Tryptophan biosynthesis in Saccharomyces cerevisiae: control of the flux through the pathway

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Abstract
Enzyme derepression and feedback inhibition of the 1st enzyme are the regulatory mechanisms demonstrated for the tryptophan pathway in S. cerevisiae. The relative contributions of the 2 mechanisms to the control of the flux through the pathway in vivo were analyzed by measuring feedback inhibition of anthranilate synthase in vivo; determining the effect of regulatory mutations on the level of the tryptophan pool and the flux through the pathway; and, varying the gene dose of individual enzymes of the pathway at the tetraploid level. Apparently the flux through the pathway is adjusted to the rate of protein synthesis by means of feedback inhibition of the 1st enzyme by the end product, tryptophan. The synthesis of the tryptophan enzymes was not repressed below a basal level by tryptophan supplementation of the media. The enzymes are present in excess. Increasing or lowering the concentration of individual enzymes had no noticeable influence on the overall flux to tryptophan. The uninhibited capacity of the pathway was observed both upon relieving feedback inhibition by tryptophan limitation and in feedback-insensitive mutants. It exceeded the rate of consumption of the amino acid on minimal medium by a factor of 3. Tryptophan limitation caused derepression of 4 of the 5 tryptophan enzymes and, as a consequence, led to a further increase in the capacity of the pathway. Because of the large reserve capacity of the repressed pathway, tryptophan limitation could not be imposed on wild-type cells without resorting to the use of analogs. Apparently derepression does not serve as an instrument for the specific regulation of the flux through the tryptophan pathway.