The seeds proteins are converted into the peptides during the germination through peptidases. These bioactive peptides are fractionated through the enzymes. The bioactive peptides are reported with the number of therapeutic effects like antioxidant, antibacterial, antidiabetic, anti-cancerous and antihypertensive etc. Synthetic drugs have number of adverse effects along with the therapeutic effects, but natural sources are safer and free from this problem. In the present research, Poncirus trifoliata seedling peptides extracts were used to determine antibacterial, antioxidant and anti-diabetic activities. The peptides extract showed substantial growth of inhibition against pathogenic strains including Escherichia coli, Staphylococcus aureus and Bacillus subtilis having the superior effect as compared to amoxicillin antibiotic as a standard drug. The IC50 value of P. trifoliata seeds peptides extract was 32.91 μg mL-1 as compared with standard (IC50) value 25.52 μg mL-1 of ascorbic acid showed good potential as antioxidant activity. The extracted germinated seeds peptides showed considerable antidiabetic activity in controlling the blood glucose level of alloxan induced diabetic rabbits to compare with standard drugs like glibenclamide. Thus, this study confirmed the antibacterial, antioxidant and antidiabetic effects of P. trifoliata seeds peptides is significantly higher as compared to pure drugs. Therefore, further complete separation and detailed investigation study is needed to isolate target peptides and revealed mechanism of action of bioactive peptides and formulation as a drug to replace the synthetics drugs with safe one. © 2021 Friends Science Publishers