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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. Incomparison with fastidious magnetotactic bacteria, At. Ferrooxidans can grow without the drastic regimes of oxidative stress, and the mass cultivation of this bacteria has been easilyavailable. 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.