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Title: Crosslinked Enzyme Aggregates in Hierarchically-Ordered Mesoporous Silica: A Simple and Effective Method for Enzyme Stabilization

Abstract

alpha-chymotrypsin (CT) and lipase (LP) were immobilized in hierarchically-ordered mesocellular mesoporous silica (HMMS) in a simple but effective way for the enzyme stabilization, which was achieved by the enzyme adsorption followed by glutaraldehyde (GA) crosslinking. This resulted in the formation of nanometer scale crosslinked enzyme aggregates (CLEAs) entrapped in the mesocellular pores of HMMS (37 nm), which did not leach out of HMMS through narrow mesoporous channels (13 nm). CLEA of alpha-chymotrypsin (CLEA-CT) in HMMS showed a high enzyme loading capacity and significantly increased enzyme stability. No activity decrease of CLEA-CT was observed for two weeks under even rigorously shaking condition, while adsorbed CT in HMMS and free CT showed a rapid inactivation due to the enzyme leaching and presumably autolysis, respectively. With the CLEA-CT in HMMS, however, there was no tryptic digestion observed suggesting that the CLEA-CT is not susceptible to autolysis. Moreover, CLEA of lipase (CLEA-LP) in HMMS retained 30% specific activity of free lipase with greatly enhanced stability. This work demonstrates that HMMS can be efficiently employed as host materials for enzyme immobilization leading to highly enhanced stability of the immobilized enzymes with high enzyme loading and activity.

Authors:
; ; ; ; ; ; ; ; ; ; ; ;
Publication Date:
Research Org.:
Pacific Northwest National Laboratory (PNNL), Richland, WA (US), Environmental Molecular Sciences Laboratory (EMSL)
Sponsoring Org.:
USDOE
OSTI Identifier:
898617
Report Number(s):
PNNL-SA-45554
24826; 24801; TRN: US200706%%231
DOE Contract Number:  
AC05-76RL01830
Resource Type:
Journal Article
Resource Relation:
Journal Name: Biotechnology and Bioenegineering, 96(2):210-218
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; ADSORPTION; AUTOLYSIS; CAPACITY; DIGESTION; ENZYMES; IMMOBILIZED ENZYMES; INACTIVATION; LEACHING; LIPASES; SILICA; STABILITY; STABILIZATION; enzyme stabilization; mesoporous silica; nanoscale enzyme reactors; nanobiotechnology; Environmental Molecular Sciences Laboratory

Citation Formats

Kim, Moon Il, Kim, Jungbae, Lee, Jinwoo, Jia, Hongfei, Na, Hyon Bin, Youn, Jongkyu, Kwak, Ja Hun, Dohnalkova, Alice, Grate, Jay W., Wang, Ping, Hyeon, Taeghwan, Park, Hyun-Gyu, and Chang, Ho Nam. Crosslinked Enzyme Aggregates in Hierarchically-Ordered Mesoporous Silica: A Simple and Effective Method for Enzyme Stabilization. United States: N. p., 2007. Web. doi:10.1002/bit.21107.
Kim, Moon Il, Kim, Jungbae, Lee, Jinwoo, Jia, Hongfei, Na, Hyon Bin, Youn, Jongkyu, Kwak, Ja Hun, Dohnalkova, Alice, Grate, Jay W., Wang, Ping, Hyeon, Taeghwan, Park, Hyun-Gyu, & Chang, Ho Nam. Crosslinked Enzyme Aggregates in Hierarchically-Ordered Mesoporous Silica: A Simple and Effective Method for Enzyme Stabilization. United States. doi:10.1002/bit.21107.
Kim, Moon Il, Kim, Jungbae, Lee, Jinwoo, Jia, Hongfei, Na, Hyon Bin, Youn, Jongkyu, Kwak, Ja Hun, Dohnalkova, Alice, Grate, Jay W., Wang, Ping, Hyeon, Taeghwan, Park, Hyun-Gyu, and Chang, Ho Nam. Thu . "Crosslinked Enzyme Aggregates in Hierarchically-Ordered Mesoporous Silica: A Simple and Effective Method for Enzyme Stabilization". United States. doi:10.1002/bit.21107.
@article{osti_898617,
title = {Crosslinked Enzyme Aggregates in Hierarchically-Ordered Mesoporous Silica: A Simple and Effective Method for Enzyme Stabilization},
author = {Kim, Moon Il and Kim, Jungbae and Lee, Jinwoo and Jia, Hongfei and Na, Hyon Bin and Youn, Jongkyu and Kwak, Ja Hun and Dohnalkova, Alice and Grate, Jay W. and Wang, Ping and Hyeon, Taeghwan and Park, Hyun-Gyu and Chang, Ho Nam},
abstractNote = {alpha-chymotrypsin (CT) and lipase (LP) were immobilized in hierarchically-ordered mesocellular mesoporous silica (HMMS) in a simple but effective way for the enzyme stabilization, which was achieved by the enzyme adsorption followed by glutaraldehyde (GA) crosslinking. This resulted in the formation of nanometer scale crosslinked enzyme aggregates (CLEAs) entrapped in the mesocellular pores of HMMS (37 nm), which did not leach out of HMMS through narrow mesoporous channels (13 nm). CLEA of alpha-chymotrypsin (CLEA-CT) in HMMS showed a high enzyme loading capacity and significantly increased enzyme stability. No activity decrease of CLEA-CT was observed for two weeks under even rigorously shaking condition, while adsorbed CT in HMMS and free CT showed a rapid inactivation due to the enzyme leaching and presumably autolysis, respectively. With the CLEA-CT in HMMS, however, there was no tryptic digestion observed suggesting that the CLEA-CT is not susceptible to autolysis. Moreover, CLEA of lipase (CLEA-LP) in HMMS retained 30% specific activity of free lipase with greatly enhanced stability. This work demonstrates that HMMS can be efficiently employed as host materials for enzyme immobilization leading to highly enhanced stability of the immobilized enzymes with high enzyme loading and activity.},
doi = {10.1002/bit.21107},
journal = {Biotechnology and Bioenegineering, 96(2):210-218},
number = ,
volume = ,
place = {United States},
year = {Thu Feb 01 00:00:00 EST 2007},
month = {Thu Feb 01 00:00:00 EST 2007}
}