Smart Core鈥揝hell Microgel Support for Acetyl Coenzyme A Synthetase: A Step Toward Efficient Synthesis of Polyketide-Based Drugs
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- 作者：Nidhi C. Dubey ; Bijay P. Tripathi ; Manfred Stamm ; Leonid Ionov
- 刊名：Biomacromolecules
- 出版年：2014
- 出版时间：July 14, 2014
- 年：2014
- 卷：15
- 期：7
- 页码：2776-2783
- 全文大小：447K
- ISSN：1526-4602
文摘
The flexibility in tuning the structure and charge properties of PNIPAm microgels during their synthesis makes them a suitable choice for various biological applications. Two-step free radical polymerization, a common method employed for synthesis of core鈥搒hell microgel has been well adopted to obtain cationic poly(N-isopropylacrylamide-aminoethyl methacrylate) (PNIPAm-AEMA) shell and PNIPAm core. Scanning electron microscopy (SEM), dynamic light scattering (DLS), zeta potential, and ninhydrin assay suggests nearly monodispersed particles of cationic nature. Amino groups on the microgel provides suitable attachment point for covalent immobilization of acetyl coenzyme A synthetase (Acs) via 1-ethyl-3-(3-N,N- dimethylaminopropyl) carbodiimide (EDC) chemistry. On immobilization, 61.55% of initial activity of Acs has been retained, while Michaelis鈥揗enten kinetics of the immobilized Acs indicates identical Km (Michaelis constant) but decrease in the Vmax (maximum substrate conversion rate) compared to free enzyme. Immobilized Acs shows an improvement in activity at wide temperature and pH range and also demonstrates good thermal, storage, and operational stability. The Acs鈥搈icrogel bioconjugate has been successfully reused for four consecutive operation cycles with more than 50% initial activity.