[Objectives] To obtain a resveratrol synthase gene of Vitis vinifera and make bioinformatics analysis. [Methods] Taking total RNA of V. vinifera as the template,by RT-PCR method,a complete c DNA sequence of resveratro...[Objectives] To obtain a resveratrol synthase gene of Vitis vinifera and make bioinformatics analysis. [Methods] Taking total RNA of V. vinifera as the template,by RT-PCR method,a complete c DNA sequence of resveratrol synthase gene was amplified from V. vinifera,and the resveratrol synthase gene was named as RS. The nucleic acid and protein sequences were analyzed using bioinformatics software.[Results]This sequence was 1179 bp in length,the similarity with reported resveratrol synthase gene reached 94%-99%,and the similarity with amino acid sequence reached 96%-99%; the RS gene encoded 392 amino acids,and amino acid sequence contained complete characteristic sequence GVLFGPGLT and active center sequence GCYAGGTVLR of stilbene synthase family; the predicted molecular weight was42. 78 k Da,the theoretical isoelectric point was 6. 57,the instability parameter was 35. 92,and it belonged to stable protein in the classification; the secondary structure was mainly α-helix,random coil and β-folding,α-helix content was 44. 13%,the random coil content was26. 53%,and β-folding content was 17. 66%. [Conclusions] The isolated RS gene is a resveratrol synthase gene from V. vinifera. This experiment is expected to lay a certain foundation for biosynthesis of resveratrol by the genetic engineering method.展开更多
Background: Resveratrol naturally occurring antioxidant in peanut (Legume: Arachis hypogaea) has phytochemical human health dietary effects associated with reduced inflammatory cancer risks. Its levels in peanut are u...Background: Resveratrol naturally occurring antioxidant in peanut (Legume: Arachis hypogaea) has phytochemical human health dietary effects associated with reduced inflammatory cancer risks. Its levels in peanut are ultra-low and variable (0 to 26 μg·g-1), which has made it difficult to market as a consistent high resveratrol produce. Objective: Understanding the regulation of resveratrol accumulation in peanut might lead to development of new techniques for optimizing and stabilizing its yield. Method: Peanuts were cultivated in horticultural field plots and treated with solutions of mineral salts (sulfate, potassium, phosphate, ammonium ion) that were optimized in stoichiometric (reactive) ratios. Peanut seed’s RNAs were subjected to Northern blot analysis for profiling the RNAs synthesized by glutamate dehydrogenase (GDH), and mRNAs encoding resveratrol synthase. The seed’s extracts were analyzed by GC-MS for determination of the resveratrol and fatty acid compositions. Result: Stoichiometric mixes of mineral ions induced the peanut GDH to synthesize some RNA that silenced the mRNAs encoding resveratrol synthase, phosphoglucomutase, isocitrate lyase, malate synthase, enolase, phosphoenolpyruvate carboxylase, malate dehydrogenase, and phosphoglycerate mutase in the control, KN-, and NPKS-treated but not in the NPPK-treated peanut. These resulted to decreased resveratrol content (6.0 μg·g-1) in the control peanut but maximized it (1.15 mg·g-1) in the NPPK-treated peanut. Therefore, resveratrol accumulation was optimized by coupling of glycolysis and citric-glyoxylic acid cycles to resveratrol biosynthesis. Fatty acid content of control (55.6 g·kg-1) was higher than the NPKS-treated (48.5 g·kg-1) and NPPK-treated peanut (44.9 g·kg-1) meaning that malonyl-CoA intermediate in both fatty acid and stilbenoid pathways was diverted to support maximum resveratrol biosynthesis in the NPPK-treated peanut. Conclusion: The functional coupling of citric-glyoxylic acid cycles and glycolysis to optimize resveratrol展开更多
基金Supported by Construction and Expression of Heterologous Biosynthesis Pathway for Resveratrol(BAK:201502bsh)
文摘[Objectives] To obtain a resveratrol synthase gene of Vitis vinifera and make bioinformatics analysis. [Methods] Taking total RNA of V. vinifera as the template,by RT-PCR method,a complete c DNA sequence of resveratrol synthase gene was amplified from V. vinifera,and the resveratrol synthase gene was named as RS. The nucleic acid and protein sequences were analyzed using bioinformatics software.[Results]This sequence was 1179 bp in length,the similarity with reported resveratrol synthase gene reached 94%-99%,and the similarity with amino acid sequence reached 96%-99%; the RS gene encoded 392 amino acids,and amino acid sequence contained complete characteristic sequence GVLFGPGLT and active center sequence GCYAGGTVLR of stilbene synthase family; the predicted molecular weight was42. 78 k Da,the theoretical isoelectric point was 6. 57,the instability parameter was 35. 92,and it belonged to stable protein in the classification; the secondary structure was mainly α-helix,random coil and β-folding,α-helix content was 44. 13%,the random coil content was26. 53%,and β-folding content was 17. 66%. [Conclusions] The isolated RS gene is a resveratrol synthase gene from V. vinifera. This experiment is expected to lay a certain foundation for biosynthesis of resveratrol by the genetic engineering method.
文摘Background: Resveratrol naturally occurring antioxidant in peanut (Legume: Arachis hypogaea) has phytochemical human health dietary effects associated with reduced inflammatory cancer risks. Its levels in peanut are ultra-low and variable (0 to 26 μg·g-1), which has made it difficult to market as a consistent high resveratrol produce. Objective: Understanding the regulation of resveratrol accumulation in peanut might lead to development of new techniques for optimizing and stabilizing its yield. Method: Peanuts were cultivated in horticultural field plots and treated with solutions of mineral salts (sulfate, potassium, phosphate, ammonium ion) that were optimized in stoichiometric (reactive) ratios. Peanut seed’s RNAs were subjected to Northern blot analysis for profiling the RNAs synthesized by glutamate dehydrogenase (GDH), and mRNAs encoding resveratrol synthase. The seed’s extracts were analyzed by GC-MS for determination of the resveratrol and fatty acid compositions. Result: Stoichiometric mixes of mineral ions induced the peanut GDH to synthesize some RNA that silenced the mRNAs encoding resveratrol synthase, phosphoglucomutase, isocitrate lyase, malate synthase, enolase, phosphoenolpyruvate carboxylase, malate dehydrogenase, and phosphoglycerate mutase in the control, KN-, and NPKS-treated but not in the NPPK-treated peanut. These resulted to decreased resveratrol content (6.0 μg·g-1) in the control peanut but maximized it (1.15 mg·g-1) in the NPPK-treated peanut. Therefore, resveratrol accumulation was optimized by coupling of glycolysis and citric-glyoxylic acid cycles to resveratrol biosynthesis. Fatty acid content of control (55.6 g·kg-1) was higher than the NPKS-treated (48.5 g·kg-1) and NPPK-treated peanut (44.9 g·kg-1) meaning that malonyl-CoA intermediate in both fatty acid and stilbenoid pathways was diverted to support maximum resveratrol biosynthesis in the NPPK-treated peanut. Conclusion: The functional coupling of citric-glyoxylic acid cycles and glycolysis to optimize resveratrol
文摘以葡萄总RNA为模板,利用RT-PCR的方法从葡萄中扩增获得一条白藜芦醇合酶基因完整的c DNA序列,命名为RS。利用生物信息学软件对其核酸和蛋白质序列进行分析,结果表明,该序列长1179 bp,与已报道的葡萄白藜芦醇合酶基因的序列相似性达到94%~99%,氨基酸序列相似性为96%~99%;RS基因编码392个氨基酸,氨基酸序列含有完整的芪合酶家族的特征序列GVLFGPGLT和活性中心序列GCYAGGTVLR;预测的分子量为42.78 k Da,理论等电点为6.57,不稳定参数为35.92,在分类上属于稳定性蛋白;二级结构主要以α-螺旋、无规则卷曲以及β-折叠为主,其中α-螺旋含量为44.13%、无规则卷曲含量为26.53%,β-折叠含量为17.66%。
