Nutritional dependencies, especially those regarding nitrogen sources, govern numerous microbial positive interactions. As for lactic acid bacteria (LAB), responsible for the sanitary, organoleptic, and health properties of most fermented products, such positive interactions have previously been studied between yogurt bacteria. However, they have never been exploited to create artificial co-cultures of LAB that would not necessarily coexist naturally, i.e from different origins. The objective of this study was to promote LAB positive interactions, based on nitrogen dependencies in co-cultures, and to investigate how these interactions affect some functional outputs, e.g. acidification rates, carbohydrate consumption, and volatile compound production. The strategy was to exploit both proteolytic activities and amino acid auxotrophies of LAB. A chemically defined medium was thus developed to specifically allow the growth of six strains used, three proteolytic and three non-proteolytic. Each of the proteolytic strains, Enterococcus faecalis CIRM-BIA2412, Lactococcus lactis NCDO2125, and CIRM-BIA244, was co-cultured with each one of the non-proteolytic LAB strains: L. lactis NCDO2111, Lactiplantibacillus plantarum CIRM-BIA465 and CIRM-BIA1524. Bacterial growth was monitored using compartmented chambers to compare growth in mono- and co-cultures. Acidification, carbohydrate consumption and volatile compound production was evaluated in direct co-cultures. Each proteolytic strain induced different types of interactions: either strongly positive, weakly positive, or no interactions, with E. faecalis CIRM-BIA2412, L. lactis NCDO2125 and L. lactis CIRM-BIA244, respectively. Strong interactions were associated with higher concentrations in tryptophan, valine, phenylalanine, leucine, isoleucine, and peptides. They led to faster acidification rates, lower pH, higher raffinose utilization and concentrations in five volatile compounds. Importance: Lactic acid bacteria (LAB) interactions are often studied in association with yeasts or propionibacteria in various fermented food products and the mechanisms underlying their interactions are being quite well characterized. Concerning interactions between LAB, they have mainly been investigated to test antagonistic interactions. Understanding how they can positively interact could be useful in multiple food-related fields: production of fermented food products with enhanced functional properties or fermentation of new food matrices. This study investigates the exploitation of the proteolytic activity of LAB strains to promote positive interactions between proteolytic and non-proteolytic strains. The results suggest that proteolytic LAB do not equally stimulate non-proteolytic LAB and that the stronger the interactions between LAB are, the more functional outputs we can expect. Thus, this study gives insight into how to create new associations of LAB strains and to guaranty their positive interactions.