Activation of inflammatory responses regulates the transmission of pain pathways through an integrated network in the peripheral and central nervous systems.The immunopotentiator thymosin alpha-1(Tal)has recently been...Activation of inflammatory responses regulates the transmission of pain pathways through an integrated network in the peripheral and central nervous systems.The immunopotentiator thymosin alpha-1(Tal)has recently been reported to have anti-inflammatory and neuroprotective functions in rodents.However,how Tα1 affects inflammatory pain remains unclear.In the present study,intraperitoneal injection of Tal attenuated complete Freund's adjuvant(CFA)-induced pain hypersensitivity,and decreased the up-regulation of pro-inflammatory cytokines(TNF-α,IL-1β,and IL-6)in inflamed skin and the spinal cord.We found that CFA-induced peripheral inflammation evoked strong microglial activation,but the effect was reversed by Tα1.Notably,Tα1 reversed the CFA-induced up-regulation of vesicular glutamate transporter(VGLUT)and down-regulated the vesicular γ-aminobutyric acid transporter(VGAT)in the spinal cord.Taken together,these results suggest that Tα1 plays a therapeutic role in inflammatory pain and in the modulation of microgliainduced pro-inflammatory cytokine production in addition to mediation of VGLUT and VGAT expression in the spinal cord.展开更多
Cross-linked enzyme aggregates(CLEAs) of nitrile hydratase(NHase) ES-NHT-118 from Escherichia coli were prepared by using ammonium sulfate as precipitating agent followed by cross-linking with dextran polyaldehyde for...Cross-linked enzyme aggregates(CLEAs) of nitrile hydratase(NHase) ES-NHT-118 from Escherichia coli were prepared by using ammonium sulfate as precipitating agent followed by cross-linking with dextran polyaldehyde for the first time. In this process, egg white was added as protein feeder for facilitating the formation of CLEAs. The optimal conditions of the immobilization process were determined. Michaelis constants(Km) of free NHase and NHase CLEAs were also determined. The NHase CLEAs exhibited increased stability at varied pH and temperature conditions compared to its free counterpart. When exposed to high concentrations of acrylamide, NHase CLEAs also exhibited effective catalytic activity.展开更多
The continuous use of chemical dyes in various industries,and their discharge into industrial effluents,results in severe problems to human life and water pollution.Laccases have the ability to decolorize dyes and tox...The continuous use of chemical dyes in various industries,and their discharge into industrial effluents,results in severe problems to human life and water pollution.Laccases have the ability to decolorize dyes and toxic chemicals in industrial effluents as green biocatalysts.Their possible industrial applications have been limited by poor reusability,low stability,and loss of free laccase action.In this research,lac-case was immobilized on zeolitic imidazolate framework coated multi-walled carbon nanotubes(Laccase@ZIF-8@MWCNTs)via metal affinity adsorption to develop an easy separable and stable enzyme.The optimum reaction conditions for immobilized laccase are at a pH of 3.0 and a temperature of 60℃.The immobilized laccase was enhanced in storage and thermal stability.The results indicated that Laccase@ZIF-8@MWCNTs still maintained 68%of its original activity after 10 times of repeated use.Most importantly,the biocatalytic system was applied for decolorization of different dyes(20 mg·L^(-1))without a mediator,and up to 97.4%for Eriochrome black T and 95.6%Acid red 88 was achieved in 25 min.Biocatalysts with these properties may be used in a variety of environmental and industrial applications.展开更多
Immobilization is an effective method to promote the application of enzyme industry for improving the stability and realizing recovery of enzyme.To some extent,the performance of immobilized enzyme depends on the choi...Immobilization is an effective method to promote the application of enzyme industry for improving the stability and realizing recovery of enzyme.To some extent,the performance of immobilized enzyme depends on the choice of carrier material.Therefore,the development of new carrier materials has been one of the key issues concerned by enzyme immobilization researchers.In this work,a novel organic–inorganic hybrid material,nickel-carnosine complex(NiCar),was synthesized for the first time by solvothermal method.The obtained NiCar exhibits spherical morphology,hierarchical porosity and abundant unsaturated coordination nickel ions,which provide excellent anchoring sites for the immobilization of proteins.His-tagged organophosphate-degrading enzyme(Opd A)and x-transaminase(ω-TA)were used as model enzymes to evaluate the performance of NiCar as a carrier.By a simple adsorption process,the enzyme molecules can be fixed on the particles of NiCar,and the stability and reusability are significantly improved.The analysis of protein adsorption on NiCar verified that the affinity adsorption between the imidazole functional group on the protein and the unsaturated coordination nickel ions on NiCar was the main force in the immobilization process,which provided an idea way for the development of new enzyme immobilization carriers.展开更多
Casein kinase 1α(CK1α) mediates the phosphorylation and degradation of interferon-α/β receptor 1(IFNAR1) in response to viral infection. However, how CK1α regulates hepatitis B virus(HBV) replication and the anti...Casein kinase 1α(CK1α) mediates the phosphorylation and degradation of interferon-α/β receptor 1(IFNAR1) in response to viral infection. However, how CK1α regulates hepatitis B virus(HBV) replication and the anti-HBV effects of IFN-α are less reported. Here we show that CK1α can interact with IFNAR1 in hepatoma carcinoma cells and increased the abundance of IFNAR1 by reducing the ubiquitination levels in the presence of HBV.Furthermore, CK1α promotes the IFN-α triggered JAK-STAT signaling pathway and consequently enhances the antiviral effects of IFN-α against HBV replication. Our results collectively provide evidence that CK1α positively regulates the anti-HBV activity of IFN-α in hepatoma carcinoma cells, which would be a promising therapeutic target to improve the effectiveness of IFN-α therapy to cure CHB.展开更多
D-allulose has very little content in nature,and it needs to be synthesized artificially and meet the purity requirements of industrial grade.The basic physical and chemical properties of D-allulose,its preparation me...D-allulose has very little content in nature,and it needs to be synthesized artificially and meet the purity requirements of industrial grade.The basic physical and chemical properties of D-allulose,its preparation methods and many different ways of isolation and purification were described.In order to achieve the goal of industrial production of D-allulose as soon as possible,the research progress of D-allulose isolation and purification technologies at home and abroad in recent years was classified and discussed,so as to provide useful reference for the practical improvement of D-allulose isolation and purification process technologies.展开更多
Chiral compounds have a huge market demand in the fields of pharmaceuticals,pesticides,and fine chemicals.Enzymatic electrosynthesis can couple enzyme catalysis,possessing high product purity,high efficiency,and mild ...Chiral compounds have a huge market demand in the fields of pharmaceuticals,pesticides,and fine chemicals.Enzymatic electrosynthesis can couple enzyme catalysis,possessing high product purity,high efficiency,and mild conditions,with electrochemical regeneration of expensive cofactor nicotinamide adenine dinucleotide(NADH),possessing easy process monitoring and simple operation for efficient chiral synthesis.In this study,hydrophobic covalent organic framework(COF)was synthesized as the immobilized carrier,which not only enhanced the enzyme catalysis through enriching substrate but also enhanced the stability and reuse of the enzyme.Besides,Rh complex was anchored on hydrophilically-modified electrode to promote the regeneration of NADH,where the anchor of Rh complex can effectively avoid the mutual deactivation from the interference between electron mediator and enzyme,and simplify the separation of products.The immobilized enzyme catalysis and the electrochemical cofactor regeneration were coupled to construct an enzymatic electrosynthesis system for the efficient asymmetric reduction to obtain chiral alcohols,with a maximum turnover frequency(TOF)of 101.1 h^(-1).Furthermore,the relevant parameters of the system were optimized,and the substrate scope was expanded.展开更多
The capture and conversion of CO_(2) to valuable products has garnered significant attention.The electroenzymatic reduction of CO_(2) is of particular interest owing to its high selectivity and yield.However,this proc...The capture and conversion of CO_(2) to valuable products has garnered significant attention.The electroenzymatic reduction of CO_(2) is of particular interest owing to its high selectivity and yield.However,this process is hindered by various challenges,such as the low CO_(2) solubility in water and the requirement for efficient cofactor regeneration.In this work,we developed a green,water-based,room-temperature synthesis of Zn-MOF-74 for enzyme immobilization.Formate dehydrogenase(FDH)and carbonic anhydrase(CA)are co-encapsulated within the Zn-MOF-74 framework to create a dual enzyme cascade system.This strategy effectively enhances enzyme proximity,stability,and reusability to subsequently promote the enzyme cascade reaction efficiency.Additionally,the adsorption of CO_(2) by CA improves the solubility of CO_(2) in water.NADH regeneration is achieved using an electron mediator,2-hydroxy-1-naphthoquinone(HNQ).The optimized electroenzymatic system achieved a formate yield of 3.01 mM within 3 h,which was 4.98 times higher than that of the pristine free enzyme system and 2.27 times higher than the free enzyme system with NADH regeneration.展开更多
Escalating electromagnetic(EM)pollution and advanced stealth technologies require next-generation microwave absorbers that combine broadband response,strong attenuation,and lightweight characteristics with high-temper...Escalating electromagnetic(EM)pollution and advanced stealth technologies require next-generation microwave absorbers that combine broadband response,strong attenuation,and lightweight characteristics with high-temperature stability.In this work,a vacancy-mediated strategy is proposed to tailor EM losses in Cr-doped lanthanum manganite perovskites(LaMn_(1-x)Cr_(x)O_(3))synthesized via a sol–gel method.The cooperative modulation between Mn-site cation vacancies and oxygen vacancies enables a well-balanced contribution of polarization loss and conduction loss,resulting in excellent absorption performance.Specifically,LaMn_(0.85)Cr_(0.15)O_(3) achieves a remarkable minimum reflection loss(RL_(min))of-75.37 dB and an effective absorption bandwidth(EAB)of 6.0 GHz at a thickness of only 2.8 mm.Structural and spectroscopic analyses reveal that Cr^(3+)substitution induces Mn vacancies and modulates oxygen vacancy concentrations,thereby generating defect dipoles and facilitating carrier migration.Density functional theory(DFT)calculations further elucidate the role of Cr-induced defect states in enhancing conduction and polarization losses.This vacancy-engineered approach not only establishes a new paradigm for designing high-efficiency perovskite-based microwave absorbers but also offers significant potential for high-temperature EM compatibility applications.展开更多
A novel integrated nanobiocatalyst system based on an enzyme@silica nanoflower@metal-organic framework (enzyme@SNF@ZIF-8) structure with improved stability is fabricated for the first time. The versatility of this s...A novel integrated nanobiocatalyst system based on an enzyme@silica nanoflower@metal-organic framework (enzyme@SNF@ZIF-8) structure with improved stability is fabricated for the first time. The versatility of this system is validated using penicillin G acylase (PGA) and catalase (CAT) as model enzymes. The microporous ZIF-8 layer can be controlled by varying the number of ZIF-8 coating cycles, which produces PGA@SNF@ZIF-8 nanobiocatalysts with different ZIF-8 layer thicknesses. After the second ZIF-8 coating cycle, a PGA@SNF@ZIF-8(2) structure with a homogeneous and well-intergrown ZIF-8 layer is formed, which possesses excellent mechanical and chemical stability. Moreover, PGA@SNF@ZIF-8(2) shows improved thermal/storage stability and reusability compared with free PGA and PGA immobilized on silica nanoflowers (PGA@SNF). The obtained CAT-based nanobiocatalysts (CAT@SNF@ZIF-8(2)) also show excellent catalytic performance.展开更多
Dendritic mesoporous silica nanoparticles(DMSNs)are a new class of solid porous materials used for enzyme immobilization support due to their intrinsic characteristics,including their unique open central-radial struct...Dendritic mesoporous silica nanoparticles(DMSNs)are a new class of solid porous materials used for enzyme immobilization support due to their intrinsic characteristics,including their unique open central-radial structures with large pore channels and their excellent biocompatibility.In this review,we review the recent progress in research on enzyme immobilization using DMSNs with different structures,namely,flower-like DMSNs and tree-branch-like DMSNs.Three DMSN synthesis methods are briefly compared,and the distinct characteristics of the two DMSN types and their effects on the catalytic performance of immobilized enzymes are comprehensively discussed.Possible directions for future research on enzyme immobilization using DMSNs are also proposed.展开更多
Main observation and conclusion Aqueous heterogeneous catalysis is a green,sustainable catalytic process that attracts increasing attention,but it often suffers from poor mass transfer,substrate adsorption and catalys...Main observation and conclusion Aqueous heterogeneous catalysis is a green,sustainable catalytic process that attracts increasing attention,but it often suffers from poor mass transfer,substrate adsorption and catalyst dispersion.Herein,we synthesized a type of amphiphilic core-shell catalysts with a hydrophilic polydopamine(PDA)shell and a hydrophobic dendritic organosilica nanoparticle(DON)core for heterogeneous catalysis in water.展开更多
The worldwide application of organophosphorus pesticides(OPs)has promoted agricultural development,but their gradual accumulation in soil and water can seriously affect the central nervous system of humans and other m...The worldwide application of organophosphorus pesticides(OPs)has promoted agricultural development,but their gradual accumulation in soil and water can seriously affect the central nervous system of humans and other mammals.Organophosphorus hydrolase(OPH)is an effective enzyme that can catalyze the degradation of the residual OPs.However,the degradation products such as p-nitrophenol(p-NP)is still toxic.Thus,it is of great significance to develop a multi-functional support that can be simultaneously used for the immobilization of OPH and the further degradation of p-NP.Herein,a visible light assisted enzyme-photocatalytic integrated catalyst was constructed by immobilizing OPH on hollow structured Au-TiO_(2)(named OPH@H-Au-TiO_(2))for the degradation of OPs.The obtained OPH@H-Au-TiO_(2)can degrade methyl parathion to p-NP by OPH and then degrade p-NP to hydroquinone with low toxicity by using H-Au-TiO_(2)under visible light.OPH molecules were immobilized on HAu-TiO_(2)through adsorption method to prepare OPH@H-Au-TiO_(2).After 2.5 h of reaction,methyl parathion is completely degraded,and about 82.64%of the generated p-NP is further degraded into hydroquinone.After reused for 4 times,the OPH@H-Au-TiO_(2)retains more than 80%of the initial degradation activity.This research presents a new insight in designing and constructing multi-functional biocatalyst,which greatly expands the application scenarios and industrial value of enzyme catalysis.展开更多
Here,we reported a cancer nanovaccine based on SiO_(2)nanoflowers with a special radial pore structure,which greatly enhanced cross-presentation and induced the production of cytotoxic T lymphocyte cells secreting gra...Here,we reported a cancer nanovaccine based on SiO_(2)nanoflowers with a special radial pore structure,which greatly enhanced cross-presentation and induced the production of cytotoxic T lymphocyte cells secreting granzymes B and interferon-γ.The antigen ovalbumin was covalently conjugated onto the as-synthesized hierarchical SiO_(2)nanoflowers,and the adjuvant cytosine-phosphate-guanine was electrostatically adsorbed into their radial pore by simple mixing before use.The nanovaccine exhibited excellent storage stability without antigen release after 27 days of incubation,negligible cytotoxicity to dendritic cells,and a high antigen loading capacity of 430±66 mg·g^(−1)support.Besides,the nanovaccine could be internalized by dendritic cells via multiple pathways.And the enhancement of antigen/adjuvant uptake and lysosome escape of antigen were observed.Noteworthy,in vitro culture of bone marrow-derived dendritic cells in the presence of nanovaccine proved the activation of dendritic cells and antigen cross-presentation as well as secretion of proinflammatory cytokines.Besides,in vivo study verified the targeting of nanovaccine to draining lymph nodes,the complete suppression of tumor in six out of ten mice,and the triggering of notable tumor growth delay.Overall,the present results indicated that the nanovaccine can be served as a potential therapeutic vaccine to treat cancer.展开更多
The hollow spherical covalent organic frameworks(COFs)have a wide application prospect thanks to their special structures.However,the controllable synthesis of uniform and stable hollow COFs is still a challenge.We he...The hollow spherical covalent organic frameworks(COFs)have a wide application prospect thanks to their special structures.However,the controllable synthesis of uniform and stable hollow COFs is still a challenge.We herein propose a self-templated method for the preparation of hollow COFs through the Ostwald ripening mechanism under ambient conditions,which avoids most disadvantages of the commonly used hard-templating and soft-templating methods.A detailed time-dependent study reveals that the COFs are transformed from initial spheres to hollow spheres because of the inside-out Ostwald ripening process.The obtained hollow spherical COFs have high crystallinity,specific surface area(2,036 m^(2)·g^(−1)),stability,and single-batch yield.Thanks to unique hollow structure,clear through holes,and hydrophobic pore environment of the hollow spherical COFs,the obtained immobilized lipase(BCL@H-COF-OMe)exhibits higher thermostability,polar organic solvent tolerance,and reusability.The BCL@H-COF-OMe also shows higher catalytic performance than the lipase immobilized on non-hollow COF and free lipase in the kinetic resolution of secondary alcohols.This study provides a simple approach for the preparation of hollow spherical COFs,and will promote the valuable research of COFs in the field of biocatalysis.展开更多
Unspecific peroxygenases exhibit high activity for the selective oxyfunctionalization of inert C(sp3)–H bonds using only H_(2)O_(2) as a clean oxidant,while also exhibiting sensitivity to H_(2)O_(2) concentration.CdS...Unspecific peroxygenases exhibit high activity for the selective oxyfunctionalization of inert C(sp3)–H bonds using only H_(2)O_(2) as a clean oxidant,while also exhibiting sensitivity to H_(2)O_(2) concentration.CdS-based semiconductors are promising for the photosynthesis of H_(2)O_(2) owing to their adequately negative potential for oxygen reduction reaction via a proton-coupled electron transfer process,however,they suffer from fast H_(2)O_(2) decomposition on the surface of pristine CdS.Therefore,[Cp*Rh(bpy)H_(2)O]2+,a highly selective proton-coupled electron transfer catalyst,was anchored onto a supramolecular polymer-grafted CdS nanoflower to construct an efficient integrated photocatalyst for generating H_(2)O_(2),mitigating the surface issue of pristine CdS,increasing light absorption,accelerating photonic carrier separation,and enhancing oxygen reduction reaction selectivity to H_(2)O_(2).This photocatalyst promoted the light driven H_(2)O_(2) generation rate up to 1345μmol·L^(-1)·g^(-1)·h^(-1),which was 2.4 times that of pristine CdS.The constructed heterojunction photocatalyst could supply H_(2)O_(2) in situ for nonspecific peroxygenases to catalyze the C–H oxyfunctionalization of ethylbenzene,achieving a yield of 81%and an ee value of 99%under optimum conditions.A wide range of substrates were converted to the corresponding chiral alcohols using this photo-enzyme catalytic system,achieving the corresponding chiral alcohols in good yield(51%–88%)and excellent enantioselectivity(90%–99%ee).展开更多
A microscale vaccine containing SiO_(2)nanoparticles loaded in CaC〇3 microparticles was constructed using the co-precipitation method.The antigen ovalbumin(OVA)was covalently conjugated with SiO_(2)nanoparticles,and ...A microscale vaccine containing SiO_(2)nanoparticles loaded in CaC〇3 microparticles was constructed using the co-precipitation method.The antigen ovalbumin(OVA)was covalently conjugated with SiO_(2)nanoparticles,and these nanoparticles and CpG were co-encapsulated into CaCO_(3)microparticles,generating a vaccine with a size of approximately 5.2μm.Scanning electron microscopy(SEM),energy-dispersive X-ray(EDX),elemental mapping,and Fourier transform infrared(FTIR)analyses confirmed the successful preparation of the microscale vaccine;the vaccine had good storage stability without sustained antigen release,and negligible cytotoxicity to dendritic cells(DCs)and macrophages.Compared to SiO_(2)nanoparticles,the microscale vaccine can significantly improve antigen/adjuvant uptake.DCs internalized the entire microscale vaccine into lysosomes via macropinocytosis,and an increase in antigen endo/lysosomal escape was observed by confocal User scanning microscopy(CLSM).Specifically,DCs pulsed with the vaccine were fully mature,expressing high levels of costimulatory molecules(CD40,CD80,and CD86),MHCⅡ,and MHCⅠand secreting high levels of proinflammatory cytokines(IL-12,TNF-α,IL-1β,and IL-6).In addition,the vaccine had good in vivo biocompatibility,could protect the antigen from rapid degradation,and increased the retention time in lymph nodes.SiO_(2)nanoparticles-in-CaCO_(3)microparticles were an excellent carrier for antigen and adjuvant delivery.Hopefully,this study can provide some information on the design of microscale carriers for vaccine delivery systems.展开更多
Core-shell structured magnetic wrinkled organosilica-based metal-enzyme integrated catalysts were synthesized,and their catalytic performances were studied in the chemoenzymatic dynamic kinetic resolution of chiral am...Core-shell structured magnetic wrinkled organosilica-based metal-enzyme integrated catalysts were synthesized,and their catalytic performances were studied in the chemoenzymatic dynamic kinetic resolution of chiral amines in an organic solvent,as well as in the chemoenzymatic synthesis of chiral alcohols in water.Structureperformance studies revealed the important influence of their tunable structure and composition on the optimization of activity,stability,and recyclability in chemoenzymatic catalysis.展开更多
The highly efficient chemoselectivity,stereoselectivity,and regioselectivity render enzyme catalysis an ideal pathway for the synthesis of various chemicals in broad applications.While the cofactor of an enzyme is nec...The highly efficient chemoselectivity,stereoselectivity,and regioselectivity render enzyme catalysis an ideal pathway for the synthesis of various chemicals in broad applications.While the cofactor of an enzyme is necessary but expensive,the conversed state of the cofactor is not beneficial for the positive direction of the reaction.Cofactor regeneration using electrochemical methods has the advantages of simple operation,low cost,easy process monitoring,and easy product separation,and the electrical energy is green and sustainable.Therefore,bioelectrocatalysis has great potential in synthesis by combining electrochemical cofactor regeneration with enzymatic catalysis.In this review,we detail the mechanism of cofactor regeneration and categorize the common electron mediators and enzymes used in cofactor regeneration.The reaction type and the recent progress are summarized in electrochemically coupled enzymatic catalysis.The main challenges of such electroenzymatic catalysis are pointed out and future developments in this field are foreseen.展开更多
Asymmetric reduction of the conjugated C=C bonds by the old yellow enzymes(OYEs)presents a promising field in the synthesis of chiral chemicals.Nevertheless,few natural OYEs have been applied in large-scale applicatio...Asymmetric reduction of the conjugated C=C bonds by the old yellow enzymes(OYEs)presents a promising field in the synthesis of chiral chemicals.Nevertheless,few natural OYEs have been applied in large-scale applications due to the requirement of costly NADPH and low operational stability.Herein,a stable and efficient fusion of YqjM from Bacillus subtilis and glucose dehydrogenase(GDH)from Bacillus megaterium was constructed to stereoselectively reduce the conjugated C=C bonds in a self-sufficient continuous process.The effects of the enzyme order and different linkers on the fusions were investigated by structural analysis and all-atom molecular dynamics simulation.The best fusion YqjM_G_GDH gave 98% conversion of 100 mmol/L 2-methylcyclopentenone with an excellent ee value(>99%)in 3 h,while the mixture of individual enzymes only obtained 68% conversion after more than 8 h.The improved substrate conversion of YqjM_G_GDH fusion was probably attributed to the increased flexibility of each fused enzyme and the shortening of the diffusion distance of NADPH regenerated.A one-pot process was designed to purify and immobilize the fusion on the Ni2t-nitrilotriacetic acid functionalized magnetic mesoporous silica nanoflowers.The resulting immobilized biocatalyst not only catalyzed the asymmetric reduction of various α,β-unsaturated ketones(20 mmol/L)continuously with only 50μmol/L NADPt to initiate the whole process,but also retained more than 82%of the initial activity after seven cycles,serving as a good candidate for the industrial applications.展开更多
基金supported by the Foundation for Distinguished Young Talents in Higher Education of Guangdong Province, China (2016KQNCX019 and 2016KQNCX027)the National Natural Science Foundation of China (31571041)+1 种基金the Guangdong Provincial Department of Education Innovating Strong National Engineering Major Project (2014GKXM031)Guangdong Provincial Universities and Colleges Pearl River Scholar Funded Scheme (2016)
文摘Activation of inflammatory responses regulates the transmission of pain pathways through an integrated network in the peripheral and central nervous systems.The immunopotentiator thymosin alpha-1(Tal)has recently been reported to have anti-inflammatory and neuroprotective functions in rodents.However,how Tα1 affects inflammatory pain remains unclear.In the present study,intraperitoneal injection of Tal attenuated complete Freund's adjuvant(CFA)-induced pain hypersensitivity,and decreased the up-regulation of pro-inflammatory cytokines(TNF-α,IL-1β,and IL-6)in inflamed skin and the spinal cord.We found that CFA-induced peripheral inflammation evoked strong microglial activation,but the effect was reversed by Tα1.Notably,Tα1 reversed the CFA-induced up-regulation of vesicular glutamate transporter(VGLUT)and down-regulated the vesicular γ-aminobutyric acid transporter(VGAT)in the spinal cord.Taken together,these results suggest that Tα1 plays a therapeutic role in inflammatory pain and in the modulation of microgliainduced pro-inflammatory cytokine production in addition to mediation of VGLUT and VGAT expression in the spinal cord.
基金Supported by the National Nature Science Foundation of China(Nos.21306039,21276060,21276062)the Natural Science Foundation of Hebei Province(B2015202082,B2016202027)the Tianjin City High School Science&Technology Fund Planning Project(20140513)
文摘Cross-linked enzyme aggregates(CLEAs) of nitrile hydratase(NHase) ES-NHT-118 from Escherichia coli were prepared by using ammonium sulfate as precipitating agent followed by cross-linking with dextran polyaldehyde for the first time. In this process, egg white was added as protein feeder for facilitating the formation of CLEAs. The optimal conditions of the immobilization process were determined. Michaelis constants(Km) of free NHase and NHase CLEAs were also determined. The NHase CLEAs exhibited increased stability at varied pH and temperature conditions compared to its free counterpart. When exposed to high concentrations of acrylamide, NHase CLEAs also exhibited effective catalytic activity.
基金supported by the National Natural Science Foundation of China(Nos.21576068,21276060,21276062,and 21306039)the Natural Science Foundation of Tianjin City(16JCY-BJC19800)+3 种基金the Natural Science Foundation of Hebei Province(B2015202082,B2016202027,and B2020202036)the Science and Technology Program Project of Tianjin(20YDTPJC00260)the Program for Top 100 Innovative Talents in Colleges and Universities of Hebei Province(SLRC2017029)Hebei High level personnel of support program(A2016002027).
文摘The continuous use of chemical dyes in various industries,and their discharge into industrial effluents,results in severe problems to human life and water pollution.Laccases have the ability to decolorize dyes and toxic chemicals in industrial effluents as green biocatalysts.Their possible industrial applications have been limited by poor reusability,low stability,and loss of free laccase action.In this research,lac-case was immobilized on zeolitic imidazolate framework coated multi-walled carbon nanotubes(Laccase@ZIF-8@MWCNTs)via metal affinity adsorption to develop an easy separable and stable enzyme.The optimum reaction conditions for immobilized laccase are at a pH of 3.0 and a temperature of 60℃.The immobilized laccase was enhanced in storage and thermal stability.The results indicated that Laccase@ZIF-8@MWCNTs still maintained 68%of its original activity after 10 times of repeated use.Most importantly,the biocatalytic system was applied for decolorization of different dyes(20 mg·L^(-1))without a mediator,and up to 97.4%for Eriochrome black T and 95.6%Acid red 88 was achieved in 25 min.Biocatalysts with these properties may be used in a variety of environmental and industrial applications.
文摘Immobilization is an effective method to promote the application of enzyme industry for improving the stability and realizing recovery of enzyme.To some extent,the performance of immobilized enzyme depends on the choice of carrier material.Therefore,the development of new carrier materials has been one of the key issues concerned by enzyme immobilization researchers.In this work,a novel organic–inorganic hybrid material,nickel-carnosine complex(NiCar),was synthesized for the first time by solvothermal method.The obtained NiCar exhibits spherical morphology,hierarchical porosity and abundant unsaturated coordination nickel ions,which provide excellent anchoring sites for the immobilization of proteins.His-tagged organophosphate-degrading enzyme(Opd A)and x-transaminase(ω-TA)were used as model enzymes to evaluate the performance of NiCar as a carrier.By a simple adsorption process,the enzyme molecules can be fixed on the particles of NiCar,and the stability and reusability are significantly improved.The analysis of protein adsorption on NiCar verified that the affinity adsorption between the imidazole functional group on the protein and the unsaturated coordination nickel ions on NiCar was the main force in the immobilization process,which provided an idea way for the development of new enzyme immobilization carriers.
基金supported by the National Key Research and Development Program of China(2018YFE0107500)the Natural Science Foundation Project of Science and Technology Agency of Chongqing YuZhong District(20200122)to Hu Yuana Natural Science Foundation Project of CQ CSTC(cstc2021jcyj-msxmX0276)to Chen YanMeng
文摘Casein kinase 1α(CK1α) mediates the phosphorylation and degradation of interferon-α/β receptor 1(IFNAR1) in response to viral infection. However, how CK1α regulates hepatitis B virus(HBV) replication and the anti-HBV effects of IFN-α are less reported. Here we show that CK1α can interact with IFNAR1 in hepatoma carcinoma cells and increased the abundance of IFNAR1 by reducing the ubiquitination levels in the presence of HBV.Furthermore, CK1α promotes the IFN-α triggered JAK-STAT signaling pathway and consequently enhances the antiviral effects of IFN-α against HBV replication. Our results collectively provide evidence that CK1α positively regulates the anti-HBV activity of IFN-α in hepatoma carcinoma cells, which would be a promising therapeutic target to improve the effectiveness of IFN-α therapy to cure CHB.
基金Shandong Province Key R&D Program(Major Innovation Project)(No.2020CXGC010603,No.2019JZZY011003)National Natural Science Foundation of China(No.31801527)Taishan Industry Leading Talent Project(No.tscy20180103).
文摘D-allulose has very little content in nature,and it needs to be synthesized artificially and meet the purity requirements of industrial grade.The basic physical and chemical properties of D-allulose,its preparation methods and many different ways of isolation and purification were described.In order to achieve the goal of industrial production of D-allulose as soon as possible,the research progress of D-allulose isolation and purification technologies at home and abroad in recent years was classified and discussed,so as to provide useful reference for the practical improvement of D-allulose isolation and purification process technologies.
基金supported by the National Key Research and Development Program of China(No.2023YFA0914500)the National Natural Science Foundation of China(Nos.22378096,22308083,and 22178083)+1 种基金the Natural Science Foundation of Hebei Province(No.B2022202014)the Educational Commission of Hebei Province(No.JZX2023012).
文摘Chiral compounds have a huge market demand in the fields of pharmaceuticals,pesticides,and fine chemicals.Enzymatic electrosynthesis can couple enzyme catalysis,possessing high product purity,high efficiency,and mild conditions,with electrochemical regeneration of expensive cofactor nicotinamide adenine dinucleotide(NADH),possessing easy process monitoring and simple operation for efficient chiral synthesis.In this study,hydrophobic covalent organic framework(COF)was synthesized as the immobilized carrier,which not only enhanced the enzyme catalysis through enriching substrate but also enhanced the stability and reuse of the enzyme.Besides,Rh complex was anchored on hydrophilically-modified electrode to promote the regeneration of NADH,where the anchor of Rh complex can effectively avoid the mutual deactivation from the interference between electron mediator and enzyme,and simplify the separation of products.The immobilized enzyme catalysis and the electrochemical cofactor regeneration were coupled to construct an enzymatic electrosynthesis system for the efficient asymmetric reduction to obtain chiral alcohols,with a maximum turnover frequency(TOF)of 101.1 h^(-1).Furthermore,the relevant parameters of the system were optimized,and the substrate scope was expanded.
基金supported by the National Key Research and Development Program of China(Grant No.2023YFA0914500)the National Natural Science Foundation of China(Grant Nos.22378096,and 22308083)Baoding City Science and Technology Bureau(Grant No.2494F016).
文摘The capture and conversion of CO_(2) to valuable products has garnered significant attention.The electroenzymatic reduction of CO_(2) is of particular interest owing to its high selectivity and yield.However,this process is hindered by various challenges,such as the low CO_(2) solubility in water and the requirement for efficient cofactor regeneration.In this work,we developed a green,water-based,room-temperature synthesis of Zn-MOF-74 for enzyme immobilization.Formate dehydrogenase(FDH)and carbonic anhydrase(CA)are co-encapsulated within the Zn-MOF-74 framework to create a dual enzyme cascade system.This strategy effectively enhances enzyme proximity,stability,and reusability to subsequently promote the enzyme cascade reaction efficiency.Additionally,the adsorption of CO_(2) by CA improves the solubility of CO_(2) in water.NADH regeneration is achieved using an electron mediator,2-hydroxy-1-naphthoquinone(HNQ).The optimized electroenzymatic system achieved a formate yield of 3.01 mM within 3 h,which was 4.98 times higher than that of the pristine free enzyme system and 2.27 times higher than the free enzyme system with NADH regeneration.
基金financial support from the Defense Industrial Technology Development Program(JCKY2023603C016).
文摘Escalating electromagnetic(EM)pollution and advanced stealth technologies require next-generation microwave absorbers that combine broadband response,strong attenuation,and lightweight characteristics with high-temperature stability.In this work,a vacancy-mediated strategy is proposed to tailor EM losses in Cr-doped lanthanum manganite perovskites(LaMn_(1-x)Cr_(x)O_(3))synthesized via a sol–gel method.The cooperative modulation between Mn-site cation vacancies and oxygen vacancies enables a well-balanced contribution of polarization loss and conduction loss,resulting in excellent absorption performance.Specifically,LaMn_(0.85)Cr_(0.15)O_(3) achieves a remarkable minimum reflection loss(RL_(min))of-75.37 dB and an effective absorption bandwidth(EAB)of 6.0 GHz at a thickness of only 2.8 mm.Structural and spectroscopic analyses reveal that Cr^(3+)substitution induces Mn vacancies and modulates oxygen vacancy concentrations,thereby generating defect dipoles and facilitating carrier migration.Density functional theory(DFT)calculations further elucidate the role of Cr-induced defect states in enhancing conduction and polarization losses.This vacancy-engineered approach not only establishes a new paradigm for designing high-efficiency perovskite-based microwave absorbers but also offers significant potential for high-temperature EM compatibility applications.
文摘A novel integrated nanobiocatalyst system based on an enzyme@silica nanoflower@metal-organic framework (enzyme@SNF@ZIF-8) structure with improved stability is fabricated for the first time. The versatility of this system is validated using penicillin G acylase (PGA) and catalase (CAT) as model enzymes. The microporous ZIF-8 layer can be controlled by varying the number of ZIF-8 coating cycles, which produces PGA@SNF@ZIF-8 nanobiocatalysts with different ZIF-8 layer thicknesses. After the second ZIF-8 coating cycle, a PGA@SNF@ZIF-8(2) structure with a homogeneous and well-intergrown ZIF-8 layer is formed, which possesses excellent mechanical and chemical stability. Moreover, PGA@SNF@ZIF-8(2) shows improved thermal/storage stability and reusability compared with free PGA and PGA immobilized on silica nanoflowers (PGA@SNF). The obtained CAT-based nanobiocatalysts (CAT@SNF@ZIF-8(2)) also show excellent catalytic performance.
基金supported by the National Natural Science Foundation of China(No.22178083)the Natural Science Foundation of Hebei Province(C2019208174 and B2022202014)+1 种基金the S&T Program of Hebei(20372802D,21372804D,and 21372805D)the Natural Science Foundation of Tianjin City(20JCYBJC00530)
文摘Dendritic mesoporous silica nanoparticles(DMSNs)are a new class of solid porous materials used for enzyme immobilization support due to their intrinsic characteristics,including their unique open central-radial structures with large pore channels and their excellent biocompatibility.In this review,we review the recent progress in research on enzyme immobilization using DMSNs with different structures,namely,flower-like DMSNs and tree-branch-like DMSNs.Three DMSN synthesis methods are briefly compared,and the distinct characteristics of the two DMSN types and their effects on the catalytic performance of immobilized enzymes are comprehensively discussed.Possible directions for future research on enzyme immobilization using DMSNs are also proposed.
基金This work was financially supported by the National Natural Science Foundation of China(Nos.21901058,21908040 and 22078081)the Science and Technology Research Project of Hebei Higher Education(ZD2019045)+2 种基金the Natural Science Foundation of Hebei province(B2017202056 and B2019202216)the Natural Science Foundation of Tianjin City(20JCYBJC00530)the Foundation of Tianjin Key Laboratory of Brine Chemical Engineering and Resource Eco-utilization(Tianjin University of Science and Technology),People's Republic of China(BCERE202001).
文摘Main observation and conclusion Aqueous heterogeneous catalysis is a green,sustainable catalytic process that attracts increasing attention,but it often suffers from poor mass transfer,substrate adsorption and catalyst dispersion.Herein,we synthesized a type of amphiphilic core-shell catalysts with a hydrophilic polydopamine(PDA)shell and a hydrophobic dendritic organosilica nanoparticle(DON)core for heterogeneous catalysis in water.
基金supported by the National Natural Science Foundation of China(Nos.21901058,21908040,and 21878068)Tianjin Enterprise Science and Technology Commissioner,China(21YDTPJC00810)+2 种基金Science Technology Research Project of Higher Education of Hebei Province,China(QN2021045)Hebei Province Postgraduate Innovation Funding Project,China(CXZZSS2021027)National College Student’s Science and Technology Innovation Project,China(202010080038)。
文摘The worldwide application of organophosphorus pesticides(OPs)has promoted agricultural development,but their gradual accumulation in soil and water can seriously affect the central nervous system of humans and other mammals.Organophosphorus hydrolase(OPH)is an effective enzyme that can catalyze the degradation of the residual OPs.However,the degradation products such as p-nitrophenol(p-NP)is still toxic.Thus,it is of great significance to develop a multi-functional support that can be simultaneously used for the immobilization of OPH and the further degradation of p-NP.Herein,a visible light assisted enzyme-photocatalytic integrated catalyst was constructed by immobilizing OPH on hollow structured Au-TiO_(2)(named OPH@H-Au-TiO_(2))for the degradation of OPs.The obtained OPH@H-Au-TiO_(2)can degrade methyl parathion to p-NP by OPH and then degrade p-NP to hydroquinone with low toxicity by using H-Au-TiO_(2)under visible light.OPH molecules were immobilized on HAu-TiO_(2)through adsorption method to prepare OPH@H-Au-TiO_(2).After 2.5 h of reaction,methyl parathion is completely degraded,and about 82.64%of the generated p-NP is further degraded into hydroquinone.After reused for 4 times,the OPH@H-Au-TiO_(2)retains more than 80%of the initial degradation activity.This research presents a new insight in designing and constructing multi-functional biocatalyst,which greatly expands the application scenarios and industrial value of enzyme catalysis.
基金the National Natural Science Foundation of China(Grant No.81972899)Natural Science Foundation of Tianjin City(Grant No.18JCQNJC14500)+1 种基金CAMS Innovation Fund for Medical Sciences(Grant No.2017-I2M-3-022)Specific Program for High-Tech Leader&Team of Tianjin Government,Tianjin innovation and promotion plan key innovation team of immunoreactive biomaterials.
文摘Here,we reported a cancer nanovaccine based on SiO_(2)nanoflowers with a special radial pore structure,which greatly enhanced cross-presentation and induced the production of cytotoxic T lymphocyte cells secreting granzymes B and interferon-γ.The antigen ovalbumin was covalently conjugated onto the as-synthesized hierarchical SiO_(2)nanoflowers,and the adjuvant cytosine-phosphate-guanine was electrostatically adsorbed into their radial pore by simple mixing before use.The nanovaccine exhibited excellent storage stability without antigen release after 27 days of incubation,negligible cytotoxicity to dendritic cells,and a high antigen loading capacity of 430±66 mg·g^(−1)support.Besides,the nanovaccine could be internalized by dendritic cells via multiple pathways.And the enhancement of antigen/adjuvant uptake and lysosome escape of antigen were observed.Noteworthy,in vitro culture of bone marrow-derived dendritic cells in the presence of nanovaccine proved the activation of dendritic cells and antigen cross-presentation as well as secretion of proinflammatory cytokines.Besides,in vivo study verified the targeting of nanovaccine to draining lymph nodes,the complete suppression of tumor in six out of ten mice,and the triggering of notable tumor growth delay.Overall,the present results indicated that the nanovaccine can be served as a potential therapeutic vaccine to treat cancer.
基金This work was supported by the National Natural Science Foundation of China(Nos.22078081,21908040,21901058,and 22178083)the Natural Science Foundation of Hebei Province(Nos.B2020202021 and B2019202216)+2 种基金Key Research and Development Program of Hebei Province(No.20372802D)Open Project Funding of the State Key Laboratory of Biocatalysis and Enzyme Engineering(No.SKLBEE2020011)the Natural Science Foundation of Tianjin(No.20JCYBJC00530).
文摘The hollow spherical covalent organic frameworks(COFs)have a wide application prospect thanks to their special structures.However,the controllable synthesis of uniform and stable hollow COFs is still a challenge.We herein propose a self-templated method for the preparation of hollow COFs through the Ostwald ripening mechanism under ambient conditions,which avoids most disadvantages of the commonly used hard-templating and soft-templating methods.A detailed time-dependent study reveals that the COFs are transformed from initial spheres to hollow spheres because of the inside-out Ostwald ripening process.The obtained hollow spherical COFs have high crystallinity,specific surface area(2,036 m^(2)·g^(−1)),stability,and single-batch yield.Thanks to unique hollow structure,clear through holes,and hydrophobic pore environment of the hollow spherical COFs,the obtained immobilized lipase(BCL@H-COF-OMe)exhibits higher thermostability,polar organic solvent tolerance,and reusability.The BCL@H-COF-OMe also shows higher catalytic performance than the lipase immobilized on non-hollow COF and free lipase in the kinetic resolution of secondary alcohols.This study provides a simple approach for the preparation of hollow spherical COFs,and will promote the valuable research of COFs in the field of biocatalysis.
基金supported by the National Natural Science Foundation of China(Grant No.22378096)the Natural Science Foundation of Hebei Province(Grant No.B2023202014)+1 种基金the Science Technology Research Project of Higher Education of Hebei Province(Grant Nos.QN2021045,and QN2023207)the Tianjin Science and Technology Project(Grant No.22KPHDRC00260).
文摘Unspecific peroxygenases exhibit high activity for the selective oxyfunctionalization of inert C(sp3)–H bonds using only H_(2)O_(2) as a clean oxidant,while also exhibiting sensitivity to H_(2)O_(2) concentration.CdS-based semiconductors are promising for the photosynthesis of H_(2)O_(2) owing to their adequately negative potential for oxygen reduction reaction via a proton-coupled electron transfer process,however,they suffer from fast H_(2)O_(2) decomposition on the surface of pristine CdS.Therefore,[Cp*Rh(bpy)H_(2)O]2+,a highly selective proton-coupled electron transfer catalyst,was anchored onto a supramolecular polymer-grafted CdS nanoflower to construct an efficient integrated photocatalyst for generating H_(2)O_(2),mitigating the surface issue of pristine CdS,increasing light absorption,accelerating photonic carrier separation,and enhancing oxygen reduction reaction selectivity to H_(2)O_(2).This photocatalyst promoted the light driven H_(2)O_(2) generation rate up to 1345μmol·L^(-1)·g^(-1)·h^(-1),which was 2.4 times that of pristine CdS.The constructed heterojunction photocatalyst could supply H_(2)O_(2) in situ for nonspecific peroxygenases to catalyze the C–H oxyfunctionalization of ethylbenzene,achieving a yield of 81%and an ee value of 99%under optimum conditions.A wide range of substrates were converted to the corresponding chiral alcohols using this photo-enzyme catalytic system,achieving the corresponding chiral alcohols in good yield(51%–88%)and excellent enantioselectivity(90%–99%ee).
基金The authors thank the National Natural Science Foundation of China(grant No.81972899)Natural Science Foundation of Tianjin City(grant No.18JCQNJC14500)+1 种基金CAMS Innovation Fund for Medical Sciences(grant No.2017-I2M-3-022)Specific Program for High-Tech Leader&Team of Tianjin Government,Tianjin Innovation and Promotion Plan Key Innovation Team of Immunoreactive Biomaterials.
文摘A microscale vaccine containing SiO_(2)nanoparticles loaded in CaC〇3 microparticles was constructed using the co-precipitation method.The antigen ovalbumin(OVA)was covalently conjugated with SiO_(2)nanoparticles,and these nanoparticles and CpG were co-encapsulated into CaCO_(3)microparticles,generating a vaccine with a size of approximately 5.2μm.Scanning electron microscopy(SEM),energy-dispersive X-ray(EDX),elemental mapping,and Fourier transform infrared(FTIR)analyses confirmed the successful preparation of the microscale vaccine;the vaccine had good storage stability without sustained antigen release,and negligible cytotoxicity to dendritic cells(DCs)and macrophages.Compared to SiO_(2)nanoparticles,the microscale vaccine can significantly improve antigen/adjuvant uptake.DCs internalized the entire microscale vaccine into lysosomes via macropinocytosis,and an increase in antigen endo/lysosomal escape was observed by confocal User scanning microscopy(CLSM).Specifically,DCs pulsed with the vaccine were fully mature,expressing high levels of costimulatory molecules(CD40,CD80,and CD86),MHCⅡ,and MHCⅠand secreting high levels of proinflammatory cytokines(IL-12,TNF-α,IL-1β,and IL-6).In addition,the vaccine had good in vivo biocompatibility,could protect the antigen from rapid degradation,and increased the retention time in lymph nodes.SiO_(2)nanoparticles-in-CaCO_(3)microparticles were an excellent carrier for antigen and adjuvant delivery.Hopefully,this study can provide some information on the design of microscale carriers for vaccine delivery systems.
基金financially supported by the National Key Research and Development Program of China(No.2021YFC2104100)the National Natural Science Foundation of China(Nos.21901058,22178083 and 22078081)+2 种基金the S&T program of Hebei(Nos.21372805D,21372804D and 20372802D)the Natural Science Foundation of Tianjin City(No.20JCYBJC00530)the Natural Science Foundation of Hebei Province(No.B2022202014).
文摘Core-shell structured magnetic wrinkled organosilica-based metal-enzyme integrated catalysts were synthesized,and their catalytic performances were studied in the chemoenzymatic dynamic kinetic resolution of chiral amines in an organic solvent,as well as in the chemoenzymatic synthesis of chiral alcohols in water.Structureperformance studies revealed the important influence of their tunable structure and composition on the optimization of activity,stability,and recyclability in chemoenzymatic catalysis.
基金supported by the National Natural Science Foundation of China(Nos.21908040,21878068,and 22178083)the Natural Science Foundation of Hebei Province(B2020202021)+2 种基金the Science and Technology Program Project of Tianjin(20YDTPJC00260)Key Research and Development Program of Hebei Province(20372802D)the Natural Science Foundation of Tianjin(20JCYBJC00530)
文摘The highly efficient chemoselectivity,stereoselectivity,and regioselectivity render enzyme catalysis an ideal pathway for the synthesis of various chemicals in broad applications.While the cofactor of an enzyme is necessary but expensive,the conversed state of the cofactor is not beneficial for the positive direction of the reaction.Cofactor regeneration using electrochemical methods has the advantages of simple operation,low cost,easy process monitoring,and easy product separation,and the electrical energy is green and sustainable.Therefore,bioelectrocatalysis has great potential in synthesis by combining electrochemical cofactor regeneration with enzymatic catalysis.In this review,we detail the mechanism of cofactor regeneration and categorize the common electron mediators and enzymes used in cofactor regeneration.The reaction type and the recent progress are summarized in electrochemically coupled enzymatic catalysis.The main challenges of such electroenzymatic catalysis are pointed out and future developments in this field are foreseen.
基金financially supported by the National Key Research and Development Program of China(No.2021YFC2104100)the National Natural Science Foundation of China(Nos.21901058,22178083 and 22078081)+3 种基金the Hebei Key Research and Development Project(Nos.21372805D,21372804D and 20372802D)the Science and Technology Research Project of Hebei Higher Education(No.ZD2019045)the Natural Science Foundation of Hebei province(Nos.B2019202216,B2017202056 and C2019208174)the Natural Science Foundation of Tianjin City(No.20JCYBJC00530).
文摘Asymmetric reduction of the conjugated C=C bonds by the old yellow enzymes(OYEs)presents a promising field in the synthesis of chiral chemicals.Nevertheless,few natural OYEs have been applied in large-scale applications due to the requirement of costly NADPH and low operational stability.Herein,a stable and efficient fusion of YqjM from Bacillus subtilis and glucose dehydrogenase(GDH)from Bacillus megaterium was constructed to stereoselectively reduce the conjugated C=C bonds in a self-sufficient continuous process.The effects of the enzyme order and different linkers on the fusions were investigated by structural analysis and all-atom molecular dynamics simulation.The best fusion YqjM_G_GDH gave 98% conversion of 100 mmol/L 2-methylcyclopentenone with an excellent ee value(>99%)in 3 h,while the mixture of individual enzymes only obtained 68% conversion after more than 8 h.The improved substrate conversion of YqjM_G_GDH fusion was probably attributed to the increased flexibility of each fused enzyme and the shortening of the diffusion distance of NADPH regenerated.A one-pot process was designed to purify and immobilize the fusion on the Ni2t-nitrilotriacetic acid functionalized magnetic mesoporous silica nanoflowers.The resulting immobilized biocatalyst not only catalyzed the asymmetric reduction of various α,β-unsaturated ketones(20 mmol/L)continuously with only 50μmol/L NADPt to initiate the whole process,but also retained more than 82%of the initial activity after seven cycles,serving as a good candidate for the industrial applications.