Engineering of 2-Cys Peroxiredoxin for Enhanced Stress-Tolerance
Engineering of 2-Cys Peroxiredoxin for Enhanced Stress-Tolerance
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DOI：
10.1007/s10059-011-1047-x
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发表时间：
2011-09-01
影响因子：
3.8
通讯作者：
Chung, Byung Yeoup
中科院分区：
文献类型：
--
作者：
An, Byung Chull;Lee, Seung Sik;Chung, Byung Yeoup
一个典型的2-半胱氨酸过氧化物氧还蛋白（2-Cys Prx）的蛋白质（PpPrx），可作为过氧化物酶或分子伴侣在恶臭假单胞菌KT 2440先前的特点。PpPrx的双重功能受活性Cys（51）和Cys（171）残基之间存在的额外Cys（112）的调节。在本研究中，将额外的Cys残基（Cys（31）、Cys（112）和Cys（192））添加到PpPrx变体中以改善其酶功能。评估了额外的Cys残基用于双重功能的最佳位置。与野生型相比，S31 C和Y192 C突变体的过氧化物酶活性增加了3至4倍，而伴侣蛋白活性保持在PpPrx的> 66%。为了研究双重功能的优化是否可以增强体内的胁迫耐受性，进行了互补研究。S31 C和Y192 C突变体在复杂的热和氧化应激条件下表现出比其他变体更大的耐受性。PpPrx的优化双重功能可以适用于生物工程系统和工业，例如开发对极端环境更具抵抗力的生物体。
A typical 2-cysteine peroxiredoxin (2-Cys Prx)-like protein (PpPrx) that alternatively acts as a peroxidase or a molecular chaperone in Pseudomonas putida KT2440 was previously characterized. The dual functions of PpPrx are regulated by the existence of an additional Cys(112) between the active Cys(51) and Cys(171) residues. In the present study, additional Cys residues (Cys(31), Cys(112), and Cys(192)) were added to PpPrx variants to improve their enzymatic function. 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 bioengineering systems and industries, such as to develop organisms that are more resistant to extreme environments.