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1
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Violaxanthin and diadinoxanthin de-epoxidation in various model lipid systems

100%
Latowski D., Goss R., Bojko M., Strzałka K.
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2012
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vol. 59
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issue 1
101-103
EN
The xanthophyll cycle is an important photoprotective process functioning in plants. One of its forms, the violaxanthin (Vx) cycle, involves interconversion between: Vx, antheraxanthin (Ax) and zeaxanthin (Zx). Another kind of the xanthophyll cycle is the diadinoxanthin (Ddx) cycle in which interconversion between Ddx and diatoxanthin (Dtx) occurs. In this study an information on molecular mechanism and regulation of these two types of the xanthophyll cycle is presented. The influence of lipids on the de-epoxidation of the xanthophyll cycle pigments was investigated, with special focus put on the significance of physical properties of the aggregates formed by inverted lipid micelles, which are necessary for activity of the xanthophyll cycle enzymes. In particular, thickness of the hydrophobic fraction of the aggregates, size of the inverted micelles, suggested by mathematical description of the structures and solubility of Vx and Ddx in various kind of lipids were studied. Obtained results show that the rate of de-epoxidation is strongly dependent on the physicochemical properties of the lipids used. The key role for enzyme activation play non-bilayer lipids and the parameters of inverted micelles such as thickness, fluidity of hydrophobic core and their diameter. The presented results show that MGDG and other non-lamellar lipids like different forms of phosphatidylethanolamine are necessary for the Vx and Ddx de-epoxidation because they provide the three-dimensional structures, which are needed for the binding of de-epoxidases and for the accessibility of Vx and Ddx to these enzymes.
2
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Expression of three diadinoxanthin de-epoxidase genes of Phaeodacylum tricornutum in Escherichia coli Origami b and BL21 strain

76%
Bojko M., Olchawa-Pajor M., Tuleja U., Kuczyńska P., Strzałka W., Latowski D., Strzałka K.
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2013
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vol. 60
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issue 4
857-860
EN
In the diadinoxanthin cycle the epoxy group is removed from diadinoxanthin and diatoxanthin is created. This conversion takes place e.g. in diatoms with the involvement of the enzyme diadinoxanthin de-epoxidase. In one of the diatom species, Phaeodactylum tricornutum (CCAP 1055/1 strain with genome sequenced) three de-epoxidase genes (PtVDE, PtVDL1, PtVDL2) have been identified, but only one of them (PtVDE) corresponds to violaxanthin de-epoxidase, an enzyme which is commonly found in higher plants. In these studies, the expression of two de-epoxidase genes of another Phaeodactylum tricornutum strain (UTEX 646), which is commonly used in diatom studies, were obtained in Origami b and BL21 E. coli strains. The molecular masses of the mature proteins are about 49 kDa and 60 kDa, respectively, for VDE and VDL2. Both enzymes are active with violaxanthin as a substrate.
3
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Temperature effect on growth, and selected parameters of Phaeodactylum tricornutum in batch cultures

64%
Bojko M., Brzostowska K., Kuczyńska P., Latowski D., Olchawa-Pajor M., Krzeszowiec W., Waloszek A., Strzałka K.
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2013
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vol. 60
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issue 4
861-864
EN
The effect of optimal and stress temperatures on the growth kinetics of the Phaeodactylum tricornutum CCAP/1055/1 strain (a model diatom with a known genome sequence) in batch cultures was examined. The analysis of the obtained results showed two phases of culture growth. There were significant positive correlations between OD increase of chlorophyll a chlorophyll c and protein concentration at different temperatures. The Fv/Fm parameter achieved a maximum level on the 6th or 7th day and then decreased to the values registered on the first day of observation. Genetic material undergoes gradual degradation 10 days after inoculation. The size of the cells was invariable.
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