Immobilization of Brassica oleracea chlorophyllase 1 (BoCLH1) and Candida rugosa lipase (CRL) in magnetic alginate beads: an enzymatic evaluation in the corresponding proteins.

Enzymes have a wide variety of applications in diverse biotechnological fields, and the immobilization of enzymes plays a key role in academic research or industrialization due to the stabilization and recyclability it confers. In this study, we immobilized the Brassica oleracea chlorophyllase 1 (BoCLH1) or Candida rugosa lipase (CRL) in magnetic iron oxide nanoparticles-loaded alginate composite beads. The catalytic activity and specific activity of the BoCLH1 and CRL entrapped in magnetic alginate composite beads were evaluated. Results show that the activity of immobilized BoCLH1 in magnetic alginate composite beads (3.36±0.469 U/g gel) was higher than that of immobilized BoCLH1 in alginate beads (2.96±0.264 U/g gel). In addition, the specific activity of BoCLH1 beads (10.90±1.521 U/mg protein) was higher than that immobilized BoCLH1 in alginate beads (8.52±0.758 U/mg protein). In contrast, the immobilized CRL in magnetic alginate composite beads exhibited a lower enzyme activity (11.81±0.618) than CRL immobilized in alginate beads (94.83±7.929), and the specific activity of immobilized CRL entrapped in magnetic alginate composite beads (1.99±0.104) was lower than immobilized lipase in alginate beads (15.01±1.255). A study of the degradation of magnetic alginate composite beads immersed in acidic solution (pH 3) shows that the magnetic alginate composite beads remain intact in acidic solution for at least 6 h, indicating the maintenance of the enzyme catalytic effect in low-pH environment. Finally, the enzyme immobilized magnetic alginate composite beads could be collected by an external magnet and reused for at least six cycles.