Abstract
Anthranilate synthase of Agmenellum quadruplicatum, a unicellular species of blue-green bacteria, consists of two nonidentical subunits. A 72,000 dalton protein has aminase activity but is incapable of reaction with glutamine (amido-transferase) unless a second protein (18,000 molecular weight) is present. The small subunit was first detected through its ability to complement a partially purified aminase subunit from Bacillus subtilis to produce a hybrid complex capable of amidotransferase function. Conditions for the function of the heterologous complex were less stringent than for the homologous A. quadruplicatum complex. A reducing agent such as dithiothreitol stabilizes the A. quadruplicatum aminase subunit and is obligatory for amidotransferase function. l-Tryptophan feedback inhibits both the aminase and amidotransferase reactions of anthranilate synthase; Ki values of 6×10−8 m for the amidotransferase activity and 2×10−6 m for the aminase activity were obtained. The Km value calculated for ammonia (2.2 mm) was more favorable than the Km value for glutamine (13 mm). Likewise, the Vmax of anthranilate was greater with ammonia than with glutamine. Starvation of a tryptophan auxotroph results in a threefold derepression of the aminase subunit, but no corresponding increase in the small 18,000 \(\bar M\) subunit occurs. While microbial anthranilate synthase complexes are remarkably similar overall, the relatively good aminase activity of the A. quadruplicatum enzyme may be of physiological significance in nature.
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This research was supported by Grant PCM 7619963 from the National Science Foundation.
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Friedman, E., Jensen, R.A. An anthranilate synthase of the extreme aminase type in a species of blue-green bacteria (algae). Biochem Genet 16, 867–881 (1978). https://doi.org/10.1007/BF00483739
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DOI: https://doi.org/10.1007/BF00483739