Akawaio penak, a new genus and species, is described from the upper Mazaruni River, Guyana. The new species is diagnosed from all other species of Hypopomidae by several anatomical traits. The phylogenetic affinities of the new genus were inferred using data from one nuclear (rag2) and two mitochondrial (COI and cyt b) genes. The phylogenetic analyses indicate that Akawaio is the sister taxon of a clade that includes Brachyhypopomus, Hypopomus, Microsternarchus and Racenisia. These results provide evidence for the phylogenetic composition of Hypopomidae supported by previous molecular studies and support the position of the Steatogenini (Hypopygus + Steatogenys) as the sister group of Rhamphichthys + Gymnorhamphichthys. The description of this new electric knifefish increases the total number of endemic genera and species in the upper Mazaruni, a region that is suffering freshwater habitat degradation as consequence of gold-mining activities.