: The microbiological transformaion of a natural taxoid 10-deacetylbaccatin III into its C-7 epimer, 10-deacetylbaccatin V with Microsphaeropsis onychiuri, a fungus isolated from the inner bark of Taxus yunnanensis is...: The microbiological transformaion of a natural taxoid 10-deacetylbaccatin III into its C-7 epimer, 10-deacetylbaccatin V with Microsphaeropsis onychiuri, a fungus isolated from the inner bark of Taxus yunnanensis is described.展开更多
AIM： To study the therapeutic effect of exogenous interleuldn-10 on CCl4-induced hepatic fibrosis in rats and its passible mechanisms. METHODS： Fourty-seven SD rats were randomly divided into control group （group N...AIM： To study the therapeutic effect of exogenous interleuldn-10 on CCl4-induced hepatic fibrosis in rats and its passible mechanisms. METHODS： Fourty-seven SD rats were randomly divided into control group （group N） and CCl4-induced hepatic fibrosis model group （group C）. After CCl4 was given for 9 wk, the model group was divided into three groups. Rats in group H were put to death immediately, rats in group T were treated with IL-10 for another three wk and then put to death, rats in group R recovered after three weeks and were then killed. The degree of hepatic fibrosis was measured by HE staining and histological activity index （HAI）. Histological activity index （HAI）, change of collagen types Ⅰ and Ⅲ were measured by Picrosirius staining. The expression of TNF-α, HHP-2 and TIMP-1 in liver tissue was measured by S-P immunohis tochemistry.RESULTS： CCl4- induced experimental rat hepatic fibrosis model was established successfully. The degree of hepatic fibrosis was markedly lower in group T than in groups H and R, and there was no difference between the two groups. The expression of collagen types I and III was significantly suppressed in group T and was slightly suppressed in groups H and R. The positive levels of TNF-α, HHP-2 and TIHP-1 in group H increased significantly compared to those in group N （P〈0.01）. The positive signals decreased significantly in groups T and R （P〈0.01）, but positive score was significantly lower in group T than in group R （P〈 0.01）. CONCLUS10N： Exogenous IL-10 can reverse CCl4-induced hepatic fibrosis in rats. IL-10 may exert its reversible effects on hepatic fibrosis by blocking CCl4-induced inflammation, inhibiting expression of HHP-2 and TIMP-1 and promoting resolution of collagen types Ⅰ and Ⅲ.展开更多
The new compound was prepared by the reaction of Cd(ClO4)2·6H2O,1,10 phenanthroline (phen)and picric acid (HPA)(ratio 1∶3∶2) in methanol aqueous solution refluxing for 4 hours. The crystals used for X ray analy...The new compound was prepared by the reaction of Cd(ClO4)2·6H2O,1,10 phenanthroline (phen)and picric acid (HPA)(ratio 1∶3∶2) in methanol aqueous solution refluxing for 4 hours. The crystals used for X ray analysis <IMG SRC="IMAGE/04260089.JPG" HEIGHT=11 WIDTH=12>was obtained by slow evaporation of the solution at room temperature. The crystal compound is in triclinic with space group with a=1.1363(4)nm,b=1.2593(2)nm,c=1.3972(4)nm,α=93.97(2)?,β=100.84(3)?,γ=106.29(2)?,Z=2,V=1.8690(9)nm3,Dc=1.715g·cm-3, μ(MoKα)=0.677mm-1,F(000)=972,R=0.0907,wR=0.2240. In the complex, the Cd? cation is octahedrally coordinated to two 1,10 phenanthroline ligands and two water molecules incis disposition. As outer sphere, picrate anions are combined with inner by hydrogen bond and static electricity interaction.展开更多
Objective To optimize the extraction technology of Taxus x media by using the contents of Paclitaxel and 10-deacetylbaccatin(10-DAB) ,two representative active diterpene alkaloids of taxane type from T.x media,as eval...Objective To optimize the extraction technology of Taxus x media by using the contents of Paclitaxel and 10-deacetylbaccatin(10-DAB) ,two representative active diterpene alkaloids of taxane type from T.x media,as evaluation standard.Methods The smashing tissue extraction(STE) of Paclitaxel and 10-DAB from T.x media,was investigated by comparing with ultrasonic extraction(UE) which was one of the modern technologies of extraction.Results STE was more efficient than UE,and the contents of 10-DAB and Paclitaxel in the extracts obtained by STE were higher than those by UE.Conclusion STE is a fast,high-performance,and energy-saving technology for the extraction of diterpene alkaloids of taxane type.STE also provides a simple,component-safe,workable,and highly efficient method for the extraction of active natural product.展开更多
10-DeacetylbaccatinⅢ(10-DAB)C10 acetylation is an indispensable procedure for Taxol semi-synthesis,which often requires harsh conditions.10-DeacetylbaccatinⅢ-10-β-O-acetyltransferase(DBAT)catalyzes the acetylation ...10-DeacetylbaccatinⅢ(10-DAB)C10 acetylation is an indispensable procedure for Taxol semi-synthesis,which often requires harsh conditions.10-DeacetylbaccatinⅢ-10-β-O-acetyltransferase(DBAT)catalyzes the acetylation but acetyl-CoA supply remains a key limiting factor.Here we refactored the innate biosynthetic pathway of acetyl-CoA in Escherichia coli and obtained a chassis with acetyl-CoA productivity over three times higher than that of the host cell.Then,we constructed a microbial cell factory by introducing DBAT gene into this chassis for efficiently converting 10-DAB into baccatinⅢ.We found that baccatinⅢcould be efficiently deacetylated into 10-DAB by DBAT with CoASH and K+under alkaline condition.Thus,we fed acetic acid to the engineered strain both for serving as a substrate of acetyl-CoA biosynthesis and for alleviating the deacetylation of baccatinⅢ.The fermentation conditions were optimized and the baccatinⅢtiters reached 2,3 and 4.6 g/L,respectively,in a 3-L bioreactor culture when 2,3 and 6 g/L of 10-DAB were supplied.Our study provides an environmentfriendly approach for the large scale 10-DAB acetylation without addition of acetyl-CoA in the industrial Taxol semi-synthesis.The finding of DBAT deacetylase activity may broaden its application in the structural modification of pharmaceutically important lead compounds.展开更多
文摘: The microbiological transformaion of a natural taxoid 10-deacetylbaccatin III into its C-7 epimer, 10-deacetylbaccatin V with Microsphaeropsis onychiuri, a fungus isolated from the inner bark of Taxus yunnanensis is described.
基金Supported by Nature Science Foundation of Fujian Province. No.2005D094 and No.C0410025
文摘AIM： To study the therapeutic effect of exogenous interleuldn-10 on CCl4-induced hepatic fibrosis in rats and its passible mechanisms. METHODS： Fourty-seven SD rats were randomly divided into control group （group N） and CCl4-induced hepatic fibrosis model group （group C）. After CCl4 was given for 9 wk, the model group was divided into three groups. Rats in group H were put to death immediately, rats in group T were treated with IL-10 for another three wk and then put to death, rats in group R recovered after three weeks and were then killed. The degree of hepatic fibrosis was measured by HE staining and histological activity index （HAI）. Histological activity index （HAI）, change of collagen types Ⅰ and Ⅲ were measured by Picrosirius staining. The expression of TNF-α, HHP-2 and TIMP-1 in liver tissue was measured by S-P immunohis tochemistry.RESULTS： CCl4- induced experimental rat hepatic fibrosis model was established successfully. The degree of hepatic fibrosis was markedly lower in group T than in groups H and R, and there was no difference between the two groups. The expression of collagen types I and III was significantly suppressed in group T and was slightly suppressed in groups H and R. The positive levels of TNF-α, HHP-2 and TIHP-1 in group H increased significantly compared to those in group N （P〈0.01）. The positive signals decreased significantly in groups T and R （P〈0.01）, but positive score was significantly lower in group T than in group R （P〈 0.01）. CONCLUS10N： Exogenous IL-10 can reverse CCl4-induced hepatic fibrosis in rats. IL-10 may exert its reversible effects on hepatic fibrosis by blocking CCl4-induced inflammation, inhibiting expression of HHP-2 and TIMP-1 and promoting resolution of collagen types Ⅰ and Ⅲ.
文摘The new compound was prepared by the reaction of Cd(ClO4)2·6H2O,1,10 phenanthroline (phen)and picric acid (HPA)(ratio 1∶3∶2) in methanol aqueous solution refluxing for 4 hours. The crystals used for X ray analysis <IMG SRC="IMAGE/04260089.JPG" HEIGHT=11 WIDTH=12>was obtained by slow evaporation of the solution at room temperature. The crystal compound is in triclinic with space group with a=1.1363(4)nm,b=1.2593(2)nm,c=1.3972(4)nm,α=93.97(2)?,β=100.84(3)?,γ=106.29(2)?,Z=2,V=1.8690(9)nm3,Dc=1.715g·cm-3, μ(MoKα)=0.677mm-1,F(000)=972,R=0.0907,wR=0.2240. In the complex, the Cd? cation is octahedrally coordinated to two 1,10 phenanthroline ligands and two water molecules incis disposition. As outer sphere, picrate anions are combined with inner by hydrogen bond and static electricity interaction.
基金E&T modern center for Natural Products of Liaoning Province of China (2008402021)
文摘Objective To optimize the extraction technology of Taxus x media by using the contents of Paclitaxel and 10-deacetylbaccatin(10-DAB) ,two representative active diterpene alkaloids of taxane type from T.x media,as evaluation standard.Methods The smashing tissue extraction(STE) of Paclitaxel and 10-DAB from T.x media,was investigated by comparing with ultrasonic extraction(UE) which was one of the modern technologies of extraction.Results STE was more efficient than UE,and the contents of 10-DAB and Paclitaxel in the extracts obtained by STE were higher than those by UE.Conclusion STE is a fast,high-performance,and energy-saving technology for the extraction of diterpene alkaloids of taxane type.STE also provides a simple,component-safe,workable,and highly efficient method for the extraction of active natural product.
基金supported by the National Key Research and Development Program of China(grant Nos.2018YFA0901900 and 2020YFA0908003)the Drug Innovation Major Project(grant No.2018ZX09711001-006-001,China)+2 种基金the National Natural Science Foundation of China(grant No.81573325)the CAMS Innovation Fund for Medical Sciences(CIFMS,(grant No.2017-I2M-2-004,2019-I2M-1-005,China)PUMC Disciplinary Development of Synthetic Biology(201920100801,China)。
文摘10-DeacetylbaccatinⅢ(10-DAB)C10 acetylation is an indispensable procedure for Taxol semi-synthesis,which often requires harsh conditions.10-DeacetylbaccatinⅢ-10-β-O-acetyltransferase(DBAT)catalyzes the acetylation but acetyl-CoA supply remains a key limiting factor.Here we refactored the innate biosynthetic pathway of acetyl-CoA in Escherichia coli and obtained a chassis with acetyl-CoA productivity over three times higher than that of the host cell.Then,we constructed a microbial cell factory by introducing DBAT gene into this chassis for efficiently converting 10-DAB into baccatinⅢ.We found that baccatinⅢcould be efficiently deacetylated into 10-DAB by DBAT with CoASH and K+under alkaline condition.Thus,we fed acetic acid to the engineered strain both for serving as a substrate of acetyl-CoA biosynthesis and for alleviating the deacetylation of baccatinⅢ.The fermentation conditions were optimized and the baccatinⅢtiters reached 2,3 and 4.6 g/L,respectively,in a 3-L bioreactor culture when 2,3 and 6 g/L of 10-DAB were supplied.Our study provides an environmentfriendly approach for the large scale 10-DAB acetylation without addition of acetyl-CoA in the industrial Taxol semi-synthesis.The finding of DBAT deacetylase activity may broaden its application in the structural modification of pharmaceutically important lead compounds.
