Lysyl oxidase is a copper-dependent enzyme that plays an important role in the cross linkages on elastin and collagen in connective tissues and other biological systems. The crystal structure of LOX has never been observed mainly due to the low solubility of the enzyme and the high levels of urea required to solubilize the enzyme. Crystallography cannot be accomplished if urea is present because the concentration of urea is much greater than that of the enzyme. To increase the solubility of LOX without the use of urea, the enzyme was co-expressed with a NusA solubility tag and purified using an aqueous buffer with the absence of urea. Overexpression of this enzyme with the solubility tag yielded 2.7mg of protein per liter of media. Total copper incorporation was calculated to be 68%. Average specific catalytic activity was calculated to be 0.1065 units/mg of enzyme.

Overexpression and characterization of NusA tagged lysyl oxidase

245th National Meeting (2013)

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