The present study aimed to scrutinize the potential of Bacillus subtilis SPB1biosurfactant, orally administered, for preventing diabetic complications in rats. The findings revealed that, Bacillus subtilis biosurfactant was an effective reducer of α-amylase activity in the plasma. Moreover, this supplement helped protect the β-cells from death and damage. Both the inhibitory action of SPB1 biosurfactant on α-amylase and the protection of the pancreas' β-cells lead to a decrease of the blood glucose levels, consequently antihyperglycemic effect. Interestingly, this lipopeptide biosurfactant modulated key enzyme related to hyperlipidemia as lipase; which leads to the regulation of the lipid profile in serum by the delay in the absorption of LDL-cholesterol and triglycerides, and a significant increase in HDL-cholesterol. Histological analyses also showed that it exerted a protective action on the pancreases and efficiently preserved the liver-kidney functions of diabetic rats, evidenced by significant decreases in aspartate transaminase, alanine transaminase, gamma-glytamyl transpeptidase and lactate deshydrogenase activities in the plasma, as well as in the creatinine and urea contents. Overall, the present study demonstrated that the hypoglycemic and antilipidemic activities exhibited by Bacillus subtilis biosurfactant were effective enough to alleviate induced diabetes in experimental rats. Therefore, SPB1biosurfactant could be considered as a potential strong candidate for the treatment and prevention of diabetes.