Rate of 11.alpha.-hydroxylation of progesterone with Rhizopus nigricans was accelerated by glucose. Glucose seemed to play an important role in the formation of cofactor because its effects on the reaction were almost same as those of electron acceptors such as NADPH and $NaIO_{4}$. Rate of glucose consumption appeared to increase in proportion as the rate of hydroxylation reaction, which enhanced with increase in the glucose concentration to level off at 0.5 g/l for mycelia and at 20 g/l for spores. However, for mycelia immobilized in polyacrylamide gel, externally added glucose did not affect the reaction rate at all because of the glucose accumulated in the gel during the cultivation period. 5.alpha.-Reduction of 11.alpha.-hydroxyprogesternoe required much higher concentration of glucose than 11.alpha.-hydroxylation of progesternoe so that high yield of 11.alpha.-hydroxyprogesternoe can be obtained by repressing the activity of 5.alpha.-reductase at low concentration of glucose.