Identification of eggshell membrane proteins and purification of ovotransferrin and beta-NAGase from hen egg white.
Exposure of selected Gram-positive and Gram-negative bacterial pathogens to egg shell membranes (ESM) significantly reduced their thermal resistance and/or inactivated cells. Although the components responsible for this antibacterial activity have not been conclusively identified, several proteins associated with the ESM activity have been identified including beta-N-acetylglucosaminidase, lysozyme and ovotransferrin, with each displaying varying degrees of antibacterial activity. Numerous attempts to purify active fractions of beta-N-acetylglucosaminidase, lysozyme and ovotransferrin from the ESM proved somewhat limited; however, hen egg white (HEW) beta-N-acetylglucosaminidase was purified using a two-step chromatographic procedure, isoelectric focusing followed by cation exchange chromatography. Pure fractions of ovotransferrin were also obtained in the process. SDS-PAGE electrophoresis and Matrix-Assisted Laser Desorption Time-of-Flight Mass Spectrometry were then used to partially characterize the individual protein components. Purified protein fractions such as these will be required in order to fully elucidate the mechanism responsible for the antimicrobial properties associated with the ESM.
Ahlborn
G J
GJ
Department of Food Science, North Carolina State University, Raleigh, NC, 27695, USA.
Clare
D A
DA
Sheldon
B W
BW
Kelly
R W
RW
D000118:Acetylglucosaminidase / Q000032:analysis / Q000302:isolation & purification*; D000818:Animals; D000900:Anti-Bacterial Agents / Q000032:analysis*; D002645:Chickens; D002852:Chromatography, Ion Exchange; D003207:Conalbumin / Q000032:analysis / Q000302:isolation & purification*; D004527:Egg Proteins / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D007525:Isoelectric Focusing; D008565:Membrane Proteins / Q000737:chemistry; D009113:Muramidase / Q000032:analysis*; D010063:Ovum / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D000900:Anti-Bacterial Agents; D004527:Egg Proteins; D008565:Membrane Proteins; D003207:Conalbumin; C421864:hen egg lysozyme; D009113:Muramidase; D000118:Acetylglucosaminidase