[Objective]The aim was to clone TapⅡchalcone isomerases（CHI1 A） from soybean specially and construct expression vector of PNZ8149-CHI1 A,and then transform it into Lactococcus Lactis NICE systers.[Method]Chalcone i...[Objective]The aim was to clone TapⅡchalcone isomerases（CHI1 A） from soybean specially and construct expression vector of PNZ8149-CHI1 A,and then transform it into Lactococcus Lactis NICE systers.[Method]Chalcone isomerases（CHI1 A） was cloned by RTPCR method,and it was sequenced after cloning into pMD18-T vectors,and recombined to expression vector PNZ8149-CHI1 A,then it was transformed into Lactococcus Lactis NZ3900[Result]The sequencing results indicated that the cloned fragment of CHI1 A contained 670 nucleotides,and shared a sequence homology of 92% with that from Genbank accession number AF595413（CHI1 A）.CHI1 A was transformed into NICE expression system successfully by identification of PCR and digestion.[Conclusion]The foundation of using the microorganism fermentation method to produce flavonoids was laid by construction of efficient induction expression vector with chalcone isomerases CHI1 A.展开更多
Prenylflavonoids are valuable natural products that have diverse biological properties, and are usually generated biologically by multiple metabolic enzymes in nature. In this study, structurally diverse prenylflavono...Prenylflavonoids are valuable natural products that have diverse biological properties, and are usually generated biologically by multiple metabolic enzymes in nature. In this study, structurally diverse prenylflavonoids were conveniently synthesized by enzymatic catalysis by combining GuILDT, a regiospecific chalcone prenyltransferase, and Gu CHI, a stereospecific chalcone isomerase that has promiscuous activity for both chalcones and prenylchalcones as substrates. Our findings provided a new approach for the synthesis of natural/unnatural bioactive prenylflavonoids, including prenylchalcones and optical prenylflavanones with chalcone origins.展开更多
Metabolite biosynthesis is regulated by gene expression,which is altered by DNA methylation in the promoter region.Chalcone isomerase(CHI)gene encodes a key enzyme in the Lithocarpus polystachyus Rehd flavonoid pathwa...Metabolite biosynthesis is regulated by gene expression,which is altered by DNA methylation in the promoter region.Chalcone isomerase(CHI)gene encodes a key enzyme in the Lithocarpus polystachyus Rehd flavonoid pathway,and the expression of L.polystachyus CHI(LpCHI)is closely related to the synthesis of flavonoid metabolites.In this study,we analyzed the DNA methylation site of the LpCHI promoter and its effect on gene expression and metabolite accumulation.The proportions of three types of LpCHI promoter DNA methylation are 7.5%,68.75%,18.75%,determined by bisulfite sequencing.Transcriptome sequencing shows that LpCHI is strongly up-regulated in LpCHI promoter methylation Type A but down-regulated in LpCHI promoter methylation Type B and Type C.The expression of LpCHI shows no significant difference between Type B and Type C.Moreover,nine kinds of differentially expressed transcription factors(DETFs)bind to seven CpG-sites of the LpCHI promoter region to regulate LpCHI expression.The results of metabolomics show that differentially accumulated flavonoids are higher in LpCHI promoter methylation Type A than in LpCHI promoter methylation Type B and Type C.Additionally,a positive correlation was found between the LpCHI expression and flavonoids accumulation.These results show that the effect of CpG site-specificity on gene transcription is great than that of overall promoter DNA methylation on gene transcription.The mechanisms of flavonoid genes regulating metabolite accumulation are further revealed.展开更多
文摘[Objective]The aim was to clone TapⅡchalcone isomerases（CHI1 A） from soybean specially and construct expression vector of PNZ8149-CHI1 A,and then transform it into Lactococcus Lactis NICE systers.[Method]Chalcone isomerases（CHI1 A） was cloned by RTPCR method,and it was sequenced after cloning into pMD18-T vectors,and recombined to expression vector PNZ8149-CHI1 A,then it was transformed into Lactococcus Lactis NZ3900[Result]The sequencing results indicated that the cloned fragment of CHI1 A contained 670 nucleotides,and shared a sequence homology of 92% with that from Genbank accession number AF595413（CHI1 A）.CHI1 A was transformed into NICE expression system successfully by identification of PCR and digestion.[Conclusion]The foundation of using the microorganism fermentation method to produce flavonoids was laid by construction of efficient induction expression vector with chalcone isomerases CHI1 A.
基金financially supported by the National Natural Science Foundation of China (Grant No. 81273405)CAMS Innovation Fund for Medical Sciences (Nos. CIFMS-2016-I2M-3012 and CIFMS-2017-I2M-3-013)
文摘Prenylflavonoids are valuable natural products that have diverse biological properties, and are usually generated biologically by multiple metabolic enzymes in nature. In this study, structurally diverse prenylflavonoids were conveniently synthesized by enzymatic catalysis by combining GuILDT, a regiospecific chalcone prenyltransferase, and Gu CHI, a stereospecific chalcone isomerase that has promiscuous activity for both chalcones and prenylchalcones as substrates. Our findings provided a new approach for the synthesis of natural/unnatural bioactive prenylflavonoids, including prenylchalcones and optical prenylflavanones with chalcone origins.
基金supported by the Natural Science Foundation of Hebei Province(H2020209033)the Science and Technology Project of Hebei Education Department(ZD2019075).
文摘Metabolite biosynthesis is regulated by gene expression,which is altered by DNA methylation in the promoter region.Chalcone isomerase(CHI)gene encodes a key enzyme in the Lithocarpus polystachyus Rehd flavonoid pathway,and the expression of L.polystachyus CHI(LpCHI)is closely related to the synthesis of flavonoid metabolites.In this study,we analyzed the DNA methylation site of the LpCHI promoter and its effect on gene expression and metabolite accumulation.The proportions of three types of LpCHI promoter DNA methylation are 7.5%,68.75%,18.75%,determined by bisulfite sequencing.Transcriptome sequencing shows that LpCHI is strongly up-regulated in LpCHI promoter methylation Type A but down-regulated in LpCHI promoter methylation Type B and Type C.The expression of LpCHI shows no significant difference between Type B and Type C.Moreover,nine kinds of differentially expressed transcription factors(DETFs)bind to seven CpG-sites of the LpCHI promoter region to regulate LpCHI expression.The results of metabolomics show that differentially accumulated flavonoids are higher in LpCHI promoter methylation Type A than in LpCHI promoter methylation Type B and Type C.Additionally,a positive correlation was found between the LpCHI expression and flavonoids accumulation.These results show that the effect of CpG site-specificity on gene transcription is great than that of overall promoter DNA methylation on gene transcription.The mechanisms of flavonoid genes regulating metabolite accumulation are further revealed.
