A few years ago and coinciding with the dilemma posed by March about the contradictions involved in exploration and exploitation learning, we saw the emergence of the “Organizational Ambidexterity” (OA) concept as a metaphor to define organizations that are able to develop exploitation and exploration learning at the same time. Despite the efforts made to discern the OA antecedents and moderating factors, a number of aspects still remain to be studied on the map of research into this topic. One of them is the role that human resources can play in ambidextrous learning. Special attention has also been paid in the field of human resource management to the link between high performance work systems (HPWS) and performance. Recent studies highlight the need to make further progress in this direction but using some organizational capacity as a mediating variable between HPWSs and performance. This article uses OA as the mediator variable in this relationship. The main objectives of this research are (a) to determine whether the utilization of an HPWS exerts a positive influence on OA, (b) to know the extent to which HPWSs and OA contribute to organizational performance, and (c) to verify the potential mediating role played by OA on the HPWS performance relationship. The theoretical model and the hypotheses proposed were tested using a sample of 100 Spanish hotels.