Functional analysis of the gene controlling hydroxylation of festuclavine in the ergot alkaloid pathway of Neosartorya fumigata.
Functional analysis of the gene controlling hydroxylation of festuclavine in the ergot alkaloid pathway of Neosartorya fumigata.
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DOI：
10.1007/s00294-016-0591-5
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发表时间：
2016-11
期刊：
影响因子：
2.5
通讯作者：
Panaccione, Daniel G.
中科院分区：
文献类型：
--
作者：
Bilovol, Yulia;Panaccione, Daniel G.
麦角生物碱是由多种真菌产生的生物活性物质，在农业和医药上具有重要的应用价值。麦角生物碱途径的大部分已被阐明，但一些步骤，包括控制毛茛麦角碱羟基化为烟曲霉素B的基因，仍然没有解决。狐茅麦角碱是条件致病菌烟奈氏菌麦角生物碱途径的烟麦角碱分支中的关键中间体，也是某些麦角菌属物种基于二水解麦角酸的麦角生物碱途径中的关键中间体。根据几条证据，N。烟曲霉基因easM是编码festuclavine-羟化酶的合理候选者。为了验证这一假设，我们通过用潮霉素抗性基因替换easM的部分编码序列并转化N。fumigata与此结构。高压液相色谱分析表明，easM缺失突变体在麦角生物碱途径中被封闭在festuclavine处，并且下游产物被消除。一个额外的生物碱，提出了一个异戊二烯化形式的festuclavine的质谱数据的基础上，也积累了较高的浓度在easM敲除。与easM基因的野生型等位基因互补恢复了真菌产生下游化合物的能力。这些结果表明easM编码烟麦角碱B合成所需的酶，可能通过羟基化festuclavine。对N. fumigata可能有助于将来对二氢麦角酸衍生物的生物合成的研究，二氢麦角酸衍生物衍生自狐茅麦角碱，并且是几种重要药物的基础。
Bioactive ergot alkaloids produced by several species of fungi are important molecules in agriculture and medicine. Much of the ergot alkaloid pathway has been elucidated, but a few steps, including the gene controlling hydroxylation of festuclavine to fumigaclavine B, remain unsolved. Festuclavine is a key intermediate in the fumigaclavine branch of the ergot alkaloid pathway of the opportunistic pathogen Neosartorya fumigata and also in the dihydrolysergic acid-based ergot alkaloid pathway of certain Claviceps species. Based on several lines of evidence, the N. fumigata gene easM is a logical candidate to encode the festuclavine-hydroxylating enzyme. To test this hypothesis we disrupted easM function by replacing part of its coding sequences with a hygromycin resistance gene and transforming N. fumigata with this construct. High pressure liquid chromatography analysis demonstrated that easM deletion mutants were blocked in the ergot alkaloid pathway at festuclavine, and downstream products were eliminated. An additional alkaloid, proposed to be a prenylated form of festuclavine on the basis of mass spectral data, also accumulated to higher concentrations in the easM knockout. Complementation with the wild-type allele of easM gene restored the ability of the fungus to produce downstream compounds. These results indicate that easM encodes an enzyme required for fumigaclavine B synthesis likely by hydroxylating festuclavine. The festuclavine-accumulating strain of N. fumigata may facilitate future investigations of the biosynthesis of dihydrolysergic acid derivatives, which are derived from festuclavine and are the basis for several important drugs.
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影响因子：
4.2
作者：
Gerhards N;Neubauer L;Tudzynski P;Li SM
通讯作者：
Li SM
影响因子：
4.4
作者：
Panaccione, DG;Coyle, CM
通讯作者：
Coyle, CM
影响因子：
4.4
作者：
Coyle, Christine M.;Cheng, Johnathan Z.;Panaccione, Daniel G.
通讯作者：
Panaccione, Daniel G.
DOI：
10.1016/s1099-4831(06)63002-2
发表时间：
2006-01-01
期刊：
ALKALOIDS: CHEMISTRY AND BIOLOGY, VOL 63
影响因子：
--
作者：
Schardl, Christopher L.;Panaccione, Daniel G.;Tudzynski, Paul
通讯作者：
Tudzynski, Paul
影响因子：
4.2
作者：
Young CA;Schardl CL;Panaccione DG;Florea S;Takach JE;Charlton ND;Moore N;Webb JS;Jaromczyk J
通讯作者：
Jaromczyk J