A novel rapid and continuous procedure for large-scale purification of magnetosomes from Magnetospirillum gryphiswaldense
A novel rapid and continuous procedure for large-scale purification of magnetosomes from Magnetospirillum gryphiswaldense
复制标题
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
DOI：
10.1007/s00253-011-3189-3
复制
发表时间：
2011-05-01
影响因子：
5
通讯作者：
Li, Jilun
中科院分区：
文献类型：
--
作者：
Guo, Fangfang;Liu, Yang;Li, Jilun
建立了一种快速、连续的大规模纯化gryphismagnetospirillum MSR-1细胞磁小体的新方法。该程序包括以下步骤：用高压均质机破坏细胞，用连续磁分离系统分离磁小体，伴随低功率超声和尿素处理，用蛋白酶K去除吸附和表面蛋白质，用电洗脱去除核酸，用蒸馏水代替PBS缓冲液。纯化后的磁小体经冻干和γ射线处理后保存于-20℃。纯化所需的时间从20-30天减少到2-5天。利用SDS-PAGE、PCR和傅里叶变换红外光谱对所得磁小体的纯度进行了评价。总体数据表明，本文提出的方法是一种简单、快速、连续和高效的大规模纯化磁小体的方法。
A new rapid and continuous procedure was developed for purifying magnetosomes from Magnetospirillum gryphiswaldense MSR-1 cells on a large scale. The procedure included these steps: disruption of cells with a high-pressure homogeniser, isolation of magnetosomes with a continuous magnetism isolation system accompanied by low-power ultrasonication and urea treatment, removal of adsorbed and surface proteins with proteinase K, removal of nucleic acids with electro-elution, and replacement of the PBS buffer with distilled water by a magnetically stirred system. The purified magnetosomes were stored at -20 °C after lyophilized and treated with γ-rays. The time required for purification was reduced from 20-30 to 2-5 days. Evaluation of the purity of the resulting magnetosomes was carried out with SDS-PAGE, PCR, and Fourier-transform infrared spectroscopy. The overall data suggest that the method presented here is a simple, rapid, continuous, and highly efficient procedure for large-scale purification of magnetosomes.