Electrophoretic method for the identification of a haze-active protein in grape seeds.
A simple, fast, and more selective approach is presented in this study for the identification of haze-active proteins. Grape seed proteins were unfolded by 1% SDS and then interacted with different amounts of tannin at 4 degrees C, followed by gel electrophoresis. It was found that the intensity of the band at 45 kDa was decreased as tannins increased. The amino acid composition of this isolated 45-kDa protein was higher in proline (9.49%) than the average proline content of total grape seed proteins (4.85%). To verify the selectivity of the proposed method, a globular protein (bovine serum albumin, BSA) and a proline-rich protein (gelatin) were selected and used in the model system. As expected, gelatin was removed as it reacted with the increasing added tannins, whereas BSA did not. These results showed that it is possible to identify haze-active proteins by modulating the accessibility of protein to tannins, suggesting this new method can be used by the beverage industry to trouble-shoot haze problems and for quality control.
Wu
Li-Chen
LC
Department of Applied Chemistry, National Chi-Nan University, Puli, Nantou, 545 Taiwan. lw25@ncnu.edu.tw
Lu
Ying-Wei
YW
D000596:Amino Acids / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D010940:Plant Proteins / Q000032:analysis* / Q000737:chemistry; D012639:Seeds / Q000737:chemistry*; D013634:Tannins / Q000378:metabolism*; D027843:Vitis / Q000737:chemistry*
D000596:Amino Acids; D010940:Plant Proteins; D013634:Tannins