A new non-radioactive assay of phytoene desaturase to evaluate bleaching herbicides.
A non-radioactive cell-free assay was developed to quantitatively determine inhibition of plant-type phytoene desaturase by bleaching herbicides. An active desaturase was prepared from an appropriately cloned E. coli transformant. Another E. coli transformant was used to produce the required phytoene. Phytofluene and zeta-carotene, the products of the desaturase reaction, were either determined by HPLC or optical absorption spectra. Enzyme kinetics and inhibition data for the bleaching tetrazole herbicide WL110547 are presented as an example.
Sandmann
G
G
Department of Botany, J.-W.-Goethe University, Frankfurt, Bundesrepublik Deutschland.
Schneider
C
C
Böger
P
P
D002338:Carotenoids / Q000032:analysis; D002474:Cell-Free System; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D000458:Cyanobacteria / Q000201:enzymology; D004926:Escherichia coli; D006540:Herbicides / Q000032:analysis* / Q000494:pharmacology*; D007700:Kinetics; D010088:Oxidoreductases / Q000032:analysis* / Q000037:antagonists & inhibitors*; D011994:Recombinant Proteins / Q000032:analysis / Q000037:antagonists & inhibitors; D013053:Spectrophotometry / Q000379:methods; D013777:Tetrazoles / Q000032:analysis
D006540:Herbicides; D011994:Recombinant Proteins; D013777:Tetrazoles; C102554:WL 110547; D002338:Carotenoids; C003140:phytofluene; D010088:Oxidoreductases; C022398:phytoene dehydrogenase