Zhang Shan, Deng Qianchun, Li Ya, Zheng Mingming, Wan Chuyun, Zheng Chang, Tang Hu, Huang Fenghong, Shi Jie
Hubei Key Laboratory of Lipid Chemistry and Nutrition, Oil Crops and Lipids Process Technology National and Local Joint Engineering Laboratory, Key Laboratory of Oilseeds Processing, Ministry of Agriculture, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, Wuhan 430062, People's Republic of China.
R Soc Open Sci. 2018 Jun 13;5(6):172368. doi: 10.1098/rsos.172368. eCollection 2018 Jun.
The high catalytic activity, specificity and stability of immobilized lipase have been attracting great interest. How to reduce the cost of support materials has always been a hot topic in this field. Herein, for the development of low-cost immobilized lipase, we demonstrate an amphiphilic polyvinylpyrrolidone (PVP) grafted on silicone particle (SP) surface materials (SP-PVP) with a rational design based on interfacial activation and solution polymerization. Meanwhile, hydrophilic pristine SP and hydrophobic polystyrene-corded silicone particles (SP-Pst) were also prepared for lipase immobilization. SP-PVP was characterized by X-ray diffraction, scanning electron microscopy, X-ray photoelectron spectroscopy, Fourier transform infrared spectroscopy and thermogravimetry. Our results indicated that the lipase loading amount on the SP-PVP composites was about 215 mg of protein per gram. In the activity assay, the immobilized lipase SP-PVP@CRL exhibited higher catalysis activity and better thermostability and reusability than SP@CRL and SP-Pst@CRL. The immobilized lipase retained more than 54% of its initial activity after 10 times of re-use and approximately trended to a steady rate in the following cycles. By introducing the interesting amphiphilic polymer to this cheap and easily obtained SP surface, the relative performance of the immobilized lipase can be significantly improved, facilitating interactions between the low-cost support materials and lipase.
固定化脂肪酶的高催化活性、特异性和稳定性一直备受关注。如何降低载体材料成本一直是该领域的热门话题。在此,为了开发低成本的固定化脂肪酶,我们基于界面活化和溶液聚合的合理设计,展示了一种接枝在硅颗粒(SP)表面材料(SP-PVP)上的两亲性聚乙烯吡咯烷酮(PVP)。同时,还制备了亲水性原始SP和疏水性聚苯乙烯接枝硅颗粒(SP-Pst)用于脂肪酶固定化。通过X射线衍射、扫描电子显微镜、X射线光电子能谱、傅里叶变换红外光谱和热重分析对SP-PVP进行了表征。我们的结果表明,SP-PVP复合材料上的脂肪酶负载量约为每克215毫克蛋白质。在活性测定中,固定化脂肪酶SP-PVP@CRL比SP@CRL和SP-Pst@CRL表现出更高的催化活性、更好的热稳定性和可重复使用性。固定化脂肪酶在重复使用10次后保留了其初始活性的54%以上,并且在随后的循环中大致趋于稳定速率。通过将这种有趣的两亲性聚合物引入到这种廉价且易于获得的SP表面,可以显著提高固定化脂肪酶的相对性能,促进低成本载体材料与脂肪酶之间的相互作用。