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Effect of glucose on the fatty acid composition of Cupriavidus necator JMP134 during 2,4-Dichlorophenoxyacetic acid degradation: Implications for lipid-based stable isotope probing methods

机译:葡萄糖对2,4-二氯苯氧基乙酸降解过程中铜氰菊酯JMP134脂肪酸组成的影响:对基于脂质的稳定同位素探测方法的启示

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摘要

Combining lipid biomarker profiling with stable isotope probing (SIP) is a powerful technique for studying specific microbial populations responsible for the degradation of organic pollutants in various natural environments. However, the presence of other easily degradable substrates may induce significant physiological changes by altering both the rate of incorporation of the target compound into the biomass and the microbial lipid profiles. In order to test this hypothesis, Cupriavidus necator JMP134, a 2,4-dichlorophenoxyacetic acid (2,4-D)-degrading bacterium, was incubated with [(13)C]2,4-D, [(13)C]glucose, or mixtures of both substrates alternatively labeled with (13)C. C. necator JMP134 exhibited a preferential use of 2,4-D over glucose. The isotopic analysis showed that glucose had only a small effect on the incorporation of the acetic chain of 2,4-D into the biomass (at days 2 and 3) and no effect on that of the benzenic ring. The addition of glucose did change the fatty acid methyl ester (FAME) composition. However, the overall FAME isotopic signature reflected that of the entire biomass. Compound-specific individual isotopic analyses of FAME composition showed that the (13)C-enriched FAME profiles were slightly or not affected when tracing the 2,4-D acetic chain or 2,4-D benzenic ring, respectively. This batch study is a necessary step for validating the use of lipid-based SIP methods in complex environments.
机译:将脂质生物标志物分析与稳定同位素探测(SIP)结合使用是一项强大的技术,可用于研究负责在各种自然环境中降解有机污染物的特定微生物种群。但是,其他易降解底物的存在可能会通过同时改变目标化合物掺入生物质的速率和微生物脂质谱来诱导明显的生理变化。为了检验该假设,将降解2,4-二氯苯氧基乙酸(2,4-D)的Cupriavidus necator JMP134与[(13)C] 2,4-D,[(13)C]孵育葡萄糖或两种底物的混合物,也可分别用(13)C标记。梭状芽孢杆菌JMP134表现出比葡萄糖优先使用2,4-D。同位素分析表明,葡萄糖对2,4-D乙酸链掺入生物质的影响很小(在第2天和第3天),而对苯环则没有影响。葡萄糖的添加确实改变了脂肪酸甲酯(FAME)的组成。但是,整个FAME同位素特征反映了整个生物质的特征。对FAME组成进行化合物特定的单独同位素分析表明,分别追踪2,4-D乙酸链或2,4-D苯环时,富含(13)C的FAME分布图受到的影响很小或没有受到影响。此批研究是验证在复杂环境中使用基于脂质的SIP方法的必要步骤。

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