Evaluation of cell disruption technologies on magnetosome chain length and aggregation behaviour from Magnetospirillum gryphiswaldense MSR-1.
Evaluation of cell disruption technologies on magnetosome chain length and aggregation behaviour from Magnetospirillum gryphiswaldense MSR-1.
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DOI：
10.3389/fbioe.2023.1172457
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发表时间：
2023
影响因子：
5.7
通讯作者：
中科院分区：
文献类型：
--
作者：
磁小体是由趋磁细菌（MTB）自然产生的生物衍生的磁性纳米颗粒（MNPs）。由于其独特的特性，如窄的尺寸分布和高的生物相容性，磁小体代表了现有的商业上可用的化学合成MNPs的一个有吸引力的替代品。然而，要从细菌中提取磁小体，需要一个细胞破坏步骤。本研究系统比较了三种破坏技术（酶处理、探针超声和高压均质）对gryphiswaldense Magnetospirillum MSR-1细胞中分离的磁小体的链长、完整性和聚集状态的影响。实验结果表明，这三种方法均显示出较高的细胞破坏率（约89%）。透射电镜（TEM）、动态光散射（DLS）和纳米流式细胞术（nFCM）首次对纯化后的磁小体制备进行了表征。透射电镜和能谱分析结果表明，高压均质处理能最大程度地保持链的完整性，而酶促处理能提高链的裂解率。获得的数据表明，nFCM最适合表征单个膜包裹的磁小体，这对于需要使用单个磁小体的应用特别有用。磁小体也成功地用荧光CellMask™深红色膜染色标记（>90%），并通过nFCM进行分析，证明了该技术作为磁小体质量保证的快速分析工具的潜力。这项工作的结果有助于未来开发一个强大的磁小体生产平台。
Magnetosomes are biologically-derived magnetic nanoparticles (MNPs) naturally produced by magnetotactic bacteria (MTB). Due to their distinctive characteristics, such as narrow size distribution and high biocompatibility, magnetosomes represent an attractive alternative to existing commercially-available chemically-synthesized MNPs. However, to extract magnetosomes from the bacteria, a cell disruption step is required. In this study, a systematic comparison between three disruption techniques (enzymatic treatment, probe sonication and high-pressure homogenization) was carried out to study their effect on the chain length, integrity and aggregation state of magnetosomes isolated from Magnetospirillum gryphiswaldense MSR-1 cells. Experimental results revealed that all three methodologies show high cell disruption yields (>89%). Transmission electron microscopy (TEM), dynamic light scattering (DLS) and, for the first time, nano-flow cytometry (nFCM) were employed to characterize magnetosome preparations after purification. TEM and DLS showed that high-pressure homogenization resulted in optimal conservation of chain integrity, whereas enzymatic treatment caused higher chain cleavage. The data obtained suggest that nFCM is best suited to characterize single membrane-wrapped magnetosomes, which can be particularly useful for applications that require the use of individual magnetosomes. Magnetosomes were also successfully labelled (>90%) with the fluorescent CellMask™ Deep Red membrane stain and analysed by nFCM, demonstrating the promising capacity of this technique as a rapid analytical tool for magnetosome quality assurance. The results of this work contribute to the future development of a robust magnetosome production platform.
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一种新型快速连续程序，用于大规模纯化磁螺菌 gryphiswaldense 中的磁小体
影响因子：
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作者：
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通讯作者：
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影响因子：
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