Low serum 25(OH)D concentrations have been reported in obese humans. Inadequate sun exposure and impaired hepatic 25‐hydroxylation have been suggested as possible reasons for obesity‐associated vitamin D deficiency; however, the underlying mechanism has not been elucidated. We investigated the effects of high fat diet‐induced obesity on vitamin D status and vitamin D metabolizing enzyme expression. Male C57BL mice (4 weeks old) were fed control diet containing 10% energy from fat (control group) or high fat diet containing 45% energy from fat (obese group) for 18 weeks. There were no differences in serum 25(OH)D concentrations between two groups, while serum 1,25(OH)2D concentrations were significantly higher in obese mice. Hepatic mRNA levels of 25‐hydroxylases (Cyp2r1, Cyp27a1, and Cyp2j3) were lower in the obese group (31, 30, and 48% lower, respectively). Renal 1α‐hydroxylase (Cyp27b1) mRNA levels were higher and 24‐hydroxylase (Cyp24) mRNA levels were lower in the obese group. Serum 1,25(OH)2D concentrations correlated positively with renal Cyp27b1 expression levels and negatively with renal Cyp24 expression levels. Serum PTH concentrations were higher in obese mice. In visceral adipose tissue, Cyp27a1, Cyp2j3, and vitamin D receptor mRNA levels were higher in obese mice. Overall, vitamin D metabolizing enzyme expression was influenced by high fat diet‐induced obesity, which might partly explain the mechanisms of the altered vitamin D endocrine system associated with obesity. Higher serum PTH and 1,25(OH)2D concentrations in obese mice suggest abnormal regulation of serum 1,25(OH)2D concentrations due to hyperparathyroidism, which might have contributed to lower hepatic 25‐hydroxylase mRNA levels. © 2015 BioFactors, 41(3):175–182, 2015