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EC Number Renatured (Commentary) Reference
Show all pathways known for 1.5.1.30Display the word mapDisplay the reaction diagram Show all sequences 1.5.1.30soluble denatured mutant enzyme E99K is mixed into 50fold volume of pre-cooled 50 mM phosphate buffer containing 0.1 M FMN and stirred on ice in the dark for 15 min for protein renaturation and the reconstitution of E99K holoenzyme 684711
Show all pathways known for 1.5.1.30Display the word mapDisplay the reaction diagram Show all sequences 1.5.1.30the recombinant FlaR is insoluble under physiological conditions and its purification can be obtained under denaturing conditions in buffer supplemented with 8 M urea. Different refolding conditions are tested for their ability to renature the protein. Dialysis of the denatured protein against buffers at different pH levels reveals that rFlaR can be solubilized at below pH 8.0. The presence of cysteine-rich sequence suggests that the protein has a transitional metal-binding ability. Thus, rFlaR is dialyzed against PBS (pH 7.3) supplemented with 2 mM salts of either transition metal or calcium and magnesium (2 mM of each). rFlaR becomes fully soluble in the presence of 2 mM FeSO4. Partial solubility can be obtained in the presence of 2 mM CoCl2 or CuSO4 765814
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