Wu Sha, Tian Jiesheng, Xue Xianle, Tang Zongwen, Huang Zekai, Hammock Bruce D, Morisseau Christophe, Li Qing X, Xu Ting
China Agricultural University.
University of California.
Res Sq. 2023 Mar 13:rs.3.rs-2630343. doi: 10.21203/rs.3.rs-2630343/v1.
Protein downstream processing remains a challenge in protein production, especially in low yields of products, in spite of ensuring effective disruption of cell and separation of target proteins. It is complicated, expensive and time-consuming. Here, we report a novel nano-bio-purification system for producing recombinant proteins of interest with automatic purification from engineered bacteria.
This system employed a complete genetic engineering downstream processing platform for proteins at low expression levels, referred to as a genetically encoded magnetic platform (GEMP). GEMP consists of four elements as follows. (1) A truncated phage lambda lysis cassette (RRz/Rz1) is controllable for lysis of MSR-1 (host cell). (2) A surface-expressed nuclease (NucA) is to reduce viscosity of homogenate by hydrolyzing long chain nucleic acids. (3) A bacteriogenic magnetic nanoparticle, known as magnetosome, allows an easy separation system in a magnetic field. (4) An intein realizes abscission of products (nanobodies against tetrabromobisphenol A) from magnetosome.
In this work, removal of most impurities greatly simplified the subsequent purification procedure. The system also facilitated the bioproduction of nanomaterials. The developed platform can substantially simplify industrial protein production and reduce its cost.
尽管能够确保有效地破碎细胞并分离目标蛋白，但蛋白质下游加工仍然是蛋白质生产中的一个挑战，尤其是在产品产量较低的情况下。它复杂、昂贵且耗时。在此，我们报道了一种新型的纳米生物纯化系统，用于从工程细菌中自动纯化生产感兴趣的重组蛋白。
该系统采用了一个完整的针对低表达水平蛋白质的基因工程下游加工平台，称为基因编码磁性平台（GEMP）。GEMP由以下四个元件组成。（1）一个截短的噬菌体λ裂解盒（RRz/Rz1）可控制MSR-1（宿主细胞）的裂解。（2）一个表面表达的核酸酶（NucA），通过水解长链核酸来降低匀浆的粘度。（3）一种细菌源性磁性纳米颗粒，即磁小体，可在磁场中实现简易的分离系统。（4）一个内含肽实现了产物（抗四溴双酚A纳米抗体）与磁小体的分离。
在这项工作中，去除大多数杂质极大地简化了后续的纯化过程。该系统还促进了纳米材料的生物生产。所开发的平台可大幅简化工业蛋白质生产并降低其成本。
