Intermittent severe energy restriction (SER) can induce substantial weight loss, but the appetite regulatory responses to SER are unknown and may dictate long-term dietary adherence. We determined the effect of 24-h SER on appetite regulation, metabolism, and energy intake. Eighteen lean men and women completed two 3-d trials in randomized, counterbalanced order. On day 1 subjects consumed standardized diets containing 100% (mean ± SD: 9.3 ± 1.3 MJ; energy balance) or 25% [2.3 ± 0.3 MJ; energy restriction (ER)] of energy requirements. On day 2, a standardized breakfast was consumed, with plasma concentrations of acylated ghrelin, glucagon-like peptide 1, insulin, glucose, and nonesterified fatty acids determined for 4 h. Ad libitum energy intake was assessed at lunch and dinner with subjective appetite and resting metabolism assessed throughout. On day 3, ad libitum energy intake was assessed at breakfast and by weighed food records. Energy intake was 7% greater on day 2 (P < 0.05) during ER but not significantly different on day 3 (P = 0.557). Subjective appetite was greater during ER on the morning of day 2 (P < 0.05) but was not significantly different thereafter (P > 0.145). During ER, postprandial concentrations of acylated ghrelin were lower (P < 0.05), whereas glucose (P < 0.05) and nonesterified fatty acids (P < 0.0001) were higher. Postprandial glucagon-like peptide 17-36 (P = 0.784) and insulin (P = 0.06) concentrations were not significantly different between trials. Energy expenditure was lower during ER in the morning (P < 0.01). In lean young adults, 24-h SER transiently elevated subjective appetite and marginally increased energy intake, but hormonal appetite markers did not respond in a manner indicative of hyperphagia. These results suggest that intermittent SER might be useful to attenuate energy intake and control body weight in this population. This trial was registered at www.clinicaltrials.gov.uk as NCT02696772.