IMPACT OF RHIZOPUS AND BOTRYTIS ON WINE FOAMING PROPERTIES

Abstract

A lot of work has been done on the impact of Botrytis on the foam of sparkling wines. This work often concerns wines produced in cool regions, where Botrytis is the dominant fungal pathogen. However, in southern countries such as Spain, in particularly hot years such as 2022, the majority fungal pathogen is sometimes Rhizopus. Like Botrytis, Rhizopus is a fungus that produces an aspartic protease. The aim of this study was therefore to determine the foaming properties of wines produced with a synthetic must contaminated by a Rhizopus or Botrytis culture. In order to confirm the identification of the fungal strain, the D1-D3 domains of the 28S rRNA gene were amplified and sequenced. BLAST search indicated 100% identity with a reference strain of Rhizopus lyococcus (CBS 320.35).

The complete experimental design presents 12 modalities (AF in triplicate, i.e. 36 bottles). The fungal isolates of Botrytis cinerea (B. c.) and Rhizopus lyococcus (R. l.) were cultured using a modified version of the method described by Gimenez et al. (2022). Alcoholic fermentations (AF) were performed in 500mL glass bottles from synthetic grape must supplemented or not with 50 mg/L of epicatechin. The yeast strain S. cerevisiae Lalvin EC1118 (Lallemand) was used for the AF process. To examine the impact of the pathoge- nic fungi, 10% (v/v) of B.c. or R.l. culture were added (separately) to the model grape juice. Furthermore, two different concentrations of L-malic acid were added to the fermentation media creating two sets of conditions : 2g/L of L-malic acid (pH=3.5) and 6 g/L of L-malic acid (pH=3). The results of the wines with fungus were compared to those of the control wines obtained without fungus.

The results of this study show that the presence of Rhizopus in the must significantly or highly significantly degrades the foamability and foam stability of the wines (foam measured with the KRUSS DFA100 equipment). The analysis of the protein composition by SDS-PAGE clearly shows a degradation of the yeast proteins by the fungal proteases of Rhizopus. Surprisingly, the Botrytis strain used did not affect the foam of the wines. These differences in proteolytic activity are confirmed by using BSA as a subs- trate: the Rhizopus culture degrades the 500 mg/L BSA in a few minutes, whereas the BSA degradation by the Botrytis culture remains considerably lower despite the longer culture of the fungus. Finally, the presence of epicatechin did not affect the wines’ foaming properties.