趋磁细菌及其应用于生物导航的研究进展

趋磁性细菌是一种由于体内含有对磁场具有敏感性的磁小体,而能够沿着磁力线运动的特殊细菌,本文综述了趋磁细菌的分布、分类、特性、磁小体研究以及趋磁细菌在生物导航方面的研究进展.     

趋磁细菌及其应用于生物导航的研究进展

趋磁性细菌是一种由于体内含有对磁场具有敏感性的磁小体,而能够沿着磁力线运动的特殊细菌,本文综述了趋磁细菌的分布、分类、特性、磁小体研究以及趋磁细菌在生物导航方面的研究进展。     

趋磁细菌及磁小体的生物医学应用研究进展

近年来,趋磁细菌及其生物自身合成的磁小体由于良好的生物安全性逐渐被人们所认识,并被用于生物工程和医学应用研究。与人工化学合成磁性纳米颗粒相比,从趋磁细菌中提取的磁小体具有生物膜包被、生物相容性高、粒径均一及磁性高等优势。趋磁细菌因磁小体在其胞内呈链状排列,具有沿磁场方向泳动的能力,也被应用于各种应用研究。因此,综述了趋磁细菌及磁小体特性,并就最近的研究进展重点综述趋磁细菌和磁小体在生物工程及医学应用等领域的最新研究进展。     

趋磁细菌及磁小体在肿瘤治疗中的应用

提高抗肿瘤药物的靶向性是肿瘤治疗、降低药物副作用的重要手段。在肿瘤组织内部由于癌细胞的快速增殖致使其形成低氧区,低氧区会对多种肿瘤治疗方案产生耐受。趋磁细菌 (Magnetotactic bacteria, MTB) 是一类能在细胞内产生外包生物膜、纳米尺寸、单磁畴磁铁矿 (Fe3O4) 或硫铁矿 (Fe3S4) 晶体颗粒-磁小体的微生物的统称。在磁场的作用下,趋磁细菌可凭借鞭毛运动至厌氧区。趋磁细菌在动物体内毒性较低且生物相容性良好,其磁小体与人工合成的磁性纳米材料相比优势显著。文中在介绍趋磁细菌及其磁小体生物学特点、理化性能的基础上,综述了趋磁细菌作为载体偶联药物进入肿瘤内部,并通过感受低氧信号定位于肿瘤低氧区,以及趋磁细菌竞争肿瘤细胞铁源的研究进展,总结了磁小体运载化疗药物、抗体、DNA疫苗靶向结合肿瘤的研究进展,分析了趋磁细菌及磁小体肿瘤治疗中面临的问题,并对趋磁细菌和磁小体在肿瘤治疗中的应用进行了展望。     

趋磁细菌的磁小体

趋磁细菌是一类对磁场有趋向性反应的细菌,其菌体能吸收外界环境中铁元素并在体内合成包裹有膜的纳米磁性颗粒Fe₃O₄或Fe₃O_{3S4晶体即磁小体。综述了趋磁细菌的磁小体生物矿化的条件,以及趋磁细菌的铁离子吸收、磁小体囊泡的形成、铁离子的转运到磁小体囊泡及囊泡中受控的Fe3O4生物矿化的分子生物学和生物化学等方面的研究进展,重点介绍了趋磁细菌磁小体合成机制的研究进展及未来研究磁小体的发展方向。}     

趋磁细菌概述及应用进展

趋磁细菌是一类具有趋磁行为的革兰氏阴性茵的统称,其趋磁特性是由于菌体内含有磁小体。磁小体是由膜包被的纳米尺寸单磁畴颗粒,在菌体内多呈链状排列。自被发现以来,趋磁细菌及磁小体已逐步成为新的生物资源被广泛研究于材料学、医学、生物学、物理学、地质学等多个学科领域,并在仿生学、生态学、医学、地质学、工业处理、卫生检验等多个领域得到应用。主要介绍了趋磁细菌的生物特征、研究发展进程,以及近年来在多个学科领域的研究与应用。     

磁泳分离细菌新方法的研究

从酸性矿坑水中富集培养分离到的嗜酸氧化亚铁硫杆菌（Acidithiobacillus ferrooxidans,A.ferrooxidans）[1-2] 菌同趋磁细菌具有一定的相似性。通过显微镜观察发现,部分浸矿细菌在外加磁场的作用下具有微弱的趋磁性,基于菌种的这种特性,设计了磁泳分离仪,对其在磁场作用下泳动（磁泳）进行分析,经磁泳后的近磁、远磁菌的生理特性有较大的差异。从用涂布平板法获得的近磁菌纯培养A. ferrooxidans菌体中,分离得到纳米磁性颗粒,能谱分析表明,其主要成分为Fe和O元素。实验结果证明,A. ferrooxidans具有微弱趋磁性,采用磁泳分离该类菌体内含有磁性颗粒的细菌是可行的,这一分离技术的进一步完善和改进将为传统的微生物菌种分离提供一种新型分离技术,也将大大促进趋磁细菌的研究,而且它与浸矿工艺的结合将大大促进我国生物冶金的研究步伐。     

趋磁细菌多样性与应用研究进展 *

趋磁细菌是一类可以沿磁场方向进行运动的微生物统称,在细胞内合成由生物膜包被、链状排列、纳米级、单磁畴的磁铁矿 (Fe₃O₄) 或胶黄铁矿 (Fe₃S₄) 的磁小体颗粒。趋磁细菌在自然界分布广泛且多样性丰富,不仅在水环境和沉积环境的铁、硫、碳、氮、磷等元素生物地球化学循环中发挥重要作用,而且在污染治理、疾病诊断和治疗等方面有较好的应用。趋磁细菌磁小体由生物膜包被并在细胞调控下合成,是一类新型的生物源磁性纳米材料。相比常规化学合成的磁性纳米颗粒,磁小体具有大小均一、生物相容性高、兼具化学修饰和基因工程修饰功能等特点,在磁性分离、固定化酶、食品检测、环境监测、医学诊断、磁共振成像、磁热疗和靶向治疗等方面具有广阔的应用前景。在介绍趋磁细菌多样性研究的基础上,综述了趋磁细菌和磁小体的制备、修饰及其应用的最新进展,并对未来的研究进行了展望。     

趋磁性细菌的研究与应用现状

趋磁性细菌是一类能够沿着磁力线运动的特殊细菌 ,其细胞内含有对磁场具有敏感性的磁小体 ,它起了导向的作用。国内外已对其分离培养、菌体特性、基因遗传等方面进行了大量研究 ,并探讨了其在传感技术、临床医药、废水处理等多方面的应用 ,大大推动了人类在生物磁学领域的研究进展。     

趋磁螺菌中磁小体链装配相关基因及其功能

趋磁细菌(MTB)依赖于体内磁小体结构在磁场中取向,多个磁小体以一定的组 织形式排列是形成菌体内生物磁罗盘的重要环节．多数趋磁细菌中磁小体成链排列,有效增加了细胞磁偶极矩,从而使菌体表现出在环境磁场中定向的能力．趋磁螺菌M. magneticum AMB－1和M. gryphiswaldense MSR－1中磁小体均沿细胞长轴形成一条磁 小体链．通过对相关基因突变体表型的研究,结合对磁小体链形成过程的实时动态观 察,人们已初步了解MamJ、MamK和MamA等基因在磁小体链装配和维护过程中的功能．本文介绍了近年来趋磁螺菌磁小体链装配过程中重要功能性基因的研究进展,并重点分析了AMB-1和MSR-1中磁小体链装配的差异．     

Some have it all: multicellularity,magnetotaxis and photokinesis in one bacterium

Bacteria developed many different ways to orient themselves in the environment. Magnetoreception with following motility along Earth's magnetic field lines and photoreception with subsequent positive or negative phototaxis allow bacteria to optimally position themselves for survival and growth. Some bacteria show both magnetotactic and photoresponsive behaviour and additionally live in a multicellular organism adding another layer of complexity. A novel study by Qian and colleagues visualized different species of multicellular magnetotactic bacteria and shed light on their reproductive as well as photoresponsive behaviour. This study paves the way towards understanding the evolutionary advantage of multicellular lifestyle of prokaryotes.     

Characterization of Bacterial Magnetotactic Behaviors by Using a Magnetospectrophotometry Assay

Magnetotactic bacteria have the unique capacity of synthesizing intracellular single-domain magnetic particles called magnetosomes. The magnetosomes are usually organized in a chain that allows the bacteria to align and swim along geomagnetic field lines, a behavior called magnetotaxis. Two mechanisms of magnetotaxis have been described. Axial magnetotactic cells swim in both directions along magnetic field lines. In contrast, polar magnetotactic cells swim either parallel to the geomagnetic field lines toward the North Pole (north seeking) or antiparallel toward the South Pole (south seeking). In this study, we used a magnetospectrophotometry (MSP) assay to characterize both the axial magnetotaxis of “Magnetospirillum magneticum” strain AMB-1 and the polar magnetotaxis of magneto-ovoid strain MO-1. Two pairs of Helmholtz coils were mounted onto the cuvette holder of a common laboratory spectrophotometer to generate two mutually perpendicular homogeneous magnetic fields parallel or perpendicular to the light beam. The application of magnetic fields allowed measurements of the change in light scattering resulting from cell alignment in a magnetic field or in absorbance due to bacteria swimming across the light beam. Our results showed that MSP is a powerful tool for the determination of bacterial magnetism and the analysis of alignment and swimming of magnetotactic bacteria in magnetic fields. Moreover, this assay allowed us to characterize south-seeking derivatives and non-magnetosome-bearing strains obtained from north-seeking MO-1 cultures. Our results suggest that oxygen is a determinant factor that controls magnetotactic behavior.Magnetotactic bacteria are morphologically, metabolically, and phylogenetically diverse prokaryotes (1, 11). They synthesize unique intracellular organelles, the magnetosomes, which are single-domain magnetic crystals of the mineral magnetite or greigite enveloped by membranes. Magnetosomes are usually organized in a chain(s) within the cell and cause the cell to align along geomagnetic field lines while it swims. The highest numbers of magnetotactic bacteria are generally found at, or just below, the oxic-anoxic transition zone (OATZ) or redoxocline in aquatic habitats (1). Early studies showed that Northern Hemisphere magnetotactic bacteria swim preferentially northward in parallel with the geomagnetic field lines (north seeking [NS]) (2) and that those from the Southern Hemisphere swim preferentially antiparallel to the geomagnetic field lines to the magnetic South Pole (south seeking [SS]) (4). The geomagnetic field is inclined downward from horizontal in the Northern Hemisphere and upward in the Southern Hemisphere, with the inclination magnitude increasing from the equator to the poles. Therefore, magnetotaxis might guide cells in each hemisphere downward to less-oxygenated regions of aquatic habitats, where they would presumably stop swimming until conditions change (1). A recent study reported the coexistence of both NS and SS magnetotactic bacteria in the Northern Hemisphere, which conflicts with the prevalent model of the adaptive value of magnetotaxis (14).Under laboratory conditions, magnetotactic bacteria form microaerophilic bands of cells in oxygen-gradient medium. In fact, magnetotaxis and aerotaxis work together in these bacteria, and the behavior observed has been referred to as “magnetoaerotaxis.” Two different magnetoaerotactic mechanisms, termed polar and axial, are found in different bacterial species (6). The magnetotactic bacteria, principally the magnetotactic cocci, that swim persistently in one direction along the magnetic field (NS or SS) are polar magnetoaerotactic. Magnetotactic bacteria, especially the freshwater spirilla, that swim in either direction along the magnetic field lines with frequent, spontaneous reversals of swimming direction without turning around are axial magnetoaerotactic. For polar magnetotactic bacteria, the magnetic field provides an axis and a direction for motility, whereas for axial magnetotactic bacteria, the magnetic field provides only an axis of motility. The two mechanisms can best be seen in flattened capillary tubes containing suspensions of cells in reduced medium in a magnetic field oriented parallel to the capillary. An oxygen gradient forms along the tube, beginning at the ends of the capillary, with one oriented parallel and the other antiparallel to the magnetic field (1). Band formation by axial magnetoaerotactic cells, such as Magnetospirillum magnetotacticum cells, occurs at both ends of the capillary. Rotation of the magnetic field by 180° after the formation of the bands causes the cells in both bands to rotate 180°, but the bands remain intact. In contrast, band formation by polar magnetoaerotactic cells, such as the marine cocci, occurs only at the end of the capillary for which the magnetic field and the oxygen concentration gradient are oriented opposite to each other. Rotation of the magnetic field by 180° after the formation of the band causes the cells in the band to rotate 180° and swim away, resulting in the dispersal of the band (1). In this study, we developed a magnetospectrophotometry (MSP) assay that provides an alternative method for the quantitative and versatile characterization of the two magnetotactic mechanisms. Using this assay, we demonstrated the effect of artificial magnetic fields on the generation of homogeneous NS or SS magnetotactic bacterial populations.     

Bacterial magnetosomes: microbiology, biomineralization and biotechnological applications

Magnetotactic bacteria orient and migrate along geomagnetic field lines. This ability is based on intracellular magnetic structures, the magnetosomes, which comprise nanometer-sized, membrane-bound crystals of the magnetic iron minerals magnetite (Fe₃O₄) or greigite (Fe₃S₄). Magnetosome formation is achieved by a mineralization process with biological control over the accumulation of iron and the deposition of the mineral particle with specific size and orientation within a membrane vesicle at specific locations in the cell. This review focuses on the current knowledge about magnetotactic bacteria and will outline aspects of the physiology and molecular biology of the biomineralization process. Potential biotechnological applications of magnetotactic bacteria and their magnetosomes as well as perspectives for further research are discussed. Received: 2 December 1998 / Received revision: 2 March 1999 / Accepted: 5 March 1999     

Study of the motion of magnetotactic bacteria

Motion of flagellate bacteria is considered from the point of view of rigid body mechanics. As a general case we consider a flagellate coccus magnetotactic bacterium swimming in a fluid in the presence of an external magnetic field. The proposed model generalizes previous approaches to the problem and allows one to access parameters of the motion that can be measured experimentally. The results suggest that the strong helical pattern observed in typical trajectories of magnetotactic bacteria can be a biological advantage complementary to magnetic orientation. In the particular case of zero magnetic interaction the model describes the motion of a non-magnetotactic coccus bacterium swimming in a fluid. Theoretical calculations based on experimental results are compared with the experimental track obtained by dark field optical microscopy. Correspondence to: H. G. P. Lins de Barros     

Reduced Efficiency of Magnetotaxis in Magnetotactic Coccoid Bacteria in Higher than Geomagnetic Fields

Magnetotactic bacteria are microorganisms that orient and migrate along magnetic field lines. The classical model of polar magnetotaxis predicts that the field-parallel migration velocity of magnetotactic bacteria increases monotonically with the strength of an applied magnetic field. We here test this model experimentally on magnetotactic coccoid bacteria that swim along helical trajectories. It turns out that the contribution of the field-parallel migration velocity decreases with increasing field strength from 0.1 to 1.5 mT. This unexpected observation can be explained and reproduced in a mathematical model under the assumption that the magnetosome chain is inclined with respect to the flagellar propulsion axis. The magnetic disadvantage, however, becomes apparent only in stronger than geomagnetic fields, which suggests that magnetotaxis is optimized under geomagnetic field conditions. It is therefore not beneficial for these bacteria to increase their intracellular magnetic dipole moment beyond the value needed to overcome Brownian motion in geomagnetic field conditions.     

A Comparison of Methods to Measure the Magnetic Moment of Magnetotactic Bacteria through Analysis of Their Trajectories in External Magnetic Fields

Magnetotactic bacteria possess organelles called magnetosomes that confer a magnetic moment on the cells, resulting in their partial alignment with external magnetic fields. Here we show that analysis of the trajectories of cells exposed to an external magnetic field can be used to measure the average magnetic dipole moment of a cell population in at least five different ways. We apply this analysis to movies of Magnetospirillum magneticum AMB-1 cells, and compare the values of the magnetic moment obtained in this way to that obtained by direct measurements of magnetosome dimension from electron micrographs. We find that methods relying on the viscous relaxation of the cell orientation give results comparable to that obtained by magnetosome measurements, whereas methods relying on statistical mechanics assumptions give systematically lower values of the magnetic moment. Since the observed distribution of magnetic moments in the population is not sufficient to explain this discrepancy, our results suggest that non-thermal random noise is present in the system, implying that a magnetotactic bacterial population should not be considered as similar to a paramagnetic material.     

Cell viability in magnetotactic multicellular prokaryotes.

A magnetotactic multicellular prokaryote (MMP) is an assembly of bacterial cells organized side by side in a hollow sphere in which each cell faces both the external environment and an internal acellular compartment in the center of the multicellular organism. MMPs swim as a unit propelled by the coordinated beating of the many flagella on the external surface of each cell. At every stage of its life cycle, MMPs are multicellular. Initially, a spherical MMP grows by enlarging the size of each of its cells, which then divide. Later, the cells separate into two identical spheres. Swimming individual cells of MMPs have never been observed. Here we have used fluorescent dyes and electron microscopy to study the viability of individual MMP cells. When separated from the MMP, the cells cease to move and they no longer respond to magnetic fields. Viability tests indicated that, although several cells could separate from a MMP before completely losing their motility and viability, all of the separated cells were dead. Our data show that the high level of cellular organization in MMPs is essential for their motility, magnetotactic behavior, and viability.     

Improved technique for the isolation of magnetotactic spirilla from a freshwater sediment and their phylogenetic characterization.

An improved technique for the isolation of magnetotactic bacteria was used for the axenic cultivation of microaerophilic magnetotactic spirilla. Magnetotactic bacteria were first separated from non-magnetic contaminants by exploiting their active migration along magnetic field lines by a capillary "racetrack" method. The purified magnetic cells were then inoculated into a two-layer isolation medium with opposing oxygen and sulfide gradients. Several strains of magnetotactic spirilla were isolated from a freshwater sediment sample using this method. Based on their morphology, physiology and comparative analysis of almost complete 16S rRNA gene sequences, all newly isolated strains were identified as members of the genus Magnetospirillum. While five of the isolates were closely related to previously described species (> 99% sequence similarity), two isolates appear to represent a third phylogenetic cluster within the genus Magnetospirillum.     

Killing of Staphylococcus aureus via Magnetic Hyperthermia Mediated by Magnetotactic Bacteria

Staphylococcus aureus is a common hospital and household pathogen. Given the emergence of antibiotic-resistant derivatives of this pathogen resulting from the use of antibiotics as general treatment, development of alternative therapeutic strategies is urgently needed. Here, we assess the feasibility of killing S. aureus cells in vitro and in vivo through magnetic hyperthermia mediated by magnetotactic bacteria that possess magnetic nanocrystals and demonstrate magnetically steered swimming. The S. aureus suspension was added to magnetotactic MO-1 bacteria either directly or after coating with anti-MO-1 polyclonal antibodies. The suspensions were then subjected to an alternating magnetic field (AMF) for 1 h. S. aureus viability was subsequently assessed through conventional plate counting and flow cytometry. We found that approximately 30% of the S. aureus cells mixed with uncoated MO-1 cells were killed after AMF treatment. Moreover, attachment between the magnetotactic bacteria and S. aureus increased the killing efficiency of hyperthermia to more than 50%. Using mouse models, we demonstrated that magnetic hyperthermia mediated by antibody-coated magnetotactic MO-1 bacteria significantly improved wound healing. These results collectively demonstrated the effective eradication of S. aureus both in vitro and in vivo, indicating the potential of magnetotactic bacterium-mediated magnetic hyperthermia as a treatment for S. aureus-induced skin or wound infections.