Fourier Transform Infrared Reflectance Microspectroscopy Study of Bacillus Subtilis Engineered without Dipicolinic Acid: The Contribution of Calcium Dipicolinate to the Mid-Infrared Absorbance of Bacillus Subtilis Endospores

Abstract

Mid-infrared spectra of spores of two strains of Bacillus subtilis, PS832 (wild-type) and FB122 (sleB spoVF), that are isogenic except for the two mutations in FB122 were obtained by Fourier transform infrared (FT-IR) reflectance microspectroscopy. The mutations in FB122 cause the spores of this strain to be devoid of dipicolinic acid (pyridine-2,6-dicarboxylic acid; DPA), a biomarker characteristic of bacterial spores. Analysis of these two strains by difference spectroscopy revealed a spectrum similar to that of calcium dipicolinate (CaDPA), a chelate salt of DPA. This difference spectrum was compared to mid-infrared spectra of both DPA and CaDPA, and was attributed to CaDPA only. This is the first report known to the authors of a genetically engineered organism being used to identify the spectral contribution of a particular cellular component.

Keywords

Bacillus subtilis Pyridine-2,6-dicarboxylic acid Dipicolinic acid Calcium dipicolinate Endospores FT-IR spectroscopy Fourier transform infrared spectroscopy

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