Chiral syntheses by use of substrate specificities of by use of recombinant prenyltransferases

通过使用重组异戊二烯基转移酶的底物特异性进行手性合成

• 批准号: 12680586
• 负责人: NAGAKI Masahiko
• 金额: $ 2.3万
• 依托单位: Hirosaki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12680586/
• 关键词: chiral synthesis; farnesyl diphospahte synthase; geranyl diphosphate; substrate specificity; prenyltransferase; ndecaprenyl diphosphate; isoprenoid; granylgeranyl diphosphate; ブレニル基転移酵素; ウンデカプレニル基転移酵素; イソプレン化学; 遺伝子組み替え体酵素; キラル化合物; キラルシントン; フ

In order to investigate substrate spcificity of Bacillus stearothermophilus farnesyl diphosphate synthase (FPS), we examined the reactivity of 4-alkyl group homologs of isopentenyl diphosphate (IPP).The enzymatic reactions of the 4-methyl homologs, (E)-3-methylpent-3-enyl diphophate (1a) and (Z)-3methylpent-3-enyl diphophate (Ib) with geranyl diphosphate (GPP) gave 4-methyl-farnesyl diphosphates (2a and 2b), respectively. The stereochemistry of each aldehyde derived from 2a or 2b was determined by CD spectrometry to be (S)- or (R)-4-methylfarnesal, respectively.Similarly, 1a reacted with dimethylallyl diphosphate (DMAPP) to give a mixture of (4S)-4-methylgeranyl diphospahte (3a) and (4S,8S)-4,8-dimethylfarnesyl diphosphate (4a). The Z-isomer 1b also reacted with DMAPP to give the corresponding enantiomers with (4R)- and (4R,8R)-configurations.Reactions of the 4-ethyl homologs, (E)--3-methylhex-3-enyl diphosphate (1c) and (Z)-3-methylhex-3enyl diphosphate (1d) with GPP gave two types of … More 4-ethylfarnesyl diphosphates. Reactions of 1c or 1d with DMAPP also gave two types of 4-ethylgeranyl- and 4,8- diethylfarnesyl diphosphates. Meanwhile, reaction of the 4-propyl homologs, (E)--3-methylhept-3-enyl diphosphate (1e) and (Z)-3-methylhept-3-enyl diphosphate (1f) with GPP gave two types of 4-propylfarnesyl diphosphates. Reactions of 1e or 1f with DMAPP gave only two types of the4-propylGPPsHowever, neither (E)-3-methyloct-3-enyl diphosphate (1g) or (Z)-3-methyloct-3-enyl diphosphate (1h), nor (E)-4-bromo-3-methylbut-3-enyl diphosphate (1i) or (Z)-4-bromo-3-methylbut-3-enyl diphosphate (1j) was acceptable as a substrate for the thermophilic FPS at all.On the other hand, in order to investigate the reactivity of the homologs of homoallylic substrates, we examined the enzymatic reactions of (E)-3-methylhex-3-enyl diphosphate or (Z)-3-methylhex-3-enyl diphosphate with FPP using the undecaprenyl diphosphate synthase (UPS) of Micrococcus luteus B-P 26. (E)-3-methylhex-3-enyl diphosphate reacted with FPP to give a chiral (4S)-(2Z,6E,10E,14E)-4-ethyl3,7,11,15-tetramethylhexadeca-2,6,10,14-tetraen-1-ol. However, the Z-isomer is not accepted as a substrate for the bacterial UPS at all. Less

为了研究嗜热脂肪芽孢杆菌法呢基二磷酸合酶（FPS）的底物特异性，我们研究了异戊烯基二磷酸（IPP）的4-烷基同系物的反应性，（E）-3-甲基-3-烯基二磷酸酯（1a）和（Z）-3-甲基-3-烯基二磷酸酯（Ib）与香叶基二磷酸（GPP）反应生成4-甲基-法呢基二磷酸（2a和2b）。2a和2b的醛经圆二色光谱测定分别为（S）-或（R）-4-甲基法呢醛，1a与二甲基烯丙基二磷酸酯（DMAPP）反应得到（4S）-4-甲基香叶基二磷酸酯（3a）和（4S，8 S）-4，8-二甲基法呢基二磷酸酯（4a）的混合物。Z-异构体1b也与DMAPP反应生成相应的（4 R）-和（4 R，8 R）-构型的对映体，4-乙基同系物（E）-3-甲基己-3-烯基二磷酸酯（1c）和（Z）-3-甲基己-3-烯基二磷酸酯（1d）与GPP反应生成两种构型的对映体 ...更多信息 4-乙基法呢基二磷酸酯。1c或1d与DMAPP反应也得到两种类型的4-乙基香叶基-和4，8-二乙基法呢基二磷酸酯。同时，4-丙基同系物（E）-3-甲基庚-3-烯基二磷酸酯（1 e）和（Z）-3-甲基庚-3-烯基二磷酸酯（1f）与GPP反应，得到两种4-丙基法呢基二磷酸酯。1 e或1f与DMAPP的反应仅得到两种类型的4-丙基GPP。然而，（E）-3-甲基辛-3-烯基二磷酸酯（1g）或（Z）-3-甲基辛-3-烯基二磷酸酯（1h），（E）-4-溴-3-甲基丁-3-烯基二磷酸酯（1 i）或（Z）-4-溴-3-甲基丁-3-烯基二磷酸酯（1 j）都不能作为嗜热FPS的底物。为了研究高烯丙基底物的同系物的反应性，我们使用藤黄微球菌B-P26的十一异戊二烯基二磷酸合酶（UPS）检测了（E）-3-甲基己-3-烯基二磷酸或（Z）-3-甲基己-3-烯基二磷酸与FPP的酶促反应。（E）-3-甲基己-3-烯基二磷酸酯与FPP反应得到手性（4S）-（2 Z，6 E，10 E，14 E）-4-乙基-3，7，11，15-四甲基十六-2，6，10，14-四烯-1-醇。然而，Z-异构体根本不被接受为细菌UPS的底物。少

项目成果

長岐正彦: "Artificial substrates of medium-chain elongating enzymes, hexaprenyl-and heptaprenyl diphosphate synthases"Bioorganic Medicinal Chemistry Letters. 11. 2157-2159 (2001)

Masahiko Nagagi：“中链延长酶、己烯基和庚烯基二磷酸合酶的人工底物”《生物有机药物化学快报》11. 2157-2159 (2001)。

Y. Maki, M. Nagaki, T. Nishino, and T. Koyama: "Usefulness of prenyltransferase for organic synthesis: Carbon-carbon Bond forming reactions by prenyltransferases and their mutated enzymes"Yuki Gosei Kagaku Kyoukaishi. (in press). (2002)

Y. Maki、M. Nagaki、T. Nishino 和 T. Koyama：“异戊二烯基转移酶在有机合成中的用途：异戊二烯基转移酶及其突变酶的碳-碳键形成反应”Yuki Gosei Kagaku Kyoukaishi。

長岐正彦: "Artificial substrate for undecaprenyl diphosphate synthase from Micrococcus luteus B-P 26"Catalysis Communications. 11. 33-35 (2000)

Masahiko Nagagi：“来自藤黄微球菌 B-P 26 的十一碳二烯基二磷酸合酶的人工底物”，《催化通讯》11. 33-35 (2000)。

M. Nagaki, S. Sato, Y. Maki, T. Nishino, and T. Koyama: "Artificial Substrates for undecaprenyl diphosphate synthase from Micrococcus luteus B-P 26"J. Mol. Catal., B: Enzymatic.. 9(1-3). 33-38 (2000)

M. Nagaki、S. Sato、Y. Maki、T. Nishino 和 T. Koyama：“来自藤黄微球菌 B-P 26 的十一碳二烯基二磷酸合酶的人工底物”J。

長岐正彦: "Substrate specificity of thermostable farnesyl diphosphate synthase with respect to 4-alkyl homologs of isopentenyl diphosphate"J.Mol.Catal.B : Enzymatic. (in press). (2002)

Masahiko Nagagi：“热稳定性法尼基二磷酸合酶相对于异戊烯基二磷酸的 4-烷基同系物的底物特异性”J.Mol.Catal.B：酶学（2002 年）。