Journal article
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.
Language: | English |
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Publisher: | Springer-Verlag |
Year: | 2013 |
Pages: | 2076-2087 |
Journal subtitle: | Part A: Enzyme Engineering and Biotechnology |
ISSN: | 15590291 and 02732289 |
Types: | Journal article |
DOI: | 10.1007/s12010-013-0122-5 |
ORCIDs: | Hobley, Timothy John |
Biochemistry, general Biotechnology Chemistry Chemistry and Materials Science Enzyme Stability Enzymes Enzymes, Immobilized Glutaral Hydrolysis Immobilization Lignin Lignocellulose hydrolysis Magnetic particles Magnetics Pretreated spruce Triazines beta-Glucosidase cyanuric chloride lignocellulose poly(glutaraldehyde)