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 97
- Downloads 0
Strategies and Approaches for Antibiotic Yield Improvement in Actinomycetes
Actinomycetes represent a valuable source of commercially significant antibiotics. However, the yields of antibiotics in wild Actinomycetes usually could not meet the commercial needs and so antibiotics yield improvement...
Reuse of Residual Biomass of Cellulose Industry for Second Generation Bioethanol Production
This study aimed at evaluating the potential of pulp mill residue (PMR) as a feedstock for ethanol production. The simultaneous saccharification and fermentation (SSF) process was operated using 8 gL -1 of a commercial s...
Involvement of the Selenium Level in Plasma Glutathione Peroxidase Activity in Newly Diagnosed Patients with Graves Disease and Hashimoto Thyroiditis
Disturbed antioxidant enzymes activities was known to be associated to Auto-Immune Thyroid Diseases (AITD), but it is noteworthy that the selenoenzyme glutathione peroxidase (GPx) showed the lowest activity and a seleniu...
PGPR Induced Differencial Expression of Defence Enzymes Regulating Resistance Against Colletotrichum Capsici in Turmeric
Plant growth promoting Bacillus amyloliquefaciens strain BaTNAU5 and Pseudomonas fluorescens strain Pf3TNAU were evaluated in turmeric for induction defence by differential expression of defence enzymes against Colletotr...
AIRLIFT BIOREACTOR FLUID-DYNAMIC CHARACTERIZATIONFOR THE CULTIVATION OF SHEAR STRESS SENSITIVEMICROORGANISMS
Airlift bioreactors are considered very efficient for aerobic cultivation of microorganisms. In particular, for the cultivation of filamentous fungi, where low shear rates may be required. In this context, the main aim o...