High hydrostatic pressure (HHP) was applied to fresh mango nectar (FMN) and sterilized mango nectar (SMN) to inactivate Escherichia coli and pectin methylesterase (PME). Pressure was applied at 275, 345 and 414 MPa. The come-up time (CUT) as well as 1, 2 and 4 min of treatment times were applied at the selected pressure to evaluate the inactivation effect on E. coli and PME. Total plate counts (TPC) were also evaluated in FMN. Results showed that mesophiles are inactivated in FMN to an important degree (up to 4 log) only with the CUT; the highest inactivation for mesophiles (7 log) was reported at 414 MPa after 4 min. Meanwhile, for E. coli 345 and 414 MPa after 2 and 1 min, respectively, were able to inactivate all viable cells in FMN. However, in SMN after 4 min at 275 MPa all cells of E. coli were also inactivated, showing the protective effect of the media between FMN and SMN. The PME showed its resistance to be inactivated with high pressure, showing the highest decrease in enzymatic activity (45%) after 4 min at 345 MPa but with an important activation at the highest pressure (414 MPa).