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

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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; CALCIUM DIPICOLINATE; DIPICOLINIC ACID; ENDOSPORES; FOURIER TRANSFORM INFRARED SPECTROSCOPY; FT-IR SPECTROSCOPY; PYRIDINE-2,6-DICARBOXYLIC ACID

Document Type: Research Article

DOI: http://dx.doi.org/10.1366/0003702054411742

Affiliations: 1: Department of Chemistry and Biochemistry, University of South Carolina, 631 Sumter St. Columbia, South Carolina 29208 2: Department of Biological Sciences, University of South Carolina, Columbia, South Carolina 29208 3: Air Force Research Laboratory at U.S. Army Edgewood Chemical Biological Center, Aberdeen Proving Ground, Maryland 21010-5424 4: Department of Molecular, Microbial and Structural Biology, University of Connecticut Health Center, Farmington, Connecticut 06030

Publication date: July 1, 2005

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