Magnetic Combined Cross-Linked Enzyme Aggregates of Ketoreductase and Alcohol Dehydrogenase: An Efficient and Stable Biocatalyst for Asymmetric Synthesis of (R)-3-Quinuclidinol with Regeneration of Coenzymes In Situ
Abstract
:1. Introduction
2. Results
2.1. Preparation and Characterization of the Magnetic Combi-CLEAs of QNR and GDH
2.2. Screening of Precipitant Agents for Magnetic Combi-CLEAs Preparation
2.3. Effect of Cross-Linker Concentration and Cross-linking Time on Activity Recovery of Magnetic Combi-CLEAs
2.4. Optimal pH and Temperature for Magnetic Combi-CLEAs Activity Recovery
2.5. Time-Course of Biosynthesis of (R)-3-quinuclidinol
3. Materials and Methods
3.1. Materials
3.2. Expression of Recombinant Enzyme QNR and GDH
3.3. Enzyme Activity Assay
3.4. Immobilization of QNR and GDH
3.4.1. Preparation of NH2-Functionalized Fe3O4 Nanoparticles (Fe3O4@SiO2-NH2)
3.4.2. Preparation of Magnetic Combi-CLEAs of QNR and GDH
3.5. Application of Magnetic Combi-CLEAs for Biosynthesis (R)-3-quinuclidinol
3.6. Reusability of Enzymes in Magnetic Combi-CLEAs
3.7. Characterization
4. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Chen, Y.; Jiang, Q.; Sun, L.; Li, Q.; Zhou, L.; Chen, Q.; Li, S.; Yu, M.; Li, W. Magnetic Combined Cross-Linked Enzyme Aggregates of Ketoreductase and Alcohol Dehydrogenase: An Efficient and Stable Biocatalyst for Asymmetric Synthesis of (R)-3-Quinuclidinol with Regeneration of Coenzymes In Situ. Catalysts 2018, 8, 334. https://doi.org/10.3390/catal8080334
Chen Y, Jiang Q, Sun L, Li Q, Zhou L, Chen Q, Li S, Yu M, Li W. Magnetic Combined Cross-Linked Enzyme Aggregates of Ketoreductase and Alcohol Dehydrogenase: An Efficient and Stable Biocatalyst for Asymmetric Synthesis of (R)-3-Quinuclidinol with Regeneration of Coenzymes In Situ. Catalysts. 2018; 8(8):334. https://doi.org/10.3390/catal8080334
Chicago/Turabian StyleChen, Yuhan, Qihua Jiang, Lili Sun, Qiang Li, Liping Zhou, Qian Chen, Shanshan Li, Mingan Yu, and Wei Li. 2018. "Magnetic Combined Cross-Linked Enzyme Aggregates of Ketoreductase and Alcohol Dehydrogenase: An Efficient and Stable Biocatalyst for Asymmetric Synthesis of (R)-3-Quinuclidinol with Regeneration of Coenzymes In Situ" Catalysts 8, no. 8: 334. https://doi.org/10.3390/catal8080334