Lysozyme and the reduction of metabisulfite addition to red wine were evaluated during a winemaking process and after malolactic fermentation (MLF). Treatment, with lysozyme, of the must from Tempranillo grapes and at the end of alcoholic fermentation (AF) caused the 100% implantation of the inoculated bacterial strain and shortened the duration of MLF by 7 days. At the end of the MLF, wines treated with lysozyme showed lower volatile acidity, color intensity, and biogenic amine content. The differences in color intensity disappeared during wine stabilization. The lysozyme addition after MLF led to lower histamine concentrations in wines. These phenomena occurred irrespective of the lactic acid bacteria (LAB) proliferation control and of the Oenococcus oeni dominant strain identified at this period. The results of this study show the significance of preventive use of lysozyme in vinification of red wine to maintain low histamine levels and ensure a successful implantation of inoculated O. oeni starters.