A New Method for the Preparation of Jack-bean Urease Involving Covalent Chromatography.

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Abstract
A simple 3-step method for the preparation of highly active urease (EC 3.5.1.5) from jack-bean meal is described which involves an ethanol extraction with subsequent covalent chromatography followed by a gel filtration step. The covalent chromatography procedure requires consecutive thiol-disulfide interchange reactions. Active urease, containing reactive thiol groups, is covalently bonded to the chromatographic material (agarose-2-pyridyl disulfide) via disulfide bridges and subsequently removed by reduction with dithiothreitol.

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