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alpha-Amylase immobilized on bulk acoustic-wave sensor by UV-curing coating
1College of Chemistry and Chemical Engineering, Hunan University, 410082, Changsha, China
Biochemical Engineering Journal
|July 26, 2000
Summary
This study introduces a novel UV-curing coating method for immobilizing alpha-amylase, enhancing its stability and reusability. The bulk acoustic-wave sensor effectively monitors the immobilized enzyme
Area of Science:
- Biochemistry
- Enzyme immobilization
- Biosensors
Background:
- Enzyme immobilization is crucial for enzyme reusability and stability.
- Traditional methods can lead to significant loss of enzymatic activity.
- Developing efficient and less damaging immobilization techniques is essential.
Purpose of the Study:
- To propose a new method for alpha-amylase immobilization using UV-curing coating.
- To evaluate the activity and kinetic parameters of the immobilized alpha-amylase.
- To demonstrate the reusability of the immobilized enzyme.
Main Methods:
- Immobilization of alpha-amylase onto a piezoelectric quartz crystal using UV-curing coating.
- Monitoring enzyme activity with a bulk acoustic-wave (BAW) sensor.
- Determining kinetic parameters (K(m), V(max)) using the Lineweaver-Burk plot.
Main Results:
- The UV-curing immobilization method is simple, convenient, and preserves enzymatic activity.
- The BAW sensor accurately reflects starch hydrolysis by the immobilized alpha-amylase.
- Optimal hydrolysis occurs at pH 7.0, similar to free alpha-amylase.
- Estimated kinetic parameters: K(m)=12.7 mg/ml, V(max)=15.9 Hz/min.
- The immobilized enzyme retained adequate activity and was reused over 50 times.
Conclusions:
- UV-curing coating provides an effective method for alpha-amylase immobilization.
- The immobilized alpha-amylase exhibits good stability, activity, and reusability.
- This technique offers a promising approach for enzyme-based biosensors and biocatalysis.