The asymmetric hydroxylation of inactive carbon atoms in organic compounds remains an important reaction in the industrial synthesis of valuable chiral compounds.Fe(II)and 2-ketoglutarate-dependent dioxygenases(Fe/2-k...The asymmetric hydroxylation of inactive carbon atoms in organic compounds remains an important reaction in the industrial synthesis of valuable chiral compounds.Fe(II)and 2-ketoglutarate-dependent dioxygenases(Fe/2-kg DOs)are the largest known subgroups of mononuclear nonheme-Fe(II)-dependent oxygenases,catalyzing various oxidation reactions of C-H bonds.Recent developments in Fe/2-kg DO-related researches have coupled concepts from bioinformatics,synthetic biology,and computational biology to establish effective biotransformation systems.The most well-studied and characterized activ-ity of the Fe/2-kg DOs is substrate hydroxylation,with regard to which mechanistic studies involving the Fe center assist in engineering the protein frameworks of these enzymes to obtain the desired catalytic enhancements.Amino acids are typical substrates of Fe/2-kg DOs and are usually converted into hydroxyl amino acids,which are widely used as intermediates in pharmaceutical and fine chemical industries.Herein,we have reviewed prior structural and mechanistic studies on Fe/2-kg DOs,as well as studies on the Fe/2-kg DO-mediated selective C-H oxidation process for selective hydroxyl amino acid synthesis,which will further our journey along the promising path of building complexity via C-H bond oxidation.Further,new bioinformatics techniques should be adopted with structure-based protein rational design to mine sequence databases and shrink mutant libraries to produce a diverse panel of functional Fe/2-kg DOs capable of catalyzing targeted reactions.展开更多
Polycyclic aromatic hydrocarbons(PAHs)are a class of persistent pollutants with adverse biological effects and pose a serious threat to ecological environments and human health.The previously isolated phenanthrene‐de...Polycyclic aromatic hydrocarbons(PAHs)are a class of persistent pollutants with adverse biological effects and pose a serious threat to ecological environments and human health.The previously isolated phenanthrene‐degrading bacterial consortium(PDMC)consists of the genera Sphingobium and Pseudomonas and can degrade a wide range of PAHs.To identify the degradation mechanism of PAHs in the consortium PDMC,metagenomic binning was conducted and a Sphingomonadales assembly genome with 100%completeness was obtained.Additionally,Sphingobium sp.SHPJ‐2,an efficient degrader of PAHs,was successfully isolated from the consortium PDMC.Strain SHPJ‐2 has powerful degrading abilities and various degradation pathways of high‐molecular‐weight PAHs,including fluoranthene,pyrene,benzo[a]anthracene,and chrysene.Two ring‐hydroxylating dioxygenases,five cytochrome P450s,and a pair of electron transfer chains associated with PAH degradation in strain SHPJ‐2,which share 83.0%–99.0%similarity with their corresponding homologous proteins,were identified by a combination of Sphingomonadales assembly genome annotation,reverse‐transcription quantitative polymerase chain reaction and heterologous expression.Furthermore,when coexpressed in Escherichia coli BL21(DE3)with the appropriate electron transfer chain,PhnA1B1 could effectively degrade chrysene and benzo[a]anthracene,while PhnA2B2 degrade fluoranthene.Altogether,these results provide a comprehensive assessment of strain SHPJ‐2 and contribute to a better understanding of the molecular mechanism responsible for the PAH degradation.展开更多
基金Financial supports from the National Natural Science Foundation of China(NSFC)(No.21676120)the 111 Project(No.111-2-06)+5 种基金the High-End Foreign Experts Recruitment Program(No.G20190010083)the Program for Advanced Talents within Six Industries of Jiangsu Province(No.2015-NY-007)the National Program for Support of Top-Notch Young Professionals,the Fundamental Research Funds for the Central Universities(No.JUSRP51504)the Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions,Top-Notch Academic Programs Project of Jiangsu Higher Education Institutions,the Jiangsu Province“Collaborative Innovation Center for Advanced Industrial Fermentation”Industry Development Program,the Postgraduate Research&Practice Innovation Program of Jiangsu Province(No.KYCX19_1833)the Program for the Key Laboratory of Enzymes of Suqian(No.M201803)the National First-Class Discipline Program of Light Industry Technology and Engineering(No.LITE2018-09)are greatly appreciated。
文摘The asymmetric hydroxylation of inactive carbon atoms in organic compounds remains an important reaction in the industrial synthesis of valuable chiral compounds.Fe(II)and 2-ketoglutarate-dependent dioxygenases(Fe/2-kg DOs)are the largest known subgroups of mononuclear nonheme-Fe(II)-dependent oxygenases,catalyzing various oxidation reactions of C-H bonds.Recent developments in Fe/2-kg DO-related researches have coupled concepts from bioinformatics,synthetic biology,and computational biology to establish effective biotransformation systems.The most well-studied and characterized activ-ity of the Fe/2-kg DOs is substrate hydroxylation,with regard to which mechanistic studies involving the Fe center assist in engineering the protein frameworks of these enzymes to obtain the desired catalytic enhancements.Amino acids are typical substrates of Fe/2-kg DOs and are usually converted into hydroxyl amino acids,which are widely used as intermediates in pharmaceutical and fine chemical industries.Herein,we have reviewed prior structural and mechanistic studies on Fe/2-kg DOs,as well as studies on the Fe/2-kg DO-mediated selective C-H oxidation process for selective hydroxyl amino acid synthesis,which will further our journey along the promising path of building complexity via C-H bond oxidation.Further,new bioinformatics techniques should be adopted with structure-based protein rational design to mine sequence databases and shrink mutant libraries to produce a diverse panel of functional Fe/2-kg DOs capable of catalyzing targeted reactions.
基金This study was supported by grants from the National Key R&D Program of China(2021YFA0909500)Shanghai Excellent Academic Leaders Program(20XD1421900)+1 种基金grants from the National Natural Science Foundation of China(32100075 and 32030004)“Shuguang Program”(17SG09)supported by the Shanghai Education Development Foundation and the Shanghai Municipal Education Commission.
文摘Polycyclic aromatic hydrocarbons(PAHs)are a class of persistent pollutants with adverse biological effects and pose a serious threat to ecological environments and human health.The previously isolated phenanthrene‐degrading bacterial consortium(PDMC)consists of the genera Sphingobium and Pseudomonas and can degrade a wide range of PAHs.To identify the degradation mechanism of PAHs in the consortium PDMC,metagenomic binning was conducted and a Sphingomonadales assembly genome with 100%completeness was obtained.Additionally,Sphingobium sp.SHPJ‐2,an efficient degrader of PAHs,was successfully isolated from the consortium PDMC.Strain SHPJ‐2 has powerful degrading abilities and various degradation pathways of high‐molecular‐weight PAHs,including fluoranthene,pyrene,benzo[a]anthracene,and chrysene.Two ring‐hydroxylating dioxygenases,five cytochrome P450s,and a pair of electron transfer chains associated with PAH degradation in strain SHPJ‐2,which share 83.0%–99.0%similarity with their corresponding homologous proteins,were identified by a combination of Sphingomonadales assembly genome annotation,reverse‐transcription quantitative polymerase chain reaction and heterologous expression.Furthermore,when coexpressed in Escherichia coli BL21(DE3)with the appropriate electron transfer chain,PhnA1B1 could effectively degrade chrysene and benzo[a]anthracene,while PhnA2B2 degrade fluoranthene.Altogether,these results provide a comprehensive assessment of strain SHPJ‐2 and contribute to a better understanding of the molecular mechanism responsible for the PAH degradation.
