Abstract
An aromatic amino acid aminotransferase (aromAT) was purified over 33 000-fold from the shoots and primary leaves of mung beans (Vigna radiata L. Wilczek). The enzyme was purified by ammonium sulfate precipitation, gel filtration and anion exchange followed by fast protein liquid chromatography using Mono Q and Phenylsuperose. The relative amino transferase activities using the most active amino acid substrates were: tryptophan 100, tyrosine 83 and phenylalanine 75, withK m values of 0.095, 0.08 and 0.07 mM, respectively. The enzyme was able to use 2-oxoglutarate, oxaloacetate and pyruvate as oxo acid substrates at relative activities of 100, 128 and 116 andK m values of 0.65, 0.25 and 0.24 mM, respectively. In addition to the aromatic amino acids the enzyme was able to transaminate alanine, arginine, aspartate, leucine and lysine to a lesser extent. The reverse reactions between glutamate and the oxo acids indolepyruvate and hydroxyphenylpyruvate occurred at 30 and 40% of the forward reactions of tryptophan and tyrosine, withK m, values of 0.1 and 0.8 mM, respectively. The enzyme was not inhibited by indoleacetic acid, although α-naphthaleneacetic acid did inhibit slightly. Addition of the cofactor pyridoxal phosphate only slightly increased the activity of the purified enzyme. The aromAT had a molecular weight of 55–59 kDa. The possible role of the aromAT in the biosynthesis of indoleacetic acid is discussed.
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Abbreviations
- AAT:
-
aspartate aminotransferase
- aromAT:
-
aromatic amino acid aminotransferase
- FPLC:
-
fast protein liquid chromatography
- IPyA:
-
indolepyruvate
- OHPhPy:
-
hydroxyphenylpyruvate
- PLP:
-
pyridoxal phosphate
- TAT:
-
tryptophan aminotransferase
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Simpson, R.M., Nonhebel, H.M. & Christie, D.L. Partial purification and characterisation of an aromatic amino acid aminotransferase from mung bean (Vigna radiata L. Wilczek). Planta 201, 71–77 (1997). https://doi.org/10.1007/BF01258682
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DOI: https://doi.org/10.1007/BF01258682