Portable Alkaline Phosphatase-Hydrogel Platform: From Enzyme Characterization to Phosphate Sensing

Area of Science:

• Biomaterials Science
• Enzyme Engineering
• Biosensor Technology

Background:

• Alkaline phosphatase (ALP) is crucial in biological processes and environmental monitoring.
• Developing stable, reusable enzyme-based biosensors remains a challenge.
• Immobilization strategies are key to maintaining enzyme activity and stability.

Purpose of the Study:

• To create transparent, room-temperature-storable hydrogels incorporating alkaline phosphatase (ALP@AETA).
• To characterize the stability and activity of immobilized ALP within a 3D polymeric network.
• To evaluate ALP@AETA as a disposable biosensor platform for detecting ALP inhibitors and phosphate.

Main Methods:

• Green protocol for enzyme immobilization into a 3D polymeric network.
• Characterization of hydrogel properties and enzyme conformation using intrinsic fluorescence and temperature-dependent studies.
• Assays for enzymatic activity inhibition and proof-of-concept testing as an optical biosensor for phosphate detection.

Main Results:

• Successfully incorporated and characterized alkaline phosphatase (ALP) into transparent hydrogels (ALP@AETA).
• Immobilized ALP retained its enzymatic activity and exhibited enhanced thermal stability within the hydrogel.
• ALP@AETA xerogels demonstrated stability at room temperature and were effective in detecting phosphate in water samples.

Conclusions:

• The developed ALP@AETA hydrogels offer a stable, room-temperature-storable platform for enzyme immobilization.
• This material shows significant potential for disposable biosensor applications in environmental monitoring and therapeutic enzyme delivery.
• The study highlights a promising approach for creating robust enzyme-based sensing devices.