TY - JOUR
T1 - Single enzyme nanoparticles in nanoporous silica
T2 - A hierarchical approach to enzyme stabilization and immobilization
AU - Kim, Jungbae
AU - Jia, Hongfei
AU - Lee, Chang won
AU - Chung, Seung wook
AU - Kwak, Ja Hun
AU - Shin, Yongsoon
AU - Dohnalkova, Alice
AU - Kim, Byung Gee
AU - Wang, Ping
AU - Grate, Jay W.
N1 - Funding Information:
This work was supported by U.S. Department of Energy (DOE) LDRD funds administered by the Pacific Northwest National Laboratory, and the DOE Office of Biological and Environmental Research under the Environmental Management Science Program. The research was performed at the W. R. Wiley Environmental Molecular Sciences Laboratory, a national scientific user facility sponsored by the U.S. Department of Energy's Office of Biological and Environmental Research and located at Pacific Northwest National Laboratory. PNNL is operated for the Department of Energy by Battelle.
PY - 2006/7/3
Y1 - 2006/7/3
N2 - Single enzyme nanoparticles of α-chymotrypsin (SEN-CT), in which each CT molecule is surrounded by a thin polymeric organic/inorganic network, stabilized the CT activity in a shaking condition as well as in a non-shaking condition. Since SEN-CT is soluble in a buffer solution and less than 10 nm in size, SEN-CT could be further immobilized in nanoporous silica with an average pore size of 29 nm. Free CT and SEN-CT were immobilized in nanoporous silica (NPS), and nanoporous silica that was first silanized with aminopropyltriethoxysilane (amino-NPS) to generate a positive surface charge. The SEN-CT adsorbed in amino-NPS was more stable than CT immobilized by either adsorption in NPS or covalent bonding to amino-NPS. In shaking conditions, nanoporous silica provided an additional stabilization by protecting SEN-CT from shear stresses. At 22 °C with harsh shaking, free, NPS-adsorbed and NPS-covalently attached CT showed half lives of 1, 62, and 80 h, respectively; whereas SEN-CT adsorbed in amino-NPS showed no activity loss within 12 days. The combination of SENs and nanoporous silica, which makes an active and stable immobilized enzyme system, represents a new structure for biocatalytic applications.
AB - Single enzyme nanoparticles of α-chymotrypsin (SEN-CT), in which each CT molecule is surrounded by a thin polymeric organic/inorganic network, stabilized the CT activity in a shaking condition as well as in a non-shaking condition. Since SEN-CT is soluble in a buffer solution and less than 10 nm in size, SEN-CT could be further immobilized in nanoporous silica with an average pore size of 29 nm. Free CT and SEN-CT were immobilized in nanoporous silica (NPS), and nanoporous silica that was first silanized with aminopropyltriethoxysilane (amino-NPS) to generate a positive surface charge. The SEN-CT adsorbed in amino-NPS was more stable than CT immobilized by either adsorption in NPS or covalent bonding to amino-NPS. In shaking conditions, nanoporous silica provided an additional stabilization by protecting SEN-CT from shear stresses. At 22 °C with harsh shaking, free, NPS-adsorbed and NPS-covalently attached CT showed half lives of 1, 62, and 80 h, respectively; whereas SEN-CT adsorbed in amino-NPS showed no activity loss within 12 days. The combination of SENs and nanoporous silica, which makes an active and stable immobilized enzyme system, represents a new structure for biocatalytic applications.
KW - Enzyme immobilization
KW - Enzyme stabilization
KW - Mesocellular foams
KW - Nanoporous silica
KW - Single enzyme nanoparticles
KW - α-Chymotrypsin
UR - http://www.scopus.com/inward/record.url?scp=33646796717&partnerID=8YFLogxK
U2 - 10.1016/j.enzmictec.2005.11.042
DO - 10.1016/j.enzmictec.2005.11.042
M3 - Article
AN - SCOPUS:33646796717
VL - 39
SP - 474
EP - 480
JO - Enzyme and Microbial Technology
JF - Enzyme and Microbial Technology
SN - 0141-0229
IS - 3
ER -