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F J Murillo

Publications and source records attributed to F J Murillo.

24 records · Page 2Linked to original sources

Substrate transfer in carotene biosynthesis in Phycomyces.

Lycopene cyclization to gamma-carotene and then to beta-carotene is partially blocked in Phycomyces by the presence of car R mutations in heterokaryosis or by the addition of 2-(4-chlorophenyl)thiotriethylamine . HCl to the medium. We have quantitatively determined the carotenes synthesized by Phycomyces under conditions of partially blocked cyclization and in the presence of either car A mutations in heterokaryosis or polyoxyethylenesorbitan monooleate (Tween-80) in the medium. We conclude that transfer of intermediate substrates between the enzyme aggregates involved in carotenogenesis does occur in the wild type but not in heterokaryons for car A mutations, and is facilitated by Tween-80.

Carotenoids↗

Carotene-superproducing strains of Phycomyces.

Production of beta-carotene by wild-type Phycomyces blakesleeanus can be stimulated by light, chemicals, regulatory mutations, and sexual interaction between mycelia of opposite sex. Through genetic manipulations, we have isolated strains which have simultaneously and constitutively incorporated several of these stimulatory effects. In the dark and in a simple medium, some of the strains produce up to 25 mg of beta-carotene per g (dry weight), or about 500 times the wild-type production under the same conditions. High lycopene-producing strains have also been isolated by using carR mutants, which are blocked in the conversion of lycopene to beta-carotene. These strains should be useful in both industrial production of these pigments and basic research related to carotenogenesis.

Carotenoids↗

Regulation of carotene synthesis in Phycomyces.

Three independent mutations of Phycomyces blakesleeanus resulting in overaccumulation of beta-carotene are recessive and belong to the same complementation group. The corresponding gene has been named carS. Evidence is presented that gene carS is not the same as gene carA, previously defined by mutations blocking carotene production. Vitamin A increases carotenogenesis in wild types and in carS mutants to about the same extent. Intersexual heterokaryosis increases carotenogenesis most prominently in carS genetic backgrounds (up to 300 times the production of the wild type in the same conditions). Vitamin A, intersexual heterokaryosis and carS mutations are thought to stimulate carotenogenesis through different mechanisms. It is suggested that the carS gene product participates in end-product regulation of the pathway.

Carotenoids↗

An enzyme complex for the dehydrogenation of phytoene in Phycomyces.

Phycomyces strain C5, carrying mutation carB10, accumulates phytoene instead of beta-carotene. Heterokaryons containing C5 nuclei and different other nuclei carrying the wild type carB allele accumulate significant amounts of phytofluene, zota-carotene and neurosporene. From quantitative analyses of carotenes and nuclear proportions in the heterokaryons we conclude that four copies of the carB gene product, assembled in an enzyme complex, act sequentially in the conversion of phytoene to lycopene.

Carotenoids↗

A carotenogenic enzyme aggregate in Phycomyces: evidence from quantitive complementation.

Wild-type Phycomyces blakesleeanus accumulates beta-carotene, while the mutant strain C2 is unable to synthesize carotenoids, and the mutant strain C9 accumulates lycopene. Heterokaryons containing a proportion, p, of C2 nuclei and (l - p) of C9 nuclei accumulate lycopene, gamma-carotene, and beta-carotene in the relative amounts (l - p), p(l - p), and p(2), respectively, for different values of p. It is shown that these results are expected from the operation of a carotenogenic enzyme aggregate that works as an assembly line and contains two copies of a cyclase, which is defective in strain C9, as well as other enzymes.

Carotenoids↗