Sun Xinzeng, Niu Huanqing, Song Jiarui, Jiang Dahai, Leng Jing, Zhuang Wei, Chen Yong, Liu Dong, Ying Hanjie
State Key Laboratory of Materials-Oriented Chemical Engineering, Nanjing Tech University, No. 5, Xinmofan Road, Nanjing 210009, P. R. China.
College of Biotechnology and Pharmaceutical Engineering, National Engineering Technique Research Center for Biotechnology, Nanjing Tech University, No. 30, Puzhu South Road, Nanjing 211816, P. R. China.
ACS Omega. 2020 Apr 27;5(17):9991-9998. doi: 10.1021/acsomega.0c00329. eCollection 2020 May 5.
In this research article, we reported a self-assembly approach to prepare a copper polyphosphate kinase 2 hybrid nanoflower and established a cofactor ADP regeneration system from AMP using the nanoflower. First, the structure of the hybrid nanoflower was confirmed by scanning electron microscopy, X-ray diffraction, Fourier transform infrared spectroscopy, and X-ray photoelectron spectroscopy, which indicated the successful loading of the enzyme in the hybrid nanoflower. Moreover, compared to the free enzyme, the hybrid nanoflower exhibited a better performance in ADP production and possessed wider catalytic pH and temperature ranges as well as improved storage stability. The hybrid nanoflower also exhibited well reusability, preserving 71.7% of initial activity after being used for ten cycles. In addition, the phosphorylation of glucose was conducted by utilizing ADP-dependent glucokinase coupled with the ADP regeneration system, in which the hybrid nanoflower was used for regenerating ADP from AMP. It was observed that the ADP regeneration system operated effectively at a very small amount of AMP. Thus, the hybrid nanoflower had great application potential in industrial catalytic processes that were coupled with ADP-dependent enzymes.
在这篇研究文章中，我们报道了一种制备聚磷酸铜激酶2杂交纳米花的自组装方法，并利用该纳米花建立了一个从AMP再生辅因子ADP的系统。首先，通过扫描电子显微镜、X射线衍射、傅里叶变换红外光谱和X射线光电子能谱对杂交纳米花的结构进行了确认，这些结果表明酶已成功负载于杂交纳米花中。此外，与游离酶相比，杂交纳米花在生成ADP方面表现出更好的性能，具有更宽的催化pH和温度范围以及更高的储存稳定性。杂交纳米花还表现出良好的可重复使用性，在使用十个循环后仍保留71.7%的初始活性。此外，利用依赖ADP的葡萄糖激酶与ADP再生系统相结合进行葡萄糖的磷酸化反应，其中杂交纳米花用于从AMP再生ADP。结果发现，ADP再生系统在极少量的AMP下就能有效运行。因此，杂交纳米花在与依赖ADP的酶相关的工业催化过程中具有巨大的应用潜力。
