Ageing is characterized by changes in several cellular processes, with dysregulation of peroxisome function being one of them. Interestingly, the most conserved function of peroxisomes, ROS homeostasis, is strongly associated with ageing and age-associated pathologies. Previous studies have identified a role for peroxisomes in the regulation of chronological lifespan in yeast. In this study, we report the effect of altered peroxisome number on the chronological lifespan of yeast in two different growth media conditions. Three mutants, pex11, pex25 and pex27, defective in peroxisome fission, have been thoroughly investigated for the chronological lifespan. Reduced chronological lifespan of all the mutants was observed in peroxisome-inducing growth conditions. Furthermore, the combined deletion pex11pex25 exhibited the most prominent reduction in lifespan. Interestingly altered peroxisomal phenotype upon ageing was observed in all the cells. Increased ROS accumulation and reduced catalase activity was exhibited by chronologically aged mutant cells. Interestingly, mutants with reduced number of peroxisomes concomitantly also exhibited an accumulation of free fatty acids and increased number of lipid droplets. Taken together, our results reveal a previously unrealized effect of fission proteins in the chronological lifespan of yeast.