Quantification of Both the Presence, and Oxidation State, of Mn in Bacillus atrophaeus Spores and its Imparting of Magnetic Susceptibility to the Spores

Bacillus atrophaeus spores were previously reported to have significant magnetic susceptibility in a magnetic field due to the presence of Mn. However, relatively little is known about the total amount and distribution of the oxidation state of Mn associated with this specific strain's spores. Using the instrument, cell tracking velocimetry (CTV) both magnetically induced velocity and settling velocity was quantitatively measured. Visual observations, and calculated diameter using previously reported densities, indicate that the spores are present in the form of clusters of approximately 3-6 mu m. Treatment of these clusters with EDTA or pH of 2.0 or below resulted in not only the disruption of the spore clusters, but also a significant decrease in magnetic susceptibility, in some cases by almost two orders of magnitude. Since the magnetic susceptibility of Mn varies significantly between the three typically reported valance states of Mn, Mn(II), Mn(III), and Mn(IV); X-Ray Photoelectron Spectroscopy, XPS, was used to determined the valance states of Mn in the spores. This XPS analysis, which penetrates up to 10 nm into the spore, returned the following fractions: 0.41, 0.38, and 0.21 for the valance states: Mn(II), Mn(III), and Mn(IV), respectively. The total mass of Mn associated with each spore cluster was determined by ICP-MS. A second, completely independent estimate of Mn mass associated with each spore cluster was made, by mathematically solving for the amount of Mn per spore cluster using the experimentally measured magnetophoretic mobility and the magnetic susceptibility of each of the three valence states from the XPS analysis. IPC-MS returned a value of 3.28 x 10(-11) g of Mn per spore cluster while the calculated estimation from mobility and XPS analysis retuned a value of 1.16 x 10(-11) g, which given the complexity of the two techniques, is a reasonable agreement. Finally, a discussion of potential applications of the magnetic properties of these spores is presented. Biotechnol. Bioeng. 2011;108: 1119-1129. (C) 2010 Wiley Periodicals, Inc.