Presentation Information
[P147]Comparative analysis of yield and properties of soy-derived emulsifying proteins isolated by different extraction methods
*Chau Thi Bao Bui1,2, Shohei Nosaki1, Mito Kokawa1 (1. University of Tsukuba Institute of Life and Environmental Sciences, 2. Japan Society for the Promotion of Science)
Keywords:
soybean,emulsifying proteins,protein purification,size-exclusion chromatography
【【Purpose】Emulsifying proteins are emerging as key natural emulsifiers in food and pharmaceutical applications. In food processing, proteins are usually isolated based on their isoelectric points using a combination of alkaline and acid precipitation, which however can severely alter the protein structure. By comparing different extraction methods, this study aimed to deepen the understandings of efficient emulsifying protein extraction and purification at the molecular level using soy proteins as a model.
【Method】Two extraction methods for protein isolation included (1) alkaline-acid precipitation (AAP) and (2) wet milling in buffer (WMB) applying the Simultaneous Milling and Extraction method (previously established by our group; Bui et al., 2022). In AAP, soy flour was defatted and subjected to pH adjustment to 9.0 and 5.5, followed by freeze-drying. WMB, on the other hand, used whole soybeans for milling in neutral HEPES buffer at pH 7.5 and crude proteins were maintained in buffer medium until analysis. The isolated crude proteins were then quantified using bicinchoninic acid (BCA) assay, separated and analyzed by SDS-PAGE and size-exclusion chromatography. The emulsifying activity of the resulting fractions were evaluated.
【Results】Significant differences were observed in protein recovery, profile and activity between methods, suggesting a complex relationship between yield, protein properties and functional performance.
【Method】Two extraction methods for protein isolation included (1) alkaline-acid precipitation (AAP) and (2) wet milling in buffer (WMB) applying the Simultaneous Milling and Extraction method (previously established by our group; Bui et al., 2022). In AAP, soy flour was defatted and subjected to pH adjustment to 9.0 and 5.5, followed by freeze-drying. WMB, on the other hand, used whole soybeans for milling in neutral HEPES buffer at pH 7.5 and crude proteins were maintained in buffer medium until analysis. The isolated crude proteins were then quantified using bicinchoninic acid (BCA) assay, separated and analyzed by SDS-PAGE and size-exclusion chromatography. The emulsifying activity of the resulting fractions were evaluated.
【Results】Significant differences were observed in protein recovery, profile and activity between methods, suggesting a complex relationship between yield, protein properties and functional performance.
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