Objective To investigate the antioxidant, anti-α-glucosidase and pancreatic β-cell protective potential of Ensete superbum (E. superbum) seeds. Methods A variety of in vitro assays including radical scavenging, reducing power potential, phenolic content determination, α-glucosidase assay and pancreatic β-cell (1.4E7 cells) viability were employed for assessing the effect of methanolic extract of E. superbum seeds. Results The radical scavenging and reducing power effects comparable with the standard rutin were obtained while the enzyme inhibitory activity of the extract was 68-fold better than the standard antidiabetic drug, acarbose. The seed extract of E. superbum was packed-full of polyphenols with mean percentage gallic acid equivalent value of (38.2 ± 1.8) (n = 3). The protection of pancreatic cells from massive onslaught of hydrogen peroxide was far superior to that obtained for rutin. Conclusions The reputed antidiabetic therapeutic uses of the seeds extract of E. superbum may be justified on the basis of inhibition of carbohydrate enzymes, antioxidant effects and pancreatic β-cell protection.