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首页> 外文期刊>Biotechnology Progress >Recombinant peptide fusion construction for protein-templated catalytic palladium nanoparticles
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Recombinant peptide fusion construction for protein-templated catalytic palladium nanoparticles

机译：蛋白质模板催化钯纳米粒子的重组肽融合结构

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Although peptide-enabled synthesis of nanostructures has garnered considerable interest for use in catalytic applications, it has so far been achieved mostly via Fmoc based solid phase peptide synthesis. Consequently, the potential of longer peptides in nanoparticle synthesis have not been explored largely due to the complexities and economic constraints of this chemical synthesis route. This study examines the potential of a 45-amino acid long peptide expressed as fusion to green fluorescence protein (GFPuv) in Escherichia coli for use in palladium nanoparticle synthesis. Fed-batch fermentation with E. coli harboring an arabinose-inducible plasmid produced a product containing three copies of Pd4 peptide fused to N-terminus of GFPuv ((Pd4)(3)-GFPuv). Using the intrinsic fluorescence of GFPuv, expression and enrichment of the fusion product was easily monitored. Crude lysate, desalted lysate, and an ion-exchange enriched fraction containing (Pd4)(3)-GFPuv were used to test the hypothesis that high purity of the biologic material used as the nanoparticle synthesis template may not be necessary. Nanoparticles were characterized using a variety of material science techniques and used to catalyze a model Suzuki-Miyaura coupling reaction. Results demonstrated that palladium nanoparticles can be synthesized using the soluble cell extract containing (Pd4)(3)-GFPuv without extensive purification or cleavage steps, and as a catalyst the crude mixture is functional.
机译：尽管使能肽的合成纳米结构的合成已经在催化应用中使用了相当大的兴趣，但到目前为止已经通过FMOC基固相肽合成来实现。因此，由于该化学合成途径的复杂性和经济限制，纳米粒子合成中较长肽的潜力尚未探讨。本研究检测了45氨基酸长肽作为融合到绿色荧光蛋白（GFPUV）的潜力，用于在大肠杆菌中用于钯纳米粒子合成。含有阿拉伯糖诱导型质粒的大肠杆菌的喂养批量发酵产生了含有三个PD4肽拷贝的产物，融合到GFPUV（（PD4）（3）-GFPUV）的N-末端。使用GFPUV的内在荧光，容易监测融合产物的表达和富集。粗裂解物，脱盐裂解物和含有（PD4）（3）-GFPUV的离子交换级分，用于测试作为纳米颗粒合成模板的高纯度的高纯度可能不是必需的。使用各种材料科学技术表征纳米颗粒，并用于催化模型铃木宫偶联偶联反应。结果表明，钯纳米颗粒可以使用含有（PD4）（3）-GFPUV的可溶性细胞萃取物而无需广泛的纯化或切割步骤，并且作为催化剂，粗混合物是功能性的。

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