(727d) Optimizing a Porous Calcium-Phosphate Supraparticle for Enzyme Immobilization
- Conference: AIChE Annual Meeting
- Year: 2017
- Proceeding: 2017 Annual Meeting
- Group: Nanoscale Science and Engineering Forum
- Time: Thursday, November 2, 2017 - 2:00pm-2:20pm
We sought to optimize the synthesis of the supraparticles to maximize enzyme loading, minimize polydispersity of the size distribution, and show immobilized enzyme activity. Supraparticles were synthesized with several calcium concentrations, rotational speeds, and fluid viscosities. After synthesis, these samples were mixed with eGFP- ZE, a fluorescent protein, and imaged using fluorescence microscopy. Image segmentation and analysis were then used to quantify the number density and size distribution of supraparticles loaded with protein. The samplesâ loading capacities were determined using isothermal titration calorimetry. After selecting the supraparticles with the most desirable characteristics, they were loaded with AmDH- ZE and FDH- ZE. The conversion of ketone to amine in the biocatalyst was determined using HPLC, and the optimal loading ratio of the two enzymes was determined to maximize conversion with respect to time. Immobilization of the enzymes in the supraparticles did not significantly reduce their specific activity. These results indicate supraparticles can immobilize multiple enzymes with high activity, laying the framework for their use in industrial enzyme cascades.
1. Bommarius, A. S. & Paye, M. F. Stabilizing biocatalysts. Chem. Soc. Rev. Chem. Soc. Rev 42,
2. Mohamad, N. R., Marzuki, N. H. C., Buang, N. A., Huyop, F. & Wahab, R. A. An overview of
technologies for immobilization of enzymes and surface analysis techniques for immobilized
enzymes. Biotechnol. Biotechnol. Equip. 29, 205â220 (2015).
3. Park, W. M. & Champion, J. A. Colloidal Assembly of Hierarchically Structured Porous
Supraparticles from Flower-Shaped ProteinâInorganic Hybrid Nanoparticles.
4. Park, W. M., Yee, C. M. & Champion, J. A. Self-assembled hybrid supraparticles that
proteolytically degrade tumor necrosis factor-a. J. Mater. Chem. B 4, 1633â1639 (2016).
5. Bommarius, B. R., SchÃ¼rmann, M. & Bommarius, A. S. A novel chimeric amine dehydrogenase
shows altered substrate specificity compared to its parent enzymes. Chem. Commun. 50, 14953â