The Peruvian upwelling system is a highly productive ecosystem with a large oxygen minimum zone (OMZ) close to the surface. In this work, we carried out a mesocosm experiment off Callao, Peru, with the addition of water masses from the regional OMZ collected at two different sites simulating two different upwelling scenarios. Here, we focus on the pelagic remineralization of organic matter by the extracellular enzyme activity of leucine aminopeptidase (LAP) and alkaline phosphatase activity (APA). After the addition of the OMZ water, dissolved inorganic nitrogen (N) was depleted, but the standing stock of phytoplankton was relatively high, even after N depletion (mostly > 4 µg chlorophyll a L−1). During the initial phase of the experiment, APA was 0.6 nmol L−1 h−1 even though the PO43- concentration was > 0.5 µmol L−1. Initially, the dissolved organic phosphorus (DOP) decreased, coinciding with an increase in the PO43- concentration that was probably linked to the APA. The LAP activity was very high, with most of the measurements in the range of 200–800 nmol L−1 h−1. This enzyme hydrolyzes terminal amino acids from larger molecules (e.g., peptides or proteins), and these high values are probably linked to the highly productive but N-limited coastal ecosystem. Moreover, the experiment took place during a rare coastal El Niño event with higher than normal surface temperatures, which could have affected enzyme activity. Using a nonparametric multidimensional scaling analysis (NMDS) with a generalized additive model (GAM), we found that biogeochemical variables (e.g., nutrient and chlorophyll-a concentrations) and phytoplankton and bacterial communities explained up to 64 % of the variability in APA. The bacterial community best explained the variability (34 %) in LAP. The high hydrolysis rates for this enzyme suggest that pelagic N remineralization, likely driven by the bacterial community, supported the high standing stock of primary producers in the mesocosms after N depletion.