BACKGROUND: Being nutricereals with recognized nutritional content, bioactive components, and therapeutic prospects, millets support agronomic relevance and dietary value objectives of sustainable development.METHOD: The present study investigated soil health and nutriology, phyto-compounds (gas chromatography-mass spectrometry [GC-MS] profiling), proximate composition, the total phenolic content (TPC), total flavonoid content (TFC), DPPH (2,2-diphenyl-1-picrylhydrazyl), and H2O2 (peroxide) scavenging activity, antimicrobial potential, of Eleusine coracana, Pennisetum glaucum, and Sorghum bicolor cultivated in Panchal, Bankura, West Bengal, India.KEY RESULTS: The soil texture is sandy clay loam in nature, and iron content was found maximum among the micronutrients present in the soil. The E. coracana and P. glaucum contain the maximum carbohydrate and protein content, respectively. GC-MS profiling millets detected the presence of hydrocarbon, fatty acid, triterpene, fatty amide, and sterol compounds. Sorghum exhibited total phenolic and flavonoid content with 231 mg GAE/100 g and 186.79 mg quercetin equivalents (QE)/100 g, respectively. However, E. coracana shows 16.04% and 23.85% more H2O2 scavenging activity than sorghum and P. glaucum, respectively, and more DPPH scavenging activity at 91.02% at a concentration of 36 µg/mL. Ethanolic extract of sorghum showed antimicrobial efficacy against Escherichia coli (ATCC 25922), Staphylococcus aureus (ATCC 6538), and Pseudomonas aeruginosa (ATCC 10145) with the zone of inhibition of 2.4 ± 0.12, 2.6 ± 0.12, and 2.1 ± 0.12 cm, respectively. The minimum inhibitory concentration (15 µg/mL) of ethanolic extract of sorghum exhibited inhibition of S. aureus by 92.64% ± 2.01%, P. aeruginosa by 96.64% ± 3.02%, and E. coli by 98.64% ± 3.02%. Further, molecular docking study found cis-stilbene with LasR protein from P. aeruginosa, Lupeol, methyl ester with SarA protein from S. aureus, 1,3-docosenamide, (Z)- with CpxR from E. coli as effective.CONCLUSION: Sorghum exhibited maximum phenolic, flavonoid content, and antimicrobial potential against P. aeruginosa (ATCC 10145). Sorghum bioactive compounds exhibited more efficiency than ciprofloxacin in biofilm eradication, rhamnolipid, pyocyanin, and elastase activity reduction in P. aeruginosa (ATCC 10145). cis-stilbene exhibited maximum binding efficacy (-8.2 kcal/mol) against the LasR protein from P. aeruginosa.