Abstract
BackgroundEggshells which consist largely of calcareous outer shell and shell membranes, constitute a significant part of poultry hatchery waste. The shell membranes (ESM) not only contain proteins that originate from egg whites but also from the developing embryos and different contaminants of microbial and environmental origins. As feed supplements, during post hatch growth, the hatchery egg shell membranes (HESM) have shown potential for imparting resistance of chickens to endotoxin stress and exert positive health effects. Considering that these effects are mediated by the bioactive proteins and peptides present in the membrane, the objective of the study was to identify the protein profiles of hatchery eggshell membranes (HESM).MethodsHatchery egg shell membranes were extracted with acidified methanol and a guanidine hydrochloride buffer then subjected to reduction/alkylation, and trypsin digestion. The methanol extract was additionally analyzed by matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS). The tryptic digests were analyzed by liquid chromatography and tandem mass spectrometry (LC-MS-MS) to identify the proteins.ResultsOur results showed the presence of several proteins that are inherent and abundant in egg white such as, ovalbumin, ovotransferrin, ovocleidin-116, and lysozyme, and several proteins associated with cytoskeletal, cell signaling, antimicrobial, and catalytic functions involving carbohydrate, nucleic acid, and protein metabolisms. There were some blood derived proteins most likely originating from the embryos and several other proteins identified with different aerobic, anaerobic, gram positive, gram negative, soil, and marine bacterial species some commensals and others zoonotic.ConclusionThe variety of bioactive proteins, particularly the cell signaling and enzymatic proteins along with the diverse microbial proteins, make the HESM suitable for nutritional and biological application to improve post hatch immunity of poultry.Electronic supplementary materialThe online version of this article (doi:10.1186/s12953-017-0112-6) contains supplementary material, which is available to authorized users.
Highlights
Eggshells which consist largely of calcareous outer shell and shell membranes, constitute a significant part of poultry hatchery waste
Several identified proteins on the basis of a single peptide were considered tentative they were identified repeatedly in replicate samples and by Skyline retention time correlation refinements. These identifications were largely occurred in methanol extract which may relate to degraded proteins
Based on our previous identification in Egg shell membrane (ESM) [17], we surmised that the m/z 4773 was the mature gallinacin 10 peptide with a corresponding sequence of “DPLFPDTVACRTQGNFCRAGACPPTFTIS GQCHGGLLNCCAKIPAQ” and the m/z 4643 was the same peptide with the truncation of glutamine (Q) from the N-terminal
Summary
Eggshells which consist largely of calcareous outer shell and shell membranes, constitute a significant part of poultry hatchery waste. As feed supplements, during post hatch growth, the hatchery egg shell membranes (HESM) have shown potential for imparting resistance of chickens to endotoxin stress and exert positive health effects. Considering that these effects are mediated by the bioactive proteins and peptides present in the membrane, the objective of the study was to identify the protein profiles of hatchery eggshell membranes (HESM). Egg shells that constitute a significant part of poultry hatchery waste, comprise largely of calcium carbonate crusts and underlying shell membranes laced with proteins originating from egg whites as well as contaminants of microbial and environmental origins [1]. The current study was conducted utilizing the HESM that were dried at room temperature in a chemical hood without additional cleanup for 2 month period in the room temperature
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