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具有超氧化物清除活性的锰-萨罗芬配合物的制备与表征

Preparation and characterization of Mn-salophen complex with superoxide scavenging activity.

作者信息

Liu Z X, Robinson G B, Gregory E M

机构信息

Department of Biochemistry and Anaerobic Microbiology, Virginia Polytechnic Institute, Blacksburg 24061-0308.

出版信息

Arch Biochem Biophys. 1994 Nov 15;315(1):74-81. doi: 10.1006/abbi.1994.1473.

Abstract

Mn(III)-salophen complex with superoxide scavenging activity was prepared from manganese(III) acetate dihydrate and salophen in ethanol. Visible absorption spectrum of the red-brown complex exhibits a shoulder at 430 nm which was absent with either salophen or manganic acetate alone. Titration of salophen with manganese(III) is consistent with a 1:1 Mn to salophen stoichiometry of the complex based on changes in the absorbance at 500 nm or of superoxide scavenging activity. The superoxide dismutase (SOD)-like activity of the complex in the xanthine-xanthine oxidase/cytochrome c assay was 1450 units/mg salophen. The SOD activity of the complex was suppressed 50% in the presence of EDTA (1 mM), but was not altered in the presence of bovine serum albumin (1 mg/ml) or crude protein extract of Escherichia coli QC779 sodA-sodB- (1 mg/ml). E. coli QC779 sodA-sodB- grew scantily after an 8-h lag phase in aerobic M63 glucose minimal medium. The aerobic growth of the E. coli SOD double mutant in glucose minimal medium was greatly enhanced in the presence of 5 or 10 microM Mn-salophen complex compared to that of control after 24 h incubation. Mn-desferal green complex (10 microM) and pink complex (5 microM) also increased growth rate of E. coli QC779 sodA-sodB- but to a lesser extent than Mn-salophen complex. However, the growth was completely inhibited by 50 microM Mn-salophen complex, 100 microM Mn-desferal green complex, or 10 microM Mn-desferal pink complex.

摘要

由二水合乙酸锰和萨罗芬在乙醇中制备了具有超氧化物清除活性的Mn(III)-萨罗芬配合物。红棕色配合物的可见吸收光谱在430 nm处有一个肩峰,单独的萨罗芬或乙酸锰都没有这个峰。基于500 nm处吸光度的变化或超氧化物清除活性,用锰(III)滴定萨罗芬与该配合物中Mn与萨罗芬的化学计量比为1:1一致。在黄嘌呤-黄嘌呤氧化酶/细胞色素c测定中,该配合物的超氧化物歧化酶(SOD)样活性为1450单位/毫克萨罗芬。在存在EDTA(1 mM)时,该配合物的SOD活性被抑制50%,但在存在牛血清白蛋白(1 mg/ml)或大肠杆菌QC779 sodA-sodB-的粗蛋白提取物(1 mg/ml)时没有改变。大肠杆菌QC779 sodA-sodB-在有氧M63葡萄糖基本培养基中经过8小时的延迟期后生长稀疏。与对照相比,在5或10 microM Mn-萨罗芬配合物存在下,大肠杆菌SOD双突变体在葡萄糖基本培养基中的有氧生长在孵育24小时后得到极大增强。Mn-去铁胺绿色配合物(10 microM)和粉红色配合物(5 microM)也提高了大肠杆菌QC779 sodA-sodB-的生长速率,但程度小于Mn-萨罗芬配合物。然而,50 microM Mn-萨罗芬配合物、100 microM Mn-去铁胺绿色配合物或10 microM Mn-去铁胺粉红色配合物完全抑制了生长。

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