Department of Biochemistry, Faculty of Biology and Environmental Protection, Nicolaus Copernicus University in Toruń, 1 Lwowska St., 87-100 Toruń, Poland.
Department of Biochemistry, Faculty of Biology and Environmental Protection, Nicolaus Copernicus University in Toruń, 1 Lwowska St., 87-100 Toruń, Poland.
Mater Sci Eng C Mater Biol Appl. 2018 Jul 1;88:130-139. doi: 10.1016/j.msec.2018.03.006. Epub 2018 Mar 14.
In this study graphene oxide (GO), carbon quantum dots (CQD) and carbon nanoonions (CNO) have been characterized and applied for the first time as a matrix for recombinant adenylate kinase (AK, EC 2.7.4.3) immobilization. AK is an enzyme fulfilling a key role in metabolic processes. This phosphotransferase catalyzes the interconversion of adenine nucleotides (ATP, ADP and AMP) and thereby participates in nucleotide homeostasis, monitors a cellular energy charge as well as acts as a component of purinergic signaling system. The AK activity in all obtained biocatalytic systems was higher as compared to the free enzyme. We have found that the immobilization on carbon nanostructures increased both activity and stability of AK. Moreover, the biocatalytic systems consisting of AK immobilized on carbon nanostructures can be easily and efficiently lyophilized without risk of desorption or decrease in the catalytic activity of the investigated enzyme. The positive action of AK-GO biocatalytic system in maintaining the nucleotide balance in in vitro cell culture was proved.
在这项研究中，氧化石墨烯（GO）、碳量子点（CQD）和碳纳米洋葱（CNO）被进行了表征，并首次被应用于固定化重组腺嘌呤激酶（AK，EC 2.7.4.3）。AK 是一种在代谢过程中起关键作用的酶。这种磷酸转移酶催化腺嘌呤核苷酸（ATP、ADP 和 AMP）的相互转化，从而参与核苷酸的动态平衡，监测细胞的能量电荷，并作为嘌呤能信号系统的组成部分。与游离酶相比，在所有获得的生物催化系统中，AK 的活性都更高。我们发现，在碳纳米结构上的固定化提高了 AK 的活性和稳定性。此外，由固定在碳纳米结构上的 AK 组成的生物催化系统可以很容易且高效地进行冷冻干燥，而不会有酶解吸或催化活性降低的风险。在体外细胞培养中，AK-GO 生物催化系统在维持核苷酸平衡方面的积极作用得到了证明。
