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Preparation of polyhydroxyalkanoate-based magnetic microspheres for carbonyl reductase purification and immobilization.
- Source :
-
International Journal of Biological Macromolecules . Dec2023:Part 4, Vol. 253, pN.PAG-N.PAG. 1p. - Publication Year :
- 2023
-
Abstract
- A polyhydroxyalkanoate (PHA) magnetic microsphere was designed for one-step purification and immobilization of a novel carbonyl reductase (RLSR5) from recombinant Escherichia coli lysate. The hydrophobic core of this microsphere was composed of a highly biocompatible polymer, poly(3-hydroxybutyrate- co -3-hydroxyhexanoate) (PHBHHx), in which magnetic Fe 3 O 4 particles were embedded during solvent evaporation. The hydrophilic shell of the fusion protein formed by PHA particle-binding protein (PhaP) and RLSR5 (PR) was expressed in recombinant E. coli. The magnetic core of Fe 3 O 4 @PHBHHx directly purified the hydrophilic shell from the E. coli lysate, and the two self-assembled to form Fe 3 O 4 @PHBHHx-PR through hydrophobic and hydrophilic interactions, eliminating the separation of the fusion protein. The microstructure, magnetic properties, morphology, size, and dispersion of Fe 3 O 4 @PHBHHx-PR were investigated by XRD, VSM, SEM, TEM, elemental mapping and DLS. It was found that Fe 3 O 4 @PHBHHx-PR correctly assembled, with a well dispersed spherical structure at the nanoscale and superparamagnetism properties. The amount of RLSR5 immobilized on PHA microspheres reached 121.9 mg/g. The Fe 3 O 4 @PHBHHx-PR was employed to synthesize (R)-tolvaptan with 99 % enantiomeric excess and 97 % bioconversion efficiency, and the catalyst maintained 78.6 % activity after 10 recovery cycles. These PHA magnetic microspheres are versatile carriers for enzyme immobilization and demonstrate improved stability and reusability of the free enzyme. [ABSTRACT FROM AUTHOR]
Details
- Language :
- English
- ISSN :
- 01418130
- Volume :
- 253
- Database :
- Academic Search Index
- Journal :
- International Journal of Biological Macromolecules
- Publication Type :
- Academic Journal
- Accession number :
- 173784086
- Full Text :
- https://doi.org/10.1016/j.ijbiomac.2023.126814