Optimisation of immobilisation conditions for chick pea p-galactosidase (CpGAL) to alkylamine glass using response surface methodology and its applications in lactose hvdrolvsis

摘要

Response surface methodology was advantageously used to optimally immobilise a p-galactosidase from chick pea onto alkylamine glass using Box-Behnken experimental design, resulting in an overall 91% immobilisation efficiency. Analysis of variance was performed to determine the adequacy and signifi-cance of the quadratic model. Immobilised enzyme showed a shift in the optimum pH; however, opti-mum temperature remained unaffected. Thermal denaturation kinetics demonstrated significant improvement in thermal stability of the enzyme after immobilisation. Galactose competitively inhibits the enzyme in both soluble and immobilised conditions. Lactose in milk whey was hydrolysed at compar-atively higher rate than that of milk. Immobilised enzyme showed excellent reusability with retention of more than 82% enzymatic activity after 15 uses. The immobilised enzyme was found to be fairly stable in both dry and wet conditions for three months with retention of more than 80% residual activity.