近日，湖南大学白玉罡团队实现了利用隔室化大肠杆菌体在真核细胞中建立人工亚细胞细胞器，助力人工金属酶介导的非生物催化。相关论文于2025年4月24日发表在《美国化学会杂志》上。

人工金属酶（ArMs）因其多方面的优势而成为介导细胞内非生物转化的优秀工具，包括其通过定向进化的适应性和作为全细胞催化剂的可用性。然而，由于蛋白质纯化和递送、金属酶稳定性和复杂催化剂制备方面的问题，ArMs作为外源性试剂在真核系统中的应用仍然具有挑战性。 

研究组提出了一种受自然界内共生过程启发的方法，可以直接使用细菌细胞内的ArMs，这些ArMs在真核细胞中作为基于全细胞的催化平台表达。这种方法利用HaloTag-SNAPTag融合蛋白作为ArM支架，该支架经过液-液相分离，在大肠杆菌中为由同一融合蛋白产生的不同ArM形成避难所。 

然后对这种分隔的大肠杆菌进行消毒，并赋予其聚合物装饰的细胞渗透性，使其可以进入真核细胞并作为人工亚细胞细胞器工作，利用体内保护良好的ArMs介导非生物转化。研究组进一步证明了这一策略在概念验证演示中的治疗应用潜力，表明这些封装的ArM可以通过在活细胞和动物中激活前药，成为消除细胞内细菌病原体和治疗癌症的可行选择。很可能，这一策略将为扩大ArM在化学生物学和生物医学中的应用提供一条不同的途径。

附：英文原文

Title: Creation of Artificial Subcellular Organelles Using Compartmentalized Escherichia coli Bodies for Artificial Metalloenzyme-Mediated Abiotic Catalysis in Eukaryotic Cells

Author: Tong Wu, Yating Fei, Yingjiao Deng, Xianhui Chen, Yuli Duan, Ying Liu, Yugang Bai

Issue&Volume: April 24, 2025

Abstract: Artificial metalloenzymes (ArMs) stand out as excellent tools mediating intracellular abiotic transformations due to their multifaceted advantages, including their adaptability through directed evolution and availability as whole-cell catalysts. However, the applications of ArMs as exogenous agents in eukaryotic systems remain challenging due to issues with protein purification and delivery, metalloenzyme stability, and complex catalyst preparation. In this article, we present a method inspired by nature’s endosymbiotic process, enabling the direct use of ArMs residing within the bacterial cells that express them as whole-cell-based catalytic platforms in eukaryotic cells. This approach utilizes HaloTag-SNAPTag fusion protein as the ArM scaffold, which undergoes liquid–liquid phase separation to form sanctuaries in Escherichia coli for different ArMs created from the same fusion protein. Such compartmentalized E. coli are then sterilized and granted cell permeability with polymer decoration so that they may enter eukaryotic cells and work as artificial subcellular organelles, mediating abiotic transformations using those well-protected ArMs residing within. We further demonstrate the potential of this strategy in therapeutic applications in proof-of-concept demonstrations, by showing that these encapsulated ArMs can be viable options for intracellular bacterial pathogen elimination and cancer therapy through prodrug activation in live cells and animals. Likely, this strategy will suggest a different pathway for expanding ArM applications in chemical biology and biomedicine.

DOI: 10.1021/jacs.5c00473

Source: https://pubs.acs.org/doi/abs/10.1021/jacs.5c00473