Zhuang Meng-Yao, Wang Cong, Xu Meng-Qiu, Ling Xiao-Min, Shen Jia-Jia, Zhang Ye-Wang
School of Pharmacy, United Institute of Jiangsu University and Shandong Tianzhilvye Biotechnology Co. Ltd., Jiangsu University, Zhenjiang, 212013, People's Republic of China.
School of Pharmacy, United Institute of Jiangsu University and Shandong Tianzhilvye Biotechnology Co. Ltd., Jiangsu University, Zhenjiang, 212013, People's Republic of China.
Int J Biol Macromol. 2017 Nov;104(Pt A):63-69. doi: 10.1016/j.ijbiomac.2017.05.150. Epub 2017 May 27.
ConcanavalinA (conA) is a protein extracted from the concanavalin, which has specific recognition through mannose components on bacterial cell surfaces. A magnetic nanocarrier with the structure of a dopamine functionalized magnetic nanoparticles was grafted with conA, and was used for immobilization of recombinant Escherichia coli harboring glycerol dehydrogenase with the specific recognition between glycoconjugates and glycoprotein. The effect of various factors on the immobilization including temperature, pH, cell concentration and immobilization time were investigated. The highest immobilization yield of 91% was obtained under the conditions: enzyme/support 1.28mg/mg, pH 8.0, immobilization time 2h and temperature 4°C. The obtained immobilized cell was characterized and exhibited higher thermal stability compared with the free cell. After ten cycles, the immobilized cell remained 62% initial activity. These results indicate that the cell immobilized onto conA-grafted nanoparticles by specific recognition of glycoconjugates and glycoprotein is a potential method for preparation of stable cell, and the immobilized cell showed perspective applications in the biocatalysis and biosensors.
伴刀豆球蛋白A(ConA)是从伴刀豆中提取的一种蛋白质,它能通过细菌细胞表面的甘露糖成分进行特异性识别。将具有多巴胺功能化磁性纳米颗粒结构的磁性纳米载体与ConA接枝,并利用糖缀合物和糖蛋白之间的特异性识别作用,用于固定化携带甘油脱氢酶的重组大肠杆菌。研究了温度、pH值、细胞浓度和固定化时间等各种因素对固定化的影响。在酶/载体为1.28mg/mg、pH值为8.0、固定化时间为2小时、温度为4℃的条件下,获得了91%的最高固定化产率。对所得固定化细胞进行了表征,结果表明其热稳定性高于游离细胞。经过十次循环后,固定化细胞仍保留62%的初始活性。这些结果表明,通过糖缀合物和糖蛋白的特异性识别将细胞固定在ConA接枝的纳米颗粒上是制备稳定细胞的一种潜在方法,并且固定化细胞在生物催化和生物传感器方面具有潜在的应用前景。