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利用缢蛏(Sinonovacula constricta)重组铁蛋白富集Fe~(3+)和Mn~(2+)制备重组Fe-铁蛋白和Mn-铁蛋白,通过扫描电镜、X射线能量色散能谱仪(EDS)和MALDI TOF/TOF质谱系统测定蛋白的表面形貌变化、金属元素的能量变化和肽段分子量。利用综合物性测量系统(PPMS)测定蛋白纳米颗粒在室温300K,外加磁场3T下的磁学性质变化。结果显示,重组Fe-铁蛋白和Mn-铁蛋白与空白相比,表面形貌发生明显变化,Fe-铁蛋白仍为小球状,Mn-铁蛋白聚集体呈片层花球状,Cd-铁蛋白聚集体呈小圆球状,Mn-铁蛋白富集Cd~(2+)后呈片层花瓣散落状。Fe-铁蛋白和Mn-铁蛋白分别检测出相应金属元素且都有其特征能量态。两种重组蛋白的肽谱图与空白组相比,除铁蛋白保守肽段外还出现各自的特征肽段,推测与铁蛋白对Fe~(3+)和Mn~(2+)的富集功能密切相关。Fe-铁蛋白和Mn-铁蛋白纳米颗粒磁滞回线形状与铁蛋白空白组基本相同,呈顺磁性特征,磁性强度随Fe~(3+)和Mn~(2+)富集量的增加而增大。通过比较Fe-铁蛋白和Mn-铁蛋白与空白组在富集Hg~(2+)、As O43–和Cd~(2+)三种重金属离子方面能力的差异,发现Fe-铁蛋白对Hg~(2+)、AsO_4~(3–)和Cd~(2+)三种重金属的富集能力是空白组的2.4倍、1.7倍和3.7倍。Mn-铁蛋白对Hg~(2+)、AsO_4~(3–)和Cd~(2+)三种重金属离子在相同条件下的富集能力也有明显提高,分别为铁蛋白空白组的1.8倍、3.0倍和4.6倍。本研究结果为Fe-铁蛋白和Mn-铁蛋白在重金属污染治理方面的应用提供了数据参考。
The recombinant Fe-ferritin and Mn-ferritin were prepared by enrichment of Fe3 + and Mn2 + by Sinonovacula constricta recombinant ferritin. The structure of the product was characterized by scanning electron microscopy, energy dispersive X-ray spectroscopy (EDS) And MALDI TOF / TOF mass spectrometry system to determine protein surface topography, energy changes of metal elements and peptide molecular weight. The change of magnetic properties of protein nanoparticles at 300 K at room temperature and 3 T under the applied magnetic field was measured by a comprehensive physical property measurement system (PPMS). The results showed that the surface morphology of the recombinant Fe-ferritin and Mn-ferritin changed significantly compared with the blank, the Fe-ferritin was still globular, the Mn-ferritin aggregates were lamellar globular, Cd-ferritin Aggregates were spheroidal, Mn-ferritin enriched Cd ~ (2+) after the sheet petal scattered. Fe-ferritin and Mn-ferritin detect the corresponding metal elements and have their characteristic energy states. Compared with the blank group, the peptide profiles of the two recombinant proteins also showed their own characteristic peptides in addition to the conserved peptides of ferritin, suggesting that they are related to the enrichment of Fe 3+ and Mn 2+ by ferritin Function is closely related. The hysteresis loop shape of Fe-ferritin and Mn-ferritin nanoparticles is basically the same as that of the ferritin blank group, showing paramagnetic characteristics. The magnetic strength increases with the increase of Fe ~ (3+) and Mn ~ (2+) Increase. By comparing the ability of Fe-ferritin and Mn-ferritin with blank group in enriching Hg 2+, As O43- and Cd 2+, it was found that Hg The enrichment abilities of ~ (2 +), AsO_4 ~ (3-) and Cd ~ (2+) were 2.4 times, 1.7 times and 3.7 times that of blank control group. Enrichment ability of Mn-ferritin on Hg 2+, AsO 4 3- and Cd 2+ also increased significantly under the same conditions, which were 1.8 times that of ferritin blank group , 3.0 times and 4.6 times. The results of this study provide data reference for the application of Fe-ferritin and Mn-ferritin in the treatment of heavy metal pollution.