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《第三届全国微生物资源学术暨国家微生物资源平台运行服务研讨会会议论文摘要集》2011年
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Bioimpatibility study of magnetosomes formed by Acidithiobacillus ferrooxidans

【摘要】:Magnetotactic bacteria(MTB) are motile prokaryotes that have the ability to propel along geomagnetic field lines.Hallmarks of MTB are intracellular magnetosomes,which are magnetic crystals of either iron oxide,magnetite(Fe3O4),or iron sulfide,greigite(Fe3S4) or iron pyrite(FeS2) enveloped by a phospholipid bilayer membrane.It has been reported that Acidithiobacillus ferrooxidans(At.ferrooxidans) is able to produce magnetosomes.In comparison with fastidious magnetotactic bacteria,At.ferrooxidans can grow without the drastic regimes of oxidative stress,and the mass cultivation of this bacteria has been easily available.Although there is extensive literature on the biocompatibility of chemosynthetic magnetite nanocrystal,much less information on the biocompatibility of biosynthetic magnetite nanocrystal is available.In the present work,the orthogonal design of experiment was used to optimize the condition of At.ferrooxidans growth and magnetosomes synthesizing.The results indicated that the optimized conditions of At.ferrooxidans growth and magnetosomes synthesizing were not identical.The optimized conditions for the growth of At.ferrooxidans were found to be:FeSO4 120 mM,(NH4)2SO4 3.0 g/L,temperature 30 oC,initial pH 1.75,and 20% loadings.However,the optimized conditions for magnetosomes synthesizing of At.ferrooxidans were found to be:FeSO4 160 mM,(NH4)2SO4 2.4 g/L,temperature 20 oC,initial pH 1.75,and 50% loadings.Initial purification of the magnetosomes can be accomplished by washing eight to ten times with HEPES buffer,along with lysozyme treatment,low-temperature ultrasonication,and magnet collection.We investigated the properties of magnetosomes produced by At.ferrooxidans in detail,such as crystalline phase,vitro cytotoxicity,blood compatibility,genotoxicity and primary functional group on the magnetosomes membrane surface.Transmission electron microscopy(TEM) revealed that the number of magnetosomes in single bacterial cell is about 2.The X-ray diffraction(XRD) assay indicated that the magnetic nanocrystal within the magnetosome is consisted of magnetite.Fourier transform infrared spectroscopy(FTIR) analysis revealed the existence of different chemical groups on the surface of the magnetosome.The cytotoxicity of the magnetosomes was evaluated by the dimethylthiazoldiphenyltetrazoliumbromide(MTT) assay.The L929 cell line showed a complete biocompatible to magnetosomes from low to high concentration.The results indicated that the magnetosomes did not cause toxicity to mouse fibroblasts over a long period of time.Hemolysis study showed that the magnetosomes had no hemolysis activity.The micronucleus test(MNT) implied that the magnetosomes cause no mutagenic and clastogenic effects.All these results suggested that the magnetosomes possess high good biocompatibility and might be applied in biomedical science and engineering in the future.
【分类号】:Q937

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