Biosynthesis of the sesquiterpene botrydial in Botrytis cinerea. Mechanism and stereochemistry of the enzymatic formation of presilphiperfolan-8beta-ol.
Biosynthesis of the sesquiterpene botrydial in Botrytis cinerea. Mechanism and stereochemistry of the enzymatic formation of presilphiperfolan-8beta-ol.
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DOI：
10.1021/ja9021649
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发表时间：
2009-06-24
影响因子：
15
通讯作者：
Cane DE
中科院分区：
文献类型：
--
作者：
Wang CM;Hopson R;Lin X;Cane DE
由来自坏死营养型植物病原体灰葡萄孢（Botrytis cinerea）的BcBOT 2基因编码的presilphiferfolan-8β-ol合酶催化法呢基二磷酸（2）多步环化为三环倍半萜醇presilphiferfolan-8β-ol（3），其是植物毒素葡萄孢（一种菌株依赖性真菌毒力因子）的前体。将（1 R）-[1- 2 H]法呢基二磷酸（2b）与重组presilphiferfolan-8β-醇合酶孵育仅产生（5 R）-[5α-2H]-3b，而将（1 S）-[1- 2 H]FPP（2c）互补孵育产生（5S）-[5β-2H]-3c。这些结果表明，法呢基二磷酸的环化涉及从C-1的二磷酸基团的位移与构型的净反转，并排除了环化中的橙花苷基二磷酸（9）的顺式构象的拟议中间体。虽然不是强制性中间体，但（3R）-橙花苷二磷酸被证明可作为底物替代物。环化[13，13，13 - 2 H3]法呢基二磷酸酯（2d）得到[14，14，14 - 2 H3]-3d，从而确定亲电攻击仅发生在2的12，13-双键的si面上。结合的结果提供了酶结合的法呢基二磷酸在presilphiferfolan-8β-醇合酶的活性位点的构象的详细图片。
Presilphiperfolan-8β-ol synthase, encoded by the BcBOT2 gene from the necrotrophic plant pathogen Botrytis cinerea, catalyzes the multistep cyclization of farnesyl diphosphate (2) to the tricyclic sesquiterpene alcohol presilphiperfolan-8β-ol (3), the preursor of the phytotoxin botrydial, a strain-dependent fungal virulence factor. Incubation of (1R)-[1-2H]farnesyl diphosphate (2b) with recombinant presilphiperfolan-8β-ol synthase gave exclusively (5R)-[5α-2H]-3b, while complementary incubation of (1S)-[1-2H]FPP (2c) gave (5S)-[5β-2H]-3c. These results established that cyclization of farnesyl diphosphate involves displacement of the diphosphate group from C-1 with net inversion of configuration and ruled out the proposed intermediacy the cisoid conformer of nerolidyl diphosphate (9) in the cyclization. While not a mandatory intermediate, (3R)-nerolidyl diphosphate was shown to act as a substrate surrogate. Cyclization of [13,13,13-2H3] farnesyl diphosphate (2d) gave [14,14,14-2H3]-3d, thereby establishing that electrophilic attack takes place exclusively on the si face of the 12,13-double bond of 2. The combined results provide a detailed picture of the conformation of enzyme-bound farnesyl diphosphate at the active site of presilphiperfolan-8β-ol synthase.
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影响因子：
4
作者：
Pinedo, Cristina;Wang, Chieh-Mei;Pradier, Jean-Marc;Dalmais, Berengere;Choquer, Mathias;Le Pecheur, Pascal;Morgant, Guillaume;Collado, Isidro G.;Cane, David E.;Viaud, Muriel
通讯作者：
Viaud, Muriel
影响因子：
5.2
作者：
Wang, Selina C.;Tantillo, Dean J.
通讯作者：
Tantillo, Dean J.
影响因子：
15
作者：
Coates, RM;Ho, ZQ;Wilson, SR
通讯作者：
Wilson, SR
影响因子：
3.5
作者：
Siewers, V;Viaud, M;Tudzynski, P
通讯作者：
Tudzynski, P
DOI：
10.1039/p19820002187
发表时间：
1982-01-01
期刊：
JOURNAL OF THE CHEMICAL SOCIETY-PERKIN TRANSACTIONS 1
影响因子：
--
作者：
BRADSHAW, APW;HANSON, JR;SADLER, IH
通讯作者：
SADLER, IH