PURPOSE: To produce 14α-hydroxy-4-androstene-3,17-dione in high efficiency from 4-androstene-3,17-dione by microorganisms.

CONSTITUTION: A microorganism which belongs to the genus Eurotium, Scopulariopsis, Amblyosporium, Cochliobolus, Zygorhynchus, Gloerocercospora, Corynespora, Mortierella, Doratomyces, Corynascus, Cladosporium, Pithomyces or Zophiella or belongs to Mucor circinelloides f. cireinelloides, Mucor dimorphosporus, Mucor javanicus or Mucor ambiguus and having ability capable of producing 14α-hydroxy-4-androstene-3,17-dione by hydroxylating 4- androstene-3,17-dione is cultured in a medium containing 4-androstene-3,17-dione and 14α-hydroxy-4-androstene-3,17-dione is harvested from the cultured mixture to produce the objective 14a-hydroxy-4-androstene-3,17-dione.