Advanced gas chromatographic-mass spectrometric studies for identification of the cellular octadecenoate isomers and changes of fatty acid composition induced by ethanol stress in Escherichia coli and Escherichia coli O157: H7

Ethanol can be introduced to foods of various origins and is commonly used for surface disinfection. Low concentrations of residual ethanol may provide an opportunity for pathogens to adapt and grow. Change of cellular fatty acid composition is one of adaptation mechanisms enabling bacteria to grow under varied stresses. Since instrumental analyses of bacterial octadecenoate isomers are sophisticated, gas chromatographic analyses of the isomers, namely trans-9-octadecenoate, trans-11-octadecenoate, cis-9-octadecenoate, and cis-11-octadecenoate, and ethanol-induced formation of trans-9-octadecenoate in Escherichia coli and E. coli O157:H7 were intensively investigated. When an HP-1, a nonpolar capillary column, was used for gas chromatographic analyses of 28 authentic bacterial acid methyl esters, resolution was satisfied for all fatty acid components except trans-9-octadecenoate and cis-11-octadecenoate, being overlapped. When the column was replaced by an RTx-2330, a polar capillary column, all of the above-mentioned octadecenoate isomers were resolved. When cells of E. coli and E. coli O157:H7 were harvested after submerged cultivation (30 degrees C, 150 rpm) in tryptic soy broth and tryptic soy broth supplemented with 5% ethanol at early stationary phase and subjected to cellular fatty acid analyses by using an HP-1 and RTx-2330 coupled with a mass detector, 12 fatty acids, i.e., trans-9-octadecenoate, 5 saturated fatty acids, 2 cyclopropane fatty acids and 4 cis-unsaturated fatty acids, were identified. Individual fatty acid contents varied depending on nature of fatty acid, strain of E. coli, and supplement of ethanol. As affected by ethanol stress for both E. coli strains, contents of trans-9-octadecenoate increased, whereas contents of cyc-9,10-methylene octadecanoate (cyc-9,10-19:0) decreased significantly (P < 0.05). Apparently, both E. coli strains have rendered necessary fatty acid adaptation to survive and grow under ethanol stress.