Kinetic variations determine the product pattern of phytoene desaturase from Rubrivivax gelatinosus.
In bacteria and fungi, the degree of carotenoid desaturation is determined by a single enzyme, the CrtI-type phytoene desaturase. In different organisms, this enzyme can carry out either three, four or even five desaturation steps. The purple bacterium Rubrivivax gelatinosus is the only known species in which reaction products of a 3-step and a 4-step desaturation (i.e. neurosporene and lycopene derivatives) accumulate simultaneously. The properties of this phytoene desaturation to catalyze neurosporene or lycopene were analyzed by heterologous complementations in Escherichia coli and by in vitro studies. They demonstrated that high enzyme concentrations or low phytoene supply favor the formation of lycopene. Under these conditions, CrtI from Rhodobacter spheroides can be forced in vitro to lycopene formation although this carotene is not synthesized in this species. All results can be explained by a model based on the competition between phytoene and neurosporene for the substrate binding site of phytoene desaturase. Mutations in CrtI from Rvi. gelatinosus have been generated resulting in increased lycopene formation in Escherichia coli. This modification in catalysis is due to increased amounts of CrtI protein.
Stickforth
Per
P
Molecular Biosciences 213, J.W. Goethe Universität, P.O. Box 111932, D-60054 Frankfurt, Germany.
Sandmann
Gerhard
G
D020562:Betaproteobacteria / Q000201:enzymology*; D002338:Carotenoids / Q000096:biosynthesis / Q000378:metabolism; D007700:Kinetics; D000077276:Lycopene; D010088:Oxidoreductases / Q000502:physiology*
D002338:Carotenoids; C008604:neurosporene; D010088:Oxidoreductases; C022398:phytoene dehydrogenase; D000077276:Lycopene