THE CLAVICEPS-PURPUREA GENE ENCODING DIMETHYLALLYLTRYPTOPHAN SYNTHASE, THE COMMITTED STEP FOR ERGOT ALKALOID BIOSYNTHESIS
THE CLAVICEPS-PURPUREA GENE ENCODING DIMETHYLALLYLTRYPTOPHAN SYNTHASE, THE COMMITTED STEP FOR ERGOT ALKALOID BIOSYNTHESIS
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DOI：
10.1006/bbrc.1995.2599
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发表时间：
1995-11-02
影响因子：
3.1
通讯作者：
SCHARDL, CL
中科院分区：
文献类型：
--
作者：
TSAI, HF;WANG, H;SCHARDL, CL
麦角生物碱生物合成的第一个途径特异性步骤是由异戊烯基转移酶4-（γ，γ-二甲基烯丙基）色氨酸合酶催化的。部分序列信息获得纯化的酶和简并寡核苷酸混合物被用来识别和扩增的基因，dmaW的片段。克隆了该基因的全长和近全长cDNA并进行了序列测定。将cDNA以相对于诱导型GAL 1启动子的有义和反义方向克隆到酵母表达载体中。具有有义构建体而非反义构建体或克隆载体的酵母转化体的提取物催化产生4-（γ，γ-二甲基烯丙基）色氨酸。dmaW及其cDNA的序列分析表明，它编码一个455个氨基酸的多肽，预测的分子量为51，824 Da和一个推定的异戊二烯基二磷酸结合基序。(C)出版社：Academic Press
The first pathway-specific step of ergot alkaloid biosynthesis in the fungus, Claviceps purpurea, is catalyzed by the prenyltransferase, 4-(gamma,gamma-dimethylallyl)tryptophan synthase. Partial sequence information was obtained for the purified enzyme and a degenerate oligonucleotide mixture was used to identify and amplify segments of the gene, dmaW. The complete gene and near-full-length cDNA were cloned and sequenced. The cDNA was cloned in a yeast expression vector in sense and antisense orientations relative to the inducible GAL1 promoter. Extracts of yeast transformants with the sense constructs, but not antisense constructs or cloning vector, catalyzed production of 4-(gamma,gamma-dimethylallyl)tryptophan. The sequence of dmaW and its cDNA indicated that it encoded a 455 amino acid polypeptide with a predicted molecular mass of 51,824 Da and a putative prenyl diphosphate binding motif. (C) 1995 Academic Press, Inc.