Preparation of DGEBA support without spacer arm in theimmobilization of lipase B by covalent attachment

The aim of this work was the use of DGEBA coated on tin plate as a support to immobilize microbiallipase commercially available as lipase B candida Antarctica (EC3.1.1.3) recombinant fromaspergillus oryzae. The benefit of this method was, direct covalent attachment of lipase B on DGEBAsupport without incorporation of any spacer arms between support and enzyme. Hence in this work,the process for activation of support was omitted. The properties of immobilized and free biocatalystwere assayed and compared with in the mutton tallow oil hydrolysis. The tallow oil used in thismethod was extracted traditionally in 80 °C. Hydrolysis performed under mild condition oftemperature 37°C and pH 7.0. The results indicated that 54 h was enough for Cal-B to be immobilizedon the DGEBA support. The covalent attachment of Cal-B on DGEBA which was coated on tin plateturned biocatalyst more active. Based on the result a time of 8 h was selected for hydrolysis. The tinplate dimension was 50 cm2 loaded by 52 mg DGEBA, the maximum protein immobilized on theprepared DGEBA tin plate was found to be 15.99 mg offering 0.308 mg protein per one mg of support.The biocatalyst retained its activity, in aqueous medium, for more than two months with threesuccessive hydrolsis reactions

کلیدواژه ها:

Covalent immobilization ، DGEBA ، Lipase B ، Spacer arm ، Tin plate ، Tallow oil