The antibiotic （-）-anisomycin was synthesized starting from D-tyrosine using Sharpless asymmetric epoxidation as a key reaction followed by formation and hydrolysis of oxazoline set up all chiral center.
Anisomycin(compound 1),a multifunctional pyrrolidine antibiotic,primarily inhibits protein biosynthesis by binding to the ribosome.Upon binding to the ribosome,the para-phenol moiety of anisomycin inserts completely i...Anisomycin(compound 1),a multifunctional pyrrolidine antibiotic,primarily inhibits protein biosynthesis by binding to the ribosome.Upon binding to the ribosome,the para-phenol moiety of anisomycin inserts completely into the hydrophobic crevice of the A-site and blocks the access of the incoming aminoacyl-tRNAs,disrupting peptide bond formation.Hence,the para-methoxyphenyl group serves as a starting point for developing novel anisomycin analogs with potent antifungal and insecticidal properties.However,the activation and condensation mechanism of phenylpyruvic acid has not yet been elucidated.In this study,genetic manipulations of aniP and its homologue siAniP confirmed their indispensable role in 1 biosynthesis.Bioinformatics analysis suggested that AniP and siAniP function as transketolase.siAniP was found to catalyzed condensation between 4-hydroxyphe-nylpyruvic acid(3)and glyceraldehyde(GA),initiating pyrrolidine synthesis.siAniP was specific for aromatic keto acids and tolerant of aliphatic and aromatic aldehydes,and was able to catalyze the asymmetric intermo-lecular condensation of two keto acids,leading to the formation of 24α-hydroxy ketone.To the best of our knowledge,siAniP is the first TK that catalyzes the transfer of a C2 ketol and symmetrical intermolecular coupling using aromatic keto acids as donor substrates.Structural analysis,docking model construction,and site-directed mutagenesis identified that I220,H275,R322 and W391 were crucial for substrate binding.Moreover,sequence similarity network(SSN)-based genome neighborhood network(GNN)analyses of AniP suggested the widespread occurrence of the AniP-like-mediated reaction in the biosynthesis of 1 and its analogs,particularly in the assembly of benzylpyrrolidine.These findings not only expand the repertoire of TKs but also provide a potent biocatalyst that could be used for the structural innovation of 1 and its derivatives.展开更多
The protein synthesis inhibitor anisomycin features a unique benzylpyrrolidine system and exhibits potent selective activity against pathogenic protozoa and fungi.It is one of the important effective components in Agr...The protein synthesis inhibitor anisomycin features a unique benzylpyrrolidine system and exhibits potent selective activity against pathogenic protozoa and fungi.It is one of the important effective components in Agricultural Antibiotic120,which has been widely used as naturally-originated agents for treatment of crop decay in China.The chemical synthesis of anisomycin has recently been reported,but the complex process with low productivity made the biosynthesis still to be a vital mainstay in efforts.The biosynthetic gene cluster(BGC)of anisomycin in Streptomyces hygrospinosus var.beijingensis has been identified in our previous work,while poor understanding of the regulatory mechanism limited the yield enhancement via regulation engineering of S.hygrospinosus var.beijingensis.In this study here,we characterized AniF as an indispensable LuxR family transcriptional regulator for the activation of anisomycin biosynthesis.The genetic manipulations of aniF and the real-time quantitative PCR(RT-qPCR)revealed that it positively regulated the transcription of the anisomycin BGC.Moreover,the overexpression of aniF contributed to the improvement of the production of anisomycin and its derivatives.Dissection of the mechanism underlying the function of AniF revealed that it directly activated the transcription of the genes aniR-G involved in anisomycin biosynthesis.Especially,one AniF-binding site in the promoter region of aniR was identified by DNase I footprinting assay and an inverted repeat sequence(5′-GGGC-3′)composed of two 4-nt half sites in the protected region was found.Taken together,our systematic study confirmed the positive regulatory role of AniF and might facilitate the future construction of engineering strains with high productivity of anisomycin and its derivatives.展开更多
Because posttraumatic stress disorder(PTSD) is a highly debilitating condition, prevention is an important research topic. This article reviews possible prevention approaches that involve the administration of drugs b...Because posttraumatic stress disorder(PTSD) is a highly debilitating condition, prevention is an important research topic. This article reviews possible prevention approaches that involve the administration of drugs before the traumatic event takes place. The considered approaches include drugs that address the sympathetic nervous system, drugs interfere with the hypothalamic-pituitary-adrenal(HPA) axis, narcotics and other psychoactive drugs, as well as modulators of protein synthesis. Furthermore, some thoughts on potential ethical implications of the use of drugs for the primary prevention of PTDS are presented. While there are many barriers to overcome in this field of study, this paper concludes with a call for additional research, as there are currently no approaches that are well-suited for regular daily use.展开更多
基金support from The First Hospital of Jilin University
文摘The antibiotic （-）-anisomycin was synthesized starting from D-tyrosine using Sharpless asymmetric epoxidation as a key reaction followed by formation and hydrolysis of oxazoline set up all chiral center.
基金supported by a grant from the National Key research and development Program of China(2021YFA0909500,2021 YFC2100100,2021YFC2100600)National Natural Science Foundation of China(32170077,32170075).
文摘Anisomycin(compound 1),a multifunctional pyrrolidine antibiotic,primarily inhibits protein biosynthesis by binding to the ribosome.Upon binding to the ribosome,the para-phenol moiety of anisomycin inserts completely into the hydrophobic crevice of the A-site and blocks the access of the incoming aminoacyl-tRNAs,disrupting peptide bond formation.Hence,the para-methoxyphenyl group serves as a starting point for developing novel anisomycin analogs with potent antifungal and insecticidal properties.However,the activation and condensation mechanism of phenylpyruvic acid has not yet been elucidated.In this study,genetic manipulations of aniP and its homologue siAniP confirmed their indispensable role in 1 biosynthesis.Bioinformatics analysis suggested that AniP and siAniP function as transketolase.siAniP was found to catalyzed condensation between 4-hydroxyphe-nylpyruvic acid(3)and glyceraldehyde(GA),initiating pyrrolidine synthesis.siAniP was specific for aromatic keto acids and tolerant of aliphatic and aromatic aldehydes,and was able to catalyze the asymmetric intermo-lecular condensation of two keto acids,leading to the formation of 24α-hydroxy ketone.To the best of our knowledge,siAniP is the first TK that catalyzes the transfer of a C2 ketol and symmetrical intermolecular coupling using aromatic keto acids as donor substrates.Structural analysis,docking model construction,and site-directed mutagenesis identified that I220,H275,R322 and W391 were crucial for substrate binding.Moreover,sequence similarity network(SSN)-based genome neighborhood network(GNN)analyses of AniP suggested the widespread occurrence of the AniP-like-mediated reaction in the biosynthesis of 1 and its analogs,particularly in the assembly of benzylpyrrolidine.These findings not only expand the repertoire of TKs but also provide a potent biocatalyst that could be used for the structural innovation of 1 and its derivatives.
基金the National Natural Science Foundation of China(31630002,31700029,31770038,31470183,21661140002 and 31170085)the Ministry of Science and Technology,China+1 种基金Shanghai Pujiang Program from the Shanghai Municipal Council of Science and Technology(12PJD021)China Postdoctoral Science Foundation(2017M620151).
文摘The protein synthesis inhibitor anisomycin features a unique benzylpyrrolidine system and exhibits potent selective activity against pathogenic protozoa and fungi.It is one of the important effective components in Agricultural Antibiotic120,which has been widely used as naturally-originated agents for treatment of crop decay in China.The chemical synthesis of anisomycin has recently been reported,but the complex process with low productivity made the biosynthesis still to be a vital mainstay in efforts.The biosynthetic gene cluster(BGC)of anisomycin in Streptomyces hygrospinosus var.beijingensis has been identified in our previous work,while poor understanding of the regulatory mechanism limited the yield enhancement via regulation engineering of S.hygrospinosus var.beijingensis.In this study here,we characterized AniF as an indispensable LuxR family transcriptional regulator for the activation of anisomycin biosynthesis.The genetic manipulations of aniF and the real-time quantitative PCR(RT-qPCR)revealed that it positively regulated the transcription of the anisomycin BGC.Moreover,the overexpression of aniF contributed to the improvement of the production of anisomycin and its derivatives.Dissection of the mechanism underlying the function of AniF revealed that it directly activated the transcription of the genes aniR-G involved in anisomycin biosynthesis.Especially,one AniF-binding site in the promoter region of aniR was identified by DNase I footprinting assay and an inverted repeat sequence(5′-GGGC-3′)composed of two 4-nt half sites in the protected region was found.Taken together,our systematic study confirmed the positive regulatory role of AniF and might facilitate the future construction of engineering strains with high productivity of anisomycin and its derivatives.
文摘Because posttraumatic stress disorder(PTSD) is a highly debilitating condition, prevention is an important research topic. This article reviews possible prevention approaches that involve the administration of drugs before the traumatic event takes place. The considered approaches include drugs that address the sympathetic nervous system, drugs interfere with the hypothalamic-pituitary-adrenal(HPA) axis, narcotics and other psychoactive drugs, as well as modulators of protein synthesis. Furthermore, some thoughts on potential ethical implications of the use of drugs for the primary prevention of PTDS are presented. While there are many barriers to overcome in this field of study, this paper concludes with a call for additional research, as there are currently no approaches that are well-suited for regular daily use.
