Plant natural products are a kind of active substance widely used in pharmaceuticals and foods.However,the current production mode based on plant culture and extraction suffer complex processes and severe concerns for...Plant natural products are a kind of active substance widely used in pharmaceuticals and foods.However,the current production mode based on plant culture and extraction suffer complex processes and severe concerns for environmental and ecological.With the increasing awareness of environmental sustainability,engineered microbial cell factories have been an alternative approach to produce natural products.Many engineering strategies have been utilized in microbial biosynthesis of complex phytochemicals such as dynamic control and substructure engineering.Meanwhile,Enzyme engineering including directed evolution and rational design has been implemented to improve enzyme catalysis efficiency and stability as well as change promiscuity to expand product spectra.In this review,we discussed recent advances in microbial biosynthesis of complex phytochemicals from the following aspects,including pathway construction,strain engineering to boost the production.展开更多
There are lots of biochemical reactions in the biosynthetic pathway without associated enzymes.Reactions predicted by retro-biosynthetic tools are not assigned gene sequences.Besides,non-natural reactions designed wit...There are lots of biochemical reactions in the biosynthetic pathway without associated enzymes.Reactions predicted by retro-biosynthetic tools are not assigned gene sequences.Besides,non-natural reactions designed with novel functions also lack suitable enzymes.All these reactions can be categorized as orphan reactions.The absence of protein-encoding genes in these orphan reactions limits their direct experimental implementation.Computational tools have been developed to find candidate enzymes for these orphan reactions.Herein,we discuss recent advances in these computational tools,including reaction similarity-based methods for calculating the substructural similarity between orphan reactions and known enzymatic reactions;sequence-based tools combine metabolic knowledge base and phenotypic information with genomic,transcriptomic,and metabolomic data to mine appropriate enzymes for orphan reactions;and approaches based on the creation of enzyme variants for orphan reactions as enzyme engineering modifications and de novo design of enzymes.We believe that our review will greatly facilitate the design of microbial cell factories and contribute to the development of the biomanufacturing field.展开更多
Modular bioreactors can provide a flexible platform for constructing complex multi-step pathways,which may be a solution for maximizing reactions and overcoming the complexity of multi-enzyme systems.Here,we selected ...Modular bioreactors can provide a flexible platform for constructing complex multi-step pathways,which may be a solution for maximizing reactions and overcoming the complexity of multi-enzyme systems.Here,we selected wood-derived cellulose scaffold as a support for enzyme immobilization and constructed the modular bioreactor.Cellulose scaffold was prepared after removing lignin from wood,followed by citric acid functionalization and the addition of glutaraldehyde finally allowed the cross-linking of enzymes.Three enzymes,horseradish peroxidase(HRP),glucose oxidase(GOD),and catalase(CAT),were separately immobilized,resulting in the immobilized enzyme amount to over 40 mg/g.The introduction of carboxyl groups from citric acid facilitated the rapid enzyme adsorption on the support surface and immobilized enzymes possess~65%expressed activity.Modular bioreactors were constructed by using the immobilized enzymes.With the immobilized HRP module,reactor showed desired catalytic performance with the phenol degradation rate of>90%.Also,a pH regulation can occur in the bioreactors for preserving enzyme activities and neutralizing acid products.In the GOD/CAT modular bioreactor,the cascade reaction with adjusting pH values can achieve a 95%yield of sodium gluconate and exhibit a favorable reusability of 5 operation cycles.展开更多
Colitis is a common disease of the colon that is very difficult to treat.Probiotic bacteria could be an effective treatment.The probiotic Escherichia coli Nissle 1917(EcN)was engineered to synthesize the ketone body(R...Colitis is a common disease of the colon that is very difficult to treat.Probiotic bacteria could be an effective treatment.The probiotic Escherichia coli Nissle 1917(EcN)was engineered to synthesize the ketone body(R)-3-hydroxybutyrate(3HB)for sustainable production in the gut lumen of mice suffering from colitis.Components of heterologous 3HB synthesis routes were constructed,expressed,optimized,and inserted into the EcN genome,combined with deletions in competitive branch pathways.The genome-engineered EcN produced the highest 3HB level of 0.6 g/L under microaerobic conditions.The live therapeutic was found to colonize the mouse gastrointestinal tract over 14 days,elevating gut 3HB and short-chain-length fatty acid(SCFA)levels 8.7-and 3.1-fold compared to those of wild-type EcN,respectively.The sustainable presence of 3HB in mouse guts promoted the growth of probiotic bacteria,especially Akkermansia spp.,to over 31%from the initial 2%of all the microbiome.As a result,the engineered EcN termed EcNL4 ameliorated colitis induced via dextran sulfate sodium(DSS)in mice.Compared to wild-type EcN or oral administration of 3HB,oral EcNL4 uptake demonstrated better effects on mouse weights,colon lengths,occult blood levels,gut tissue myeloperoxidase activity and proinflammatory cytokine concentrations.Thus,a promising live bacterium was developed to improve colonic microenvironments and further treat colitis.This proof-of-concept design can be employed to treat other diseases of the colon.展开更多
In the fermentation process of biorefinery,industrial strains are normally subjected to adverse environmental stresses,which leads to their slow growth,yield decline,a substantial increase in energy consumption,and ot...In the fermentation process of biorefinery,industrial strains are normally subjected to adverse environmental stresses,which leads to their slow growth,yield decline,a substantial increase in energy consumption,and other negative consequences,which ultimately seriously hamper the development of biorefinery.How to minimize the impact of stress on microorganisms is of great significance.This review not only reveals the damaging effects of different environmental stresses on microbial strains but also introduces commonly used strategies to improve microbial tolerance,including adaptive evolution,reprogramming of the industrial host based on genetic circuits,global transcription machinery engineering(gTME)and bioprocess integration.Furthermore,by integrating the advantages of these strategies and reducing the cost of system operation,the tolerance of industrial strains,combined with production efficiency and process stability,will be greatly improved,and the development prospects of biorefinery will be more widespread.展开更多
Metal nanoparticles have been used as antibacterial agents widely,and the combined use of enzymes and metal nanoparticles promotes antibacterial activity,achieving a synergistic effect.Additionally,enzymes decrease th...Metal nanoparticles have been used as antibacterial agents widely,and the combined use of enzymes and metal nanoparticles promotes antibacterial activity,achieving a synergistic effect.Additionally,enzymes decrease the amounts of metals and increase biocompatibility,thereby reducing toxicity of metals.However,the efficiency of enzymes is hindered when coupled with metals,which causes deactivation in the function of enzymes.How can a balance be struck between metals and enzymes?Although the antibacterial mechanism of metal nanoparticles is relatively clear,how enzyme-metal nanocomposites work against bacteria is not conclusive.Here,we describe several examples on the synthesis of enzyme-metal nanocomposites via co-immobilization or in situ reduction and summarize how enzyme-metal nanocomposites combat microorganisms.展开更多
Maintainance of sulfomucin is a key end point in the treatment of diarrhea and inflammatory bowel disease(IBD).However,the mechanisms underlying the microbial sense to sulfomucin are poorly understood,and to date,ther...Maintainance of sulfomucin is a key end point in the treatment of diarrhea and inflammatory bowel disease(IBD).However,the mechanisms underlying the microbial sense to sulfomucin are poorly understood,and to date,there are no therapies targeting the secretion and maturation of sulfomucin in IBD.Herein,we biosynthesized poly-β-hydroxybutyrate(PHB)and found that PHB could alleviate inflammation caused by diarrhea and colitis by enhancing the differentiation of sulfomucin.Microbiota transplantation and clearance together demonstrate that PHB promoting sulfomucin is mediated by Lactobacillus johnsonii(L.johnsonii).Further studies revealed that PHB provides a favorable niche for L.johnsonii biofilm formation to resist disturbance and support its growth.L.johnsonii-biofilm alleviates colitis by regulating fucose residues to promote goblet cell differentiation and subsequent sulfomucin maturation.Importantly,PHB alleviates colitis by enhancing sulfomucin secretion and maturation in a L.johnsonii-dependent manner.PHB represents a class of guardians,acting as a safe probiotic-biofilm delivery system that significantly promotes probiotic proliferation.Altogether,this study adds weight to the possible role of probiotics and functional materials in the treatment of intestinal inflammation.The application of PHB and biofilm self-coating L.johnsonii carries high translational potential and may be of clinical relevance.展开更多
Synthetic biology aims to design or assemble existing bioparts or bio-components for useful bioproperties.During the past decades,progresses have been made to build delicate biocircuits,standardized biological buildin...Synthetic biology aims to design or assemble existing bioparts or bio-components for useful bioproperties.During the past decades,progresses have been made to build delicate biocircuits,standardized biological building blocks and to develop various genomic/metabolic engineering tools and approaches.Medical and pharmaceutical demands have also pushed the development of synthetic biology,including integration of heterologous pathways into designer cells to efficiently produce medical agents,enhanced yields of natural products in cell growth media to equal or higher than that of the extracts from plants or fungi,constructions of novel genetic circuits for tumor targeting,controllable releases of therapeutic agents in response to specific biomarkers to fight diseases such as diabetes and cancers.Besides,new strategies are developed to treat complex immune diseases,infectious diseases and metabolic disorders that are hard to cure via traditional approaches.In general,synthetic biology brings new capabilities to medical and pharmaceutical researches.This review summarizes the timeline of synthetic biology developments,the past and present of synthetic biology for microbial productions of pharmaceutics,engineered cells equipped with synthetic DNA circuits for diagnosis and therapies,live and auto-assemblied biomaterials for medical treatments,cell-free synthetic biology in medical and pharmaceutical fields,and DNA engineering approaches with potentials for biomedical applications.展开更多
Global society requires new technologies for manufacturing essential chemicals for modern life that are decoupled from fossil fuels.Therefore,Green Chemical Engineering focuses on the areas of green and sustain-able d...Global society requires new technologies for manufacturing essential chemicals for modern life that are decoupled from fossil fuels.Therefore,Green Chemical Engineering focuses on the areas of green and sustain-able development of chemistry and chemical engineering,among which biomanufacturing is an attractive topic.展开更多
Glycyrrhizin(GL)and Glycyrrhetic Acid 3-O-mono-β-D-glucuronide(GAMG)are the typical triterpenoid glycosides found in the root of licorice,a popular medicinal plant that exhibits diverse physiological effects and phar...Glycyrrhizin(GL)and Glycyrrhetic Acid 3-O-mono-β-D-glucuronide(GAMG)are the typical triterpenoid glycosides found in the root of licorice,a popular medicinal plant that exhibits diverse physiological effects and pharmacological manifestations.However,only few reports are available on the glycosylation enzymes involved in the biosynthesis of these valuable compounds with low conversion yield so far.In mammals,glycosyltransferases are involved in the phase II metabolism and may provide new solutions for us to engineer microbial strains to produce high valued compounds due to the substrate promiscuity of these glycosyltransferases.In this study,we mined the genomic databases of mammals and evaluated 22 candidate genes of O-glycosyltransferases by analyzing their catalytic potential for O-glycosylation of the native substrate,glycyrrhetinic acid(GA)for its glycodiversification.Out of 22 selected glycosyltransferases,only UGT1A1 exhibited high catalytic performance for biosynthesis of the key licorice compounds GL and GAMG.Molecular docking results proposed that the enzymatic activity of UGT1A1 was likely owing to the stable hydrogen bonding interactions and favorite conformations between the amino acid residues around substrate channels(P82~R85)and substrates.Furthermore,the complete biosynthesis pathway of GL was reconstructed in Saccharomyces cerevisiae for the first time,resulting in the production of 5.98±0.47 mg/L and 2.31±0.21 mg/L of GL and GAMG,respectively.展开更多
Photosynthesis,as an efficient pathway for solar energy capture and utilization,has supported aerobionts for billions of years.The imitation of photosynthesis to construct artificial photo-enzymatic-coupling catalysis...Photosynthesis,as an efficient pathway for solar energy capture and utilization,has supported aerobionts for billions of years.The imitation of photosynthesis to construct artificial photo-enzymatic-coupling catalysis system has become a pow-erful means to solve energy and environmental problems.After years of in-depth research on this coupled system,through ingenious and rational design,the synergistic effect of photo-and enzymatic catalyses has played a significant role in many different fields,including solar-driven fuel production,chiral chemical synthesis and carbon dioxide fixation.Furthermore,light in enzymatic catalysis could also endow enzyme new possibilities.Photo-induced radical cofactor could bring catalytic promiscuity to enzymes,making them catalyze reactions that natural enzymes cannot.This review summarizes the advances in photo-enzymatic-coupling catalysis system and introduces its essential components,their integration and application.The possibilities presented by photo-induced catalytic promiscuity and its significance for expanding the toolbox of enzymes are also discussed.展开更多
Enzyme-metal hybrid catalysts bridge the gap between enzymatic and heterogeneous catalysis,which is significant for expanding biocatalysis to a broader scope.Previous studies have demonstrated that the enzyme-metal hy...Enzyme-metal hybrid catalysts bridge the gap between enzymatic and heterogeneous catalysis,which is significant for expanding biocatalysis to a broader scope.Previous studies have demonstrated that the enzyme-metal hybrid catalysts exhibited considerably higher catalytic efficiency in cascade reactions,compared with that of the combination of separated enzyme and metal catalysts.However,the precise mechanism of this phenomenon remains unclear.Here,we investigated the diffusion process in enzyme-metal hybrid catalysts using Pd/lipase-Pluronic conjugates and the combination of immobilized lipase(Novozyme 435)and Pd/C as models.With reference to experimental data in previous studies,the Weisz-Prater parameter and efficiency factor of internal diffusion were calculated to evaluate the internal diffusion limitations in these catalysts.Thereafter,a kinetic model was developed and fitted to describe the proximity effect in hybrid catalysts.Results indicated that the enhanced catalytic efficiency of hybrid catalysts may arise from the decreased internal diffusion limitation,size effect of Pd clusters and proximity of the enzyme and metal active sites,which provides a theoretical foundation for the rational design of enzyme-metal hybrid catalysts.展开更多
Microbial cell factories (MCFs) are typical and widely used platforms in biomanufacturing for designing and constructingsynthesis pathways of target compounds in microorganisms. In MCFs, transporter engineering is esp...Microbial cell factories (MCFs) are typical and widely used platforms in biomanufacturing for designing and constructingsynthesis pathways of target compounds in microorganisms. In MCFs, transporter engineering is especially significant forimproving the biomanufacturing efficiency and capacity through enhancing substrate absorption, promoting intracellular masstransfer of intermediate metabolites, and improving transmembrane export of target products. This review discusses thecurrent methods and strategies of mining and characterizing suitable transporters and presents the cases of transporterengineering in the production of various chemicals in MCFs.展开更多
To improve the performance of yeast cell factories for industrial production,extensive CRISPR-mediated genome editing systems have been applied by artificially creating double-strand breaks(DSBs)to introduce mutations...To improve the performance of yeast cell factories for industrial production,extensive CRISPR-mediated genome editing systems have been applied by artificially creating double-strand breaks(DSBs)to introduce mutations with the assistance of intracellular DSB repair.Diverse strategies of DSB repair are required to meet various demands,including precise editing or random editing with customized gRNAs or a gRNA library.Although most yeasts remodeling techniques have shown rewarding performance in laboratory verification,industrial yeast strain manipulation relies only on very limited strategies.Here,we comprehensively reviewed the molecular mechanisms underlying recent industrial applications to provide new insights into DSB cleavage and repair pathways in both Saccharomyces cerevisiae and other unconventional yeast species.The discussion of DSB repair covers the most frequently used homologous recombination(HR)and nonhomologous end joining(NHEJ)strategies to the less well-studied illegitimate recombination(IR)pathways,such as single-strand annealing(SSA)and microhomology-mediated end joining(MMEJ).Various CRISPR-based genome editing tools and corresponding gene editing efficiencies are described.Finally,we summarize recently developed CRISPR-based strategies that use optimized DSB repair for genome-scale editing,providing a direction for further development of yeast genome editing.展开更多
基金This work was supported by the National Key Research and Development Program of China(2018YFA0901800)the National Natural Science Foundation of China(No.21736002).
文摘Plant natural products are a kind of active substance widely used in pharmaceuticals and foods.However,the current production mode based on plant culture and extraction suffer complex processes and severe concerns for environmental and ecological.With the increasing awareness of environmental sustainability,engineered microbial cell factories have been an alternative approach to produce natural products.Many engineering strategies have been utilized in microbial biosynthesis of complex phytochemicals such as dynamic control and substructure engineering.Meanwhile,Enzyme engineering including directed evolution and rational design has been implemented to improve enzyme catalysis efficiency and stability as well as change promiscuity to expand product spectra.In this review,we discussed recent advances in microbial biosynthesis of complex phytochemicals from the following aspects,including pathway construction,strain engineering to boost the production.
基金supported by the National Natural Science Foundation of China(22138006).
文摘There are lots of biochemical reactions in the biosynthetic pathway without associated enzymes.Reactions predicted by retro-biosynthetic tools are not assigned gene sequences.Besides,non-natural reactions designed with novel functions also lack suitable enzymes.All these reactions can be categorized as orphan reactions.The absence of protein-encoding genes in these orphan reactions limits their direct experimental implementation.Computational tools have been developed to find candidate enzymes for these orphan reactions.Herein,we discuss recent advances in these computational tools,including reaction similarity-based methods for calculating the substructural similarity between orphan reactions and known enzymatic reactions;sequence-based tools combine metabolic knowledge base and phenotypic information with genomic,transcriptomic,and metabolomic data to mine appropriate enzymes for orphan reactions;and approaches based on the creation of enzyme variants for orphan reactions as enzyme engineering modifications and de novo design of enzymes.We believe that our review will greatly facilitate the design of microbial cell factories and contribute to the development of the biomanufacturing field.
基金supported by the National Key Research and Development Program of China(2021YFC2102804)the Beijing Natural Science Foundation(No.2202034)the National Natural Science Foundation of China(No.21978024)。
文摘Modular bioreactors can provide a flexible platform for constructing complex multi-step pathways,which may be a solution for maximizing reactions and overcoming the complexity of multi-enzyme systems.Here,we selected wood-derived cellulose scaffold as a support for enzyme immobilization and constructed the modular bioreactor.Cellulose scaffold was prepared after removing lignin from wood,followed by citric acid functionalization and the addition of glutaraldehyde finally allowed the cross-linking of enzymes.Three enzymes,horseradish peroxidase(HRP),glucose oxidase(GOD),and catalase(CAT),were separately immobilized,resulting in the immobilized enzyme amount to over 40 mg/g.The introduction of carboxyl groups from citric acid facilitated the rapid enzyme adsorption on the support surface and immobilized enzymes possess~65%expressed activity.Modular bioreactors were constructed by using the immobilized enzymes.With the immobilized HRP module,reactor showed desired catalytic performance with the phenol degradation rate of>90%.Also,a pH regulation can occur in the bioreactors for preserving enzyme activities and neutralizing acid products.In the GOD/CAT modular bioreactor,the cascade reaction with adjusting pH values can achieve a 95%yield of sodium gluconate and exhibit a favorable reusability of 5 operation cycles.
基金This research was financially supported by a grant from the Chunfeng Foundation(2020Z99CFG002)of Tsinghua UniversityOther support includes the National Natural Science Foundation of China(Grant Nos.31870859,21761132013,31771886,and 31971170).
文摘Colitis is a common disease of the colon that is very difficult to treat.Probiotic bacteria could be an effective treatment.The probiotic Escherichia coli Nissle 1917(EcN)was engineered to synthesize the ketone body(R)-3-hydroxybutyrate(3HB)for sustainable production in the gut lumen of mice suffering from colitis.Components of heterologous 3HB synthesis routes were constructed,expressed,optimized,and inserted into the EcN genome,combined with deletions in competitive branch pathways.The genome-engineered EcN produced the highest 3HB level of 0.6 g/L under microaerobic conditions.The live therapeutic was found to colonize the mouse gastrointestinal tract over 14 days,elevating gut 3HB and short-chain-length fatty acid(SCFA)levels 8.7-and 3.1-fold compared to those of wild-type EcN,respectively.The sustainable presence of 3HB in mouse guts promoted the growth of probiotic bacteria,especially Akkermansia spp.,to over 31%from the initial 2%of all the microbiome.As a result,the engineered EcN termed EcNL4 ameliorated colitis induced via dextran sulfate sodium(DSS)in mice.Compared to wild-type EcN or oral administration of 3HB,oral EcNL4 uptake demonstrated better effects on mouse weights,colon lengths,occult blood levels,gut tissue myeloperoxidase activity and proinflammatory cytokine concentrations.Thus,a promising live bacterium was developed to improve colonic microenvironments and further treat colitis.This proof-of-concept design can be employed to treat other diseases of the colon.
基金The authors acknowledge funding support from the National Natural Science Foundation of China(21736002,21576027,21425624).
文摘In the fermentation process of biorefinery,industrial strains are normally subjected to adverse environmental stresses,which leads to their slow growth,yield decline,a substantial increase in energy consumption,and other negative consequences,which ultimately seriously hamper the development of biorefinery.How to minimize the impact of stress on microorganisms is of great significance.This review not only reveals the damaging effects of different environmental stresses on microbial strains but also introduces commonly used strategies to improve microbial tolerance,including adaptive evolution,reprogramming of the industrial host based on genetic circuits,global transcription machinery engineering(gTME)and bioprocess integration.Furthermore,by integrating the advantages of these strategies and reducing the cost of system operation,the tolerance of industrial strains,combined with production efficiency and process stability,will be greatly improved,and the development prospects of biorefinery will be more widespread.
基金supported by the National Key Research and Development Plan of China(2016YFA0204300)the National Natural Science Foundation of China(21878174,21911540467)the Beijing Natural Science Foundation(JQ18006).
文摘Metal nanoparticles have been used as antibacterial agents widely,and the combined use of enzymes and metal nanoparticles promotes antibacterial activity,achieving a synergistic effect.Additionally,enzymes decrease the amounts of metals and increase biocompatibility,thereby reducing toxicity of metals.However,the efficiency of enzymes is hindered when coupled with metals,which causes deactivation in the function of enzymes.How can a balance be struck between metals and enzymes?Although the antibacterial mechanism of metal nanoparticles is relatively clear,how enzyme-metal nanocomposites work against bacteria is not conclusive.Here,we describe several examples on the synthesis of enzyme-metal nanocomposites via co-immobilization or in situ reduction and summarize how enzyme-metal nanocomposites combat microorganisms.
基金supported by the National Key Research and Development Program of China(2022YFD1300400)the National Natural Science Foundation of China(31930106 and 31829004)+3 种基金the Chinese Universities Scientific Fund(2022TC033)the 2115 Talent Development Program of China Agricultural University(1041-00109019)the Henan Province Public Benefit Research Foundation(201300111200-05)the 111 Project(B16044)。
文摘Maintainance of sulfomucin is a key end point in the treatment of diarrhea and inflammatory bowel disease(IBD).However,the mechanisms underlying the microbial sense to sulfomucin are poorly understood,and to date,there are no therapies targeting the secretion and maturation of sulfomucin in IBD.Herein,we biosynthesized poly-β-hydroxybutyrate(PHB)and found that PHB could alleviate inflammation caused by diarrhea and colitis by enhancing the differentiation of sulfomucin.Microbiota transplantation and clearance together demonstrate that PHB promoting sulfomucin is mediated by Lactobacillus johnsonii(L.johnsonii).Further studies revealed that PHB provides a favorable niche for L.johnsonii biofilm formation to resist disturbance and support its growth.L.johnsonii-biofilm alleviates colitis by regulating fucose residues to promote goblet cell differentiation and subsequent sulfomucin maturation.Importantly,PHB alleviates colitis by enhancing sulfomucin secretion and maturation in a L.johnsonii-dependent manner.PHB represents a class of guardians,acting as a safe probiotic-biofilm delivery system that significantly promotes probiotic proliferation.Altogether,this study adds weight to the possible role of probiotics and functional materials in the treatment of intestinal inflammation.The application of PHB and biofilm self-coating L.johnsonii carries high translational potential and may be of clinical relevance.
基金supported by grants from the Ministry of Science and Technology of China[Grant number 2018YFA0900200]National Natural Science Foundation of China[Grant number 32130001]+1 种基金Center of Life Sciences of Tsinghua-Peking University,the Shuimu Tsinghua Scholar Program and Chunfeng Foundation.This project is also funded by the National Natural Science Foundation of China[Grant numbers 31961133017,31961133018]part of MIX-UP,a joint NSFC and EU H2020 collaboration.In Europe,MIX-UP has received funding from the European Union's Horizon 2020 research and innovation program[grant agreement Number870294].
文摘Synthetic biology aims to design or assemble existing bioparts or bio-components for useful bioproperties.During the past decades,progresses have been made to build delicate biocircuits,standardized biological building blocks and to develop various genomic/metabolic engineering tools and approaches.Medical and pharmaceutical demands have also pushed the development of synthetic biology,including integration of heterologous pathways into designer cells to efficiently produce medical agents,enhanced yields of natural products in cell growth media to equal or higher than that of the extracts from plants or fungi,constructions of novel genetic circuits for tumor targeting,controllable releases of therapeutic agents in response to specific biomarkers to fight diseases such as diabetes and cancers.Besides,new strategies are developed to treat complex immune diseases,infectious diseases and metabolic disorders that are hard to cure via traditional approaches.In general,synthetic biology brings new capabilities to medical and pharmaceutical researches.This review summarizes the timeline of synthetic biology developments,the past and present of synthetic biology for microbial productions of pharmaceutics,engineered cells equipped with synthetic DNA circuits for diagnosis and therapies,live and auto-assemblied biomaterials for medical treatments,cell-free synthetic biology in medical and pharmaceutical fields,and DNA engineering approaches with potentials for biomedical applications.
文摘Global society requires new technologies for manufacturing essential chemicals for modern life that are decoupled from fossil fuels.Therefore,Green Chemical Engineering focuses on the areas of green and sustain-able development of chemistry and chemical engineering,among which biomanufacturing is an attractive topic.
基金supported by the National Key Research and Development Program of China(2018YFA0901800)the Key Research and Development Program of Hebei Province(21374301D)+2 种基金the Natural Science Foundation of China(No.22078171)the Natural Science Foundation of Hebei Province(No.C2019105055)the Scientific Research Foundation of Tangshan Normal University(No.2021B34).
文摘Glycyrrhizin(GL)and Glycyrrhetic Acid 3-O-mono-β-D-glucuronide(GAMG)are the typical triterpenoid glycosides found in the root of licorice,a popular medicinal plant that exhibits diverse physiological effects and pharmacological manifestations.However,only few reports are available on the glycosylation enzymes involved in the biosynthesis of these valuable compounds with low conversion yield so far.In mammals,glycosyltransferases are involved in the phase II metabolism and may provide new solutions for us to engineer microbial strains to produce high valued compounds due to the substrate promiscuity of these glycosyltransferases.In this study,we mined the genomic databases of mammals and evaluated 22 candidate genes of O-glycosyltransferases by analyzing their catalytic potential for O-glycosylation of the native substrate,glycyrrhetinic acid(GA)for its glycodiversification.Out of 22 selected glycosyltransferases,only UGT1A1 exhibited high catalytic performance for biosynthesis of the key licorice compounds GL and GAMG.Molecular docking results proposed that the enzymatic activity of UGT1A1 was likely owing to the stable hydrogen bonding interactions and favorite conformations between the amino acid residues around substrate channels(P82~R85)and substrates.Furthermore,the complete biosynthesis pathway of GL was reconstructed in Saccharomyces cerevisiae for the first time,resulting in the production of 5.98±0.47 mg/L and 2.31±0.21 mg/L of GL and GAMG,respectively.
基金This work was supported by the National Key Research and Development Plan of China(2016YFA0204300)the National Natural Science Foundation of China(21878174,21911540467)the Beijing Natural Science Foundation(JQ18006).
文摘Photosynthesis,as an efficient pathway for solar energy capture and utilization,has supported aerobionts for billions of years.The imitation of photosynthesis to construct artificial photo-enzymatic-coupling catalysis system has become a pow-erful means to solve energy and environmental problems.After years of in-depth research on this coupled system,through ingenious and rational design,the synergistic effect of photo-and enzymatic catalyses has played a significant role in many different fields,including solar-driven fuel production,chiral chemical synthesis and carbon dioxide fixation.Furthermore,light in enzymatic catalysis could also endow enzyme new possibilities.Photo-induced radical cofactor could bring catalytic promiscuity to enzymes,making them catalyze reactions that natural enzymes cannot.This review summarizes the advances in photo-enzymatic-coupling catalysis system and introduces its essential components,their integration and application.The possibilities presented by photo-induced catalytic promiscuity and its significance for expanding the toolbox of enzymes are also discussed.
基金the Beijing Natural Science Foundation(No.JQ18006)the National Natural Science Foundation of China(Grant Nos.21878174,2191101041).
文摘Enzyme-metal hybrid catalysts bridge the gap between enzymatic and heterogeneous catalysis,which is significant for expanding biocatalysis to a broader scope.Previous studies have demonstrated that the enzyme-metal hybrid catalysts exhibited considerably higher catalytic efficiency in cascade reactions,compared with that of the combination of separated enzyme and metal catalysts.However,the precise mechanism of this phenomenon remains unclear.Here,we investigated the diffusion process in enzyme-metal hybrid catalysts using Pd/lipase-Pluronic conjugates and the combination of immobilized lipase(Novozyme 435)and Pd/C as models.With reference to experimental data in previous studies,the Weisz-Prater parameter and efficiency factor of internal diffusion were calculated to evaluate the internal diffusion limitations in these catalysts.Thereafter,a kinetic model was developed and fitted to describe the proximity effect in hybrid catalysts.Results indicated that the enhanced catalytic efficiency of hybrid catalysts may arise from the decreased internal diffusion limitation,size effect of Pd clusters and proximity of the enzyme and metal active sites,which provides a theoretical foundation for the rational design of enzyme-metal hybrid catalysts.
基金the National Basic Research Program of China(973 Program)(2018YFA0901800)the National Natural Science Foundation of China(22138006,21736002,and 22078020).
文摘Microbial cell factories (MCFs) are typical and widely used platforms in biomanufacturing for designing and constructingsynthesis pathways of target compounds in microorganisms. In MCFs, transporter engineering is especially significant forimproving the biomanufacturing efficiency and capacity through enhancing substrate absorption, promoting intracellular masstransfer of intermediate metabolites, and improving transmembrane export of target products. This review discusses thecurrent methods and strategies of mining and characterizing suitable transporters and presents the cases of transporterengineering in the production of various chemicals in MCFs.
基金supported by the National Key Research and Development Program of China(No.2021YFC2101203)the General project of National Natural Science Foundation of China(No.22078021).
文摘To improve the performance of yeast cell factories for industrial production,extensive CRISPR-mediated genome editing systems have been applied by artificially creating double-strand breaks(DSBs)to introduce mutations with the assistance of intracellular DSB repair.Diverse strategies of DSB repair are required to meet various demands,including precise editing or random editing with customized gRNAs or a gRNA library.Although most yeasts remodeling techniques have shown rewarding performance in laboratory verification,industrial yeast strain manipulation relies only on very limited strategies.Here,we comprehensively reviewed the molecular mechanisms underlying recent industrial applications to provide new insights into DSB cleavage and repair pathways in both Saccharomyces cerevisiae and other unconventional yeast species.The discussion of DSB repair covers the most frequently used homologous recombination(HR)and nonhomologous end joining(NHEJ)strategies to the less well-studied illegitimate recombination(IR)pathways,such as single-strand annealing(SSA)and microhomology-mediated end joining(MMEJ).Various CRISPR-based genome editing tools and corresponding gene editing efficiencies are described.Finally,we summarize recently developed CRISPR-based strategies that use optimized DSB repair for genome-scale editing,providing a direction for further development of yeast genome editing.
