Covalent immobilization of β-1,4-glucosidase from Agaricus arvensis onto functionalized silicon oxide nanoparticles
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Covalent immobilization of β-1,4-glucosidase from Agaricus arvensis onto functionalized silicon oxide nanoparticles. / Singh, Raushan Kumar; Zhang, Ye Wang; Nguyen, Ngoc Phuong Thao; Jeya, Marimuthu; Lee, Jung Kul.
I: Applied Microbiology and Biotechnology, Bind 89, Nr. 2, 2011, s. 337-344.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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TY - JOUR
T1 - Covalent immobilization of β-1,4-glucosidase from Agaricus arvensis onto functionalized silicon oxide nanoparticles
AU - Singh, Raushan Kumar
AU - Zhang, Ye Wang
AU - Nguyen, Ngoc Phuong Thao
AU - Jeya, Marimuthu
AU - Lee, Jung Kul
PY - 2011
Y1 - 2011
N2 - An efficient β-1,4-glucosidase (BGL) secreting strain, Agaricus arvensis, was isolated and identified. The relative molecular weight of the purified A. arvensis BGL was 98 kDa, as determined by sodium dodecylsulfate polyacrylamide gel electrophoresis, or 780 kDa by size exclusion chromatography, indicating that the enzyme is an octamer. Using a crude enzyme preparation, A. arvensis BGL was covalently immobilized onto functionalized silicon oxide nanoparticles with an immobilization efficiency of 158%. The apparent V max (k cat) values of free and immobilized BGL under standard assay conditions were 3,028 U mg protein-1 (4,945 s -1) and 3,347 U mg protein-1 (5,466 s-1), respectively. The immobilized BGL showed a higher optimum temperature and improved thermostability as compared to the free enzyme. The half-life at 65 °C showed a 288-fold improvement over the free BGL. After 25 cycles, the immobilized enzyme still retained 95% of the original activity, thus demonstrating its prospects for commercial applications. High specific activity, high immobilization efficiency, improved stability, and reusability of A. arvensis BGL make this enzyme of potential interest in a number of industrial applications.
AB - An efficient β-1,4-glucosidase (BGL) secreting strain, Agaricus arvensis, was isolated and identified. The relative molecular weight of the purified A. arvensis BGL was 98 kDa, as determined by sodium dodecylsulfate polyacrylamide gel electrophoresis, or 780 kDa by size exclusion chromatography, indicating that the enzyme is an octamer. Using a crude enzyme preparation, A. arvensis BGL was covalently immobilized onto functionalized silicon oxide nanoparticles with an immobilization efficiency of 158%. The apparent V max (k cat) values of free and immobilized BGL under standard assay conditions were 3,028 U mg protein-1 (4,945 s -1) and 3,347 U mg protein-1 (5,466 s-1), respectively. The immobilized BGL showed a higher optimum temperature and improved thermostability as compared to the free enzyme. The half-life at 65 °C showed a 288-fold improvement over the free BGL. After 25 cycles, the immobilized enzyme still retained 95% of the original activity, thus demonstrating its prospects for commercial applications. High specific activity, high immobilization efficiency, improved stability, and reusability of A. arvensis BGL make this enzyme of potential interest in a number of industrial applications.
KW - β-1-4-Glucosidase
KW - Covalent bonding
KW - Immobilization
KW - Silicon oxide nanoparticles
U2 - 10.1007/s00253-010-2768-z
DO - 10.1007/s00253-010-2768-z
M3 - Journal article
C2 - 20811797
AN - SCOPUS:78651086201
VL - 89
SP - 337
EP - 344
JO - Applied Microbiology and Biotechnology
JF - Applied Microbiology and Biotechnology
SN - 0175-7598
IS - 2
ER -
ID: 229901266