The effects of various nitrogen soruces on the expression of nif gene were investigated using nif-lac fusants of Klebsiella pneumoniae. K. pneumoniae UK 2979 was infected with Mudl lysate prepared by heat induction of K. pneumoniae UK 4482. About 80 nif-lac fusants were greatly repressed. Amino acids, such as serine, glutamine and asparagine, were found to support the growth of K. pneumoniae M5al quite well, and showed a repressive effect on .betha.-galactosidase activities of nif-lac fusants LX-9 and LX-22 in NFHM. Glutamic acid, histidine and arginine rendered poor growth but high activities of .betha.-galactosidase. Good cell growth and high enzyme activity were observed when complex nitrogen sources, such as casitone, proteose pepone, were employed. .betha.-Galactosidase activities of LX-9 and LX-22 in nitrogen free minimal medium increased sharply within first 4 hours.