Expression of deacetylvindoline-4-O-acetyltransferase in Catharanthus roseus hairy roots
Expression of deacetylvindoline-4-O-acetyltransferase in Catharanthus roseus hairy roots
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DOI：
10.1016/j.phytochem.2007.04.037
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发表时间：
2007-07-01
期刊：
影响因子：
3.8
通讯作者：
De Luca, Vincenzo
中科院分区：
文献类型：
--
作者：
Magnotta, Mary;Murata, Jun;De Luca, Vincenzo
马达加斯加长春花[Catharanthus roseus（L.）GDon]是具有园艺价值的泛热带植物，其产生强效抗癌药物长春碱和长春新碱，其衍生自单萜吲哚生物碱（MIA）、文多灵和长春质碱的二聚体。本研究描述了文多灵生物合成的末端步骤，脱乙酰文多灵-4-O-乙酰转移酶（DAT）的基因工程和表达在长春花毛状根培养。生化分析表明，几个毛状根线表达高水平的DAT酶活性相比，对照毛状根培养物表达β-古洛糖醛酸酶活性（GUS）的活性。代谢产物分析，采用高效液相色谱法建立毛状根提取物有一个改变的生物碱配置文件相对于DAT表达线相比，控制线horhammericine积累。对表达高DAT活性的一种毛状根培养物的进一步分析表明DAT表达和horhammericine的积累相关（9）。可以得出结论，DAT在毛状根中的表达改变了它们的MIA配置文件，并表明文多灵途径基因的进一步表达可能导致生物碱配置文件的显着变化。提供的证据表明，horhammericine（9）通过DAT与根特异性米诺长春碱-19-O-乙酰基转移酶（MAT）的相互作用而积累，该作用抑制MAT介导的hammericine（9）转化为19-O-乙酰基-horhammericine（12）。(c)2007爱思唯尔有限公司保留所有权利。
Madagascar periwinkle [Catharanthus roseus (L.) G Don] is a pantropical plant of horticultural value that produces the powerful anticancer drugs vinblastine and vincristine that are derived from the dimerization of the monoterpenoid indole alkaloids (MIAs), vindoline and catharanthine. The present study describes the genetic engineering and expression of the terminal step of vindoline biosynthesis, deacetylvindoline-4-O-acetyltransferase (DAT) in Catharanthus roseus hairy root cultures. Biochemical analyses showed that several hairy root lines expressed high levels of DAT enzyme activity compared to control hairy root cultures expressing beta-gulucuronidase activity (GUS) activity. Metabolite analysis using high performance liquid chromotagraphy established that hairy root extracts had an altered alkaloid profile with respect to horhammericine accumulation in DAT expressing lines in comparison to control lines. Further analyses of one hairy root culture expressing high DAT activity suggested that DAT expression and accumulation of horhammericine (9) were related. It is concluded that expression of DAT in hairy roots altered their MIA profile and suggests that further expression of vindoline pathway genes could lead to significant changes in alkaloid profiles. Evidence is provided that horhammericine (9) accumulates via a DAT interaction with the root specific minovincinine-19-O-acetyltransferase (MAT) that inhibits the MAT mediated conversion of hammericine (9) into 19-O-acetyl-horhammericine (12). (c) 2007 Elsevier Ltd. All rights reserved.