Application of tris(hydroxymethyl)phosphine as a coupling agent for β‐galactosidase immobilized on chitosan to produce galactooligosaccharides

Abstract β‐Galactosidase was immobilized on chitosan using tris(hydroxymethyl)phosphine (THP) as a coupling agent to produce galactooligosaccharides (GOS) from lactose. Both the THP‐immobilized and the free enzymes were maximally achieved at pH 5.0 and the optimal temperature was 55 °C. The residual activities for the THP‐immobilized enzyme and the free enzyme were 75 and 25%, respectively, after being incubated in 0.1 mol dm−3 sodium acetate buffer (pH 5.0) at 55 °C for 13 days. The formation of GOS was catalyzed by free and THP‐immobilized β‐galactosidase from lactose. The yield of GOS produced by the free enzyme from the lactose solution (36%, w/v) at 55 °C was 43% on a dry weight basis, which was similar to the 41% GOS yield produced by the THP‐immobilized enzyme system.