Enhancement in thermal stability and resistance to denaturants of lipase encapsulated in mesoporous silica with alkyltrimethylammonium (CTAB).
We assembled a highly durable conjugate with both a high-density accumulation and a regular array of lipase, by encapsulating it in mesoporous silica (FSM) with alkyltrimethylammonium (CTAB) chains on the surface. The activity for hydrolyzing esters of the lipase immobilized in mesoporous silica was linearly related to the concentration of lipase, whereas that of non-immobilized lipase showed saturation due to self-aggregation at a high concentration. The lipase conjugate also had increased resistance to heating when stayed in the silica coupling with CTAB. In addition, encapsulating the enzyme with FSM coupled CTAB caused the lipase to remain stable even in the presence of urea and trypsin, suggesting that the encapsulation prevented dissociation and denaturing. This conjugate had much higher activity and much higher stability for hydrolyzing esters when compared to the native lipase. These results show that FSM provides suitable support for the immobilization and dispersion of proteins in mesopores with disintegration of the aggregates.
Itoh
Tetsuji
T
National Institute of Advanced Industrial Science and Technology (AIST), Nigatake 4-2-1, Miyagino-ku, Sendai 983-8551, Japan. t-itoh@ni.aist.go.jp
Ishii
Ryo
R
Matsuura
Shun-ichi
S
Mizuguchi
Junko
J
Hamakawa
Satoshi
S
Hanaoka
Taka-aki
TA
Tsunoda
Tatsuo
T
Mizukami
Fujio
F
D000327:Adsorption / Q000187:drug effects; D000077286:Cetrimonium; D002593:Cetrimonium Compounds / Q000378:metabolism*; D004795:Enzyme Stability / Q000187:drug effects; D008049:Lipase / Q000378:metabolism*; D009584:Nitrogen / Q000378:metabolism; D016062:Porosity / Q000187:drug effects; D011489:Protein Denaturation / Q000187:drug effects*; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D012822:Silicon Dioxide / Q000378:metabolism*; D013696:Temperature; D014357:Trypsin / Q000494:pharmacology*; D014508:Urea / Q000494:pharmacology*
D002593:Cetrimonium Compounds; D012822:Silicon Dioxide; D014508:Urea; D008049:Lipase; D014357:Trypsin; D009584:Nitrogen; D000077286:Cetrimonium