The Protein Journal

, 25:71 | Cite as

Identification of Eggshell Membrane Proteins and Purification of Ovotransferrin and β-NAGase from Hen Egg White

  • G. J. Ahlborn
  • D. A. Clare
  • B. W. Sheldon
  • R. W. Kelly
Article

Abstract

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 β-N-acetylglucosaminidase, lysozyme and ovotransferrin, with each displaying varying degrees of antibacterial activity. Numerous attempts to purify active fractions of β-N-acetylglucosaminidase, lysozyme and ovotransferrin from the ESM proved somewhat limited; however, hen egg white (HEW) β-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.

Keywords

β-N-acetylglucosaminidase ovotransferrin isoelectric focusing SDS-PAGE eggshell membrane hen egg white 

Abbreviations

β-NAGase

β-N-acetylglucosaminidase

ESM

eggshell membrane

HEW

hen egg white

WL

White Leghorn

P-9-L

polyethylene 9-lauryl sulfate

GauHCl

guanadine HCl

SDS

sodium dodecyl sulfate

LDS

lithium dodecyl sulfate

BCA

bicinchoninic acid

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Copyright information

© Springer Science+Business Media, Inc. 2006

Authors and Affiliations

  • G. J. Ahlborn
    • 1
  • D. A. Clare
    • 1
  • B. W. Sheldon
    • 1
    • 2
  • R. W. Kelly
    • 3
  1. 1.Department of Food ScienceNorth Carolina State UniversityRaleighUSA
  2. 2.Department of Poultry ScienceNorth Carolina State UniversityRaleighUSA
  3. 3.Department of Chemical Engineering and BiotechnologyNorth Carolina State UniversityRaleighUSA

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