Construction of A Chemical Library of Unnatural Natural Products by Using Biosynthetic Enzymes

利用生物合成酶构建非天然天然产物化学库

• 批准号: 14580613
• 负责人: ABE Ikuro
• 金额: $ 2.37万
• 依托单位: University of Shizuoka
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14580613/
• 关键词: Unnatural products; Plant polyketide Synthase; Chalcone Synthase; Squalene Cyclase; Cyclic polypruoids; Biosynthetic enzymes; Library; Enzyme reaction; 酵素構造機能相関; 酵素機能改変; カルコン構成酵素; スクレアン環化酵素

1.Squalene cyclizing enzymes.(1)Newly synthesized C_<35> polyprene was enzymatically converted to unnatural hexacyclic polyprenoid by squalene : hopene cyclase from Alicyclobacillus acidocaldarius. This is the first demonstration of the remarkable ability of the squalene cyclizing enzyme to perform construction of unnatural hexacyclic skeleton. Further, methylidene-extended squalene analogues were converted to unnatural pentacyclic novel polyprenoides by the enzyme. (2)Recombinant β-amyrin synthase from Pisum sativum converted 22,23-dihydro-2,3-oxidosqualene into bacchar-12-en-3β-ol ; the first demonstration of the enzymatic formation of the baccharene skeleton with a six-membered D-ring. It is remarkable that the formation of the anti-Markovnikov six-membered D-ring did not depend on he participation of the termianl π-electrons. Furthermore, the recombinant enzyme also converted 24,30-bisnor-2,3-oxidosqualene into 29,30-bisnor-β-amyrin.2.Plant polyketide synthases (PKSs).(1)Substrate … More specificities of plant polyketide synthases (PKSs) were investigated using analogs of malonyl-CoA and 4-coumaroyl-CoA, plant PKSs afforded an unnatural C_6-C_5 aromatic polyketide. In the chalcone synthase (CHS) reaction, it was shown that both the starter molecule and the extension unit of the poyketide chain elongation reaction could be simultaneously replaced with non-physiological substrates.(2)One of the most characteristic features of benzalacetone synthase (BAS) is that it lacks active site Phe215 ; the residues ^<214>LF conserved in type III PKSs are uniquely replaced by IL. Our observation that BAS I214L/L215F mutant exhibited chalcone forming activity in pH dependent manner supported a hypothesis that the absence of Phe 215 in BAS accounts for the interruption of the polyketide chain elongation at the diketide stage. These results confirmed the critical role of Phe215 in the polyketide formation reactions and provided structural basis for understanding the structure-function relationship of the plant type III PKSs. Less

1.角鲨烯环化酶。（1）利用酸热脂环酸芽孢杆菌（Alicyclobacillusacidocaldarius）的角鲨<35>烯：霍烯环化酶（Squalene：Hopene cyclase）将新合成的C_ polyprene转化为非天然的六环类聚异戊二烯。这是首次证明角鲨烯环化酶具有构建非天然六环骨架的显著能力。此外，亚甲基扩展的角鲨烯类似物被转化为非天然的五环新的聚异戊二烯的酶。（2）来自豌豆（Pisum sativum）的重组β-香树脂素合酶将22，2,3-二氢-2，3-氧化角鲨烯转化为bacchar-12-en-3β-ol ;首次证明了具有六元D环的baccharene骨架的酶促形成。值得注意的是，反马氏六元D环的形成并不依赖于端π电子的参与.此外，该重组酶还能将24，30-双去甲-2，3-氧化角鲨烯转化为29，30-双去甲-β-amphaline。（1）基材 ...更多信息 利用丙二酰辅酶A和4-香豆酰辅酶A的类似物研究了植物聚酮酶（PKSs）的特异性，植物PKSs产生非天然C_6-C_5芳香聚酮。在查耳酮合酶（CHS）反应中，它表明，无论是起始分子和聚酮链延伸反应的延伸单元，可以同时取代非生理底物。（2）苄叉丙酮合酶（BAS）的最大特征之一是缺少活性位点Phe 215，<214>III型PKS中保守的残基^ LF被IL唯一取代。我们观察到BAS I214 L/L215 F突变体表现出pH依赖性的查耳酮形成活性，这支持了一个假设，即BAS中Phe 215的缺失解释了聚酮链在双酮阶段的延伸中断。这些结果证实了Phe 215在聚酮形成反应中的关键作用，并为理解植物III型PKS的结构与功能关系提供了结构基础。少

项目成果

I.Abe, H.Tanaka, H.Noguchi: "Enzymatic Formation of an Unnatural Hexacyclic C35 Polyprenoid by Bacterial Squalene Cyclase"J.Am.Chem.Soc.. 124. 14514-14515 (2002)

I.Abe、H.Tanaka、H.Noguchi：“通过细菌角鲨烯环化酶酶促形成非天然六环 C35 聚戊二烯”J.Am.Chem.Soc.. 124. 14514-14515 (2002)

阿部郁朗: "オキシドスクアレン閉環素の生物有機化学的研究"薬学研究の進歩. 18. 1-11 (2002)

阿部郁郎：《氧化角鲨烯闭环元素的生物有机化学研究》药物研究进展18. 1-11 (2002)。

Ikuro Abe, Yoriko Utsumi, Satoshi Oguro, Hiroshi Noguchi: "The First Plant Type III Polyketide Synthase Catalyzing Formation of Aromatic Heptaketide."FEBS Lett.. 562. 171-176 (2004)

Ikuro Abe、Yoriko Utsumi、Satoshi Oguro、Hiroshi Noguchi：“第一个催化芳香七肽形成的植物 III 型聚酮合酶。”FEBS Lett.. 562. 171-176 (2004)

I.Abe, Y.Sakano, H.Tanaka, W.Lou, H.Noguchi, M.Shibuya, Y.Ebizuka: "Enzymatic Cyclization of 22,23-Dihydro-2,3-oxidosqualene into Euph-7-en-3β-ol and Bacchar-12-en-3β-ol by Recombinant β-Amyrin Synthase"J.Am.Chem.Soc.. 126. 34326-3427 (2004)

I.Abe、Y.Sakano、H.Tanaka、W.Lou、H.Noguchi、M.Shibuya、Y.Ebizuka：“22,23-二氢-2,3-氧化角鲨烯酶促环化成 Euph-7-en-通过重组 β-香树脂醇合酶合成 3β-ol 和 Bacchar-12-en-3β-ol”J.Am.Chem.Soc.. 126. 34326-3427 (2004)

I.Abe, Y.Utsumi, S.Oguro, H.Noguchi: "The First Plant Type III Polyketide Synthase Catalyzing Formation of Aromatic Heptaketide"FEBS Lett.. 562. 171-176 (2004)

I.Abe、Y.Utsumi、S.Oguro、H.Noguchi：“第一个催化芳香族七肽形成的植物 III 型聚酮合酶”FEBS Lett.. 562. 171-176 (2004)