The superfamily of cytochrome P450(CYP)enzymes plays key roles in plant evolution and metabolic diversification.This review provides a status on the CYP Iandscape within green algae and land plants.The 11 conserved CY...The superfamily of cytochrome P450(CYP)enzymes plays key roles in plant evolution and metabolic diversification.This review provides a status on the CYP Iandscape within green algae and land plants.The 11 conserved CYP clans known from vascular plants are all present in green algae and several green algaespecific clans are recognized.Clan 71,72,and 85 remain the largest CYP clans and include many taxaspecific CYP(sub)families reflecting emergence of linage-specific pathways.Molecular features and dynamics of CYP plasticity and evolution are discussed and exemplified by selected biosynthetic pathways.High substrate promiscuity is commonly observed for CYPs from large families,favoring retention of gene duplicates and neofunctionalization,thus seeding acquisition of new functions.Elucidation of biosynthetic pathways producing metabolites with sporadic distribution across plant phylogeny reveals multiple exampies of convergent evolution where CYPs have been independently recruited from the same or different CYP families,to adapt to similar environmental challenges or ecological niches.Sometimes only a single or a few mutations are required for functional interconversion.A compilation of functionally characterized plant CYPs is provided online through the Plant P450 Database(erda.dk/public/vgrid/PlantP450/).展开更多
Enzymatic glycosylation catalyzed by glycosyltransferases (GTs) has great potential in creating diverse novel and bioactive glycosides. Herein, three new GTs (UGT84 A33, UGT71 AE1 and UGT90 A14) from Carthamus tinctor...Enzymatic glycosylation catalyzed by glycosyltransferases (GTs) has great potential in creating diverse novel and bioactive glycosides. Herein, three new GTs (UGT84 A33, UGT71 AE1 and UGT90 A14) from Carthamus tinctorius exhibited robust catalytic promiscuity to benzylisoquinoline alkaloids, and were used as enzymatic tools in glycosylation of bioactive benzylisoquinoline alkaloids. Seven novel benzylisoquinoline alkaloids O-glycosides were synthesized with high efficiency. These studies indicate the significant potential of promiscuous GTs in synthesis of benzylisoquinoline alkaloids glycosides for drug discovery.展开更多
Esterase BioH,which is obligatory for biotin synthesis in Escherichia coli,was found to exhibit a promiscuous ability to catalyse Aldol and Knoevenagel reactions with moderate to good yields.The reaction conditions in...Esterase BioH,which is obligatory for biotin synthesis in Escherichia coli,was found to exhibit a promiscuous ability to catalyse Aldol and Knoevenagel reactions with moderate to good yields.The reaction conditions including organic solvent,molar ratio of ketone to aldehyde,enzyme amount,and reaction time were investigated to evaluate the effect of different reaction conditions on yield.Target compounds were afforded in the best yield of 91.2％ for Aldol reaction and 54.7％ for Knoevenagel reaction.In addition,because the enzyme could be prepared with a low cost,this protocol could provide an economic route to conduct Aldol and Knoevenagel reactions,which expand the field of enzymatic promiscuity.展开更多
Enzymatic malonylation of natural glycosides provides a promising alternative method for drug-like malonylated glycosides supply.However,the catalytic potential and structural basis of plant malonyltransferase are far...Enzymatic malonylation of natural glycosides provides a promising alternative method for drug-like malonylated glycosides supply.However,the catalytic potential and structural basis of plant malonyltransferase are far from being fully elucidated.This work identified a new malonyltransferase CtMaT1 from Cistanche tubulosa.It displayed unprecedented mono-and/or di-malonylation activity toward diverse glucosides with different aglycons.A“one-pot”system by CtMaT1 and a malonyl-CoA synthetase was established to biosynthesize nine new malonylated glucosides.Structural investigations revealed that CtMaT1 possesses an adequately spacious acyl-acceptor pocket capable of accommodating diverse glucosides.Additionally,it recognizes malonyl-CoA through strong electrotactic and hydrogen interactions.QM/MM calculation revealed the H167-mediated SN2 reaction mechanism of CtMaT1,while dynamic simulations detected the formation of stable hydrogen bonds between the glucose-6-OH group and H167,resulting in its high malonylation regiospecificity.Calculated energy profiles of two isomeric glycosides highlighted lower reaction energy barriers towards glucoside substrates,emphasizing CtMaT1's preference for glucosides.Furthermore,a mutant CtMaT1H36A with notably increased di-malonylation activity was obtained.The underlying molecular mechanism was illuminated through MM/GBSA binding free energy calculation.This study significantly advances the understanding of plant acyltransferases from both functional and protein structural perspectives,while also providing a versatile tool for enzymatic malonylation applications in pharmacology.展开更多
Lignocellulosic grass biomass is potential substrate for economical and sustainable bioethanol production.However,the processing cost of bioethanol that majorly includes the hydrolysis of cellulose by cellulases is st...Lignocellulosic grass biomass is potential substrate for economical and sustainable bioethanol production.However,the processing cost of bioethanol that majorly includes the hydrolysis of cellulose by cellulases is still a major concern for its industrial production.Thus,knowledge on the sequence to the structural study of cellulase enzyme with consideration of its catalytic region can give important information for effective enzyme engineering and consequently towards enhanced bioethanol production from Pennisetum sp.Therefore,in this study,sequence conservativeness of different cellulosic site among a group of endoglucanase family of cellulase from previously isolated Aspergillus species has been determined.Furthermore,comparative molecular modeling of the endoglucanase from eight different Aspergillus species including Aspergillus fumigatus was conducted and the obtained structures revealed a high degree of difference in their conformational folds.Analysis from InterProScan revealed that the modeled endoglucanase has similar types of domains and share homology with protein family,such as glycoside hydrolase family-61 and fungal cellulose binding domain.Furthermore,molecular docking and interaction studies demonstrated the presence of residues in the endoglucanase of A.fumigatus viz.His20,His88,Asp96,Ala99,Ser100,Ser101,His102,His169,Glu170,Arg173,Glu178,and Tyr218 that are responsible in forming the substrate interaction.An interesting molecular phenomenon,i.e.,catalytic promiscuity has been noted for all the substrate bound complexes of A.fumigatus endoglucanase which also depicts the degree of ligand binding efficacy of the studied enzyme.The molecular interaction study,binding energy analysis and molecular dynamics simulation,demonstrated that heteromeric substrate XylGlc3 is more strongly interacting with the receptor enzyme.Overall,the present findings revealed that important amino acid residues can help in increasing the specificity of endoglucanase from A.fumigatus towards hydrolysis of Pennisetum sp.an展开更多
Objective:In order to obtain new glycosyltransferases with highly efficient catalysis,the glycosyltransferases from Carthamus tinctorius which contains diverse types of glycosides were mined.Methods:A new glycosyltran...Objective:In order to obtain new glycosyltransferases with highly efficient catalysis,the glycosyltransferases from Carthamus tinctorius which contains diverse types of glycosides were mined.Methods:A new glycosyltransferase gene(UGT88B2)with full length was obtained by PCR and further transformed into Escherichia coli for heterologous expression.The catalytic activity of recombinant UGT88B2 was determined by HPLC-MSn.The structures of representative catalytic products were elucidated by MS and NMR.Results:UGT88B2 exhibited catalytic promiscuity and various patterns in glycosylation of flavonoids with high efficiency.Conclusion:A new glycosyltransferase named UGT88B2 was successfully mined and can be employed as enzymatic tools in glycosylation of flavonoids.展开更多
An unprecedented enzyme-catalyzed asymmetric domino aza-Michael/aldol reaction of 2-aminobenzaldehyde and a,b-unsaturated aldehydes is achieved. Pepsin from porcine gastric mucosa provided mild and efficient access to...An unprecedented enzyme-catalyzed asymmetric domino aza-Michael/aldol reaction of 2-aminobenzaldehyde and a,b-unsaturated aldehydes is achieved. Pepsin from porcine gastric mucosa provided mild and efficient access to diverse substituted 1,2-dihydroquinolines in yields of 38%–97% with 6%–24%enantiomeric excess（ee）. This work not only provides a novel method for the synthesis of dihydroquinoline derivatives, but also promotes the development of enzyme catalytic promiscuity.展开更多
＂Amano＂ lipase AS（lipase from Aspergillus niger）, which naturally hydrolyzes triglycerides, was found promiscuously to catalyze multi-component reactions of aromatic aldehydes with malononitrile and β-naphthol to...＂Amano＂ lipase AS（lipase from Aspergillus niger）, which naturally hydrolyzes triglycerides, was found promiscuously to catalyze multi-component reactions of aromatic aldehydes with malononitrile and β-naphthol to prepare naphthopyran derivatives in anhydrous organic solvents in moderate to good yields.展开更多
A simple and efficient method was developed for the synthesis of 2.2＇-arylmethylene dicyclohexane-1,3- dione derivatives via the Knoevenagel-Michael cascade reactions of aromatic aldehydes and 1,3-cyclic diketones ca...A simple and efficient method was developed for the synthesis of 2.2＇-arylmethylene dicyclohexane-1,3- dione derivatives via the Knoevenagel-Michael cascade reactions of aromatic aldehydes and 1,3-cyclic diketones catalyzed by ＂Amano＂ lipase DF, which expands the application field of enzyme catalytic promiscuity. This protocol provides several advantages over the traditional chemical synthesis, such as simple work-up procedure, high yields Cup to 94%） and environmental friendliness.展开更多
The direct asymmetric aldol reaction of aromatic aldehydes with cyclic or acyclic ketones was catalyzed by proteinase from Aspergillus melleus (AMP) in acetonitrile in the presence of water. A wide range of substrates...The direct asymmetric aldol reaction of aromatic aldehydes with cyclic or acyclic ketones was catalyzed by proteinase from Aspergillus melleus (AMP) in acetonitrile in the presence of water. A wide range of substrates could be transformed into the corresponding aldol products in yields up to 89%, enantioselectivities up to 91% ee and diastereoselectivities up to >99:1 (anti/syn). This work provided an example of enzyme catalytic promiscuity that widens the applicability of this biocatalyst in organic synthesis without the need for additional cofactors or special equipment.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
A new method for the synthesis of 1,4-dihydropyridine(1,4-DHP)calcium channel antagonists felodipme, nitrendipine and their derivatives via papain-catalyzed three-component reactions of aldehyde,methyl acetoacetate an...A new method for the synthesis of 1,4-dihydropyridine(1,4-DHP)calcium channel antagonists felodipme, nitrendipine and their derivatives via papain-catalyzed three-component reactions of aldehyde,methyl acetoacetate and ethyl 3-aminocrotonate was developed.Operational simplicity,mild reaction conditions and eco-friendliness are the key features of this protocol.展开更多
基金supported by a PhD fellowship provided through a Villum Foundation Young Investigator Program fellowship granted to Elizabeth H.J.Neils on(grant number 13167)supported by the VILLUM Center for Plant Plasticity(VKR023054)(B.L.M.)+1 种基金a European Research Council Advanced Grant(ERC-2012-ADG_20120314)the Novo Nordisk Foundation Distinguished Investigator 2019 Grant(NNF 0054563,The Black Holes in the Plant Universe).
文摘The superfamily of cytochrome P450(CYP)enzymes plays key roles in plant evolution and metabolic diversification.This review provides a status on the CYP Iandscape within green algae and land plants.The 11 conserved CYP clans known from vascular plants are all present in green algae and several green algaespecific clans are recognized.Clan 71,72,and 85 remain the largest CYP clans and include many taxaspecific CYP(sub)families reflecting emergence of linage-specific pathways.Molecular features and dynamics of CYP plasticity and evolution are discussed and exemplified by selected biosynthetic pathways.High substrate promiscuity is commonly observed for CYPs from large families,favoring retention of gene duplicates and neofunctionalization,thus seeding acquisition of new functions.Elucidation of biosynthetic pathways producing metabolites with sporadic distribution across plant phylogeny reveals multiple exampies of convergent evolution where CYPs have been independently recruited from the same or different CYP families,to adapt to similar environmental challenges or ecological niches.Sometimes only a single or a few mutations are required for functional interconversion.A compilation of functionally characterized plant CYPs is provided online through the Plant P450 Database(erda.dk/public/vgrid/PlantP450/).
基金financially supported by the National Natural Science Foundation of China (No. 81573317)CAMS Innovation Fund for Medical Sciences(Nos. CIFMS-2016-I2M-3-012 and CIFMS-2016-I2M-2-002)
文摘Enzymatic glycosylation catalyzed by glycosyltransferases (GTs) has great potential in creating diverse novel and bioactive glycosides. Herein, three new GTs (UGT84 A33, UGT71 AE1 and UGT90 A14) from Carthamus tinctorius exhibited robust catalytic promiscuity to benzylisoquinoline alkaloids, and were used as enzymatic tools in glycosylation of bioactive benzylisoquinoline alkaloids. Seven novel benzylisoquinoline alkaloids O-glycosides were synthesized with high efficiency. These studies indicate the significant potential of promiscuous GTs in synthesis of benzylisoquinoline alkaloids glycosides for drug discovery.
基金Supported by the National Natural Science Foundation of China（No.21176215）.
文摘Esterase BioH,which is obligatory for biotin synthesis in Escherichia coli,was found to exhibit a promiscuous ability to catalyse Aldol and Knoevenagel reactions with moderate to good yields.The reaction conditions including organic solvent,molar ratio of ketone to aldehyde,enzyme amount,and reaction time were investigated to evaluate the effect of different reaction conditions on yield.Target compounds were afforded in the best yield of 91.2％ for Aldol reaction and 54.7％ for Knoevenagel reaction.In addition,because the enzyme could be prepared with a low cost,this protocol could provide an economic route to conduct Aldol and Knoevenagel reactions,which expand the field of enzymatic promiscuity.
基金This work was financially supported by National Key Research and Development Program Special Project of Synthetic Biology(Grant No.2023YFA0914100/2023YFA0914103)National Natural Science Foundation of China(Grant No.82173922,81402809)+3 种基金Beijing Natural Science Foundation(Grant No.7192112)Fundamental Research Funds for the Central Universities(Grant No.2023-JYB-JBQN-054,China)Young Elite Scientists Sponsorship Program by CAST(Grant No.CACM-2018-QNRC1-02,China)State Key Laboratory of Natural and Biomimetic Drugs Foundation(Grant No.K202119,China).
文摘Enzymatic malonylation of natural glycosides provides a promising alternative method for drug-like malonylated glycosides supply.However,the catalytic potential and structural basis of plant malonyltransferase are far from being fully elucidated.This work identified a new malonyltransferase CtMaT1 from Cistanche tubulosa.It displayed unprecedented mono-and/or di-malonylation activity toward diverse glucosides with different aglycons.A“one-pot”system by CtMaT1 and a malonyl-CoA synthetase was established to biosynthesize nine new malonylated glucosides.Structural investigations revealed that CtMaT1 possesses an adequately spacious acyl-acceptor pocket capable of accommodating diverse glucosides.Additionally,it recognizes malonyl-CoA through strong electrotactic and hydrogen interactions.QM/MM calculation revealed the H167-mediated SN2 reaction mechanism of CtMaT1,while dynamic simulations detected the formation of stable hydrogen bonds between the glucose-6-OH group and H167,resulting in its high malonylation regiospecificity.Calculated energy profiles of two isomeric glycosides highlighted lower reaction energy barriers towards glucoside substrates,emphasizing CtMaT1's preference for glucosides.Furthermore,a mutant CtMaT1H36A with notably increased di-malonylation activity was obtained.The underlying molecular mechanism was illuminated through MM/GBSA binding free energy calculation.This study significantly advances the understanding of plant acyltransferases from both functional and protein structural perspectives,while also providing a versatile tool for enzymatic malonylation applications in pharmacology.
文摘Lignocellulosic grass biomass is potential substrate for economical and sustainable bioethanol production.However,the processing cost of bioethanol that majorly includes the hydrolysis of cellulose by cellulases is still a major concern for its industrial production.Thus,knowledge on the sequence to the structural study of cellulase enzyme with consideration of its catalytic region can give important information for effective enzyme engineering and consequently towards enhanced bioethanol production from Pennisetum sp.Therefore,in this study,sequence conservativeness of different cellulosic site among a group of endoglucanase family of cellulase from previously isolated Aspergillus species has been determined.Furthermore,comparative molecular modeling of the endoglucanase from eight different Aspergillus species including Aspergillus fumigatus was conducted and the obtained structures revealed a high degree of difference in their conformational folds.Analysis from InterProScan revealed that the modeled endoglucanase has similar types of domains and share homology with protein family,such as glycoside hydrolase family-61 and fungal cellulose binding domain.Furthermore,molecular docking and interaction studies demonstrated the presence of residues in the endoglucanase of A.fumigatus viz.His20,His88,Asp96,Ala99,Ser100,Ser101,His102,His169,Glu170,Arg173,Glu178,and Tyr218 that are responsible in forming the substrate interaction.An interesting molecular phenomenon,i.e.,catalytic promiscuity has been noted for all the substrate bound complexes of A.fumigatus endoglucanase which also depicts the degree of ligand binding efficacy of the studied enzyme.The molecular interaction study,binding energy analysis and molecular dynamics simulation,demonstrated that heteromeric substrate XylGlc3 is more strongly interacting with the receptor enzyme.Overall,the present findings revealed that important amino acid residues can help in increasing the specificity of endoglucanase from A.fumigatus towards hydrolysis of Pennisetum sp.an
基金Tfinancially supported by the National Natural Science Foundation of China(No.81573317)。
文摘Objective:In order to obtain new glycosyltransferases with highly efficient catalysis,the glycosyltransferases from Carthamus tinctorius which contains diverse types of glycosides were mined.Methods:A new glycosyltransferase gene(UGT88B2)with full length was obtained by PCR and further transformed into Escherichia coli for heterologous expression.The catalytic activity of recombinant UGT88B2 was determined by HPLC-MSn.The structures of representative catalytic products were elucidated by MS and NMR.Results:UGT88B2 exhibited catalytic promiscuity and various patterns in glycosylation of flavonoids with high efficiency.Conclusion:A new glycosyltransferase named UGT88B2 was successfully mined and can be employed as enzymatic tools in glycosylation of flavonoids.
基金supported by the National Natural Science Foundation of China (Nos. 21276211 and 21472152)
文摘An unprecedented enzyme-catalyzed asymmetric domino aza-Michael/aldol reaction of 2-aminobenzaldehyde and a,b-unsaturated aldehydes is achieved. Pepsin from porcine gastric mucosa provided mild and efficient access to diverse substituted 1,2-dihydroquinolines in yields of 38%–97% with 6%–24%enantiomeric excess（ee）. This work not only provides a novel method for the synthesis of dihydroquinoline derivatives, but also promotes the development of enzyme catalytic promiscuity.
基金Supported by the Program for Zhejiang Leading Team of Science ＆ Technology Innovation, China（No.2011R50007）, the National Natural Science Foundation of China（Nos.21176215, 21176102） and the Outstanding Young Scholar Grant of Zhejiang University, China（No.R4110092）.
文摘＂Amano＂ lipase AS（lipase from Aspergillus niger）, which naturally hydrolyzes triglycerides, was found promiscuously to catalyze multi-component reactions of aromatic aldehydes with malononitrile and β-naphthol to prepare naphthopyran derivatives in anhydrous organic solvents in moderate to good yields.
基金financially supported by the Outstanding Young Scholar Grant of Zhejiang University (No. R4110092)the National Natural Science Foundation of China (No. 21176215/21176102)the Program for Zhejiang leading team of S&T Innovation (No. 2011R50007)
文摘A simple and efficient method was developed for the synthesis of 2.2＇-arylmethylene dicyclohexane-1,3- dione derivatives via the Knoevenagel-Michael cascade reactions of aromatic aldehydes and 1,3-cyclic diketones catalyzed by ＂Amano＂ lipase DF, which expands the application field of enzyme catalytic promiscuity. This protocol provides several advantages over the traditional chemical synthesis, such as simple work-up procedure, high yields Cup to 94%） and environmental friendliness.
基金2011 Select Project in Scientific and Technological Activities for Returned Scholars of Chongqing Personnel Bureau, and the Doctoral Foundation ofSouthwest University (SWU112019)
文摘The direct asymmetric aldol reaction of aromatic aldehydes with cyclic or acyclic ketones was catalyzed by proteinase from Aspergillus melleus (AMP) in acetonitrile in the presence of water. A wide range of substrates could be transformed into the corresponding aldol products in yields up to 89%, enantioselectivities up to 91% ee and diastereoselectivities up to >99:1 (anti/syn). This work provided an example of enzyme catalytic promiscuity that widens the applicability of this biocatalyst in organic synthesis without the need for additional cofactors or special equipment.
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 National Natural Science Foundation of China(No.21706236)the China Postdoctoral Science Foundation (No.2016M592012)the Foundation for Selected Postdoctoral Project of Zhejiang Province,China(No.BSH1502150).
文摘A new method for the synthesis of 1,4-dihydropyridine(1,4-DHP)calcium channel antagonists felodipme, nitrendipine and their derivatives via papain-catalyzed three-component reactions of aldehyde,methyl acetoacetate and ethyl 3-aminocrotonate was developed.Operational simplicity,mild reaction conditions and eco-friendliness are the key features of this protocol.
