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1

Violaxanthin conversion by recombinant diatom and plant de-epoxidases, expressed in Escherichia coli - comparative analysis

100%
Olchawa-Pajor M., Bojko M., Strzalka W., Strzalka K., Latowski D.
Acta Biochimica Polonica
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2019
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tom 66
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nr 3
2

Purification of biologically active violaxanthin de-epoxidase from Escherichia coli

100%
Olchawa-Pajor M., Latowski D., Kuczynska P., Bojko M., Strzalka K.
Acta Biologica Cracoviensia. Series Botanica. Supplement
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2011
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tom 53
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nr 1
3
Artykuł dostępny w postaci pełnego tekstu - kliknij by otworzyć plik
Content available

The effect of temperature on growth of Thalassiosira pseudonana in vitro system

86%
Bojko M., Wanda P., Latowski D., Kuczynska P., Olchawa-Pajor M., Strzalka K.
BioTechnologia. Journal of Biotechnology Computational Biology and Bionanotechnology
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2013
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tom 94
|
nr 3
4

Zeaxanthin epoxidation - an in vitro approach

86%
Kuczynska P., Latowski D., Olchawa-Pajor M., Jahns P., Gruszecki W. I., Strzalka K.
Acta Biochimica Polonica
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2012
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tom 59
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nr 1
Zeaxanthin epoxidase (ZE) is an enzyme operating in the violaxanthin cycle, which is involved in photoprotective mechanisms. In this work model systems to study zeaxanthin (Zx) epoxidation were developed. Two assay systems are presented in which epoxidation of Zx was observed. In these assays two mutants of Arabidopsis thaliana which have active only one of the two xanthophyll cycle enzymes were used. The npq1 mutant possesses an active ZE and is thus able to convert Zx to violaxanthin (Vx) but the violaxanthin de-epoxidase (VDE) is inactive, so that Vx cannot be converted to Zx. The other mutant, npq2, possesses an active VDE and can convert exogenous Vx to Zx under strong light conditions but reverse reaction is not possible. The first assay containing thylakoids from npq1 and npq2 mutants of A. thaliana gave positive results and high efficiency of epoxidation reaction was observed. The amount of Zx was reduced by 25%. To optimize high efficiency of epoxidation reaction additional factors facilitating both fusion of the two types of thylakoids and incorporation of Zx to their membranes were also studied. The second kind of assay contained npq1 mutant thylakoids of A. thaliana supplemented with exogenous Zx and monogalactosyldiacylglycerol (MGDG). Experiments with different proportions of Zx and MGDG showed that their optimal ratio is 1:60. In such system, due to epoxidation, the amount of Zx was reduced by 38% of its initial level. The in vitro systems of Zx epoxidation described in this paper enable analysis some properties of the ZE without necessity of its isolation.
5

Expression of three diadinoxanthin de-epoxidase genes of Phaeodacylum tricornutum in Escherichia coli Origami b and BL21 strain

86%
Bojko M., Olchawa-Pajor M., Tuleja U., Kuczynska P., Strzalka W., Latowski D., Strzalka K.
Acta Biochimica Polonica
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2013
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tom 60
|
nr 4
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.
6
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Determination of epoxides in Escherichia coli expressing recombinant zeaxanthin epoxidase

86%
Kuczynska P., Leskiewicz S., Strzalka W., Olchawa-Pajor M., Bojko M., Latowski D., Strzalka K.
BioTechnologia. Journal of Biotechnology Computational Biology and Bionanotechnology
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2013
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tom 94
|
nr 3
7

Temperature effect on growth, and selected parameters of Phaeodactylum tricornutum in batch cultures

72%
Bojko M., Brzostowska K., Kuczynska P., Latowski D., Olchawa-Pajor M., Krzeszowiec W., Waloszek A., Strzalka K.
Acta Biochimica Polonica
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2013
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tom 60
|
nr 4
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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