Biotechnology Branch , US Army Research Laboratory , 2800 Powder Mill Road , Adelphi , Maryland 20783 , United States.
General Technical Services , 1451 Route 34 South , Wall Township , New Jersey 07727 , United States.
Langmuir. 2018 May 22;34(20):5837-5848. doi: 10.1021/acs.langmuir.8b00114. Epub 2018 May 2.
In this study, we investigated the preparation of living bacteria-nanoparticle hybrids mediated by surface-displayed peptides. The assembly of metallic nanoparticles on living bacteria has been achieved under mild conditions utilizing metal-peptide interactions, whereas the viability of the bacterial cells was greatly preserved. Escherichia coli was engineered with inducible gene circuits to control the display of peptides with desired sequences. Several designed peptide sequences as well as known gold-binding peptides were expressed on the cell surface using enhanced circularly permuted outer membrane protein X (eCPX) scaffolds. Driven by metal-peptide affinity, "biofriendly" citrate-stabilized gold nanoparticles were self-assembled onto the surface of bacteria with displayed peptides, which required overcoming the repulsive force between negatively charged nanoparticles and negatively charged cells. The bacteria/Au nanoparticle hybrids were highly viable and maintained the ability to grow and divide, which is a crucial step toward the creation of living material systems. Further activity and preservation of the bacterial hybrid assembly was demonstrated. The method described herein enables the conjugation of bacterial surfaces with diverse metal-rich nanoparticles in an inducible, and therefore easily controlled, manner. The expressed peptide sequences can be easily modified to alter the binding affinity and specificity for a wide variety of materials to form on-demand, high-density living biohybrids.
在这项研究中,我们研究了通过表面展示肽介导的活细菌-纳米颗粒杂化的制备。利用金属-肽相互作用,在温和条件下实现了金属纳米颗粒在活细菌上的组装,而细菌细胞的活力得到了很好的保留。大肠杆菌被设计成带有诱导型基因回路,以控制具有所需序列的肽的展示。使用增强型循环截短的外膜蛋白 X(eCPX)支架,在细胞表面表达了几种设计的肽序列和已知的金结合肽。受金属-肽亲和力的驱动,“生物友好”的柠檬酸稳定的金纳米颗粒自组装到带有展示肽的细菌表面上,这需要克服带负电荷的纳米颗粒和带负电荷的细胞之间的排斥力。细菌/Au 纳米颗粒杂化物具有很高的活力,并保持生长和分裂的能力,这是朝着创建活材料系统迈出的关键一步。进一步展示了细菌杂化组装的活性和保留。本文描述的方法能够以诱导的方式(因此可以轻松控制)将细菌表面与各种富含金属的纳米颗粒偶联。表达的肽序列可以很容易地修改,以改变与各种材料的结合亲和力和特异性,从而按需形成高密度的活生物杂化物。