Evaluation of lysozyme stability in young red wine and model systems by a validated HPLC method

Hen’s egg lysozyme (LYZ) is added to red wine for preventing malolactic activity. Sulphur dioxide and polyphenolic compounds can depress LYZ solubility and effectiveness, likely by sulphonation of enzyme disulphide bonds or protein binding, respectively. These phenomena were evaluated in either young red wine or model wine systems. A reliable and validated HPLC method was first developed in order to quantify soluble LYZ in wine (r_95% = 10.7 mg l⁻¹). The amount of insolubilized LYZ in red wine was related to the content of non-anthocyan flavonoids of low molecular weight and enzyme precipitation occurred in few minutes when flavonoid concentration exceeded 50 mg l⁻¹. Interaction between sulphur dioxide and LYZ occurred in polyphenol-free model wine systems and for the first time the formation of mono-thiosulphonated LYZ was demonstrated. This reaction was favoured by increasing pH value and sulphur dioxide concentration. Nevertheless, sulphonation alone did not fully explain LYZ instability in model wine systems.