Multienzyme cascades enable the sequential synthesis of complex chemicals by combining multiple catalytic processes in one pot,offering considerable time and cost savings compared to a series of separate batch reactio...Multienzyme cascades enable the sequential synthesis of complex chemicals by combining multiple catalytic processes in one pot,offering considerable time and cost savings compared to a series of separate batch reactions.However,challenges related to coordination and regulatory interplay among multiple enzymes reduce the catalytic efficiency of such cascades.Herein,we genetically programmed a scaffold framework that selectively and orthogonally recruits enzymes as designed.The system was then used to generate multienzyme complexes of D-allulose 3-epimerase(DAE),ribitol dehydrogenase(RDH),and formate dehydrogenase(FDH)for rare sugar production.This scaffolded multienzymatic assembly achieves a 10.4-fold enhancement in the catalytic performance compared to its unassembled counterparts,obtaining allitol yield of more than 95%.Molecular dynamics simulations revealed that shorter distances between neighboring enzymes in scaffold-mounted complexes facilitated the transfer of reaction intermediates.A dual-module catalytic system incorporating(1)scaffold-bound complexes of DAE,RDH,and FDH and(2)scaffold-bound complexes of alcohol dehydrogenase and NADH oxidase expressed intracellularly in E.coli was used to synthesize D-allulose from D-fructose.This system synthesized 90.6%D-allulose from 300 g L^(−1)D-fructose,with a space-time yield of 13.6 g L^(−1)h^(−1).Our work demonstrates the programmability and versatility of scaffold-based strategies for the advancement of multienzyme cascades.展开更多
Background:Lipid metabolic reprogramming has been increasingly recognized as a key factor contributing to tumor immune evasion,therapeutic resistance,and plasticity,which collectively compromise the efficacy of target...Background:Lipid metabolic reprogramming has been increasingly recognized as a key factor contributing to tumor immune evasion,therapeutic resistance,and plasticity,which collectively compromise the efficacy of targeted radionuclide therapy(TRT).Overcoming the immunosuppressive and hypoxic tumor microenvironment(TME)while interfering with tumor lipid metabolism may offer a promising strategy to potentiate TRT outcomes.Methods:In this report,a radiopharmaceutical with multienzymatic catalysis activities is developed,wherein tumor cell membrane-coated manganese single-atom nanozymes(Mn/SAE@M)as supports deliver iodine-131(^(131)I)to the tumor.The Mn/SAE nanozyme core was synthesized in situ within hollow mesoporous zeolitic imidazolate frame-8(ZIF-8)nanoparticles,then coated with homologous tumor cell membranes for targeted delivery and subsequently labeled with^(131)I using the Chloramine-T method.A series of in vitro and in vivo experiments was performed in nonsmall cell lung cancer(NSCLC)models to evaluate therapeutic efficacy and immune activation.Results:131I-Mn/SAE@M exhibited efficient tumor targeting and internalization mediated by membrane camouflage.Within the TME,the radiopharmaceuticals initiated abundant oxygen(O_(2))release through catalase(CAT)-like catalysis,thereby mitigating a hypoxic microenvironment.In particular,it produced and enriched more reactive oxygen species(ROS)through oxidase(OXD)-,peroxidase(POD)-,and glutathione oxidase(GSHOx)-like catalytic processes.Importantly,^(131)I-Mn/SAE@M activated the cGAS-STING pathway,interfered with the lipid metabolic homeostasis of tumor cells,and induced ferroptosis,which is unraveled to take responsibility for the potentiated antitumor immunity.In bilateral NSCLC tumor-bearing mice,the treatment suppressed both the first and the second tumors,indicating the generation of systemic antitumor immune responses and immunological memory.Conclusion:Such SAE-based radiopharmaceuticals provide a candidate platform to elevate TRT efficiency,and the proof-of-concept rationale of disrupting lipid metabolic homeostasis through multienzyme-mimicking cascade reactions also provides a general avenue to improve TRT and synergistically magnify antitumor immunity.展开更多
There are several natural materials which have evolved functional gradients,ingeniously attaining maximal efficacy from limited components.Herein,we utilized the spatiotemporal distribution of initiator acetylacetone ...There are several natural materials which have evolved functional gradients,ingeniously attaining maximal efficacy from limited components.Herein,we utilized the spatiotemporal distribution of initiator acetylacetone to regulate the multienzyme polymerization and fabricate a chitosan-polymer hydrogel.The temporal priority order of acetylacetone was higher than phenolmodified chitosan by density functional theory calculation.The acetylacetone within the gelatin could gradually diffuse spatially into the chitosan hydrogel to fabricate the composite hydrogel with gradient network structure.The gradient hydrogel possessed a transferring topography from the two-dimensional pattern.A continuously decreased modulus along with acetylacetone diffusion was confirmed by atomic force microscope-based force mapping experiment.The water-retaining ability of various regions was confirmed by low-field nuclear magnetic resonance(NMR)and thermogravimetric analysis(TG)analysis,which led to the spontaneous actuation of gradient hydrogel with maximum 1821°/h curling speed and 227°curling angle.Consequently,the promising gradient hydrogels could be applied as intelligent actuators and flexible robots.展开更多
文摘Multienzyme cascades enable the sequential synthesis of complex chemicals by combining multiple catalytic processes in one pot,offering considerable time and cost savings compared to a series of separate batch reactions.However,challenges related to coordination and regulatory interplay among multiple enzymes reduce the catalytic efficiency of such cascades.Herein,we genetically programmed a scaffold framework that selectively and orthogonally recruits enzymes as designed.The system was then used to generate multienzyme complexes of D-allulose 3-epimerase(DAE),ribitol dehydrogenase(RDH),and formate dehydrogenase(FDH)for rare sugar production.This scaffolded multienzymatic assembly achieves a 10.4-fold enhancement in the catalytic performance compared to its unassembled counterparts,obtaining allitol yield of more than 95%.Molecular dynamics simulations revealed that shorter distances between neighboring enzymes in scaffold-mounted complexes facilitated the transfer of reaction intermediates.A dual-module catalytic system incorporating(1)scaffold-bound complexes of DAE,RDH,and FDH and(2)scaffold-bound complexes of alcohol dehydrogenase and NADH oxidase expressed intracellularly in E.coli was used to synthesize D-allulose from D-fructose.This system synthesized 90.6%D-allulose from 300 g L^(−1)D-fructose,with a space-time yield of 13.6 g L^(−1)h^(−1).Our work demonstrates the programmability and versatility of scaffold-based strategies for the advancement of multienzyme cascades.
基金supported by the National Natural Science Foundation of China(82272030,82022033,82071956)the Construction Project of Shanghai Key Laboratory of Molecular Imaging(18DZ2260400)the Sichuan Provincial Science and Technology Program(2024NSFJQ0048,2024NSFTD0024)。
文摘Background:Lipid metabolic reprogramming has been increasingly recognized as a key factor contributing to tumor immune evasion,therapeutic resistance,and plasticity,which collectively compromise the efficacy of targeted radionuclide therapy(TRT).Overcoming the immunosuppressive and hypoxic tumor microenvironment(TME)while interfering with tumor lipid metabolism may offer a promising strategy to potentiate TRT outcomes.Methods:In this report,a radiopharmaceutical with multienzymatic catalysis activities is developed,wherein tumor cell membrane-coated manganese single-atom nanozymes(Mn/SAE@M)as supports deliver iodine-131(^(131)I)to the tumor.The Mn/SAE nanozyme core was synthesized in situ within hollow mesoporous zeolitic imidazolate frame-8(ZIF-8)nanoparticles,then coated with homologous tumor cell membranes for targeted delivery and subsequently labeled with^(131)I using the Chloramine-T method.A series of in vitro and in vivo experiments was performed in nonsmall cell lung cancer(NSCLC)models to evaluate therapeutic efficacy and immune activation.Results:131I-Mn/SAE@M exhibited efficient tumor targeting and internalization mediated by membrane camouflage.Within the TME,the radiopharmaceuticals initiated abundant oxygen(O_(2))release through catalase(CAT)-like catalysis,thereby mitigating a hypoxic microenvironment.In particular,it produced and enriched more reactive oxygen species(ROS)through oxidase(OXD)-,peroxidase(POD)-,and glutathione oxidase(GSHOx)-like catalytic processes.Importantly,^(131)I-Mn/SAE@M activated the cGAS-STING pathway,interfered with the lipid metabolic homeostasis of tumor cells,and induced ferroptosis,which is unraveled to take responsibility for the potentiated antitumor immunity.In bilateral NSCLC tumor-bearing mice,the treatment suppressed both the first and the second tumors,indicating the generation of systemic antitumor immune responses and immunological memory.Conclusion:Such SAE-based radiopharmaceuticals provide a candidate platform to elevate TRT efficiency,and the proof-of-concept rationale of disrupting lipid metabolic homeostasis through multienzyme-mimicking cascade reactions also provides a general avenue to improve TRT and synergistically magnify antitumor immunity.
基金supported by the National Natural Science Foundation of China(51873156,51773155)the National Key Research and Development Program(2016YFA0100800)。
文摘There are several natural materials which have evolved functional gradients,ingeniously attaining maximal efficacy from limited components.Herein,we utilized the spatiotemporal distribution of initiator acetylacetone to regulate the multienzyme polymerization and fabricate a chitosan-polymer hydrogel.The temporal priority order of acetylacetone was higher than phenolmodified chitosan by density functional theory calculation.The acetylacetone within the gelatin could gradually diffuse spatially into the chitosan hydrogel to fabricate the composite hydrogel with gradient network structure.The gradient hydrogel possessed a transferring topography from the two-dimensional pattern.A continuously decreased modulus along with acetylacetone diffusion was confirmed by atomic force microscope-based force mapping experiment.The water-retaining ability of various regions was confirmed by low-field nuclear magnetic resonance(NMR)and thermogravimetric analysis(TG)analysis,which led to the spontaneous actuation of gradient hydrogel with maximum 1821°/h curling speed and 227°curling angle.Consequently,the promising gradient hydrogels could be applied as intelligent actuators and flexible robots.
