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Cheng Wang,Gang-Lin Yan,Shao-Wu Lü,Chun-Hong Sui,Yang Zhao,Ya-Wei Xu,Gang Zhao,Jun-jie Xu,Ping-Sheng Gong,Gui-Min Luo,Ying Mu 한국생물공학회 2013 Biotechnology and Bioprocess Engineering Vol.18 No.1
Glutathione peroxidase (GPX) is one of the important members of the antioxidant enzyme family. It can catalyze the reduction of hydroperoxides with glutathione to protect cells against oxidative damage. Single-chain variable fragment (scFv) can be converted into seleniumcontaining single-chain variable fragment (Se-scFv) by chemical modification of the hydroxyl groups in scFv, thus Se-scFv possesses GPX activity and becomes a prodrug. To improve the expression of scFv and simplify its purification steps, Single-protein production (SPP) system was used to express scFv and chemical modification was used to synthesize Se-scFv. Therefore, we must construct a new scFv-WCD1-lessACA gene, which can express its mRNA not containing any ACA sequences and express its amino acid sequence of target protein (scFv) being same to scFv-WCD1. In this way, the scFv-WCD1-lessACA can be only expressed in SPP system and no other background proteins in the cells could be expressed. The expression results showed that high level of scFv-WCD1-lessACA synthesis was at least sustained for 96 h in the virtual absence of background protein synthesis. Then, selenocysteine (Sec) was incorporated into the scFv-WCD1-lessACA by chemical modification and resulted in Se-scFv-WCD1-lessACA. The enzymatic characteristics of Se-scFv-WCD1-lessACA were determined. GPX activity was 2,563 U/μmol,its binding constant for GSH was 0.687 ×105/mol. Moreover,Se-scFv-WCD1-lessACA was confirmed to have a strong antioxidant ability to protect mitochondria against oxidative damage induced by Vc/Fe2+ (mitochondrial damage model),suggesting that Se-scFv-WCD1-lessACA has potential application for protection of mitochondrial damage induced by reactive oxygen species (ROS).
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Preparation of HMX by Catalytic Nitrolysis of DPT in AIL-N_2O_5-HNO_3 System
Zhi-yong He,Jun Luo,Chun-xu Lü,Ping Wang,Rong Xu,Jin-shan Li 대한화학회 2011 Bulletin of the Korean Chemical Society Vol.32 No.8
Direct nitrolysis of 3,7-dinitro-1,3,5,7-tetraazabicyclo[3,3,1]nonane (DPT) is a feasible way to synthesize HMX, and it has multiple practical applications. In this paper, a new nitrolysis process involving the use of an N_2O_5-HNO_3 system catalyzed by acidic ionic liquids (AILs) was developed. The results show that [Et_3NH]TsO was the best catalyst among the 28 AILs used and that HMX was formed at a higher yield of 61%, compared to 45% without any AIL. Moreover, with the addition of N_2O_5, the yield was further increased to a maximum value of 69%. The AILs were also efficiently recovered by simple extractions without any apparent loss of catalytic activity, even after five runs.
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