Aims/hypothesis Post-translational attachment of a small ubiquitin-like modifier (SUMO) to the lysine (K) residue(s) of target proteins (SUMOylation) is an evolutionary conserved regulatory mechanism. This modification has previously been demonstrated to be implicated in the control of a remarkably versatile regulatory mechanism of cellular processes. However, the exact regulatory role and biological actions of the E2 SUMO-conjugating enzyme (UBC9)-mediated SUMOylation function in pancreatic beta cells has remained elusive. Methods Inducible beta cell-specific Ubc9 (also known as Ube2i) knockout (KO; Ubc9(Delta beta)) and transgenic (Ubc9(Tg)) mice were employed to address the impact of SUMOylation on beta cell viability and functionality. Ubc9 deficiency or overexpression was induced at 8 weeks of age using tamoxifen. To study the mechanism involved, we closely examined the regulation of the transcription factor nuclear factor erythroid 2-related factor 2 (NRF2) through SUMOylation in beta cells. Results Upon induction of Ubc9 deficiency, Ubc9(Delta beta) islets exhibited a 3.5-fold higher accumulation of reactive oxygen species (ROS) than Ubc9(f/f) control islets. Islets from Ubc9(Delta beta) mice also had decreased insulin content and loss of beta cell mass after tamoxifen treatment. Specifically, at day 45 after Ubc9 deletion only 40% of beta cell mass remained in Ubc9(Delta beta) mice, while 90% of beta cell mass was lost by day 75. Diabetes onset was noted in some Ubc9(Delta beta) mice 8weeks after induction of Ubc9 deficiency and all mice developed diabetes by 10 weeks following tamoxifen treatment. In contrast, Ubc9(Tg) beta cells displayed an increased antioxidant ability but impaired insulin secretion. Unlike Ubc9(Delta beta) mice, which spontaneously developed diabetes, Ubc9(Tg) mice preserved normal non-fasting blood glucose levels without developing diabetes. It was noted that SUMOylation of NRF2 promoted its nuclear expression along with enhanced transcriptional activity, thereby preventing ROS accumulation in beta cells. Conclusions/interpretation SUMOylation function is required to protect against oxidative stress in beta cells; this mechanism is, at least in part, carried out by the regulation of NRF2 activity to enhance ROS detoxification. Homeostatic SUMOylation is also likely to be essential for maintaining beta cell functionality.