Extended Data Fig. 6: In vitro performance evaluation of HS–ArMs in hostile environments.
From: Artificial metalloenzyme assembly in cellular compartments for enhanced catalysis
a, Performance comparison of small molecular ruthenium complex (HG–Ru–Cl) and Ru–HS–ArMs, when mediating the fluorogenic RCM reaction of HVPA, in PBS or wild-type E. coli cell lysate. b, Performance comparison of small molecular copper complex (Cu–BTTAA) and Cu–HS–ArMs in mediating the fluorogenic CuAAC between AzHCou and 4-EA, in PBS or wild-type E. coli cell lysate. For a and b, the reaction schemes are provided on the bottom left, and the yields of the products were calculated based on the detected fluorescence, using the solution of the pure product at the concentration of presumed 100% yield. The product solutions were prepared according to the media and reagents used for each experiment to eliminate the impact of the changes in the product’s fluorescence intensity in different environments. For both experiments, standalone small molecular catalysts showed significantly hindered activity in wild-type E. coli lysate, while the assembled HS–ArMs showed much higher resistance despite slower kinetics. c, Evaluation of the GSH resistance of HG–Ru–Cl (small-molecule catalyst) and Ru–HS–ArM (aggregating macromolecular catalyst). HG–Ru–Cl showed completely no resistance toward GSH, while the resistance to GSH increased significantly for Ru–HS–ArM in an in vitro phase-separating condition. All data are presented as mean ± s.d. (n = 3 independent experiments). ***t ≤ 0.001, **t ≤ 0.01 and *t ≤ 0.05 in two-tailed Student’s t-test.
