Publication: Improving enzymatic activities and thermostability of a tri-functional enzyme with SOD, catalase and cell-permeable activities
Issued Date
2017-04-10
Resource Type
ISSN
18734863
01681656
01681656
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2-s2.0-85014883108
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Mahidol University
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SCOPUS
Bibliographic Citation
Journal of Biotechnology. Vol.247, (2017), 50-59
Suggested Citation
Piriya Luangwattananun, Warawan Eiamphungporn, Napat Songtawee, Leif Bülow, Chartchalerm Isarankura Na Ayudhya, Virapong Prachayasittikul, Sakda Yainoy Improving enzymatic activities and thermostability of a tri-functional enzyme with SOD, catalase and cell-permeable activities. Journal of Biotechnology. Vol.247, (2017), 50-59. doi:10.1016/j.jbiotec.2017.03.001 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/41922
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Title
Improving enzymatic activities and thermostability of a tri-functional enzyme with SOD, catalase and cell-permeable activities
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Abstract
© 2017 Elsevier B.V. Synergistic action of major antioxidant enzymes, e.g., superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) is known to be more effective than the action of any single enzyme. Recently, we have engineered a tri-functional enzyme, 6His-MnSOD-TAT/CAT-MnSOD (M-TAT/CM), with SOD, CAT and cell-permeable activities. The protein actively internalized into the cells and showed superior protection against oxidative stress-induced cell death over native enzymes fused with TAT. To improve its molecular size, enzymatic activity and stability, in this study, MnSOD portions of the engineered protein were replaced by CuZnSOD, which is the smallest and the most heat resistant SOD isoform. The newly engineered protein, CAT-CuZnSOD/6His-CuZnSOD-TAT (CS/S-TAT), had a 42% reduction in molecular size and an increase in SOD and CAT activities by 22% and 99%, respectively. After incubation at 70 °C for 10 min, the CS/S-TAT retained residual SOD activity up to 54% while SOD activity of the M-TAT/CM was completely abolished. Moreover, the protein exhibited a 5-fold improvement in half-life at 70 °C. Thus, this work provides insights into the design and synthesis of a smaller but much more stable multifunctional antioxidant enzyme with ability to enter mammalian cells for further application as protective/therapeutic agent against oxidative stress-related conditions.