Wort, derived from malt saccharification, contains a myriad of vital nutrients, such as polyphenols, flavonoids, and sugars. Beer, a time-honored libation originating from malt fermentation, graces the palate with its rich flavor and purported health benefits. Unfortunately, due to its high alcohol content, consuming beer is considered inappropriate for pregnant women and children. Hence, there is a need to develop a low-alcohol or alcohol-free fermented beverage derived from wort. This research utilized water kefir grains to produce a low-alcohol fermented beverage from wort and examined the effects of water kefir grains fermentation on various attributes of the wort, including physicochemical properties, organic acid composition, sensory characteristics, volatile compounds, and microbial diversity. Remarkably, after fermentation, a new type of low-alcohol or alcohol-free wort beverage is produced. Moreover, the fermentation process generated numerous volatile components that enhanced the sensory profile of the beverage. The findings presented in this study serve as a valuable guide for the development of innovative low-alcohol or non-alcohol fermented beverages from wort. •The impact of water kefir grain fermentation on wort was studied.•Fermentation produces a new low-alcohol or alcohol-free wort beverage.•Fermentation increased the total flavonoid content in wort.•GC-MS and GC-IMS investigated flavor differences of wort.