Department of Agronomy, Food, Natural Resources, Animals and Environment (DAFNAE), University of Padova, Viale dell'Università 16, 35020 Padova, Italy.
Centre for Research in Viticulture and Enology (CIRVE), Viale XXVIII Aprile 14, 31015 Conegliano, Italy.
Molecules. 2018 Nov 23;23(12):3070. doi: 10.3390/molecules23123070.
Mannoproteins (MPs) are cell wall proteoglycans released in wine by yeast during fermentation and ageing on lees, a procedure used for the production of several wines to enrich them in these components with consequences from both a technological and sensory point of view. Given the significance that wine MPs have for wine quality, winemakers would welcome a simple and accurate method for their quantification, as this would allow them to have a better control of this aspect at different winemaking stages. This study develops and validates a novel, simple and accurate method for MPs quantification in white wines based on a competitive indirect enzyme-linked lectin sorbent assay (CI-ELLSA), using the highly mannosylated yeast invertase as the standard. The method utilizes the lectin concanavalin A (ConA) as the immobilized ligand for MPs, and peroxidase, an enzyme rich in mannose, as the competitor for ConA. After addition of the peroxidase substrate, the intensity of the signal produced by the activity of this enzyme (absorbance at 450 nm) is inversely proportional to the amount of mannosylated proteins in the sample. Results have been validated on several wine styles including still, sparkling and sweet wines.
甘露糖蛋白（MPs）是酵母在发酵和酒泥陈酿过程中释放到葡萄酒中的细胞壁糖蛋白，这一过程用于生产多种葡萄酒，以使这些葡萄酒富含这些成分，从技术和感官角度来看都有影响。鉴于葡萄酒 MPs 对葡萄酒质量的重要性，酿酒师们将欢迎一种简单而准确的定量方法，因为这将使他们能够在不同的酿酒阶段更好地控制这一方面。本研究开发并验证了一种基于竞争性间接酶联凝集素吸附测定法（CI-ELLSA）的新型、简单、准确的白葡萄酒 MPs 定量方法，该方法使用高度甘露糖化的酵母转化酶作为标准。该方法利用凝集素伴刀豆球蛋白 A（ConA）作为 MPs 的固定配体，而过氧化物酶，一种富含甘露糖的酶，作为 ConA 的竞争物。加入过氧化物酶底物后，该酶活性产生的信号强度（在 450nm 处的吸光度）与样品中甘露糖化蛋白的量成反比。该方法已在包括静止、起泡和甜酒在内的多种葡萄酒风格上得到验证。
