Molecular structure and enzymatic function of lycopene cyclase from the cyanobacterium Synechococcus sp strain PCC7942.
A gene encoding the enzyme lycopene cyclase in the cyanobacterium Synechococcus sp strain PCC7942 was mapped by genetic complementation, cloned, and sequenced. This gene, which we have named crtL, was expressed in strains of Escherichia coli that were genetically engineered to accumulate the carotenoid precursors lycopene, neurosporene, and zeta-carotene. The crtL gene product converts the acyclic hydrocarbon lycopene into the bicyclic beta-carotene, an essential component of the photosynthetic apparatus in oxygen-evolving organisms and a source of vitamin A in human and animal nutrition. The enzyme also converts neurosporene to the monocyclic beta-zeacarotene but does not cyclize zeta-carotene, indicating that desaturation of the 7-8 or 7'-8' carbon-carbon bond is required for cyclization. The bleaching herbicide 2-(4-methylphenoxy)triethylamine hydrochloride (MPTA) effectively inhibits both cyclization reactions. A mutation that confers resistance to MPTA in Synechococcus sp PCC7942 was identified as a point mutation in the promoter region of crtL. The deduced amino acid sequence of lycopene cyclase specifies a polypeptide of 411 amino acids with a molecular weight of 46,125 and a pI of 6.0. An amino acid sequence motif indicative of FAD utilization is located at the N terminus of the polypeptide. DNA gel blot hybridization analysis indicated a single copy of crtL in Synechococcus sp PCC7942. Other than the FAD binding motif, the predicted amino acid sequence of the cyanobacterial lycopene cyclase bears little resemblance to the two known lycopene cyclase enzymes from nonphotosynthetic bacteria. Preliminary results from DNA gel blot hybridization experiments suggest that, like two earlier genes in the pathway, the Synechococcus gene encoding lycopene cyclase is homologous to plant and algal genes encoding this enzyme.
Cunningham
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Department of Botany, University of Maryland, College Park 20742.
Sun
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Chamovitz
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Hirschberg
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Gantt
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D000595:Amino Acid Sequence; D001483:Base Sequence; D002338:Carotenoids / Q000096:biosynthesis* / Q000378:metabolism*; D002874:Chromosome Mapping; D000458:Cyanobacteria / Q000201:enzymology / Q000235:genetics*; D004351:Drug Resistance / Q000235:genetics; D004885:Erwinia / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D005021:Ethylamines / Q000494:pharmacology; D005798:Genes, Bacterial / Q000235:genetics*; D005816:Genetic Complementation Test; D019753:Intramolecular Lyases; D007535:Isomerases / Q000235:genetics*; D000077276:Lycopene; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010944:Plants / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D019207:beta Carotene
D005021:Ethylamines; D011994:Recombinant Proteins; D019207:beta Carotene; C082524:2-(4-methylphenoxy)triethylamine; D002338:Carotenoids; D007535:Isomerases; D019753:Intramolecular Lyases; C070925:lycopene cyclase-isomerase; D000077276:Lycopene