TY - JOUR
T1 - Covalent immobilization of phenylalanine dehydrogenase on cellulose membrane for biosensor construction
AU - Villalonga, Reynaldo
AU - Fujii, Akira
AU - Shinohara, Hiroaki
AU - Tachibana, Shinjiro
AU - Asano, Yasuhisa
N1 - Funding Information:
This research was supported by grants from The Japan Society for the Promotion of Sciences to R. Villalonga and Y. Asano (Grant S-04257), and from Toyama Medical-Bio Cluster (The Ministry of Education, Culture, Sports, Science and Technology, Japan) to Y. Asano. Financial support to R. Villalonga from the International Foundation for Science, Stockholm, Sweden, and the Organisation for the Prohibition of Chemical Weapons, The Hague, The Netherlands (Grant F/3004-2) is also acknowledged.
PY - 2008/1/29
Y1 - 2008/1/29
N2 - Bacillus badius phenylalanine dehydrogenase was covalent immobilized on amino-activated cellulose membrane by cross-linking with glutaraldehyde. The amount of immobilized enzyme was estimated as 440 μg/cm2, and the biocatalyst retained 82% of the initial PheDH activity. The functionalized membrane was further used as coating material for a glassy carbon electrode, which was employed for the construction of an amperometric biosensor toward l-phenylalanine. The enzyme electrode (poised at +700 mV versus Ag/AgCl) showed a linear amperometric response up to 9.1 mM l-phenylalanine with a detection limit of 25 μM. The biosensor reached 95% of steady-state current in about 25 s and its sensitivity was 177 μA/M cm2. The electrode retained full electrocatalytic activity after 16 days of storage at 4 °C in 50 mM sodium phosphate buffer pH 7.0.
AB - Bacillus badius phenylalanine dehydrogenase was covalent immobilized on amino-activated cellulose membrane by cross-linking with glutaraldehyde. The amount of immobilized enzyme was estimated as 440 μg/cm2, and the biocatalyst retained 82% of the initial PheDH activity. The functionalized membrane was further used as coating material for a glassy carbon electrode, which was employed for the construction of an amperometric biosensor toward l-phenylalanine. The enzyme electrode (poised at +700 mV versus Ag/AgCl) showed a linear amperometric response up to 9.1 mM l-phenylalanine with a detection limit of 25 μM. The biosensor reached 95% of steady-state current in about 25 s and its sensitivity was 177 μA/M cm2. The electrode retained full electrocatalytic activity after 16 days of storage at 4 °C in 50 mM sodium phosphate buffer pH 7.0.
KW - Biosensor
KW - Cellulose
KW - Enzyme immobilization
KW - Phenylalanine dehydrogenase
UR - http://www.scopus.com/inward/record.url?scp=38149099279&partnerID=8YFLogxK
U2 - 10.1016/j.snb.2007.07.097
DO - 10.1016/j.snb.2007.07.097
M3 - 学術論文
AN - SCOPUS:38149099279
SN - 0925-4005
VL - 129
SP - 195
EP - 199
JO - Sensors and Actuators, B: Chemical
JF - Sensors and Actuators, B: Chemical
IS - 1
ER -