| Summary: | Repurposing by-products to alternative applications has become a vital part of food research. Barley rootlets (BRs) are a side-stream of malting and brewing industries. This study focuses on processing BRs into food ingredients, using fermentation with five lactic acid bacteria (LAB) as a valorisation technique. The strains used were <i>Lactiplantibacillus plantarum</i> FST 1.7, <i>Lactobacillus amylovorus</i> FST2.11, <i>Weissella cibaria</i> MG1, <i>Leuconostoc citreum</i> TR116 and <i>Limosilactobacillus reuteri</i> R29. The influence of fermentation on sugar/FODMAP/acid compositions and microbial metabolites in BRs was analysed. A variety of techno-functional properties were also evaluated. Results showed BRs were a suitable substrate for LAB, particularly for <i>Lactiplantibacillus plantarum</i> FST 1.7 and <i>Lactobacillus amylovorus</i> FST2.11. Sugar, acid and the FODMAP composition of the fermented BRs demonstrated various traits imparted by LABs, including high mannitol production from <i>Leuconostoc citreum</i> TR116 and <i>Limosilactobacillus reuteri</i> R29. <i>Limosilactobacillus reuteri</i> R29 also produced fructans using BRs as a substrate. A techno-functional analysis of BRs showed a significant reduction in α-amylase activity post sterilisation and fermentation. Fermentation reduced water-binding capacity and significantly increased oil-binding capacity. The LAB used displayed great potential in improving the functionality of BRs as a food ingredient while also showcasing LAB fermentation as a viable processing aid for BR valorisation.
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