Biocatalysis with nicotinamide adenine dinucleotide phosphate(NADP)-dependent oxidoreductases faces a challenge in improving the efficiency of the costly cofactor utilization.Although enzyme fusion can offer cofactor ...Biocatalysis with nicotinamide adenine dinucleotide phosphate(NADP)-dependent oxidoreductases faces a challenge in improving the efficiency of the costly cofactor utilization.Although enzyme fusion can offer cofactor regeneration,the high-volume input and limited cofactor recyclability still make the enzymatic processes unsustainable.Therefore,it is of great significance to reduce cofactor input in a fusion enzyme(FuE)system,but no successful practice has been reported.Herein,we design a decapeptide bridge,RRRQRRRARR(R10),with high affinity for NADPH to construct fusion oxidoreductases(phenylacetone monooxygenase and phosphite dehydrogenase)for ester synthesis and NADP recycling.The peptide bridge enables electrostatic cofactor channeling that transports NADPH/NADP^(+)across the peptide between the enzymes’NADP-binding pockets,so the fusion enzyme(FuE-R10)presents 2.1-folds and 2.0-folds higher conversions than mixed free enzymes and a flexible linker(GGGGSGGGGS)-fused enzyme,respectively,at NADPH/FuE of 0.1.The fusion enzyme,FuE-R5,bridged by a half-shortened linker,is proved more effective in facilitating cofactor channeling;compared to the mixed free enzymes,FuE-R5 exhibits two orders of magnitude reduction of NADPH input in ester synthesis.The work has thus demonstrated the potential of the cofactor bridging strategy in the development of sustainable cofactor-dependent cascade biocatalysis.展开更多
Pre-harvest sprouting(PHS)or vivipary is a major problem affecting cereal quality and grain quantity and is primarily linked to the dysregulation of abscisic acid(ABA)biosynthesis in plants.Therefore,elucidating the m...Pre-harvest sprouting(PHS)or vivipary is a major problem affecting cereal quality and grain quantity and is primarily linked to the dysregulation of abscisic acid(ABA)biosynthesis in plants.Therefore,elucidating the molecular mechanisms governing seed dormancy is crucial for developing strategies to improve crop productivity.In this study,we identified a novel viviparous maize mutant,viviparous-like 5(vp-like5),which exhibits precocious germination in developing seeds.Through map-based cloning,we discovered that ZmCNX6,which encodes a small subunit of molybdopterin synthase essential for molybdenum cofactor(MoCo)biosynthesis,is the causal gene responsible for the vp-like5 phenotype.Biochemical assays have demonstrated significantly reduced activities of MoCo-dependent enzymes,including aldehyde oxidase(AO),xanthine dehydrogenase(XDH),and nitrate reductase(NR),in vplike5.AO is essential for the ABA biosynthesis,and the observed ABA deficiency in vp-like5 likely drives the viviparous phenotype.Expression analysis showed that ZmCNX6 was stably expressed during seed development,indicating its significant role in seed development.Furthermore,overexpression of ZmCNX6 not only enhanced the activities of MoCo-dependent enzymes but also improved drought tolerance in maize.Collectively,our study revealed ZmCNX6 as a multifunctional hub coordinating MoCo metabolism,ABA-dependent dormancy regulation,and abiotic stress responses,offering a potential target for simultaneously mitigating vivipary and improving drought resistance in maize.展开更多
Cytochrome P450 enzymes catalyze diverse oxidative transformations at the expense of reduced nicotinamide adenine dinucleotide phosphate(NADPH),however,their applications remain limited largely because NADPH is cost-p...Cytochrome P450 enzymes catalyze diverse oxidative transformations at the expense of reduced nicotinamide adenine dinucleotide phosphate(NADPH),however,their applications remain limited largely because NADPH is cost-prohibitive for biocatalysis at scale yet tightly regulated in host cells.A highly challenging task for P450 catalysis has been to develop an alternative and biocompatible electrondonating system.Here we engineered P450 BM3 to favor reduced nicotinamide cytosine dinucleotide(NCDH)and created non-natural cofactor-dependent P450 catalysis.Two outstanding mutants were identified with over 640-fold NCDH preference improvement and good catalytic efficiencies of over15,000 M^(-1)s^(-1)for the oxidation of the fatty acid probe 12-(para-nitrophenoxy)-dodecanoate.Molecular docking analysis indicated that these mutants bear a compacted cofactor entrance.Upon fusing with an NCD-dependent formate dehydrogenase,fused proteins functioned as NCDH-specific P450catalysts by using formate as the electron donor.Importantly,these mutants and fusions catalyzed NCDH-dependent hydroxylation of fatty acids with similar chain length preference to those by natural P450 BM3 in the presence of NADPH and also similar regioselectivity for subterminal hydroxylation of lauric acid.As P450 BM3 and its variants are catalytically powerful to take diverse substrates and convey different reaction paths,our results offer an exciting opportunity to devise advanced cell factories that convey oxidative biocatalysis with an orthogonal reducing power supply system.展开更多
The iron molybdenum cofactor(FeMoco) of nitrogenase MoFe protein from Azotobacter vinelandii OP was extracted by N methylformamide(NMF). The effects of FeMoco(in NMF) on electronic spectrum and fluorescence...The iron molybdenum cofactor(FeMoco) of nitrogenase MoFe protein from Azotobacter vinelandii OP was extracted by N methylformamide(NMF). The effects of FeMoco(in NMF) on electronic spectrum and fluorescence intensity of fluorescein dimercury acetate(FDMA)(in 1 mol/L NaOH) were investigated by fluorophotometric titrations and compared with those of (NH 4) 2MoS 4 and complexes of (NH 4) 2MoS 4 with Na 2S or Na 2S 2 or (NH 4) 2S x on the relative properties of FDMA. It was found that the electronic spectrum of FDMA displayed hypsochromic shift(17 nm) in the presence of FeMoco just like that in the presence of other inorganic sulfides and that the titration curve for the quench of FDMA with FeMoco is very similar to that for the quench of FDMA with complex of (NH 4) 2MoS 4 with Na 2S 2(mole ratio is 1∶3). The results showed that FeMoco(N) probably contained S—S bonds and its structure was found to be changed compared with Kim Rees structural model. This change should profit the formation of the polymer.展开更多
Two k-hyperconnection expressions of a general k-order cofactor Y<sub>（i,j）</sub> are presentedfor the indefinite parameter matrix Y of a linear system by applying directed hypergraph theory,and based on...Two k-hyperconnection expressions of a general k-order cofactor Y<sub>（i,j）</sub> are presentedfor the indefinite parameter matrix Y of a linear system by applying directed hypergraph theory,and based on it a decomposition theorem of Y<sub>（i,j）</sub> is derived.By this theorem,the multi-leveltearing and analysis can be carried out easily for any linear large system.This is a new mul-tilevel topological analysis method.Using proposed method the scale of systems which can betopologically analysed by a computer will be enlarged.展开更多
Microtubules play important roles in neuronal morphogenesis, including cellular polarization, neurite growth, and branching. A microtubule is a polymer of a- and β-tubulin heterodimers that are formed by a multistep ...Microtubules play important roles in neuronal morphogenesis, including cellular polarization, neurite growth, and branching. A microtubule is a polymer of a- and β-tubulin heterodimers that are formed by a multistep process assisted by at least five tubulin-fold- ing cofactors （TBCA-E） （Lopez-Fanarraga et al., 2001）. Newly syn- thesized α- and β-tubulins associate with the cytosolic chaperonin complex （CCT）, and then the quasi-native tubulins interact with five tubulin-folding cofactors. The α- and β-tubulins first interact with TBCB and TBCA, which are further transferred to TBCE and TBCD, respectively. The α- and β-tubulins, TBCC, TBCD, and TBCE form a supercomplex, and guanosine triphosphate （GTP） hydrolysis occurs within the supercomplex, releasing a tubulin heterodimer. In addition to their functions in tubulin folding and tubulin heterodimer formation, excess amounts of TBCD or TBCE may interact with tubulin heterodimers, leading to their degrada- tion. Thus, tubulin-folding cofactors may play a role in both the synthesis and degradation of tubulin heterodimers.展开更多
Introduction: High grade dysplasia of the cervix has a high incidence and can progress to cervical cancer. The aim was to study cofactors associated with high-grade cervical dysplasia. Methodology: This was a retrospe...Introduction: High grade dysplasia of the cervix has a high incidence and can progress to cervical cancer. The aim was to study cofactors associated with high-grade cervical dysplasia. Methodology: This was a retrospective case-control study without matching. Women with high grade dysplasia were the cases while those with a normal screening test represented the controls. The study took place at the Gabriel Touré University Hospital Center in Bamako. We included 351 cases and 420 controls. The capture and analysis were performed using the SPSS 20 software. A univariate and multivariate logistic regression analysis was performed for the analysis of risk cofactors. The statistical tests used were the odds ratio and its confidence interval and the statistical significance threshold was set at p Results: In univariate analysis, the co-factors statistically significantly associated with the occurrence of high-grade dysplasia were parity 0.6 (0.5 - 0.9), gestational 0.7 (0.5 - 0.9), smoking of the spouse 3.4 (1.1 - 11.3), the non-schooling 1.4 (1.2 - 2.1). In multivariate analysis after adjusting for confounding factors, two co-factors have significantly increased the risk of high-grade dysplasia: lack of schooling 1.4 (1.2 - 2.0) and polygamy 1.5 (1.4 - 2.5). Conclusion: At the end of this study, polygamy and lack of schooling were the main risk factors. The prevention of cervical cancer will go through the education of girls and women as well as communication for behavioral change and social change.展开更多
In literature, neutral polyethylene oxide (PEO) flocculated fines at low shear rates, while with cofactor (CF) addition, the formed CF-PEO complex showed larger ability to bridge fines, producing flocs. In this work, ...In literature, neutral polyethylene oxide (PEO) flocculated fines at low shear rates, while with cofactor (CF) addition, the formed CF-PEO complex showed larger ability to bridge fines, producing flocs. In this work, some process factors were found having significant effects on fines flocculation. Increases in CF to PEO ratio at constant PEO enhanced the bridging bonds, causing increases in flocculation initial rate (efficiency), amplitude (floc size), and fastness (a decrease in characteristic time). On the other hand, an increase in stirring rate (shear rate) in flocculation vessel caused decreases in initial rate and amplitude, and an increase in the fastness. All runs showed transient flocculation;the amplitude increased with time, reached maximum at equilibrium, and then started to decrease showing deflocculation. In brief, the CF to PEO ratio and the shear rate were found important parameters in mill operation, having significant effects on flocculation efficiency, fastness, and floc size.展开更多
Dicarboxylates are valuable platform compounds with a broad range of applications.The in vitro biosynthetic system used to produce dicarboxylates fromω-amino acids via the natural pathway requires costly cofactors an...Dicarboxylates are valuable platform compounds with a broad range of applications.The in vitro biosynthetic system used to produce dicarboxylates fromω-amino acids via the natural pathway requires costly cofactors and co-substrates,which restricts its economic feasibility of use.In this study,we designed a cofactor-and co-substrate-free artificial pathway for the production of dicarboxylates fromω-amino acids.Only three enzymes(viz.,amine oxidase from Kluyveromyces marxianus DMKU3-1042,xanthine oxidase from bovine milk,and cata-lase from Aspergillus niger)were required for dicarboxylate production.Succinate(0.79 g g^(-1)),glutarate(0.83 g g^(-1)),and adipate(0.77 g g^(-1))were produced in high yields from the correspondingω-amino acids through the in vitro biosynthetic system with the artificial pathway.Glutarate could also be produced from l-lysine by further introducing l-lysine monooxygenase and 5-aminovaleramide amidohydrolase from Pseudomonas putida KT2440 into the in vitro biosynthetic system,with the cofactor-and co-substrate-free system achieving a product yield of 0.63 g g^(-1).Considering its desirable characteristics,this artificial pathway-based in vitro biosynthetic system may be a promising alternative for dicarboxylate production from biotechnologically producedω-amino acids.展开更多
Chemoenzymatic catalysis can give full play to the advantages of versatile reactivity of chemocatalysis and excellent chemo-,regio-,and stereoselectivities of biocatalysis.These chemoenzymatic methods can not only sav...Chemoenzymatic catalysis can give full play to the advantages of versatile reactivity of chemocatalysis and excellent chemo-,regio-,and stereoselectivities of biocatalysis.These chemoenzymatic methods can not only save resource,cost,and operating time but also reduce the number of reaction steps,and avoid separating unstable intermediates,leading to the generation of more products under greener circumstances and thereby playing an indispensable role in the fields of medicine,materials and fine chemicals.Although incompatible challenges between chemocatalyst and biocatalyst remain,strategies such as biphasic system,artificial metalloenzymes,immobilization or supramolecular host,and protein engineering have been designed to overcome these issues.In this review,chemoenzymatic catalysis according to different chemocatalysis types was classifiably described,and in particular,the classic dynamic kinetic resolutions(DKR)and cofactor regeneration were summarized.Finally,the bottlenecks and development of chemoenzymatic catalysis were summarized,and future development was prospected.展开更多
DNAzyme machines play critical roles in the fields of cell imaging,disease diagnosis,and cancer therapy.However,the applications of DNAzyme machines are limited by the nucleases-induced degradation,non-specific bindin...DNAzyme machines play critical roles in the fields of cell imaging,disease diagnosis,and cancer therapy.However,the applications of DNAzyme machines are limited by the nucleases-induced degradation,non-specific binding of proteins,and insufficient provision of cofactors.Herein,protected DNAzyme machines with different cofactor designs(referred to as Pro Ds)were nanoengineered by the construction of multifunctional metal-phenolic nanoshells to deactivate the interferential proteins,including nucleases and non-specific binding proteins.Moreover,the nanoshells not only facilitate the cellular internalization of Pro Ds but provide specific metal ions acting as cofactors of the designed DNAzymes.Cellular imaging results demonstrated that Pro Ds could effectively and simultaneously monitor multiple tumor-related micro RNAs in living cells.This facile and rapid strategy that encapsulates DNAzyme machines into the protective metal-phenolic nanoshells is anticipated to extend to a wide range of functional nucleic acidsbased biomedical applications.展开更多
The mechanical background of the bivariate spline space of degree 2 and smoothness 1 on rectangular partition is presented constructively. Making use of mechanical analysis method, by acting couples along the interior...The mechanical background of the bivariate spline space of degree 2 and smoothness 1 on rectangular partition is presented constructively. Making use of mechanical analysis method, by acting couples along the interior edges with suitable evaluations, the deflection surface is divided into piecewise form, therefore, the relation between a class of bivariate splines on rectangular partition and the pure bending of thin plate is established. In addition, the interpretation of smoothing cofactor and conformality condition from the mechanical point of view is given. Furthermore, by introducing twisting moments, the mechanical background of any spline belong to the above space is set up.展开更多
A contiguous derivation of radius and center of the insphere of a general tetrahedron is given. Therefore a linear system is derived. After a transformation of it the calculation of radius and center can be separated ...A contiguous derivation of radius and center of the insphere of a general tetrahedron is given. Therefore a linear system is derived. After a transformation of it the calculation of radius and center can be separated from each other. The remaining linear system for the center of the insphere can be solved after discovering the inverse of the corresponding coefficient matrix. This procedure can also be applied in the planar case to determine radius and center of the incircle of a triangle.展开更多
文摘Biocatalysis with nicotinamide adenine dinucleotide phosphate(NADP)-dependent oxidoreductases faces a challenge in improving the efficiency of the costly cofactor utilization.Although enzyme fusion can offer cofactor regeneration,the high-volume input and limited cofactor recyclability still make the enzymatic processes unsustainable.Therefore,it is of great significance to reduce cofactor input in a fusion enzyme(FuE)system,but no successful practice has been reported.Herein,we design a decapeptide bridge,RRRQRRRARR(R10),with high affinity for NADPH to construct fusion oxidoreductases(phenylacetone monooxygenase and phosphite dehydrogenase)for ester synthesis and NADP recycling.The peptide bridge enables electrostatic cofactor channeling that transports NADPH/NADP^(+)across the peptide between the enzymes’NADP-binding pockets,so the fusion enzyme(FuE-R10)presents 2.1-folds and 2.0-folds higher conversions than mixed free enzymes and a flexible linker(GGGGSGGGGS)-fused enzyme,respectively,at NADPH/FuE of 0.1.The fusion enzyme,FuE-R5,bridged by a half-shortened linker,is proved more effective in facilitating cofactor channeling;compared to the mixed free enzymes,FuE-R5 exhibits two orders of magnitude reduction of NADPH input in ester synthesis.The work has thus demonstrated the potential of the cofactor bridging strategy in the development of sustainable cofactor-dependent cascade biocatalysis.
基金supported by the Open Project of Key Laboratory,Xinjiang Uygur Autonomous Region(2023D04070)the Agricultural Science and Technology Innovation Program of CAAS(CAAS-CSCB-202403)。
文摘Pre-harvest sprouting(PHS)or vivipary is a major problem affecting cereal quality and grain quantity and is primarily linked to the dysregulation of abscisic acid(ABA)biosynthesis in plants.Therefore,elucidating the molecular mechanisms governing seed dormancy is crucial for developing strategies to improve crop productivity.In this study,we identified a novel viviparous maize mutant,viviparous-like 5(vp-like5),which exhibits precocious germination in developing seeds.Through map-based cloning,we discovered that ZmCNX6,which encodes a small subunit of molybdopterin synthase essential for molybdenum cofactor(MoCo)biosynthesis,is the causal gene responsible for the vp-like5 phenotype.Biochemical assays have demonstrated significantly reduced activities of MoCo-dependent enzymes,including aldehyde oxidase(AO),xanthine dehydrogenase(XDH),and nitrate reductase(NR),in vplike5.AO is essential for the ABA biosynthesis,and the observed ABA deficiency in vp-like5 likely drives the viviparous phenotype.Expression analysis showed that ZmCNX6 was stably expressed during seed development,indicating its significant role in seed development.Furthermore,overexpression of ZmCNX6 not only enhanced the activities of MoCo-dependent enzymes but also improved drought tolerance in maize.Collectively,our study revealed ZmCNX6 as a multifunctional hub coordinating MoCo metabolism,ABA-dependent dormancy regulation,and abiotic stress responses,offering a potential target for simultaneously mitigating vivipary and improving drought resistance in maize.
基金supported by the National Key R&D Program of China(2019YFA0904900)the National Natural Science Foundation of China(21877112,21837002,21721004)。
文摘Cytochrome P450 enzymes catalyze diverse oxidative transformations at the expense of reduced nicotinamide adenine dinucleotide phosphate(NADPH),however,their applications remain limited largely because NADPH is cost-prohibitive for biocatalysis at scale yet tightly regulated in host cells.A highly challenging task for P450 catalysis has been to develop an alternative and biocompatible electrondonating system.Here we engineered P450 BM3 to favor reduced nicotinamide cytosine dinucleotide(NCDH)and created non-natural cofactor-dependent P450 catalysis.Two outstanding mutants were identified with over 640-fold NCDH preference improvement and good catalytic efficiencies of over15,000 M^(-1)s^(-1)for the oxidation of the fatty acid probe 12-(para-nitrophenoxy)-dodecanoate.Molecular docking analysis indicated that these mutants bear a compacted cofactor entrance.Upon fusing with an NCD-dependent formate dehydrogenase,fused proteins functioned as NCDH-specific P450catalysts by using formate as the electron donor.Importantly,these mutants and fusions catalyzed NCDH-dependent hydroxylation of fatty acids with similar chain length preference to those by natural P450 BM3 in the presence of NADPH and also similar regioselectivity for subterminal hydroxylation of lauric acid.As P450 BM3 and its variants are catalytically powerful to take diverse substrates and convey different reaction paths,our results offer an exciting opportunity to devise advanced cell factories that convey oxidative biocatalysis with an orthogonal reducing power supply system.
文摘The iron molybdenum cofactor(FeMoco) of nitrogenase MoFe protein from Azotobacter vinelandii OP was extracted by N methylformamide(NMF). The effects of FeMoco(in NMF) on electronic spectrum and fluorescence intensity of fluorescein dimercury acetate(FDMA)(in 1 mol/L NaOH) were investigated by fluorophotometric titrations and compared with those of (NH 4) 2MoS 4 and complexes of (NH 4) 2MoS 4 with Na 2S or Na 2S 2 or (NH 4) 2S x on the relative properties of FDMA. It was found that the electronic spectrum of FDMA displayed hypsochromic shift(17 nm) in the presence of FeMoco just like that in the presence of other inorganic sulfides and that the titration curve for the quench of FDMA with FeMoco is very similar to that for the quench of FDMA with complex of (NH 4) 2MoS 4 with Na 2S 2(mole ratio is 1∶3). The results showed that FeMoco(N) probably contained S—S bonds and its structure was found to be changed compared with Kim Rees structural model. This change should profit the formation of the polymer.
文摘Two k-hyperconnection expressions of a general k-order cofactor Y<sub>（i,j）</sub> are presentedfor the indefinite parameter matrix Y of a linear system by applying directed hypergraph theory,and based on it a decomposition theorem of Y<sub>（i,j）</sub> is derived.By this theorem,the multi-leveltearing and analysis can be carried out easily for any linear large system.This is a new mul-tilevel topological analysis method.Using proposed method the scale of systems which can betopologically analysed by a computer will be enlarged.
基金supported by grants from the Ministry of Education,Culture,Sports,Science and Technology in Japan to MM and TCthe Japan Society for the Promotion of Science to MO,MM,and TCthe Japan Science and Technology Agency to M.M.and TC
文摘Microtubules play important roles in neuronal morphogenesis, including cellular polarization, neurite growth, and branching. A microtubule is a polymer of a- and β-tubulin heterodimers that are formed by a multistep process assisted by at least five tubulin-fold- ing cofactors （TBCA-E） （Lopez-Fanarraga et al., 2001）. Newly syn- thesized α- and β-tubulins associate with the cytosolic chaperonin complex （CCT）, and then the quasi-native tubulins interact with five tubulin-folding cofactors. The α- and β-tubulins first interact with TBCB and TBCA, which are further transferred to TBCE and TBCD, respectively. The α- and β-tubulins, TBCC, TBCD, and TBCE form a supercomplex, and guanosine triphosphate （GTP） hydrolysis occurs within the supercomplex, releasing a tubulin heterodimer. In addition to their functions in tubulin folding and tubulin heterodimer formation, excess amounts of TBCD or TBCE may interact with tubulin heterodimers, leading to their degrada- tion. Thus, tubulin-folding cofactors may play a role in both the synthesis and degradation of tubulin heterodimers.
文摘Introduction: High grade dysplasia of the cervix has a high incidence and can progress to cervical cancer. The aim was to study cofactors associated with high-grade cervical dysplasia. Methodology: This was a retrospective case-control study without matching. Women with high grade dysplasia were the cases while those with a normal screening test represented the controls. The study took place at the Gabriel Touré University Hospital Center in Bamako. We included 351 cases and 420 controls. The capture and analysis were performed using the SPSS 20 software. A univariate and multivariate logistic regression analysis was performed for the analysis of risk cofactors. The statistical tests used were the odds ratio and its confidence interval and the statistical significance threshold was set at p Results: In univariate analysis, the co-factors statistically significantly associated with the occurrence of high-grade dysplasia were parity 0.6 (0.5 - 0.9), gestational 0.7 (0.5 - 0.9), smoking of the spouse 3.4 (1.1 - 11.3), the non-schooling 1.4 (1.2 - 2.1). In multivariate analysis after adjusting for confounding factors, two co-factors have significantly increased the risk of high-grade dysplasia: lack of schooling 1.4 (1.2 - 2.0) and polygamy 1.5 (1.4 - 2.5). Conclusion: At the end of this study, polygamy and lack of schooling were the main risk factors. The prevention of cervical cancer will go through the education of girls and women as well as communication for behavioral change and social change.
文摘In literature, neutral polyethylene oxide (PEO) flocculated fines at low shear rates, while with cofactor (CF) addition, the formed CF-PEO complex showed larger ability to bridge fines, producing flocs. In this work, some process factors were found having significant effects on fines flocculation. Increases in CF to PEO ratio at constant PEO enhanced the bridging bonds, causing increases in flocculation initial rate (efficiency), amplitude (floc size), and fastness (a decrease in characteristic time). On the other hand, an increase in stirring rate (shear rate) in flocculation vessel caused decreases in initial rate and amplitude, and an increase in the fastness. All runs showed transient flocculation;the amplitude increased with time, reached maximum at equilibrium, and then started to decrease showing deflocculation. In brief, the CF to PEO ratio and the shear rate were found important parameters in mill operation, having significant effects on flocculation efficiency, fastness, and floc size.
基金supported by grants from the National Nat-ural Science Foundation of China(32300045,32300029)Youth Program of Natural Science Foundation of Shandong Province(ZR2022QC092,ZR2023QC237)+6 种基金China Postdoctoral Science Founda-tion(2023M742085)Youth Program of Natural Science Foundation of Qingdao City(23-2-1-31-zyyd-jch)Postdoctoral Innovation Program of Shandong Province(SDCX-ZG-202400150)Qingdao Postdoctoral Re-search Project(QDBSH20230201011)State Key Laboratory of Micro-bial Technology Open Projects Fund(M2023-02)Open Funding Project of State Key Laboratory of Microbial Metabolism(MMLKF24-07),China Postdoctoral Science Foundation(2024M751800)State Key Labo-ratory of Microbial Technology(SKLMTFCP-2023-03)。
文摘Dicarboxylates are valuable platform compounds with a broad range of applications.The in vitro biosynthetic system used to produce dicarboxylates fromω-amino acids via the natural pathway requires costly cofactors and co-substrates,which restricts its economic feasibility of use.In this study,we designed a cofactor-and co-substrate-free artificial pathway for the production of dicarboxylates fromω-amino acids.Only three enzymes(viz.,amine oxidase from Kluyveromyces marxianus DMKU3-1042,xanthine oxidase from bovine milk,and cata-lase from Aspergillus niger)were required for dicarboxylate production.Succinate(0.79 g g^(-1)),glutarate(0.83 g g^(-1)),and adipate(0.77 g g^(-1))were produced in high yields from the correspondingω-amino acids through the in vitro biosynthetic system with the artificial pathway.Glutarate could also be produced from l-lysine by further introducing l-lysine monooxygenase and 5-aminovaleramide amidohydrolase from Pseudomonas putida KT2440 into the in vitro biosynthetic system,with the cofactor-and co-substrate-free system achieving a product yield of 0.63 g g^(-1).Considering its desirable characteristics,this artificial pathway-based in vitro biosynthetic system may be a promising alternative for dicarboxylate production from biotechnologically producedω-amino acids.
基金The authors thank the financial support by National Natural Science Foundation of China(21776132,21878142)Jiangsu Province Natural Science Foundation for Distinguished Young Scholars(BK20190035)+2 种基金National Key Research and Development Program of China(2019YFD1101202)Jiangsu Province Natural Science Foundation for Youths(BK20200685)China Postdoctoral Science Foundation(2019M660113).
文摘Chemoenzymatic catalysis can give full play to the advantages of versatile reactivity of chemocatalysis and excellent chemo-,regio-,and stereoselectivities of biocatalysis.These chemoenzymatic methods can not only save resource,cost,and operating time but also reduce the number of reaction steps,and avoid separating unstable intermediates,leading to the generation of more products under greener circumstances and thereby playing an indispensable role in the fields of medicine,materials and fine chemicals.Although incompatible challenges between chemocatalyst and biocatalyst remain,strategies such as biphasic system,artificial metalloenzymes,immobilization or supramolecular host,and protein engineering have been designed to overcome these issues.In this review,chemoenzymatic catalysis according to different chemocatalysis types was classifiably described,and in particular,the classic dynamic kinetic resolutions(DKR)and cofactor regeneration were summarized.Finally,the bottlenecks and development of chemoenzymatic catalysis were summarized,and future development was prospected.
基金supported by National Talents Program,Double First Class University Plan of Sichuan University,State Key Laboratory of Polymer Materials Engineering(No.sklpme 2020-0301)Natural Science Foundation of Sichuan Province(Nos.2022NSFSC1735,2023NSFSC1097)+5 种基金Fundamental Research Funds for the Central Universities(No.ZYN2022094)National Natural Science Foundation of China(Nos.22178233,22208228)China Postdoctoral Science Foundation(No.2020TQ0209)Fundamental Research Funds for the Central Universities(No.YJ201959)Science and Technology Support Program of Sichuan Province(No.2021YJ0414)Project of Chengdu Science and Technology Bureau(No.2021YF05-02110-SN)。
文摘DNAzyme machines play critical roles in the fields of cell imaging,disease diagnosis,and cancer therapy.However,the applications of DNAzyme machines are limited by the nucleases-induced degradation,non-specific binding of proteins,and insufficient provision of cofactors.Herein,protected DNAzyme machines with different cofactor designs(referred to as Pro Ds)were nanoengineered by the construction of multifunctional metal-phenolic nanoshells to deactivate the interferential proteins,including nucleases and non-specific binding proteins.Moreover,the nanoshells not only facilitate the cellular internalization of Pro Ds but provide specific metal ions acting as cofactors of the designed DNAzymes.Cellular imaging results demonstrated that Pro Ds could effectively and simultaneously monitor multiple tumor-related micro RNAs in living cells.This facile and rapid strategy that encapsulates DNAzyme machines into the protective metal-phenolic nanoshells is anticipated to extend to a wide range of functional nucleic acidsbased biomedical applications.
基金Project supported by the National Natural Science Foundation of China(Nos.60533060,69973010 and 10271022)
文摘The mechanical background of the bivariate spline space of degree 2 and smoothness 1 on rectangular partition is presented constructively. Making use of mechanical analysis method, by acting couples along the interior edges with suitable evaluations, the deflection surface is divided into piecewise form, therefore, the relation between a class of bivariate splines on rectangular partition and the pure bending of thin plate is established. In addition, the interpretation of smoothing cofactor and conformality condition from the mechanical point of view is given. Furthermore, by introducing twisting moments, the mechanical background of any spline belong to the above space is set up.
文摘A contiguous derivation of radius and center of the insphere of a general tetrahedron is given. Therefore a linear system is derived. After a transformation of it the calculation of radius and center can be separated from each other. The remaining linear system for the center of the insphere can be solved after discovering the inverse of the corresponding coefficient matrix. This procedure can also be applied in the planar case to determine radius and center of the incircle of a triangle.
