Background: Fibroblast growth factor 19 (FGF19) is an enteric hormone regulating bile acid de novo synthesis by sensing ileal bile acid flux. However, the role of FGF19 in cholelithiasis has not yet been elucidated and therefore is investigated in the present study. Methods: Total mRNA and protein were isolated from ileal biopsies and used for tissue expression analysis. FGF19, 7 alpha-hydroxycholesterol (7 alpha-OH-Chol), 27-hydroxycholesterol (27-OH-Chol), and different bile acids were determined in the blood samples. Results: FGF19 serum levels did not differ between gallstone carriers and controls but were significantly decreased in the overweight individuals (-32%, p = 0.0002), irrespective of gallstone status (normalweight to overweight controls -29%, p = 0.0017; normalweight to overweight gallstone carriers -44%, p = 0.0338), and correlated inversely with bodyweight (p < 0.0001, rho = -0.3317). Compared to non-overweight controls, apical sodium-dependent bile acid transporter expression was significantly diminished in the non-overweight gallstone carriers (-42%, P-mRNA = 0.0393; -52%, p(protein) = 0.0169) as well as in the overweight controls (-24%, P-mRNA = 0.0148; -43%, p(protein) = 0.0017). FGF19 expression varied widely and was similar in all groups. A significant negative correlation was noted between 7 alpha-OH-Chol, 27-OH-Chol, and FGF19 serum levels (p < 0.01; rho(7 alpha-OH-Chol) = -0.2155; rho(27-OH-Chol) = -0.2144) in obesity. Conclusion: Upregulation of hepatic bile acid synthesis via FGF 19 is defective in gallstone disease but functional in overweight individuals.