Crosslinked enzyme aggregates in hierarchically-ordered mesoporous silica: a simple and effective method for enzyme stabilization.
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 2 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.
Kim
Moon Il
MI
Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 305-701, Republic of Korea.
Kim
Jungbae
J
Lee
Jinwoo
J
Jia
Hongfei
H
Na
Hyon Bin
HB
Youn
Jong Kyu
JK
Kwak
Ja Hun
JH
Dohnalkova
Alice
A
Grate
Jay W
JW
Wang
Ping
P
Hyeon
Taeghwan
T
Park
Hyun Gyu
HG
Chang
Ho Nam
HN
D002918:Chymotrypsin / Q000737:chemistry; D003432:Cross-Linking Reagents / Q000737:chemistry*; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry*; D008049:Lipase / Q000737:chemistry; D046529:Microscopy, Electron, Transmission; D008954:Models, Biological; D016062:Porosity; D012822:Silicon Dioxide / Q000737:chemistry*
D003432:Cross-Linking Reagents; D004800:Enzymes, Immobilized; D012822:Silicon Dioxide; D008049:Lipase; D002918:Chymotrypsin; C117608:alpha-chymotrypsin