Yellow enzyme purification

Procion Rubine MX-B, Procion Yellow H-A, and Turquoise MX-G. These dyes have found use over the last few decades in the purification of a broad range of proteins and enzymes, including albumin, decarboxylases, glycolytic enzymes, hydrolases, lyases, nucleases, oxidoreductases, synthetases, and transferases [77,78], The first use of dye-ligand affinity chromatography was described by Staal et al. in 1971 [79], Since that time, it has become an extremely popular tool for enzyme and protein purification, with hundreds of such compounds having been isolated by this technique [3-6,76-79],... 

The reaction mixture was filtered to remove the enzyme particles and the solvent evaporated on a rotary evaporator yielding a yellow oil. The oil was dissolved in 100 mL diethyl ether and the low molecular weight phenolic products were extracted into an equal volume of 5% NaOH solution. The residual ether soluble fraction contained primarily Pummerer s ketone and higher molecular weight phenolics while the base-soluble fraction contained the p-cresol and the biscresol. Further purification was performed by preparative thin layer chromatography (TLC) (1 mm silica gel G plates, Analtech, Newark, DE) with a solvent system of diethyl ether heptane (2 1). Rf values were 0.81 for biscresol and 0.63 for Pummerer s ketone. 

Two forms of a-D-mannosidase from yellow wax beans Phaseolus vulgaris) have been purified by immunoadsorption. A high degree of immunological identity has been demonstrated between a-D-mannosidase I and II from P. vulgaris. This made possible the purification of both forms of the enzyme by chromatography on an immunoadsorbent prepared from anti-fa-D-mannosidase I) antiserum. 

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