单个菌株的细胞数量是否与其对微生物群落中底物总周转率的贡献相关?

IF 4.3 3区 材料科学 Q1 ENGINEERING, ELECTRICAL & ELECTRONIC ACS Applied Electronic Materials Pub Date : 2024-08-01 DOI:10.1039/D4EW00511B
Daniel Buchner, Johannes Scheckenbach, Philipp R. Martin and Stefan B. Haderlein
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摘要

通常使用量化菌株特异性 16S rRNA 或功能基因的 qPCR 数据来评估混合微生物群落中单个细菌菌株对特定化合物总体周转率的贡献。在这里,我们将基于 qPCR 的方法与特定化合物稳定同位素分析(CSIA)的结果进行了比较,后者依赖于与化合物生物转化相关的特定菌株动力学同位素分馏幅度。为此,我们使用含有两种不同脱硫杆菌菌株(Desulfitobacterium hafniense 菌株 Y51;εC,PCE = -5.8‰和 Desulfitobacterium dehalogenans 菌株 PCE1;εC,PCE = -19.7‰)的合成二元培养物进行了四氯乙烯(PCE)转化实验。通过功能基因的 qPCR 分析细胞丰度,并与通过碳同位素分馏得出的菌株特异性 PCE 转化率进行比较。在最初以菌株 Y51 为主导的合成二元培养物中重复添加 PCE,导致菌株 PCE1 对 PCE 转化率的贡献率稳步上升(εC,initial = -5.6 ± 0.6‰ to εC,final = -18.0 ± 0.6‰),而细胞丰度的变化没有或仅有微弱的反映。CSIA 数据进一步表明,在两个菌株细胞丰度相似的情况下,菌株特异性 PCE 转化率的变化超过 75%。因此,CSIA 方法为合成二元培养物中单株菌株代谢活性的演变提供了意想不到的新见解,并表明菌株特异性底物周转似乎是由菌株的生理和酶特性而非细胞丰度控制的。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

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Does the number of cells of individual strains correlate with their contribution to the total substrate turnover within a microbial community?†

The contribution of individual bacterial strains within a mixed microbial community to the overall turnover of a specific compound is often assessed using qPCR data quantifying strain-specific 16S rRNA or functional genes. Here we compare the results of a qPCR based approach with those of compound specific stable isotope analysis (CSIA), which relies on strain-specific magnitudes of kinetic isotope fractionation associated with the biotransformation of a compound. To this end, we performed tetrachloroethylene (PCE) transformation experiments using a synthetic binary culture containing two different Desulfitobacterium strains (Desulfitobacterium hafniense strain Y51; εC,PCE = −5.8‰ and Desulfitobacterium dehalogenans strain PCE1; εC,PCE = −19.7‰). Cell abundances were analyzed via qPCR of functional genes and compared to strain-specific PCE turnover derived via carbon isotope fractionation. Repeated spiking of an initially strain Y51 dominated synthetic binary culture with PCE led to a steadily increasing contribution of strain PCE1 to PCE turnover (εC,initial = −5.6 ± 0.6‰ to εC,final = −18.0 ± 0.6‰) which was not or only weakly reflected in the changes of the cell abundances. The CSIA data further indicate that strain-specific PCE turnover varied by more than 75% at similar cell abundances of the two strains. Thus, the CSIA approach provided new and unexpected insights into the evolution of the metabolic activity of the single strains within a synthetic binary culture and indicates that strain-specific substrate turnover appears to be controlled by physiological and enzymatic properties of the strains rather than their cell abundance.

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