Infrared signatures of Bacillus bacteria: clear IR distinctions between sporulated and vegetative cells with chemical assignments

Timothy J. Johnson; Stephen D. Williams; Nancy B. Valentine; Yin-Fong Su; Helen W. Kreuzer-Martin; Karen Wahl; Joel B. Forrester

doi:10.1117/12.823178

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8 May 2009 Infrared signatures of Bacillus bacteria: clear IR distinctions between sporulated and vegetative cells with chemical assignments

Timothy J. Johnson, Stephen D. Williams, Nancy B. Valentine, Yin-Fong Su, Helen W. Kreuzer-Martin, Karen Wahl, Joel B. Forrester

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Proceedings Volume 7304, Chemical, Biological, Radiological, Nuclear, and Explosives (CBRNE) Sensing X; 730407 (2009) https://doi.org/10.1117/12.823178
Event: SPIE Defense, Security, and Sensing, 2009, Orlando, Florida, United States

Abstract

This paper highlights the distinctions between the infrared (IR) absorption spectra of vegetative versus sporulated Bacillus bacteria. It is observed that there are unique signatures clearly associated with either the sporulated or the vegetative state, and that vegetative cells (and associated debris) can contribute to the spore spectra. A distinct feature at ~1739 cm^-1 appears to be unique to vegetative cell spectra, and can also be used as an indicator of vegetative cells or cell debris in the spore spectra. The data indicate the band is caused by a phospholipid carbonyl bond and are consistent with, but do not prove it to be, either phosphatidyl ethanolamine (PE) or phosphatidyl glycerol (PG), the two major classes of phospholipids found in vegetative cells of Bacillus species. The endospore spectra show characteristic peaks at 1441, 1277, and 1015 cm^-1 along with a distinct quartet of peaks at 766, 725, 701, and 659 cm^-1. These are clearly associated with calcium dipicolinate trihydrate, CaDP•3H₂O. We emphasize that the spore peaks, especially the quartet, arise from the calcium dipicolinate trihydrate and not from dipicolinic acid or other dipicolinate hydrate salts. The CaDP•3H₂O vibrational peaks and the effects of hydration were studied using quantum chemistry in the PQS software package. The quartet is associated with many motions including contributions from the Ca²⁺ counterion and hydration waters including Ca-O-H bends, H₂O-Ca-O torsions and O-C-O bends. The 1441 and 1015 cm^-1 modes are planar pyridine modes with the 1441 mode primarily a ring C-N stretch and the 1015 mode primarily a ring C-C stretch.

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Timothy J. Johnson, Stephen D. Williams, Nancy B. Valentine, Yin-Fong Su, Helen W. Kreuzer-Martin, Karen Wahl, and Joel B. Forrester "Infrared signatures of Bacillus bacteria: clear IR distinctions between sporulated and vegetative cells with chemical assignments", Proc. SPIE 7304, Chemical, Biological, Radiological, Nuclear, and Explosives (CBRNE) Sensing X, 730407 (8 May 2009); https://doi.org/10.1117/12.823178

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