Covalent immobilization of β-glucosidase on magnetic particles for lignocellulose hydrolysis.
β-Glucosidase hydrolyzes cellobiose to glucose and is an important enzyme in the consortium used for hydrolysis of cellulosic and lignocellulosic feedstocks. In the present work, β-glucosidase was covalently immobilized on non-porous magnetic particles to enable re-use of the enzyme. It was found that particles activated with cyanuric chloride and polyglutaraldehyde gave the highest bead-related immobilized enzyme activity when tested with p-nitrophenyl-β-D-glucopyranoside (104.7 and 82.2 U/g particles, respectively). Furthermore, the purified β-glucosidase preparation from Megazyme gave higher bead-related enzyme activities compared to Novozym 188 (79.0 and 9.8 U/g particles, respectively). A significant improvement in thermal stability was observed for immobilized enzyme compared to free enzyme; after 5 h (at 65 °C), 36 % of activity remained for the former, while there was no activity in the latter. The performance and recyclability of immobilized β-glucosidase on more complex substrate (pretreated spruce) was also studied. It was shown that adding immobilized β-glucosidase (16 U/g dry matter) to free cellulases (8 FPU/g dry matter) increased the hydrolysis yield of pretreated spruce from ca. 44 % to ca. 65 %. In addition, it was possible to re-use the immobilized β-glucosidase in the spruce and retain activity for at least four cycles. The immobilized enzyme thus shows promise for lignocellulose hydrolysis.
Institute for Food, Technical University of Denmark, Building 221, Søltofts Plads, 2800 Lyngby, Denmark.
Johan
J
Alftrén
Timothy John
TJ
Hobley
D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism*; D005976:Glutaral / Q000031:analogs & derivatives / Q000737:chemistry; D006868:Hydrolysis; D008031:Lignin / Q000378:metabolism*; D008280:Magnetics; D014227:Triazines / Q000737:chemistry; D001617:beta-Glucosidase / Q000378:metabolism*
D004800:Enzymes, Immobilized; D014227:Triazines; C036909:lignocellulose; C022841:poly(glutaraldehyde); C019309:cyanuric chloride; D008031:Lignin; D001617:beta-Glucosidase; D005976:Glutaral