To remain competitive, brewers must innovate by incorporating novel elements beyond traditional styles. Thus, exploring triticale as a modern substitute for barley malt is promising, especially given its higher amylolytic activity compared to barley. This study aimed to assess the impact of substituting up to 50% of barley malt with unmalted triticale on green beer quality, encompassing multiple stages from wort production to primary fermentation at a laboratory scale. Triticale-based worts (ratios 10-50%) had lower extract content than 100% barley malt. However, incorporating 10% of triticale led to only a 1% decrease in extract content compared to the all-malt wort. Shearzyme® 500L, an endo-1,4-β-xylanase with β-glucanase side activity, effectively addressed wort viscosity by breaking down arabinoxylans and β-glucans in triticale cell walls. All triticale-based beers exhibited lower ethanol content compared to reference beer, as is typical when using adjuncts. In green beer, a 50% triticale ratio lowered ethanol content by 16% (without enzyme) and 19% (with enzyme) compared to 100% malt beer. However, green beer with 10% triticale had satisfactory levels of total polyphenol and vicinal diketone content, among other parameters. Commercial enzyme application significantly enhanced proteolytic activity within the grain. Fermentations of enzyme-treated worts showed higher amino acid levels, further confirming the increased proteolytic activity facilitated by the chosen enzyme. Overall, this study provides a comprehensive analysis of the brewing process using native triticale. Building on this foundation, future studies will focus on optimizing mashing conditions to enhance the fermentation profile of the wort. © 2024 Society of Chemical Industry.