title: Regulation of phenylalanine biosynthesis in Rhodotorula glutinis.
abstract: The phenylalanine biosynthetic pathway in the yeast Rhodotorula glutinis was examined, and the following results were obtained. (i) 3-Deoxy-D-arabinoheptulosonate-7-phosphate (DAHP) synthase in crude extracts was partially inhibited by tyrosine, tryptophan, or phenylalanine. In the presence of all three aromatic amino acids an additive pattern of enzyme inhibition was observed, suggesting the existence of three differentially regulated species of DAHP synthase. Two distinctly regulated isozymes inhibited by tyrosine or tryptophan and designated DAHP synthase-Tyr and DAHP synthase-Trp, respectively, were resolved by DEAE-Sephacel chromatography, along with a third labile activity inhibited by phenylalanine tentatively identified as DAHP synthase-Phe. The tyrosine and tryptophan isozymes were relatively stable and were inhibited 80 and 90% by 50 microM of the respective amino acids. DAHP synthase-Phe, however, proved to be an extremely labile activity, thereby preventing any detailed regulatory studies on the partially purified enzyme. (ii) Two species of chorismate mutase, designated CMI and CMII, were resolved in the same chromatographic step. The activity of CMI was inhibited by tyrosine and stimulated by tryptophan, whereas CMII appeared to be unregulated. (iii) Single species of prephenate dehydratase and phenylpyruvate aminotransferase were observed. Interestingly, the branch-point enzyme prephenate dehydratase was not inhibited by phenylalanine or affected by tyrosine, tryptophan, or both. (iv) The only site for control of phenylalanine biosynthesis appeared to be DAHP synthase-Phe. This is apparently sufficient since a spontaneous mutant, designated FP9, resistant to the growth-inhibitory phenylalanine analog p-fluorophenylalanine contained a feedback-resistant DAHP synthase-Phe and cross-fed a phenylalanine auxotroph of Bacillus subtilis.
mesh_terms: D010723:3-Deoxy-7-Phosphoheptulonate Synthase / Q000302:isolation & purification / Q000378:metabolism*; D000446:Aldehyde-Lyases / Q000378:metabolism*; D002826:Chorismate Mutase / Q000302:isolation & purification / Q000378:metabolism*; D006836:Hydro-Lyases / Q000378:metabolism*; D007535:Isomerases / Q000378:metabolism*; D007700:Kinetics; D003904:Mitosporic Fungi / Q000378:metabolism*; D010649:Phenylalanine / Q000096:biosynthesis*; D011302:Prephenate Dehydratase / Q000302:isolation & purification / Q000378:metabolism*; D012248:Rhodotorula / Q000378:metabolism*
chemical_list: D010649:Phenylalanine; D010723:3-Deoxy-7-Phosphoheptulonate Synthase; D000446:Aldehyde-Lyases; D006836:Hydro-Lyases; D011302:Prephenate Dehydratase; D007535:Isomerases; D002826:Chorismate Mutase