Application of polymaleimidostyrene as a convenient immobilization reagent of enzyme in biosensor.
Sulfhydryl groups of glucose oxidase (GOD) were reacted with maleimide groups of polymaleimidostyrene (PMS) which was coated onto the porous carbon sheet, and the carbon sheet immobilized by GOD was combined with an oxygen electrode to fabricate a glucose sensor. The activity of thiolated GOD immobilized to PMS is much larger than that of native GOD immobilized to PMS. The good linear relationship of glucose and oxygen current response was obtained in a concentration range from 0.1 to 2 mM and upper limit of linear range was found to be 3.0 mM. The immobilized GOD activity is highly dependent on pH at immobilization and the maximum activity was obtained at pH 5.5, probably because the SH groups of GOD that are indispensable for generation of enzyme activity is not exposed at this pH. It was found that PMS is very effective reagent to immobilize enzyme strongly via covalent bond, because high density of maleimide groups of PMS can catch not only exposed SH groups but also buried SH groups.
Uchiyama
Shunichi
S
Department of Materials Science and Technology, Graduate School of Engineering, Saitama Institute of technology, 1690 Fusaiji, Okabe, Saitama 369-0293 Japan. uchiyama@sit.ac.jp
Tomita
Ryoichi
R
Sekioka
Naoyuki
N
Imaizumi
Emi
E
Hamana
Hiroshi
H
Hagiwara
Tokio
T
D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D004563:Electrochemistry / Q000295:instrumentation* / Q000379:methods; D004800:Enzymes, Immobilized / Q000737:chemistry; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005947:Glucose / Q000032:analysis* / Q000737:chemistry*; D005949:Glucose Oxidase / Q000737:chemistry*; D008301:Maleimides / Q000737:chemistry*; D008839:Microelectrodes; D011137:Polystyrenes / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D004800:Enzymes, Immobilized; D008301:Maleimides; D011137:Polystyrenes; C512149:polymaleimidostyrene; D005949:Glucose Oxidase; D005947:Glucose