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. 2018 Nov 23;23(12):3070.
doi: 10.3390/molecules23123070.

A Novel Method for the Quantification of White Wine Mannoproteins by a Competitive Indirect Enzyme-Linked Lectin Sorbent Assay (CI-ELLSA)

Matteo Marangon  1 , Mara Vegro  2 , Simone Vincenzi  3   4 , Giovanna Lomolino  5 , Alberto De Iseppi  6 , Andrea Curioni  7
Affiliations

A Novel Method for the Quantification of White Wine Mannoproteins by a Competitive Indirect Enzyme-Linked Lectin Sorbent Assay (CI-ELLSA)

Matteo Marangon et al. Molecules. .

Abstract

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.

Keywords: competitive indirect enzyme-linked lectin sorbent assay (CI-ELLSA); mannoproteins; method; quantification; wine; yeast invertase.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Variation in absorbance at 450 nm of for increasing concentrations of yeast invertase, fetuin and bovine serum albumin (BSA).
Figure 2
Figure 2
Variation in absorbance at 450 nm of serial dilutions of: ultra-filtered (UF) white wine spiked with increasing amounts of yeast invertase, precipitated with ethanol and dissolved in binding buffer (blue line); yeast invertase dissolved directly in binding buffer (green line).
Figure 3
Figure 3
Schematic representation of the protocol of analysis of mannoproteins (MPs) by competitive indirect enzyme-linked lectin sorbent assay (CI-ELLSA).
See this image and copyright information in PMC

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