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Metabolism and Enzymology

Azolla-Anabaena Relationship ¹

XIII. Fixation of [¹³N]N₂

Department of Bacteriology, University of California, Davis, California 95616, Battelle-C.F. Kettering Research Laboratory, 150 East South College Street, Yellow Springs, Ohio 45387

The major radioactive products of the fixation of [¹³N]N₂ by Azolla caroliniana Willd.-Anabaena azollae Stras. were ammonium, glutamine, and glutamate, plus a small amount of alanine. Ammonium accounted for 70 and 32% of the total radioactivity recovered after fixation for 1 and 10 minutes, respectively. The presence of a substantial pool of [¹³N]N₂-derived ¹³NH₄⁺ after longer incubation periods was attributed to the spatial separation between the site of N₂-fixation (Anabaena) and a second, major site of assimilation (Azolla). Initially, glutamine was the most highly radioactive organic product formed from [¹³N]N₂, but after 10 minutes of fixation glutamate had 1.5 times more radiolabel than glutamine. These kinetics of radiolabeling, along with the effects of inhibitors of glutamine synthetase and glutamate synthase on assimilation of exogenous and [¹³N]N₂-derived ¹³NH₄⁺, indicate that ammonium assimilation occurred by the glutamate synthase cycle and that glutamate dehydrogenase played little or no role in the synthesis of glutamate by Azolla-Anabaena.

³ Present address: Department of Agronomy and Range Science, University of California, Davis, CA 95616.

¹ Supported in part by United States Department of Agriculture, Competitive Research Grants Office grants 83-CRCR-1-1295 to J. C. M. and 83-CRCR-1-1296 to G. A. P.

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