The present study evaluated the effects of feeding frequency during the grow‐out phase of Litopenaeus vannameiin biofloc system. The experiment comprised of four treatments with three replicates each, corresponding to the following feeding frequencies: one (8:00 a.m.), two (8:00 a.m. and 4:00 p.m.), three (8:00 a.m., 12:00 p.m. and 4:00 p.m.) and four times a day (8:00 a.m., 10:00 a.m., 12:00 p.m. and 4:00 p.m.). Zootechnical performance and digestive enzyme activity were monitored for 63 days. At the end of the experiment, the highest survival rates were found for the individuals fed three and four times a day. Wet weight gain (6.63 g) and specific growth rate (3.46% per day) of the individuals fed three times a day were significantly higher compared to animals fed one and four times a day, but did not differ significantly to the group fed twice a day (6.15 g and 3.46% per day, respectively). The digestive enzyme activities were not affected by the feeding frequency, but negative correlations were found between these activities and shrimp weight. It is recommended that L. vannamei should be fed three times a day during the grow‐out phase in a biofloc system under the conditions used in this study.