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Mol. Cells 2011; 32(3): 257-264

Published online September 30, 2011

https://doi.org/10.1007/s10059-011-1047-x

© The Korean Society for Molecular and Cellular Biology

Engineering of 2-Cys Peroxiredoxin for Enhanced Stress-Tolerance

Byung Chull An, Seung Sik Lee, Jae Taek Lee, Sung Hyun Hong, Seung Gon Wi1, and Byung Yeoup Chung*

Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute, Jeongeup 580-185, Korea, 1Bio-Energy Research Institute, Chonnam National University, Gwangju 500-757, Korea

Correspondence to : *Correspondence: bychung@kaeri.re.kr

Received: March 11, 2011; Revised: June 13, 2011; Accepted: June 13, 2011

Abstract

A typical 2-cysteine peroxiredoxin (2-Cys Prx)-like protein (PpPrx) that alternatively acts as a peroxidase or a mole-cular chaperone in Pseudomonas putida KT2440 was previously characterized. The dual functions of PpPrx are regulated by the existence of an additional Cys112 between the active Cys51 and Cys171 residues. In the present study, additional Cys residues (Cys31, Cys112, and Cys192) were added to PpPrx variants to improve their enzymatic func-tion. The optimal position of the additional Cys residues for the dual functionality was assessed. The peroxidase activities of the S31C and Y192C mutants were increased 3- to 4-fold compared to the wild-type, while the chaperone activity was maintained at > 66% of PpPrx. To investigate whether optimization of the dual functions could enhance stress-tolerance in vivo, a complementation study was performed. The S31C and Y192C mutants showed a much greater tolerance than other variants under a complex condition of heat and oxidative stresses. The optimized dual functions of PpPrx could be adapted for use in bioen-gineering systems and industries, such as to develop or-ganisms that are more resistant to extreme environments.

Keywords dual function engineering, dual functions, extreme stress, peroxiredoxin, site-directed mutagenesis

Article

Research Article

Mol. Cells 2011; 32(3): 257-264

Published online September 30, 2011 https://doi.org/10.1007/s10059-011-1047-x

Copyright © The Korean Society for Molecular and Cellular Biology.

Engineering of 2-Cys Peroxiredoxin for Enhanced Stress-Tolerance

Byung Chull An, Seung Sik Lee, Jae Taek Lee, Sung Hyun Hong, Seung Gon Wi1, and Byung Yeoup Chung*

Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute, Jeongeup 580-185, Korea, 1Bio-Energy Research Institute, Chonnam National University, Gwangju 500-757, Korea

Correspondence to:*Correspondence: bychung@kaeri.re.kr

Received: March 11, 2011; Revised: June 13, 2011; Accepted: June 13, 2011

Abstract

A typical 2-cysteine peroxiredoxin (2-Cys Prx)-like protein (PpPrx) that alternatively acts as a peroxidase or a mole-cular chaperone in Pseudomonas putida KT2440 was previously characterized. The dual functions of PpPrx are regulated by the existence of an additional Cys112 between the active Cys51 and Cys171 residues. In the present study, additional Cys residues (Cys31, Cys112, and Cys192) were added to PpPrx variants to improve their enzymatic func-tion. The optimal position of the additional Cys residues for the dual functionality was assessed. The peroxidase activities of the S31C and Y192C mutants were increased 3- to 4-fold compared to the wild-type, while the chaperone activity was maintained at > 66% of PpPrx. To investigate whether optimization of the dual functions could enhance stress-tolerance in vivo, a complementation study was performed. The S31C and Y192C mutants showed a much greater tolerance than other variants under a complex condition of heat and oxidative stresses. The optimized dual functions of PpPrx could be adapted for use in bioen-gineering systems and industries, such as to develop or-ganisms that are more resistant to extreme environments.

Keywords: dual function engineering, dual functions, extreme stress, peroxiredoxin, site-directed mutagenesis

Mol. Cells
Nov 30, 2022 Vol.45 No.11, pp. 763~867
COVER PICTURE
Naive (cyan) and axotomized (magenta) retinal ganglion cell axons in Xenopus tropicalis (Choi et al., pp. 846-854).

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