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Modification of EDC method for increased labeling efficiency and characterization of low-content protein in gum acacia using asymmetrical flow field-flow fractionation coupled with multiple detectors

Zhang, Meiyu ; Nilsson, Lars ; Lee, Seungho ; Choi, Jaeyeong

Analytical and bioanalytical chemistry, 2021-10, Vol.413 (25), p.6313-6320 [Periódico revisado por pares]

Berlin/Heidelberg: Springer Berlin Heidelberg

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Título:
Modification of EDC method for increased labeling efficiency and characterization of low-content protein in gum acacia using asymmetrical flow field-flow fractionation coupled with multiple detectors
Autor: Zhang, Meiyu ; Nilsson, Lars ; Lee, Seungho ; Choi, Jaeyeong
Assuntos: Analysis ; Analytical Chemistry ; Annan teknik ; Arabinogalactan ; Asymmetrical flow field-flow fractionation (AF4) ; Asymmetry ; Biochemistry ; Carbodiimide ; Characterization and Evaluation of Materials ; Chemical Fractionation - methods ; Chemistry ; Chemistry and Materials Science ; EDC method ; Engineering and Technology ; Ethyldimethylaminopropyl Carbodiimide - chemistry ; Fluorescence ; Fluorescence spectroscopy ; Fluorescence-labeling ; Food ; Food analysis ; Food Engineering ; Food Science ; Fractionation ; Gum acacia (GA) ; Gum arabic ; Gum Arabic - chemistry ; Gums ; Hydrogen-Ion Concentration ; Identification and classification ; Labeling ; Labelling ; Laboratory Medicine ; Livsmedelsteknik ; Low-content proteinaceous matters ; Mass distribution ; Methods ; Monitoring/Environmental Analysis ; Other Engineering and Technologies ; Peptides ; pH effects ; Physiochemistry ; Polysaccharides ; Properties ; Proteins ; Proteins - chemistry ; Research Paper ; Saccharides ; Sensitivity and Specificity ; Separation (Technology) ; Serum Albumin, Bovine - chemistry ; Spectroscopy ; Spectrum analysis ; Teknik
É parte de: Analytical and bioanalytical chemistry, 2021-10, Vol.413 (25), p.6313-6320
Notas: ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
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Descrição: 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) is widely used as a crosslinker for fluorescence labeling of protein in the fields of biochemistry and food analysis. Many natural polysaccharides often contain some proteins or peptides that are very low in content but play a vital role in their biological function as well as technical applications. Determination of these low-content proteinaceous matters requires a highly sensitive and selective method. In this study, a methodological approach for investigations of the presence of proteinaceous material over the molar mass distribution ( MD ) of polysaccharides was developed using gum acacia (GA) as a model polysaccharide. EDC fluorescence-labeling method was modified by changing the pH (7, 9, and 11) of the solution for the analysis of low-content protein in food materials. Fluorescence spectroscopy and asymmetrical flow field-flow fractionation (AF4) were employed for characterizing the labeling efficiency and physiochemical properties of unlabeled and fluorescence-labeled GA. AF4 provided molar mass ( M ) and the radius of gyration ( r G ) of arabinogalactan (AG) and arabinogalactan protein complex (AGP) and determined the presence of proteinaceous matter over the MD . The labeling efficiencies of GA at pH 7, 9, and 11 determined by fluorescence spectroscopy were 56.5, 68.4, and 72.0%, respectively, with an increment of 15.5% when pH was increased from 7 to 11. The modified EDC fluorescence-labeling method allows highly sensitive and selective analysis of low-content proteinaceous matters and their distribution in natural polysaccharides. Graphical abstract
Editor: Berlin/Heidelberg: Springer Berlin Heidelberg
Idioma: Inglês

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Modification of EDC method for increased labeling efficiency and characterization of low-content protein in gum acacia using asymmetrical flow field-flow fractionation coupled with multiple detectors

Zhang, Meiyu ; Nilsson, Lars ; Lee, Seungho ; Choi, Jaeyeong

Berlin/Heidelberg: Springer Berlin Heidelberg

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