We have identified a new enzyme, skin L-tryptophan-2,3-dioxygenase (skin TDO), that catalyzes the degradation of L-tryptophan into formylkynurenine in rats. The rate of this degradation peaks in all rats at 5 to 6 weeks after birth, and also, among rats depilated at 8 weeks old, at 10 to 11 weeks after birth. We have also observed that the properties of this enzyme are closer to those of hepatic L-tryptophan-2,3-dioxygenase (hepatic TDO) than to indoleamine 2,3-dioxygenase. Although an intraperitoneal injection of L-tryptophan increased the activity of skin TDO to approximately 2.2 times greater than control values, an intraperitoneal injection of hydrocortisone and alpha-methyl-DL-tryptophan, both compounds known to affect hepatic TDO activity, had no effect on skin TDO activity. The molecular weight of skin TDO was estimated to be 16.0 kDa, which is close to the molecular weight of hepatic TDO, yet a much larger molecule than indoleamine-2,3-dioxygenase. Increased hair growth rates paralleled increased levels of skin TDO activity in 5- to 6-week-old rats, and marked increases in the activity of skin TDO occurred 2 or 3 weeks after depilation. Enzyme activity was also greatest 2 days before the time of maximum hair root length. Therefore, skin TDO may play an important role in the initiation or suppression of rat hair growth.