Intestinal mucin synthesis is sensitive to dietary threonine supply, which suggests that the gut's requirement for threonine may comprise a significant proportion of the whole body requirement. We used a continuously perfused gut loop model and intraluminal flooding dose technique in 6 young pigs to study the acute effects of varying luminal availability of threonine on intestinal protein and mucin syntheses. A complete amino acid mixture containing 0, 21, or 56 mg threonine/g total amino acids (whole body requirement ratio is approximately 28 mg/g) was continuously perfused in isolated loops for 120 min, including a 30-min (3)H-phenylalanine flooding dose. We measured fractional synthesis rates of total mucosal protein and mucin by analyzing (3)H-phenylalanine incorporation. Fractional rates of total mucosal protein synthesis were higher in loops perfused with solutions containing threonine at 56 mg/g (66 +/- 4%/d) compared with 0 mg/g (42 +/- 9%/d) and 21 mg/g (53 +/- 6%/d) (P < 0.05). For mucin, fractional rates of synthesis differed between 0 mg/g (323 +/- 72%/d), 21 mg/g (347 +/- 49%/d), and 56 mg/g (414 +/- 31%/d) (P < 0.05). In addition, total proline and threonine concentrations in the protein hydrolysates increased with luminal threonine concentration (P < 0.05), indicating an increase in threonine- and proline-rich proteins. De novo synthesis of mucosal and mucin proteins is acutely sensitive to luminal threonine concentration, which demonstrates the importance of dietary amino acid supply to gut protein metabolism.