Recent studies in southern Africa have revealed a wealth of novel freshwater crab species in high mountainous regions. In the present study, phylogeographic affinities between two sister mountain-living freshwater crab species (Potamonautes brincki and P. parvicorpus) were examined for novel lineages. Seventy-six crab specimens were collected throughout the Western Cape Province of South Africa and sequenced for the COI locus. Evolutionary relationships were analysed using Bayesian inference, maximum likelihood and maximum parsimony, a haplotype network and analyses of molecular variance (AMOVA). Results revealed three divergent clades. Clade A comprised specimens of P. brincki restricted to the Hottentots Holland; sister to which was Clade B from the Overberg, while Clade C comprised specimens of P. parvicorpus from the Cape Peninsula and adjacent interior. Haplotype networks and AMOVA provide evidence for the absence of gene flow whilst morphology of the male gonopods and the mandibular palp revealed subtle but consistent differences between the three clades. Since Clades A and C represent two described species, P. brincki and P. parvicorpus, respectively, Clade B is herein described as a new species, P. tuerkayi, sp. nov. These results highlight the importance of continued sampling of mountain habitats to document aquatic invertebrate diversity.