Involvement of L-tryptophan aminotransferase in indole-3-acetic acid biosynthesis in Enterobacter cloacae.
L-Tryptophan aminotransferase (L-tryptophan:2-oxoglutarate aminotransferase; EC 2.6.1.27) from Enterobacter cloacae was purified 62-fold and characterized to determine its role in indole-3-acetic acid biosynthesis. The enzyme reversibly catalyzed the transamination of L-tryptophan with 2-oxoglutarate as the amino acceptor to yield indole-3-pyruvic acid and L-glutamate, and the Km values for L-tryptophan and indole-3-pyruvic acid were 3.3 mM and 24 microM, respectively. In the indole-3-acetaldehyde synthesis experiments in vitro, 94% of L-tryptophan was efficiently converted to indole-3-acetaldehyde by the purified L-tryptophan aminotransferase plus indolepyruvate decarboxylase. Furthermore, the amounts of L-tryptophan decreased with increases in the indolepyruvate decarboxylase activity, while the amounts of indole-3-acetaldehyde increased with increases in this activity. In genetic experiments, the amounts of L-tryptophan produced by Enterobacter and Pseudomonas strains harboring the gene for indolepyruvate decarboxylase were lower than those produced by these same strains without the gene, while the amounts of indole-3-acetic acid produced by Enterobacter and Pseudomonas strains harboring the gene for indolepyruvate decarboxylase were higher than those produced by these same strains without the gene. These results clearly show that L-tryptophan aminotransferase is involved in the indole-3-acetic acid biosynthesis and that indolepyruvate decarboxylase is the rate-limiting step in this pathway.
Koga
J
J
Bio Science Laboratories, Meiji Seika Kaisha, Ltd., Saitama, Japan.
Syono
K
K
Ichikawa
T
T
Adachi
T
T
D002262:Carboxy-Lyases / Q000235:genetics; D016972:Enterobacter cloacae / Q000378:metabolism*; D005798:Genes, Bacterial; D006863:Hydrogen-Ion Concentration; D007210:Indoleacetic Acids / Q000378:metabolism*; D000637:Transaminases / Q000302:isolation & purification / Q000378:metabolism*; D014364:Tryptophan / Q000378:metabolism; D051279:Tryptophan Transaminase
D007210:Indoleacetic Acids; C030737:indoleacetic acid; D014364:Tryptophan; D000637:Transaminases; D051279:Tryptophan Transaminase; C068832:indolepyruvate decarboxylase; D002262:Carboxy-Lyases