Ultraviolet Resonance Raman Spectra of Escherichia Coli with 222.5–251.0 nm Pulsed Laser Excitation

1988 ◽  
Vol 42 (5) ◽  
pp. 782-788 ◽  
Author(s):  
K. A. Britton ◽  
R. A. Dalterio ◽  
W. H. Nelson ◽  
D. Britt ◽  
J. F. Sperry

Resonance Raman spectra of the gram-negative organism, Escherichia coli, have been obtained with 222.5-, 230.6-, and 251.0-nm excitation, and the results have been compared with those reported earlier for 242.4-nm excitation. Major changes in bacterial spectra have been observed with changes in exciting wavelength. The origins of the major peaks in each spectrum have been explained primarily in terms of contributions of nucleic acid bases and aromatic amino acids. As an aid in making assignments, spectra of aromatic amino acids, nucleosides, and mixtures of the two have been obtained at each wavelength used to excite bacterial spectra. Background fluorescence has been observed to be negligible below 251 nm. Selective excitation of bacterial nucleic acid and protein components has been done with ease. Results suggest that an extension of the exciting wavelength range to 190–220 nm will allow the selective excitation of additional cell components.

1993 ◽  
Vol 47 (1) ◽  
pp. 38-43 ◽  
Author(s):  
S. Chadha ◽  
R. Manoharan ◽  
P. Moënne-Loccoz ◽  
W. H. Nelson ◽  
W. L. Peticolas ◽  
...  

Resonance Raman spectra have been obtained with 218-nm excitation for Escherichia coli and Staphylococcus epidermidis. Large intensity differences seen in the tryptophan-associated 1556-cm−1 peak appear to be strongly related to Gram type. Unlike E. coli, S. epidermidis possess a very intense peak at 1658 cm−1 which varies in intensity with cultural conditions. Spectra excited from E. coli and Enterobacter aerogenes with 200-nm light show peaks which strongly reflect nucleic acid composition, unlike spectra excited at 218 nm. Purified, separated ribosomes of E. coli produce spectra which are dominated by nucleic acid vibrations when excited at 242 nm, but have peaks belonging nearly exclusively to protein aromatic amino acids when excited at 222 nm. The relative weakness of bacterial RNA modes excited at 222 nm from whole cells and ribosomes is attributed to nucleic acid hypochromism.


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