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標題: Develop a low power microwave-assisted enzyme digestion platform for efficient detection of bioactive peptide using mass spectrometry
建立低功率微波輔助酵素水解平台於蛋白 Glycinin 之活性胜肽 VVYP 釋放分析
作者: 蔡創韋
Chuang-Wei Tsai
關鍵字: 微波;VVYP;保健食品;microwave;VVYP;dietary supplements
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The bioactive peptide Val-Val-Tyr-Pro (VVYP) can be genetically modified into dietary supplements which functions in reducing the triglyceride in blood and thus aid in preventing obesity, coronary heart disease and cardiovascular diseases. Dietary supplements are usually digested by various enzymes in the intestinal tract hence cause the production of functional peptides. Previous studies showed microwave-assisted system can highly improve the hydrolysis efficiency and it is timesaving. Besides, microwave-assisted method with high power usually produces lots of unpredicted miss-cleavage peptides resulting higher sequence coverage arises from longer peptides. However, this situation causes the predicted target peptides cannot be quantified. Therefore, the analysis of genetically modified VVYP storage protein of soy bean glycinin (Gy) using strategy of bi-enzyme coupled with low power microwave-assisted system is introduced to develop the best analysis platform via HPLC-ESI-SRM. By using trypsin and carboxypeptidase B (CPB), Gy1 and Gy5 will release 7 sets and 10 sets of VVYP respectively. With this strategy, a platform with high efficiency and high recovery rate for the analysis of VVYP in Gy1 and Gy5 is expected. In the future, this platform can be applied on detecting and quantifying health foods as well as protein drugs.
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