Screening of marine cyanobacteria for high palmitoleic acid production

Abstract Screening of fatty acid composition in 150 strains of marine microalgae, cyanobacteria and green algae was carried out, and 20 strains showed relatively high contents of palmitoleic acid. Among them, two cyanobacteria, Phormidium sp. NKBG 041105 and Oscillatoria sp. NKBG 091600, showed an unusually high cis -palmitoleic acid content (54.5% and 54.4% of total fatty acid, respectively). Phormidium sp. NKBG 041105 had the highest cis -palmitoleic acid content per biomass (46.3 mg (g dry cell weight)⁻¹), and cis -palrnitoleic acid composition was found to be constant with varying temperature. These results indicate that this cyanobacterium could be considered as a new source for palmitoleic acid.     

Antioxidant activity of vegetable oils with different omega-6/omega-3 fatty acids ratio

The antioxidant activity and the oxidative stability of flax, sesame, silybum oils and seed blend oils with different ω-6/ω-3 fatty acid ratios have been investigated. The antioxidant content (AO) in crude oils and their reactivity towards peroxyl radicals were studied using the kinetic method based on oil addition to the model reaction of cumene oxidation. There were correlations between the ratio polyunsaturated fatty acids (PUFA)/ω-9 and thermal stability (50°C); between the effect of γ-tocopherol content and oil resistance to oxidative changes during long-term storage at (10 ± 2)°C.     

NITROGEN LIMITATION EFFECTS OF DIFFERENT NITROGEN SOURCES ON NUTRITIONAL QUALITY OF TWO FRESHWATER ORGANISMS, SCENEDESMUS QUADRICAUDA (CHLOROPHYCEAE) AND SYNECHOCOCCUS SP. (CYANOPHYCEAE)

Food quality for grazers has been related to mineral (nitrogen, phosphorus) and biochemical (amino acids, fatty acids) constituents. The aim of the study was to examine the influence of different nitrogen sources on these constituents in two organisms, the green alga Scenedesmus quadricauda Turp. and the cyanobacterium Synechococcus sp., commonly used in feeding experiments. The two organisms were grown in continuous cultures at different growth rates. Nitrate or ammonium salts were used as nitrogen sources under both replete and limited conditions. Carbon content (mg·g⁻¹ dry weight) was stable in both organisms independent of nitrogen source, nitrogen limitation, and growth rate. Nitrogen content decreased with limitation and growth rate in Scenedesmus and to a lesser degree in Synechococcus , whereas changes in phosphorus content were not statistically significant. The relative proportions of amino acids (% of total amino acids) were relatively stable in both organisms, whereas the proportions of fatty acids varied with growth rate and limitation. Fatty acid content was much lower in Synechococcus than in Scenedesmus . At N limitation, polyunsaturated fatty acids (PUFAs) showed lower levels in both organisms. The change occurred in the ω3 PUFA (linolenic acid) of the green alga and in the ω6 PUFA (linoleic acid) of the cyanobacterium. The difference in the response of N limitation in the two organisms may be traced to the different composition of the chloroplast membranes (the prokaryotic way) and the microsomal membranes (the eukaryotic way) where the desaturation takes place.     

Thermoadaptation and fatty acid composition of main phospholipids of the small-scaled redfin <Emphasis Type="Italic">Tribolodon brandti</Emphasis> under natural and experimental conditions

The fatty acid (FA) composition of the main membrane phospholipids, phosphatidylcholine (PC) and phosphatidylethanolamine (PE), was investigated in the muscle, gills, and liver of the small-scaled redfin Tribolodon brandti (Dybowskii, 1872) at different temperatures under natural and experimental conditions. It was established that a water temperature decrease in the natural habitat was accompanied by an increase in polyunsaturated fatty acid contents and the unsaturation index, mainly at the expense of FAs of the ω3 series (20:5ω3 and 22:6ω3), and by a decrease in saturated fatty acid levels in PC and PE. A similar, but less pronounced tendency was revealed in experiments with a rapid lowering of water temperature. These findings suggest the weak adaptation ability of the small-scaled redfin to a drastic shift of environmental temperature. Temperature changes produced the greatest alterations in the FA composition of phospholipids in the liver and the smallest changes occurred in muscle tissue.     

The synthesis and accumulation of stearidonic acid in transgenic plants: a novel source of 'heart-healthy' omega-3 fatty acids

Dietary omega-3 polyunsaturated fatty acids have a proven role in reducing the risk of cardiovascular disease and precursor disease states such as metabolic syndrome. Although most studies have focussed on the predominant omega-3 fatty acids found in fish oils (eicosapentaenoic acid and docosahexaenoic acid), recent evidence suggests similar health benefits from their common precursor, stearidonic acid. Stearidonic acid is a Δ6-unsaturated C18 omega-3 fatty acid present in a few plant species (mainly the Boraginaceae and Primulaceae ) reflecting the general absence of Δ6-desaturation from higher plants. Using a Δ6-desaturase from Primula vialii , we generated transgenic Arabidopsis and linseed lines accumulating stearidonic acid in their seed lipids. Significantly, the P. vialii Δ6-desaturase specifically only utilises α-linolenic acid as a substrate, resulting in the accumulation of stearidonic acid but not omega-6 γ-linolenic acid. Detailed lipid analysis revealed the accumulation of stearidonic acid in neutral lipids such as triacylglycerol but an absence from the acyl-CoA pool. In the case of linseed, the achieved levels of stearidonic acid (13.4% of triacylglycerols) are very similar to those found in the sole natural commercial plant source ( Echium spp.) or transgenic soybean oil. However, both those latter oils contain γ-linolenic acid, which is not normally present in fish oils and considered undesirable for heart-healthy applications. By contrast, the stearidonic acid-enriched linseed oil is essentially devoid of this fatty acid. Moreover, the overall omega-3/omega-6 ratio for this modified linseed oil is also significantly higher. Thus, this nutritionally enhanced linseed oil may have superior health-beneficial properties.     

Changes in the nutritional quality of decaying leaf litter in a stream based on fatty acid content

We examined the nutritional quality of decaying leaf litter in a third-order forested stream, using measurements of fatty acid (FA) composition over time. We measured changes in concentrations of total, polyunsaturated, microalgal, and microbial marker FAs in mixed-species leaf packs in spring and autumn and effects of including/excluding macroinvertebrates. Initial concentrations of total FAs in litter were significantly less in spring (5.2 mg/g) than in autumn (6.9 mg/g; F = 6.3; P = 0.03), but total FA concentrations in litter placed in the stream declined significantly over 120 days in both spring (62%; F = 10.9; P < 0.001) and autumn (56%; F = 19.4; P = 0.0001). Quantities of most FAs declined at a greater rate than that of bulk leaf matter. The presence or absence of macroinvertebrates (5 mm vs. 250 μm mesh) had no effect on FA concentration or composition of decomposing litter. Omega-3 polyunsaturated FAs were either nearly absent (20:5ω3) or depleted preferentially over other FAs (18:3ω3). During decomposition the polyunsaturated FA linoleic acid (18:2ω6, common in fungi), declined in concentration more rapidly than other FAs in the spring, but in autumn declined at slower rates, perhaps suggesting greater fungal activity in autumn. Quantities of bacterial (e.g., 16:1ω7) and fungal (e.g., 18:1ω9) FA markers increased over time in autumn (and 16:1ω7 also in spring). Our data provide no evidence for increasing nutritional FA quality of litter during decay and microbial colonization, based on total and polyunsaturated FAs, despite measured increases in bacterial and fungal FA over time. Routine measurements of FA composition of litter could provide insights into the nutrition of allochthonous matter and the importance of fungi and bacteria during decomposition.     

Comparative anti-inflammatory effects of plant- and marine-derived omega-3 fatty acids explored in an endothelial cell line

Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) lower risk of cardiovascular disease. The primary source of EPA and DHA is fatty fish. Plant-derived alpha linolenic acid (ALA) and stearidonic acid (SDA) could provide sustainable land-based alternatives, but their functionality is underexplored. Omega-3 fatty acids (n-3 FAs) may influence atherogenic processes through changing endothelial cell (EC) function and lowering inflammation. This study compared effects of marine- and plant-derived n-3 FAs on EC inflammatory responses. EA.hy926 cells were exposed to ALA, SDA, EPA or DHA prior to stimulation with tumor necrosis factor (TNF)-α. All FAs were shown to be incorporated into ECs in a dose-dependent manner. SDA (50 μM) decreased both production and cell-surface expression of intercellular adhesion molecule (ICAM)-1; however EPA and DHA resulted in greater reduction of ICAM-1 production and expression. EPA and DHA also significantly lowered production of monocyte chemoattractant protein 1, interleukin (IL)-6 and IL-8. ALA, SDA and DHA (50 μM) all reduced adhesion of THP-1 monocytes to EA.hy926 cells. DHA significantly decreased nuclear factor kappa-light-chain-enhancer of activated B cells (NFκB)p105 gene expression and phosphorylated NFκBp65 protein. Both EPA and DHA (50 μM) significantly decreased cyclooxygenase (COX)-2 protein. Thus, both marine-derived n-3 FAs, particularly DHA, had potent anti-inflammatory effects in this EC model. Of the plant-derived n-3 FAs, SDA showed the greatest inhibition of inflammation. Although neither ALA nor SDA reproduced the anti-inflammatory effects of EPA and DHA in this model, there is some potential for SDA to be a sustainable anti-inflammatory alternative to the marine n-3 FAs.     

Genetically engineering cyanobacteria to convert CO2, water,and light into the long-chain hydrocarbon farnesene

Genetically engineered cyanobacteria offer a shortcut to convert CO₂ and H₂O directly into biofuels and high value chemicals for societal benefits. Farnesene, a long-chained hydrocarbon (C₁₅H₂₄), has many applications in lubricants, cosmetics, fragrances, and biofuels. However, a method for the sustainable, photosynthetic production of farnesene has been lacking. Here, we report the photosynthetic production of farnesene by the filamentous cyanobacterium Anabaena sp. PCC 7120 using only CO₂, mineralized water, and light. A codon-optimized farnesene synthase gene was chemically synthesized and then expressed in the cyanobacterium, enabling it to synthesize farnesene through its endogenous non-mevalonate (MEP) pathway. Farnesene excreted from the engineered cyanobacterium volatilized into the flask head space and was recovered by adsorption in a resin column. The maximum photosynthetic productivity of farnesene was 69.1?±?1.8 μg·L^?1·O.D.^?1·d^?1. Compared to the wild type, the farnesene-producing cyanobacterium also exhibited a 60 % higher PSII activity under high light, suggesting increased farnesene productivity in such conditions. We envision genetically engineered cyanobacteria as a bio-solar factory for photosynthetic production of a wide range of biofuels and commodity chemicals.     

Genetic and biochemical evidence for involvement of HOTHEAD in the biosynthesis of long-chain α-,ω-dicarboxylic fatty acids and formation of extracellular matrix

In plants, extracellular matrix polymers built from polysaccharides and cuticular lipids have structural and protective functions. The cuticle is found to be ten times thinner in Arabidopsis thaliana (L.) Heynh than in many other plants, and there is evidence that it is unusual in having a high content of α-,ω-dicarboxylic fatty acids (FAs) in its polyesters. We designated the new organ fusion mutant hth-12 after it appeared to be allelic to adhesion of calyx edges (ace) and hothead (hth), upon molecular cloning of the gene by transposon tagging. This mutant is deficient in its ability to oxidize long-chain ω-hydroxy FAs to ω-oxo FAs, which results in leaf polyesters in decreased α-,ω-dicarboxylic FAs and increased ω-hydroxy FAs. These chemical phenotypes lead to disorder of the cuticle membrane structure in hth-12. ACE/HTH is a single-domain protein showing sequence similarity to long-chain FA ω-alcohol dehydrogenases from Candida species, and we hypothesize that it may catalyze the next step after cytochrome P450 FA ω-hydroxylases in the ω-oxidation pathway. We show that ACE/HTH is specifically expressed in epidermal cells. It appears very likely therefore that the changes in the amount of α-,ω-dicarboxylic FAs in hth-12 reflect the different composition of cuticular polyesters. The ACE/HTH gene is also expressed in root epidermal cells which do not form a polyester membrane on the exterior surface, thereby making it possible that the end products of the pathway, α-,ω-dicarboxylic FAs, are generally required for the cross-linking that ensures the integrity of the outer epidermal cell wall.     

Carnitine palmitoyl transferase activity and whole muscle oxidation rates vary with fatty acid substrate in avian flight muscles

Birds primarily fuel migratory flights with fat, and the composition of that fat has the potential to affect overall lipid oxidation rates. We measured the whole muscle lipid oxidation rates in extensor digitorum communis muscles from white-throated sparrows (Zonotrichia albicollis Gmelin) incubated for 20 min at 20°C with radiolabeled stearate (18:0), oleate (18:1ω9), or linoleate (18:2ω6). Lipid oxidation rates were ~40% higher with linoleate than oleate (oleate: 36 ± 8.54 μmol CO₂ g⁻¹ h⁻¹), and ~75% lower with stearate compared with oleate, indicating that maximal lipid oxidation rates can indeed be affected by the type of fatty acid supplied to the muscle. Additionally, we investigated the activity of the mitochondrial fatty acid transport-associated enzyme carnitine palmitoyl transferase (CPT) in pectoralis muscles of 5 bird species (Zonotrichia albicollis, Philomachus pugnax, Sturnus vulgaris, Taeniopygia guttata, Passer domesticus). Activity was measured in homogenized samples using various fatty acyl-CoA substrates (16:0, 16:1, 18:0, 18:1ω9, 18:2ω6, 18:3ω3, 18:3ω6, 20:0, 20:4ω6, 22:6ω3) in a spectrophotometric assay. CPT activity increased with the degree of unsaturation and decreased with chain length. CPT activity did not differ between ω3 and ω6 isomers of 18:3, nor was the pattern of CPT substrate preference different between captive white-throated sparrows in a migratory (i.e., displaying Zugunruhe) or non-migratory state. These findings can explain previously observed differences in peak performance induced by dietary fat composition and suggest that lipid supply is limiting to maximal exercise performance in birds.     

Conjugative gene transfer in marine cyanobacteria: Synechococcus sp., Synechocystis sp. and Pseudanabaena sp.

Summary Versatility of gene transfer by transconjugation in marine cyanobacteria was demonstrated. In this study, seven different marine cyanobacteria were used as recipient cells. First, transconjugation was carried out using the mobilizable transposon (Tn5) carrying plasmid pSUP1021. Transconjugants were observed in all marine cyanobacteria tested. Second, the broad-host-range vector pKT230 (IncQ) was tested for transconjugation. pKT230 has been successfully transferred in a marine cyanobacterium Synechococcus sp. NKBG15041C, and replicated as an autonomous replicon without alteration in the restriction enzyme pattern. A maximum transfer efficiency of 5.2 × 10^–4 transconjugants/recipient cell was observed, when mating was performed on agar plates containing low salinity (0.015 m NaCl) medium. This is the first study to demonstrate gene transfer in marine cyanobacteria via transconjugation. Correspondence to: K. Sode     

Lipids of the meristems of the main coniferous edificators from central siberia under low-temperature adaptation: 1. The characteristics of the fatty acid composition of phospholipids from winter meristems of Larix sibirica Ledeb., Picea obovata L., and Pinus sylvestris L.

Trienoic fatty acids (FAs) have been found to be the dominant group of unsaturated fatty acids in the composition of phospholipids (PLs) from Larix sibirica Ledeb., whereas dienoic FAs have been established to prevail in PLs from Picea obovata L. and Pinus sylvestris L. Bud swelling in spring is accompanied by a decrease in the content of unsaturated FAs by approximately 30%. The analysis of the change in the content of individual C₁₈ FAs in the PL structure upon the transition of the tree from winter dormancy to vegetation has revealed the important functional role of acyl-lipid ω₆ desaturase, which is responsible for the synthesis of linoleic acid (C_{18: 2}), in the formation of the cryoprotected state of meristem cell membranes of Larix sibirica Ledeb., Picea obovata L., and Pinus sylvestris L. In addition, the particular significance of acyl-lipid ω₃ desaturase for the cryoadaptation of meristem cells in Larix sibirica Ledeb., as it catalyzes the transformation of linoleic acid (C_{18: 2}) to linolenic acid (C_{18: 3}) in PLs of larch 1.5–3 times more intensively than in the case of spruce and pine, is shown.     

Seasonal distribution and fatty acid composition of littoral microalgae in the Yenisei River

We studied fatty acid (FA) composition of littoral microalgae in the fast-flowing oligotrophic river, the Yenisei, Siberia, monthly for 3 years. Seasonal dynamics of species composition had similar patterns in all the studied years. In springs, a pronounced dominance of filamentous green algae occurred, in summer and autumn diatoms were abundant, and in late autumn and winter epilithic biofilms consisted primarily of cyanobacteria and detritus. In general, FA composition of the algal periphytic community was dominated by 16:0, 16:1ω7, 20:5ω3, 14:0, and 18:3ω3 throughout the studied period. Several groups of FAs, which had peculiar seasonal dynamics, were differentiated by statistical analysis based on a method of correlation graphs. The seasonal changes in FA composition could be partly explained by the seasonal succession of species composition of the community. Besides, we found that populations of both diatom and green algae grown in summer at a higher water temperature were lower in polyunsaturated fatty acids than those in spring, at a lower temperature. Hence, we suppose that the regular seasonal dynamics of FA composition of the studied littoral microalgae was driven both by changes in species composition and by temperature adaptations of the algal populations. The highest content of essential polyunsaturated FAs, eicosapentaenoic and docosahexaenoic acids, in the spring “psychrophilic” populations of diatoms could make them of the higher nutritive value for zoobenthic primary consumers.     

The characteristics of desaturation of Trichoderma sp. AM076 and formation of 9,12,15-hexadecatrienoic acid (16 : 3ω1) through Δ15 desaturation of 9,12-hexadecadienoic acid

We investigated the characteristics of desaturation in Trichoderma sp. AM076. Although 6,9,12-octadecatrienoic acid (18 : 3ω6) was detected when Trichoderma sp. AM076 was cultivated in the presence of 6,9-octadecadienoic acid (18 : 2ω9), the desaturation products of 6,9,12-octadecatrienoic acid (18 : 3ω6) and 6-octadecenoic acid (18 : 1Δ6) were not detected. These results suggest that the double bonds at the Δ6 position of 18 : 3ω6 and 18 : 1Δ6 disturb their Δ15 and Δ9 desaturation, respectively. This fungus also introduced a double bond at the Δ15 position of 9,12-hexadecadienoic acid (16 : 2ω4), thereby yielding a novel C16 polyunsaturated fatty acid (PUFA) identified as 9,12,15-hexadecatrienoic acid (16 : 3ω1). Further investigations revealed that the mutant having enhanced accumulation of linolenic aid (18 : 3ω3) accumulates 16 : 3ω1 as one of the major PUFAs, together with 9,12-octadecadienoic acid (16 : 2ω4), when grown with palmitoleic acid (16 : 1ω7). These results suggest that, in this strain, the reaction that catalyzes the conversion of linoleic acid to linolenic acid, similar to the conversion of 16 : 2ω4 to 16 : 3ω1, is not ω3 desaturation but Δ15 desaturation.     

Seasonal dynamics of long-chain polyunsaturated fatty acids in littoral benthos in the upper Yenisei river

We studied composition and concentrations of fatty acids (FAs) in benthos from pebbly littoral region of the Yenisei River in a sampling site near Krasnoyarsk city (Siberia, Russia) for 1 year from March 2003 to February 2004. Special attention was paid to major long-chain polyunsaturated fatty acids (PUFAs) of the ω3 family: eicosapentaenoic acid (EPA, 20:5ω3) and docosahexaenoic acids (DHA, 22:6ω3). In phytobenthos, which was dominated by diatoms, the annual maxima of EPA and DHA pool occurred in spring and early summer. In zoobenthos, EPA and DHA pool peaked in autumn, due mainly to an increase of the biomass of dominant taxa (gammarids) and to a moderate increase of the PUFA content per body weight. Seasonal peaks of EPA in overwintering insect larvae (chironomids and caddisflies) generally coincided with those of biomass of these larvae, while there was no such trend for amphipods and oligochaetes. In spring and early summer, the main part of ω3 PUFA, 40–97% of total amount, in the littoral region was contained in biomass of producers, i.e., benthic microalgae, and in autumn it was transferred to primary consumers—benthic invertebrates, which contained ∼76–93% of total ω3 PUFAs.     

Functional complementation of a periila ω3 fatty acid desaturase under the seed-specific SeFAD2 promoter

Functional characterization of the fatty acid desaturase genes and seed-specific promoters is prerequisite for altering the unsaturated fatty acid content of oilseeds by genetic manipulation. The ω-6 fatty acid desaturase (FAD2) and ω-3 fatty acid desaturase (FAD3) catalyze extra-plastidial desaturation of oleic acid to linoleic acid and linoleic acid to linolenic acid, respectively. These are major constituents in seed storage oils. Here, we report the complementation of a perilla linoleic acid desaturase (PrFAD3) cDNA under the seed-specific sesame FAD2 (SeFAD2) promoter in the Arabidopsis fad3 mutant. PrFAD3 is functionally active and the SeFAD2 promoter is applicable for modifying fatty acid composition in developing seeds. Transient expression of the GUS gene under that promoter in the developing seeds and leaves of sesame, soybean, and corn via microprojectile bombardment indicated that the SeFAD2 promoter likely will be useful for altering the seed phenotypes of dicot and monocot crops.     

The Effect of Tobacco Plant Transformation with a Gene for Acyl-Lipid Δ9-Desaturase from <Emphasis Type="Italic">Synechococcus vulcanus</Emphasis> on Plant Chilling Tolerance

Tobacco plants with the introduced desC gene for acyl-lipid Δ9-desaturase from the thermophilic cyanobacterium Synechococcus vulcanus were cultivated on agar-solidified Murashige and Skoog nutrient medium supplemented with ferulic acid and antibiotics at 22°C and a 16-h photoperiod. Control plants were transformed with an empty pGA482 vector. The analysis of fatty acids (FAs) showed that, in transgenic plants, the level of 16:0 and 18:0 FAs decreased substantially, whereas the levels of di- and trienoic FAs increased. Transformed plants were more cold-tolerant. The tolerance to chilling was evaluated from electrolyte leakage from tissues damaged by cold treatments and from the accumulation of a product of lipid peroxidation, malondialdehyde. It was concluded that acyl-lipid Δ9-desaturase was actively expressed in transgenic tobacco plants and converted stearic acid into oleic acid, thus producing a substrate for further synthesis of di- and trienoic FAs. An increased proportion of polyunsaturated FAs in membrane lipids resulted in improved tobacco plant tolerance to chilling.     

Co-expression of the borage Δ6 desaturase and the Arabidopsis Δ15 desaturase results in high accumulation of stearidonic acid in the seeds of transgenic soybean

Two relatively rare fatty acids, γ-linolenic acid (GLA) and stearidonic acid (STA), have attracted much interest due to their nutraceutical and pharmaceutical potential. STA, in particular, has been considered a valuable alternative source for omega-3 fatty acids due to its enhanced conversion efficiency in animals to eicosapentaenoic acid when compared with the more widely consumed omega-3 fatty acid, α-linolenic acid (ALA), present in most vegetable oils. Exploiting the wealth of information currently available on in planta oil biosynthesis and coupling this information with the tool of genetic engineering it is now feasible to deliberately perturb fatty acid pools to generate unique oils in commodity crops. In an attempt to maximize the STA content of soybean oil, a borage Δ⁶ desaturase and an Arabidopsis Δ¹⁵ desaturase were pyramided by either sexual crossing of transgenic events, re-transformation of a Δ⁶ desaturase event with the Δ¹⁵ desaturase or co-transformation of both desaturases. Expression of both desaturases in this study was under the control of the seed-specific soybean β-conglycinin promoter. Soybean events that carried only the Δ¹⁵ desaturase possessed a significant elevation of ALA content, while events with both desaturases displayed a relative STA abundance greater than 29%, creating a soybean with omega-3 fatty acids representing over 60% of the fatty acid profile. Analyses of the membrane lipids in a subset of the transgenic events suggest that soybean seeds compensate for enhanced production of polyunsaturated fatty acids by increasing the relative content of palmitic acid in phosphatidylcholine and other phospholipids.     

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.