Moosavi Faezeh, Ahrari Faezeh, Ahmadian Gholamreza, Mohammadi Mehdi
Bioprocess Engineering Department, Institute of Industrial and Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), P. O. Box: 14965/161, Tehran, Iran.
Department of Pure Chemistry, Faculty of Chemistry, Shahid Beheshti University, G.C., Tehran, Iran.
Biotechnol Rep (Amst). 2022 Apr 21;34:e00733. doi: 10.1016/j.btre.2022.e00733. eCollection 2022 Jun.
In this study, lipase B (CalB) was covalently immobilized on the surface of graphene oxide (GO) nanoparticles by sortase-mediated immobilization as well as a chemical attachment approach. Sortase is a transpeptidase that provides one-step purification and targeted immobilization of CalB from one specific site, presenting oriented attachment of the enzyme to a solid support. Chemical immobilization, on the other hand, is considered as a random immobilization, in which the protein can bind to the support from different regions of the protein surface. In this approach, amine-functionalized GO was further modified with glutaraldehyde to facilitate the covalent binding of CalB via its amine residues. The applied methods produced 60% and 100% immobilization yields and presented 0.106 U/mg and 0.085 U/mg of specific activities for the oriented and random immobilization, respectively. The stabilized enzyme with the sortase-mediated approach retained approximately 80% of its initial activity at 50°C.
在本研究中，脂肪酶B（CalB）通过分选酶介导的固定化方法以及化学连接方法共价固定在氧化石墨烯（GO）纳米颗粒表面。分选酶是一种转肽酶，可从一个特定位点对CalB进行一步纯化和靶向固定，使酶定向附着于固体载体。另一方面，化学固定被认为是一种随机固定，其中蛋白质可从蛋白质表面的不同区域与载体结合。在这种方法中，胺功能化的GO用戊二醛进一步修饰，以促进CalB通过其胺残基进行共价结合。所应用的方法产生了60%和100%的固定化产率，并且分别为定向固定和随机固定呈现出0.106 U/mg和0.085 U/mg的比活性。采用分选酶介导方法稳定化的酶在50°C下保留了其初始活性的约80%。
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