For the purpose of reducing pollution and the reutilization of salted egg whites, which are byproducts of the manufacturing process of salted egg yolks and normally treated as waste, an aqueous two-phase flotation (ATPF) composed of polyethylene glycols (PEG 1000) and (NH4)2SO4 was applied to develop a simple, inexpensive and efficient process for the separation of ovalbumin (OVA) from salted egg whites. The effects of the concentration of PEG, the concentration of (NH4)2SO4, the flow rate and the flotation time on the flotation efficiency (Y) and purity (P) of OVA were investigated. A response surface method (RSM) experiment was carried out on the basis of a single-factor experiment. An efficient separation was achieved using ATPF containing 5 mL of 80% PEG 1000 (w/w), 28 mL of 28% (NH4)2SO4 (w/w), 35 mL/min of the flow rate and 30 min of the flotation time, while 2 mL of the salted egg white solution (salted eggs white (v): water (v) = 1:4) was loaded. Under the optimal conditions, Y and P of OVA could reach 82.15 ± 0.24% and 92.98 ± 0.68%, respectively. The purified OVA was characterized by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), reverse phase high-performance liquid chromatography (RP-HPLC), liquid chromatography-nano electrospray ionisation mass spectrometry (Nano LC-ESI-MS/MS), ultraviolet spectrum (UV), fluorescence spectrum (FL) and fourier transform infrared spectroscopy (FT-IR). The results indicated that the purity of OVA obtained by ATPF was satisfactory and there was no obvious difference in the structure of the OVA separated by ATPF and the standard. The results of the functional properties revealed no significant differences between OVA obtained by ATPF and the standard in oil binding capacity, viscosity, emulsibility and foam capacity.

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At present, peptides are separated by molecular exclusion chromatography and liquid chromatography. A separation method is needed in any case, which can be scaled up for industrial scale. In this study, aqueous two-phase extraction (ATPE) and aqueous two-phase flotation (ATPF) were applied to separate and enrich antioxidant peptides from trypsin hydrolysates of whey protein isolates (WPI). The best experimental conditions were investigated, and the results were evaluated using the 2,2′-Azinobis-(3-ethylbenzthiazoline-6-sulphonate) (ABTS) free radical scavenging activity of the peptides-per-unit concentration and the recovery rate (Y) of peptides in the top phase of both ATPE and ATPF. Under optimal conditions, the Y and ABTS free radical scavenging activity per unit concentration in top phase of ATPE could reach 38.75% and 12.94%, respectively, and in ATPF could reach 11.71% and 29.18%, respectively. The purified peptides were characterized by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) and reversed-phase high-performance liquid chromatography (RP-HPLC). PeptideCutter and PeptideMass were applied to analyze and calculate the peptide sequencing. KILDKVGINYWLAHK, VGINYWLAHKALCSEK, and TPEVDDEALEKFDKALK sequences having antioxidant activity were detected in the top phase of ATPE, and VGINYWLAHKALCSEK, KILLDKVGINYWLAHK, ILLDKVGINYWLAHK, IIAEKTKIPAVFK, KIIAEKTKIPAVFK, and VYVEELKPTPEGDLEILLQK sequences having antioxidant activity were detected in the top phase of ATPF. In conclusion, antioxidant peptides were successfully separated from the WPI hydrolysate by ATPE and ATPF; compared with ATPE, ATPF has superior specificity in separating antioxidant peptides.

For the purpose of reducing pollution and the rational use of salted egg white, which is a byproduct of the manufacturing process of salted egg yolk, an aqueous two-phase system (ATPS) composed of polyethylene glycols (PEG 1000) and (NH4)2SO4 was investigated to selectively separate ovalbumin (OVA) from salted egg white. With the aim of optimizing the selective separation of OVA using ATPS, a response surface method (RSM) experiment was carried out on the basis of a single-factor experiment. The OVA was characterized by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS–PAGE), reversed-phase high-performance liquid chromatography (RP-HPLC), liquid chromatography-nano electrospray ionization mass spectrometry (Nano LC-ESI-MS/MS), and Fourier transform infrared spectroscopy (FT-IR). Under the optimal conditions, the recovery yield of OVA through ATPS (Y) and the purity of OVA (P) could reach 89.25% and 96.28%, respectively. In conclusion, OVA was successfully separated from the salted egg white by PEG/(NH4)2SO4 ATPS.