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黄芩中酪氨酸转氨酶和对羟基苯丙酮酸还原酶的分子克隆与特性分析以及迷迭香酸的积累

Molecular cloning and characterization of tyrosine aminotransferase and hydroxyphenylpyruvate reductase, and rosmarinic acid accumulation in Scutellaria baicalensis.

作者信息

Kim Yeon Bok, Uddina Md Romij, Kim YeJi, Park Chun Geon, Park Sang Un

出版信息

Nat Prod Commun. 2014 Sep;9(9):1311-4.

Abstract

Rosmarinic acid (a-O-caffeoyl-3,4-dihydroxyphenylacetic acid, RA) is a caffeoyl ester widely distributed in plants. cDNA clones encoding tyrosine aminotransferase (TAT1 and 2) and hydroxyphenylpyruvate reductase (HPPR) have been isolated from Scutellaria baicalensis. The open reading frames (ORFs) of SbTAT1 and 2 were 1230 and 1272 bp long and encoded 409 and 423 amino acid residues, respectively. HPPR corresponded to a 942-bp ORF and 313 amino acid residues of translated protein. To study the molecular mechanisms of TAT and HPPR and investigate RA accumulation in S. baicalensis, we examined the transcript levels of TAT isoforms and HPPR with quantitative real-time PCR and analyzed the RA content in different organs by using high-performance liquid chromatography. The transcript levels of SbTATI SbTAT2, and SbHPPR in the flowers were higher than those in other organs. RA was also highly accumulated in the flowers and with a trace amount in the roots. No RA was detected in the leaves and stems of S. baicalensis. The amount of accumulated RA in the flowers was 28.7 times higher than that in the roots. Our results will be helpful in elucidating the mechanisms of RA biosynthesis in S. baicalensis.

摘要

迷迭香酸(α - O - 咖啡酰 - 3,4 - 二羟基苯乙酸,RA)是一种广泛分布于植物中的咖啡酰酯。已从黄芩中分离出编码酪氨酸转氨酶(TAT1和2)和对羟基苯丙酮酸还原酶(HPPR)的cDNA克隆。SbTAT1和2的开放阅读框(ORF)分别为1230和1272 bp长,分别编码409和423个氨基酸残基。HPPR对应于一个942 bp的ORF和313个翻译蛋白的氨基酸残基。为了研究TAT和HPPR的分子机制并探究黄芩中RA的积累情况,我们用定量实时PCR检测了TAT同工型和HPPR的转录水平,并使用高效液相色谱分析了不同器官中的RA含量。花中SbTAT1、SbTAT2和SbHPPR的转录水平高于其他器官。RA在花中也高度积累,在根中含量微量。黄芩的叶和茎中未检测到RA。花中积累的RA量比根中高28.7倍。我们的结果将有助于阐明黄芩中RA生物合成的机制。

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