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Summary: Spirillum lipoferum, an N2-fixing organism, was grown at constant concentrations of dissolved O2. When supplied with NH4 + aerobically, its doubling time was 1 h; when it fixed N2 microaerophilically, its doubling time was 5·5 to 7 h and the optimal P o2 for growth was 0·005 to 0·007 atm. At its optimal P o2 for growth on N2, S. lipofernum assimilated 8 to 10 mg nitrogen/g carbon substrate used; its efficiency was less at higher P O2 levels. Nitrogenase in cell-free extracts required Mg2+ and Mn2+, and the Fe-protein was activated by Rhodospirillum rubrum activating factor. The nitrogenase had an optimal pH of 7·1 to 7·4 and an apparent K m for acetylene of 0·0036 atm. Extracts of S. lipoferum lost their nitrogenase activity on storage at −18 °C, and activity was restored by adding purified Feprotein from other N2-fixing bacteria.
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