electron impact mass spectra indicated the compound to be 2-acetamidophenol (AAP). Measured by reversed-phase HPLC, the accumulations of AAP and PCA in cultures of strain 2-79 reached 0.05 g/l and 1 g/l, respectively. The accumulations of AAP and PCA in liquid cultures were linearly correlated (P < 0.001), as shown by studies of cultures stimulated to yield varying levels of PCA by controlling levels of oxygen transfer, pH, and growth medium composition. In this study, oxygen limitation, a defined amino-acid-free medium, and neutral pH stimulated maximal production of both AAP and PCA. Furthermore, a transposon mutant of 2-79 [2A40 2-79 (phz-)] unable to produce PCA did not accumulate AAP. These findings indicate that ABP and PCA are likely to share a common segment of biosynthetic pathway. This is the first report of BAP production by a strain of P. fluorescens. Possible routes of AAP production are discussed relative to current knowledge of the phenazine biosynthetic pathway of strain 2-79. The pertinence of AAP to the design of commercial seed inoculants of phenazine-producing bacteria for controlling wheat take-all is also considered. [References: 22]
机译：荧光假单胞菌菌株2-79（NRRL B-15132）是一种经典的生物防治剂，已知会产生吩嗪-1-羧酸（PCA）作为其抑制小麦全株病的主要手段。除PCA外，在用于生产生物防治剂的液体培养物中还发现了未知的代谢产物。当前研究的目的是分离，鉴定和评估该化合物在生产文化中的积累。在对生产培养物进行离心分离时，对细胞提取物和无细胞上清液的薄层色谱（TLC）分析表明该化合物主要存在于上清液中。通过提取培养上清液，然后对提取物和制备型TLC进行快速色谱纯化，从而获得纯化的化合物。 H-1和C-13核磁共振和电子冲击质谱表明该化合物为2-乙酰氨基苯酚（AAP）。通过反相HPLC测量，菌株2-79的培养物中AAP和PCA的积累分别达到0.05g / l和1g / l。液体培养物中AAP和PCA的积累呈线性相关（P <0.001），如通过控制氧气转移，pH和生长培养基组成而刺激产生不同水平PCA的培养物研究表明。在这项研究中，氧气限制，无氨基酸的特定培养基和中性pH刺激了AAP和PCA的最大产量。此外，不能产生PCA的2-79转座子突变体[2A40 2-79（phz-）]没有积累​​AAP。这些发现表明，ABP和PCA可能共享生物合成途径的共同部分。这是荧光假单胞菌菌株产生BAP的首次报道。相对于菌株2-79的吩嗪生物合成途径的当前知识，讨论了AAP产生的可能途径。还考虑了AAP与设计用于控制小麦通吃的吩嗪生产细菌的商业种子接种剂的相关性。 [参考：22]

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