Raman Spectra of Endospores of Bacillus Subtilis by Alkali Stress
DONG Rong1, 2, LU Ming-qian1, 3, LI Feng1, 4, SHI Gui-yu2, HUANG Shu-shi1, 3*
1. Laboratory of Biophysics, Guangxi Academy of Sciences, Nanning 530007, China 2. College of Life Science, Guangxi Normal University, Guilin 541004, China 3. Biotechnology Research Center, Kunming University of Science and Technology, Kunming 650224, China 4. Department of Biological and Chemical Engineering, Guangxi University of Technology, Liuzhou 545006, China
Abstract：To research the lethal mechanism of spores stressed by alkali, laser tweezers Raman spectroscopy (LTRS) combined with principal components analysis (PCA) was used to study the physiological process of single spore with alkali stress. The results showed that both spores and germinated spores had tolerance with alkali in a certain range, but the ability of spores was obviously lower than that of spores due to the release of their Ca2+-DPA which plays a key role in spores resistance as well as spores resistance to many stresses; A small amount of Ca2+-DPA of spores was observed to release after alkali stress, however, the behavior of release was different with the normal Ca2+-DPA release behavior induced by L-alanine; The data before and after alkali stress of the spores and g.spores with PCA reflected that alkali mainly injured the membrane of spores, and alkali could be easily enter into the inner structure of spores to damage the structure of protein backbone and injure the nucleic acid of spores. We show that the alkali could result in the small amount of Ca2+-DPA released by destroying the member channel of spores.
[1] Piggot P J, Hilbert D W. Current Opinion in Microbiology, 2004, 7: 579. [2] Stragier P, Losick R. Annual Review of Genetics, 1996, 30: 297. [3] Storz G, Hengge-Aronis R. American Society for Microbiology, 2000: 217. [4] Nicholson W L, Fajardo-Cavazos P. Recent Research Developments in Microbiology, 1997, 1: 125. [5] Nicholson W L, Munakata N, Horneck G, et al. Microbiology and Molecular Biology Reviews, 2000, 64: 548. [6] Setlow P. Annual Review of Microbiology, 1995, 49: 29. [7] Setlow P. American Society for Microbiology, 2000. 217. [8] Johnstone K. Journal of Applied Bacteriology-Symposium Supplement, 1994, 76: 17S. [9] Murrell W G. Advances in Microbial Physiology, 1967, 1: 133. [10] Murrell W G, Warth A D. American Society for Microbiology, 1965, 1. [11] Paidhungat M, Setlow B, Driks A, ed al. Journal of Bacteriol, 2000, 182: 5505. [12] Setlow B, Loshon C A, Genest P C. Journal of Applied Microbiology, 2002, 92: 362. [13] Huang Shushi, Chen De, Pelczar Patricia L, et al. Journal of Bacteriology, 2007, 189(13): 4681. [14] CHENG Qin，DONG Rong，HUANG Shu-shi，et al(程 琴, 董 蓉, 黄庶识, 等). Chinese Journal of Lasers(中国激光), 2011, 38(10): 1015002. [15] Magge A, Granger A C, Wahome P G, et al. Journal of Bacteriology, 2008, 190: 4798. [16] XU Yi-ming(许以明). Raman Spectroscopy in Application of Structure Biology(拉曼光谱及其在结构生物学中的应用). Beijing: Chemical Industry Press(北京：化学工业出版社), 2005. 11.
