The Process For The Preparation Of Purified Penicillinase From Crude Extract Using Dye Ligand Affinity Chromatography

Abstract: Purified penicillinase, an important industrial enzyme is used as a marker in ELISA based diagnostic kits. The macroporous beaded crosslinked allyl glycidyl ether copolymers with different crosslinkers such as ethylene glycol dimethacrylate (EGDM), pentaerythritol trimethacrylate (PETMA), divinylbenzene (DVB) and trimethylol propane triacrylate (TMPTA) derivatised with butylamine, benzylamine and phenylethylamine are being explored for the purification of penicillinase. These ligands are either analogs of side chain of penicillin or structures that possess the degree of hydrophobicity in the range equivalent to the hydrophobicity of side chain moiety of penicillin. The primary amino groups present in these molecules can react with the glycidyl functional group on the macroporous beaded crosslinked copolymer to form affinity support. The 2.84 fold purification was achived with benzylamine derivatised with allyl glycidyl etherpentaerythritol trimethacrylate (PETMA) copolymer.

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