At growth temperatures above 37 degrees C, Klebsiella pneumoniae does not grow in a medium containing N2 or NO3- as nitrogen sources. However, both the growth in the presence of other nitrogen sources as well as the in vitro nitrogenase activity are not affected at this temperature. The inability to fix N2 at high temperature is due to the failure of the cells to synthesize nitrogenase and other nitrogen fixation (nif) gene encoded proteins. When cells grown under nitrogen fixing conditions at 30 degrees C were shifted to 39 degrees C, there was a rapid decrease of the rate of de novo biosynthesis of nitrogenase (component 1), nitrogenase reductase (component 2), and the nifJ gene product. There was no degradation of nitrogenase at the elevated temperature since preformed enzyme remained stable over a period of at least 3 h at 39 degrees C. Thus, temperature seems to represent a third control system, besides NH4+ and O2, governing the expression of nif genes of K. pneumoniae.