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首页> 外文期刊>RSC Advances >Metabolic characterisation of Magnetospirillum gryphiswaldense MSR-1 using LC-MS-based metabolite profiling
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Metabolic characterisation of Magnetospirillum gryphiswaldense MSR-1 using LC-MS-based metabolite profiling

机译：使用基于LC-MS的代谢物分析的磁孢子Gryphiswaldense MSR-1的代谢表征

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Magnetosomes are nano-sized magnetic nanoparticles with exquisite properties that can be used in a wide range of healthcare and biotechnological applications. They are biosynthesised by magnetotactic bacteria (MTB), such as Magnetospirillum gryphiswaldense MSR-1 ( Mgryph ). However, magnetosome bioprocessing yields low quantities compared to chemical synthesis of magnetic nanoparticles. Therefore, an understanding of the intracellular metabolites and metabolic networks related to Mgryph growth and magnetosome formation are vital to unlock the potential of this organism to develop improved bioprocesses. In this work, we investigated the metabolism of Mgryph using untargeted metabolomics. Liquid chromatography-mass spectrometry (LC-MS) was performed to profile spent medium samples of Mgryph cells grown under O _(2) -limited ( n = 6) and O _(2) -rich conditions ( n = 6) corresponding to magnetosome- and non-magnetosome producing cells, respectively. Multivariate, univariate and pathway enrichment analyses were conducted to identify significantly altered metabolites and pathways. Rigorous metabolite identification was carried out using authentic standards, the Mgryph -specific metabolite database and MS/MS mzCloud database. PCA and OPLS-DA showed clear separation and clustering of sample groups with cross-validation values of R ~(2) X = 0.76, R ~(2) Y = 0.99 and Q ~(2) = 0.98 in OPLS-DA. As a result, 50 metabolites linked to 45 metabolic pathways were found to be significantly altered in the tested conditions, including: glycine, serine and threonine; butanoate; alanine, aspartate and glutamate metabolism; aminoacyl-tRNA biosynthesis and; pyruvate and citric acid cycle (TCA) metabolisms. Our findings demonstrate the potential of LC-MS to characterise key metabolites in Mgryph and will contribute to further understanding the metabolic mechanisms that affect Mgryph growth and magnetosome formation.
机译：磁性体是纳米大小的磁性纳米粒子，具有精致的性能，可用于各种医疗保健和生物技术应用。它们是通过磁通细菌（MTB）的生物合成，例如磁截面Gryphiswaldense MSR-1（Murgorph）。然而，与磁性纳米粒子的化学合成相比，磁体生物处理产生低量。因此，理解与Mgryph生长和磁体形成有关的细胞内代谢物和代谢网络对于解锁这种生物体的潜力来开发改善的生物处理至关重要。在这项工作中，我们研究了使用未标准的代谢组学的MGRYPH的新陈代谢。进行液相色谱 - 质谱（LC-MS）对概述（2）下培养的MGRYPH细胞的介质样品进行剖面，相对应的（n = 6）和o _（2） - 中等条件（n = 6）磁体和非磁体产生细胞。进行多元，单变量和途径富集分析以确定显着改变的代谢物和途径。使用真实标准，MgryPh-特异性代谢物数据库和MS / MS MZCloud数据库进行严格的代谢物识别。 PCA和OPLS-DA显示OPLS-DA中的R〜（2）x = 0.76，R〜（2）y = 0.99和q〜（2）= 0.98的交叉验证值清晰分离和聚类样品组。结果，发现50种与45个代谢途径相关的代谢物在测试的条件下显着改变，包括：甘氨酸，丝氨酸和苏氨酸;丁酸盐;丙氨酸，天冬氨酸和谷氨酸代谢;氨基酰基 - TRNA生物合成和;丙酮酸和柠檬酸循环（TCA）代谢。我们的研究结果证明了LC-MS的潜力，以在MGRYPH中表征关键代谢物，并有助于进一步理解影响MGRYPH生长和磁体形成的代谢机制。

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