Characterization, Properties and Crystallization of Malate Dehydrogenase from Streptomyces coelicolor

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  • D. Mikulášová Department of Biochemistry, Faculty of Natural Sciences, Comenius University, Bratislava, Slovak Republic
  • M. Koháryová Department of Biochemistry, Faculty of Natural Sciences, Comenius University, Bratislava, Slovak Republic
  • P. Štefanková Department of Biochemistry, Faculty of Natural Sciences, Comenius University, Bratislava, Slovak Republic
  • M. Kollárová Department of Biochemistry, Faculty of Natural Sciences, Comenius University, Bratislava, Slovak Republic

Abstrakt

Malate dehydrogenase (MDH) isolated from Streptomyces coelicolor, A3(2) strain, electrophoretically homogenous, was crystallized in the absence or in the presence of NADH or NADPH coenzymes by the hanging-drop vapor-diffusion method. For simple isolation in a sufficient yield, the above MDH was cloned and expressed in Escherichia coli. Recombinant MDH with His-tag sequence did not crystallize whereas that without His-tag sequence crystallized using the above method. The crystals were grown in a mixture of 0.1 M sodium citrate buffer (pH 5.6), 0.2 M ammonium acetate, 27% (w/v) poly(ethylene glycol) 4000, and 5 mM NADH coenzyme or in a mixture of 0.1 M sodium citrate buffer (pH 4.8), 0.2 M ammonium acetate, 100 μM oxaloacetic acid, and 27% (w/v) poly(ethylene glycol) 4000.

Publikováno

15.10.2010

Jak citovat

Mikulášová, D., Koháryová, M., Štefanková, P., & Kollárová, M. (2010). Characterization, Properties and Crystallization of Malate Dehydrogenase from Streptomyces coelicolor. Chemické Listy, 104(9). Získáno z http://ww.w.chemicke-listy.cz/ojs3/index.php/chemicke-listy/article/view/1259

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