Hou Chen, Qi Zhigang, Zhu Hao
State Key Laboratory of Applied Organic Chemistry, Key Laboratory of Nonferrous Metal Chemistry and Resources Utilization of Gansu Province, College of Chemistry and Chemical Engineering, Institute of Biochemical Engineering and Environmental Technology, Lanzhou University, Lanzhou 730000, China.
State Key Laboratory of Applied Organic Chemistry, Key Laboratory of Nonferrous Metal Chemistry and Resources Utilization of Gansu Province, College of Chemistry and Chemical Engineering, Institute of Biochemical Engineering and Environmental Technology, Lanzhou University, Lanzhou 730000, China.
Colloids Surf B Biointerfaces. 2015 Apr 1;128:544-551. doi: 10.1016/j.colsurfb.2015.03.007. Epub 2015 Mar 7.
A flexible, biocompatible and bioadhesive enzyme immobilizing material, which was synthesized based on the covalent assembly of biomimetic polymer and oxidized polysaccharide on magnetic nanoparticles (NPs), has been developed in this feasibility study. In this work, the bio-inspired polymer, polydopamine (PDA), was used to modify the well-monodispersed Fe3O4 NPs (mPDA NPs) with a controllable thickness via a dip-coating process, then the alginate di-aldehyde (ADA) was covalently assembled on the mPDA NPs and employed as a naturally occurring linking agent for Candida rugosa lipase (CRL) immobilization. The resulting support material was characterized by means of the transmission electron microscope (TEM), Fourier transform infrared spectra (FT-IR), X-ray diffraction (XRD), thermogravimetry (TG) analyser, and vibrating sample magnetometer (VSM). It was verified that the prepared mPDA NPs possessed distinct core-shell structure with uniform size and high saturation magnetization. For further application, the mPDA NPs was utilized in CRL immobilizing procedures and demonstrated can facilitate improving the enzyme activities. The optimum amount of lipase was 200 mg g(-1) support, the optimal pH and temperature for the catalyse condition of the immobilized CRL was 7.0 and 40°C, respectively. Moreover, the immobilized CRL kept the high activity at 77% after 12 times of recycling for batch hydrolysis of olive oil emulsion. This magnetic bioadhesive composite with functionalized properties and adhesion strength presents a general strategy for the immobilization of macromolecules.
在本可行性研究中,已开发出一种柔性、生物相容性和生物粘附性的酶固定化材料,该材料基于仿生聚合物和氧化多糖在磁性纳米颗粒(NPs)上的共价组装合成。在这项工作中,利用受生物启发的聚合物聚多巴胺(PDA)通过浸涂工艺对单分散性良好的Fe3O4 NPs(mPDA NPs)进行可控厚度的修饰,然后将海藻酸二醛(ADA)共价组装在mPDA NPs上,并用作固定化皱落假丝酵母脂肪酶(CRL)的天然连接剂。通过透射电子显微镜(TEM)、傅里叶变换红外光谱(FT-IR)、X射线衍射(XRD)、热重分析仪(TG)和振动样品磁强计(VSM)对所得载体材料进行了表征。证实所制备的mPDA NPs具有明显的核壳结构,尺寸均匀且饱和磁化强度高。为进一步应用,将mPDA NPs用于CRL固定化过程,并证明其有助于提高酶活性。脂肪酶的最佳用量为200 mg g(-1)载体,固定化CRL催化条件的最佳pH值和温度分别为7.0和40°C。此外,固定化CRL在对橄榄油乳液进行12次分批水解循环后,仍保持77%的高活性。这种具有功能化特性和粘附强度的磁性生物粘附复合材料为大分子的固定化提供了一种通用策略。