Improving the anti-oxidation of glucose oxidase with computer-aided structure optimization
Journal Title: JOURNAL OF ADVANCES IN BIOTECHNOLOGY - Year 2015, Vol 5, Issue 3
Abstract
Glucose oxidase (GOD) can be inactivated by hydrogen peroxide (H2O2) produced during glucose oxidation; thus, H2O2 is a competitive inhibitor of GOD. In this mechanism, methionine (Met), a sulfur-containing amino acid, can be oxidized by H2O2and converted into methionine sulfoxide, thereby inactivating GOD. In this study, to block GOD oxidation, the two-dimensional structures of three mutants were designed with computer-aided analysis, and the complex structures of GOD mutants and their substrate glucose were determined using the CDOCKER algorithm. Three mutant GODs and the wild-type GOD were expressed in Pichia pastoris. After purification, the activities and anti-oxidation capacities were evaluated. The activity of GOD was decreased substantially from 3.24 U/μg in the wild-type protein to 0.05 U/μg in the GOD-M523L-M524L mutant; in contrast, the activity of GOD-M524L-M528L was not different from that of the wild-type protein. Because the activity of GOD-M523L-M524L was negligible, we did not evaluate its anti-oxidative effects. However, the mutant GOD-M524L-M528L had better anti-oxidation capacity than the wild-type GOD. These results were consistent with the results of computer-aided analysis, suggesting that this method may be useful for enzyme structure optimization.
Authors and Affiliations
Zheng-Bing Jiang, Hui-Ting Song, Wen-Jing Xiao, Yi-Min Yang, Yong Zhao, Yuan Gao, Shi-Hui Liu, Zi-Lu Liu, Wu-Cheng Xia, Rong Li, Nuo-Nan Li
Keywords
Related Articles
- EP ID EP653886
- DOI 10.24297/jbt.v5i3.1483
- Views 100
- Downloads 0
Yacon Flour and Corn Steep Liquor as Substrate for Inulinase and Biomass Production by Kluyveromyces Marxianus NRRL Y-7571
Yacon is a perennial plant native of the Andesthat stores inulin-type polyfructans as reserve carbohydrates in its tubers. The enzymes that hydrolyze the inulin β-2,1 fructose linkages are called inulinases and are much...
Mutational Analysis of DNASE I Gene in Diabetic Patients
The main aim is to analyze the mutations of DNASE I gene in diabetic patients.A total of 120 diabetes patients and 120 controls were sampled. The total number of male diabetic patients included in the study was 79 (66%)...
Review on Dermatophytes Highlighting the Status at National and International Level: A Critical Appraisal.
Dermatophytes as the name suggest are the fungus that feed on skin. The chief source of their growth is keratin which is widely available in skin, nails and hairs and causes disease in animals and humans due to their abi...
Datura leaf curl betastellite associated for the first time with Datura inoxia leaf curl syndrome in India.
Begomovirus is one of the largest group of Bemisia tabaci transmitted plant viruses (family Geminiviridae) containing single-stranded circular DNA that encapsidated in geminate particles and prevalent in the tropical and...
A graph-theoretic model of single point mutations in the cystic fibrosis transmembrane conductance regulator
Cystic fibrosis is one of the most prevalent inherited diseases. This disease is caused by a mutation in a membrane protein, the cystic fibrosis transmembrane conductance regulator (CFTR). CFTR is known to function as a...