Microbial cultivation in microcompartments using water-in-oil(w/o)emulsion droplets has greatly advanced the study of strict anaerobic bacteria.However,a major challenge lies in the separation and recovery of the bact...Microbial cultivation in microcompartments using water-in-oil(w/o)emulsion droplets has greatly advanced the study of strict anaerobic bacteria.However,a major challenge lies in the separation and recovery of the bacterial liquid.In this study,we address this challenge by localizing strict anaerobes within w/o Pickering emulsion droplets stabilized by modified microgel particles.This approach demonstrates remarkable efficiency in bacterial microencapsulation,growth and separation.The microgel particles are modified in-situ with hydrophobic silica allowing for stabilization of w/o emulsions at the optimum cultivation temperature of 37℃ while the continuous oil phase prevents oxygen ingress thereby maximizing anaerobic fermentation.Additionally,a temperature decrease triggers the microgel particles to swell and become hydrophilic,causing an immediate inversion of the emulsion to oil-in-water type.This facilitates complete separation and collection of the bacterial suspension without any damage or loss.The intelligent phase inversion of the emulsion presents exciting possibilities for various applications in anaerobic fermentation,synthetic biology and biomanufacturing.展开更多
Chemical warfare agents represent a severe threat to mankind and their efficient decontamination is a global necessity.However,traditional disposal strategies have limitations,including high energy consumption,use of ...Chemical warfare agents represent a severe threat to mankind and their efficient decontamination is a global necessity.However,traditional disposal strategies have limitations,including high energy consumption,use of aggressive reagents and generation of toxic byproducts.Here,inspired by the compartmentalized architecture and detoxification mechanism of bacterial microcompartments,we constructed oil-in-water Pickering emulsion droplets stabilized by hydrogen-bonded organic framework immobilized cascade enzymes for decontaminating mustard gas simulant(2-chloroethyl ethyl sulfide,CEES)under sweet conditions.Two exemplified droplet systems were developed with two-enzyme(glucose oxidase/chloroperoxidase)and threeenzyme(invertase/glucose oxidase/chloroperoxidase)cascades,both achieving over 6-fold enhancement in decontamination efficiency compared with free enzymes and>99% selectivity towards non-toxic sulfoxide.We found that the favored mass transfer of sugars and CEES from their respective phases to approach the cascade enzymes located at the droplet surface and the facilitated substrate channeling between proximally immobilized enzymes were key factors in augmenting the decontamination efficacy.More importantly,the robustness of immobilized enzymes enabled easy reproduction of both the droplet formation and detoxification performance over 10 cycles,following long-term storage and in far-field locations.展开更多
基金financial support from the National Natural Science Foundation of China(grant no.22202084)Natural Science Foundation of Jiangsu Province(grant no.BK20221059)+1 种基金the Fundamental Research Funds for the Central Universities(grant no.JUSRP122017)the Research Matching Grant Scheme at CUHK(grant no.8601309).
摘要Microbial cultivation in microcompartments using water-in-oil(w/o)emulsion droplets has greatly advanced the study of strict anaerobic bacteria.However,a major challenge lies in the separation and recovery of the bacterial liquid.In this study,we address this challenge by localizing strict anaerobes within w/o Pickering emulsion droplets stabilized by modified microgel particles.This approach demonstrates remarkable efficiency in bacterial microencapsulation,growth and separation.The microgel particles are modified in-situ with hydrophobic silica allowing for stabilization of w/o emulsions at the optimum cultivation temperature of 37℃ while the continuous oil phase prevents oxygen ingress thereby maximizing anaerobic fermentation.Additionally,a temperature decrease triggers the microgel particles to swell and become hydrophilic,causing an immediate inversion of the emulsion to oil-in-water type.This facilitates complete separation and collection of the bacterial suspension without any damage or loss.The intelligent phase inversion of the emulsion presents exciting possibilities for various applications in anaerobic fermentation,synthetic biology and biomanufacturing.
基金supported by the National Key Research and Development Program of China(2020YFA0210800)the National Natural Science Foundation of China(22334004,22027805,22277011,22107019,22176035)the Major Project of Science and Technology of Fujian Province(2020HZ06006)。
摘要Chemical warfare agents represent a severe threat to mankind and their efficient decontamination is a global necessity.However,traditional disposal strategies have limitations,including high energy consumption,use of aggressive reagents and generation of toxic byproducts.Here,inspired by the compartmentalized architecture and detoxification mechanism of bacterial microcompartments,we constructed oil-in-water Pickering emulsion droplets stabilized by hydrogen-bonded organic framework immobilized cascade enzymes for decontaminating mustard gas simulant(2-chloroethyl ethyl sulfide,CEES)under sweet conditions.Two exemplified droplet systems were developed with two-enzyme(glucose oxidase/chloroperoxidase)and threeenzyme(invertase/glucose oxidase/chloroperoxidase)cascades,both achieving over 6-fold enhancement in decontamination efficiency compared with free enzymes and>99% selectivity towards non-toxic sulfoxide.We found that the favored mass transfer of sugars and CEES from their respective phases to approach the cascade enzymes located at the droplet surface and the facilitated substrate channeling between proximally immobilized enzymes were key factors in augmenting the decontamination efficacy.More importantly,the robustness of immobilized enzymes enabled easy reproduction of both the droplet formation and detoxification performance over 10 cycles,following long-term storage and in far-field locations.