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Sexual Crosses with the Mucoromycete Phycomyces blakesleeanus

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Substrate transfer in carotene biosynthesis in Phycomyces

F J Murillo, S Torres-Martinez, C M Aragon

European Journal of Biochemistry

|October 1, 1981

Summary

Carotenoid cyclization in Phycomyces is blocked by specific mutations or chemicals. This study shows that intermediate transfer occurs in wild-type but not mutated strains, and is aided by Tween-80.

Area of Science:

Biochemistry
Molecular Biology
Mycology

Background:

Carotenoids are vital pigments synthesized through complex pathways.
Lycopene cyclization is a key step, producing gamma- and beta-carotene.
Phycomyces is a model organism for studying carotenogenesis.

Purpose of the Study:

To investigate the effects of blocked lycopene cyclization on carotenoid synthesis in Phycomyces.
To determine if intermediate substrate transfer occurs between enzyme aggregates during carotenogenesis.
To examine the role of car A mutations and Tween-80 in this process.

Main Methods:

Quantitative analysis of carotenes synthesized by Phycomyces.
Utilizing car R and car A mutations in heterokaryosis to block cyclization.

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Administering 2-(4-chlorophenyl)thiotriethylamine . HCl and polyoxyethylenesorbitan monooleate (Tween-80) to the medium.

Main Results:

Partial blockage of lycopene cyclization was achieved using car R mutations or a specific chemical.
Carotenoid synthesis was analyzed under conditions of blocked cyclization with car A mutations or Tween-80.
Evidence suggests intermediate substrate transfer occurs in wild-type Phycomyces but is absent in car A heterokaryons.

Conclusions:

Enzyme aggregate-mediated substrate transfer is a feature of wild-type carotenogenesis in Phycomyces.
This transfer mechanism is disrupted in car A mutated heterokaryons.
Tween-80 facilitates intermediate substrate transfer, suggesting a role in enzyme complex organization or accessibility.