College of Life Science and Technology, Beijing University of Chemical Technology, 100029, Beijing, China.
Protein J. 2010 Oct;29(7):516-23. doi: 10.1007/s10930-010-9282-0.
Inorganic-binding peptides termed as genetically engineered polypeptides for inorganics (GEPIs), are small peptide sequences selected via combinatorial biology-based protocols of phage or cell surface display technologies. Recent advances in nanotechnology and molecular biology allow the engineering of these peptides with specific affinity to inorganics, often used as molecular linkers or assemblers, to facilitate materials synthesis, which provides a new insight into the material science and engineering field. As a case study on this biomimetic application, here we report a novel biosynthetic ZnO binding protein and its application in promoting bio-inorganic materials synthesis. In brief, the gene encoding a ZnO binding peptide(ZBP) was genetically fused with His(6)-tag and GST-tag using E.coli expression vector pET-28a (+) and pGEX-4T-3. The recombinant protein GST-His-ZBP was expressed, purified with Ni-NTA system, identified by SDS-PAGE electrophoresis and Western blot analysis and confirmed by liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) analysis. Affinity adsorption test demonstrated that the fusion protein had a specific avidity for ZnO nanoparticles (NPs). Results from the bio-inorganic synthesis experiment indicated that the new protein played a promoting part in grain refinement and accelerated precipitation during the formation of the ultra-fine precursor powders in the Zn(OH)(2) sol. X-ray diffraction (XRD) analysis on the final products after calcining the precursor powders showed that hexagonal wurtzite ZnO crystals were obtained. Our work suggested a novel approach to the application about the organic-inorganic interactions.
无机结合肽被称为用于无机物的基因工程多肽(GEPI),是通过噬菌体或细胞表面展示技术的组合生物学方法选择的小肽序列。纳米技术和分子生物学的最新进展允许这些肽具有与无机物的特定亲和力,通常用作分子接头或组装体,以促进材料合成,这为材料科学和工程领域提供了新的见解。作为这种仿生应用的案例研究,我们在这里报告了一种新型的生物合成 ZnO 结合蛋白及其在促进生物-无机材料合成中的应用。简而言之,编码 ZnO 结合肽(ZBP)的基因使用大肠杆菌表达载体 pET-28a(+)和 pGEX-4T-3 通过基因融合与 His(6)-标签和 GST-标签融合。表达重组蛋白 GST-His-ZBP,用 Ni-NTA 系统纯化,通过 SDS-PAGE 电泳和 Western blot 分析鉴定,并通过液相色谱-质谱/质谱(LC-MS/MS)分析确认。亲和吸附试验表明,融合蛋白对 ZnO 纳米颗粒(NPs)具有特异性亲和力。生物-无机合成实验结果表明,该新蛋白在 Zn(OH)(2)溶胶中超细前体粉末形成过程中对晶粒细化和加速沉淀起促进作用。对前体粉末煅烧后的最终产物进行 X 射线衍射(XRD)分析表明,得到了六方纤锌矿 ZnO 晶体。我们的工作为有机-无机相互作用的应用提供了一种新方法。