TY - JOUR
T1 - Successful encapsulation of β-glucosidase during the synthesis of siliceous mesostructured materials
AU - Gascón, Victoria
AU - Márquez-Álvarez, Carlos
AU - Blanco, Rosa María
N1 - Publisher Copyright:
© 2018 Society of Chemical Industry
PY - 2018/9
Y1 - 2018/9
N2 - BACKGROUND: The biocatalysis field demands ‘universal’ supports able to encapsulate enzymes with a straightforward methodology, and at the same time, capable of retaining their catalytic activity. The employment of siliceous materials for such a purpose is a big challenge because drastic synthesis conditions are required and improved functionalization is needed to increase affinities towards the targeted enzyme. In this work, a compromise between the development of a well-formed mesostructured support and an acceptable enzymatic activity was attempted via the in-situ immobilization approach. RESULTS: The immobilization of β-glucosidase (EC 3.2.1.21) from Aspergillus niger was approached using different strategies. After trying to immobilize β-glucosidase with a post-synthesis approach, nonhigh loadings were achieved both with covalent linkage (using epoxy activated supports; 3.5 mgE g−1) and with noncovalent bonding (using amine-functionalized materials; 7.6 mgE g−1). However, when the in-situ approach was attempted, success in reaching the highest enzyme loading (close to 200 mgE g−1) was achieved. CONCLUSION: In this work, the support cages around the in-situ encapsulated enzyme fully prevented its release through the narrow windows connecting cages, achieving a less than 5% release of the initially desorbed protein, as well as a further total absence of leaching. This enabled the biocatalyst to be reused at least eight times more without any loss in activity.
AB - BACKGROUND: The biocatalysis field demands ‘universal’ supports able to encapsulate enzymes with a straightforward methodology, and at the same time, capable of retaining their catalytic activity. The employment of siliceous materials for such a purpose is a big challenge because drastic synthesis conditions are required and improved functionalization is needed to increase affinities towards the targeted enzyme. In this work, a compromise between the development of a well-formed mesostructured support and an acceptable enzymatic activity was attempted via the in-situ immobilization approach. RESULTS: The immobilization of β-glucosidase (EC 3.2.1.21) from Aspergillus niger was approached using different strategies. After trying to immobilize β-glucosidase with a post-synthesis approach, nonhigh loadings were achieved both with covalent linkage (using epoxy activated supports; 3.5 mgE g−1) and with noncovalent bonding (using amine-functionalized materials; 7.6 mgE g−1). However, when the in-situ approach was attempted, success in reaching the highest enzyme loading (close to 200 mgE g−1) was achieved. CONCLUSION: In this work, the support cages around the in-situ encapsulated enzyme fully prevented its release through the narrow windows connecting cages, achieving a less than 5% release of the initially desorbed protein, as well as a further total absence of leaching. This enabled the biocatalyst to be reused at least eight times more without any loss in activity.
KW - enzyme immobilization
KW - in-situ
KW - ordered mesoporous silica materials
KW - post-synthesis
KW - β-glucosidase
UR - http://www.scopus.com/inward/record.url?scp=85045758962&partnerID=8YFLogxK
U2 - 10.1002/jctb.5616
DO - 10.1002/jctb.5616
M3 - Article
AN - SCOPUS:85045758962
SN - 0268-2575
VL - 93
SP - 2625
EP - 2634
JO - Journal of Chemical Technology and Biotechnology
JF - Journal of Chemical Technology and Biotechnology
IS - 9
ER -