Institute of Food Chemistry and Food Biotechnology, Justus Liebig University Giessen, Heinrich-Buff-Ring 17, 35392 Giessen, Germany.
Institute of Inorganic and Analytical Chemistry, Justus Liebig University Giessen, Heinrich-Buff-Ring 17, 35392 Giessen, Germany.
Biomolecules. 2023 Feb 28;13(3):451. doi: 10.3390/biom13030451.
Thermolabile grape berry proteins such as thaumatin-like proteins (TLPs) and chitinases (CHIs) promote haze formation in bottled wines if not properly fined. As a natural grapevine pest, the spotted-wing fly is a promising source of peptidases that break down grape berry proteins because the larvae develop and feed inside mature berries. Therefore, we produced recombinant TLP and CHI as model thermolabile wine haze proteins and applied a peptidomics strategy to investigate whether larval peptidases were able to digest them under acidic conditions (pH 3.5), which are typically found in winemaking practices. The activity of the novel peptidases was confirmed by mass spectrometry, and cleavage sites within the wine haze proteins were visualized in 3D protein models. The combination of recombinant haze proteins and peptidomics provides a valuable screening tool to identify optimal peptidases suitable for clarification processes in the winemaking industry.
热不稳定的葡萄浆果蛋白,如硫素相关蛋白(TLPs)和几丁质酶(CHIs),如果不适当澄清,会在瓶装葡萄酒中促进浑浊的形成。作为一种天然的葡萄藤害虫,豹纹斑翅果蝇是一种有前途的肽酶来源,可以分解葡萄浆果蛋白,因为幼虫在成熟的浆果内发育和进食。因此,我们生产了重组 TLP 和 CHI 作为模型热不稳定葡萄酒浑浊蛋白,并应用肽组学策略来研究幼虫肽酶是否能够在酸性条件(pH 3.5)下消化它们,这在酿酒实践中是典型的。通过质谱证实了新的肽酶的活性,并在 3D 蛋白模型中可视化了葡萄酒浑浊蛋白内的切割位点。重组浑浊蛋白和肽组学的结合提供了一种有价值的筛选工具,可用于鉴定最适合酿酒工业澄清过程的最佳肽酶。