Identification of strong aggregating regions in soy glycinin upon enzymatic hydrolysis.
Upon hydrolysis with chymotrypsin, soy glycinin has a strong tendency to aggregate. The regions of glycinin from which the aggregating peptides originate were identified by accumulative-quantitative peptide mapping. To this end, the aggregating peptides were further hydrolyzed with trypsin to obtain peptides of which the sequence can be identified using RP-HPLC-MS/MS. This resulted in a hydrolysate in which 90% of the proteinaceous material was dissolved. The soluble fraction was analyzed using the method of accumulative-quantitative peptide mapping: fractionation using ion exchange chromatography, followed by identification of peptides by RP-HPLC-MS/MS, quantification based on the absorbance at 214 nm, and finally peptide mapping. For the peptide mapping the proportions in which each of the five glycinin subunits are present, as determined by Edman degradation, were taken into account. The results showed that mainly the basic polypeptide and a part of the acidic polypeptide, close to the location of the disulfide bridge between the basic and acidic polypeptides, are present in the aggregating peptide fraction. On the basis of the results obtained, an aggregation mechanism was proposed. The hydrophilic acidic polypeptides shield the hydrophobic basic polypeptides, and the former are preferentially degraded upon hydrolysis. This results in a net increase in hydrophobicity of the remaining material, which mainly consists of the basic polypeptide fragments. This increase in hydrophobicity is proposed to be the driving force in the aggregation of chymotrypsin-derived peptides of glycinin.
Kuipers
Bas J H
BJ
Department of Agrotechnology and Food Sciences, Laboratory of Food Chemistry, Wageningen University, P.O. Box 8129, 6700 EV Wageningen, The Netherlands.
Gruppen
Harry
H
D004220:Disulfides / Q000737:chemistry; D005916:Globulins / Q000737:chemistry* / Q000378:metabolism*; D006358:Hot Temperature; D006868:Hydrolysis; D010449:Peptide Mapping; D010928:Placental Lactogen; D020836:Protein Structure, Quaternary; D021122:Protein Subunits / Q000737:chemistry; D030262:Soybean Proteins / Q000737:chemistry* / Q000378:metabolism*; D013025:Glycine max / Q000737:chemistry*; D053719:Tandem Mass Spectrometry; D014357:Trypsin / Q000378:metabolism*
D004220:Disulfides; D005916:Globulins; D021122:Protein Subunits; D030262:Soybean Proteins; C011474:glycinin; D010928:Placental Lactogen; D014357:Trypsin