epi-isozizaene

• 分子结构分类: 有机化合物 - 脂质和类脂质分子 - 异戊烯醇脂质
• 英文名称: epi-isozizaene
• 英文别名: (+)-Epi-isozizaene；(1R,2S,8S)-2,6,7,7-tetramethyltricyclo[6.2.1.01,5]undec-5-ene
• 化学式: C₁₅H₂₄
• 分子量: 204.356
• InChiKey: CYLSPJUZBPWJGC-ITDIGPHOSA-N

计算性质

• 辛醇/水分配系数(LogP)：
  4.5
• 重原子数：
  15
• 可旋转键数：
  0
• 环数：
  3.0
• sp3杂化的碳原子比例：
  0.87
• 拓扑面积：
  0
• 氢给体数：
  0
• 氢受体数：
  0

反应信息

• 作为反应物：
  描述：

  epi-isozizaene 在 氧气 作用下， 生成 ALBAFLAVENONE
  参考文献：
  名称：

  Lin, Xin; Cane, David E., Journal of the American Chemical Society, 2009, vol. 131, p. 6332 - 6333

  摘要：

  DOI：
• 作为产物：
  描述：

  farnesyl pyrophosphate 在 recombinant Streptomyces coelicolor epi-isozizaene synthase 作用下， 以 various solvent(s) 为溶剂， 反应 18.0h， 以4 mg的产率得到epi-isozizaene
  参考文献：
  名称：

  Genome Mining in Streptomyces coelicolor:  Molecular Cloning and Characterization of a New Sesquiterpene Synthase

  摘要：

  The terpene synthase encoded by the SCO5222 (SC7E4.19) gene of Streptomyces coelicolor was cloned by PCR and expressed in Escherichia coli as an N-terminal-His6-tag protein. Incubation of the recombinant protein, SCO5222p, with farnesyl diphosphate (1, FPP) in the presence of Mg(II) gave a new sesquiterpene, (+)-epi-isozizaene (2), whose structure and stereochemistry were determined by a combination of 1H, 13C, COSY, HMQC, HMBC, and NOESY NMR. The steady-state kinetic parameters were kcat 0.049 +/- 0.001 s-1 and a Km (FPP) of 147 +/- 14 nM. Individual incubations of recombinant epi-isozizaene synthase with [1,1-2H2]FPP (1a), (1R)-[1-2H]-FPP (1b), and (1S)-[1-2H]-FPP (1c) and NMR analysis of the resulting deuterated epi-isozizaenes supported an isomerization-cyclization-rearrangement mechanism involving the intermediacy of (3R)-nerolidyl diphosphate (3).

  DOI：

  10.1021/ja061292s

文献信息

• Structure of Epi-Isozizaene Synthase from <i>Streptomyces coelicolor</i> A3(2), a Platform for New Terpenoid Cyclization Templates^,
  作者：Julie A. Aaron、Xin Lin、David E. Cane、David W. Christianson

  DOI：10.1021/bi902088z

  日期：2010.3.2

  binding and catalysis in a bacterial terpenoid cyclase. Moreover, the binding interactions of BTAC may mimic those of a carbocation intermediate in catalysis. Accordingly, the aromatic rings of F95, F96, and F198 appear to be well-oriented to stabilize carbocation intermediates in the cyclization cascade through cation−π interactions. Mutagenesis of aromatic residues in the enzyme active site results in the

  重组表异齐扎烯合酶 (EIZS) 是一种来自天蓝色链霉菌A3(2) 的倍半萜环化酶，其 X 射线晶体结构已在 1.60 Å 分辨率下确定。具体而言，野生型 EIZS 的结构是其与三个 Mg 2+离子、无机焦磷酸盐 (PP i), 和苄基三乙基铵阳离子 (BTAC)。此外，D99N EIZS 的结构已在 1.90 Å 分辨率下以开放的无配体构象确定。这两种结构的比较提供了细菌萜类环化酶中底物结合和催化所需的构象变化的第一个视图。此外，BTAC 的结合相互作用可能模仿催化中碳阳离子中间体的结合相互作用。因此，F95、F96 和 F198 的芳环似乎很好地定向以通过阳离子-π 相互作用稳定环化级联中的碳阳离子中间体。由于碳阳离子中间体的稳定模式改变以及法尼基二磷酸环化的模板改变，酶活性位点中芳香族残基的诱变导致替代倍半萜产物阵列的产生。因此，F198A EIZS 在具有三个 Mg 的复合物中的 1
• A Catalytically Competent Terpene Synthase Inferred Using Ancestral Sequence Reconstruction Strategy
  作者：Daniele Guzzetti、Aurélien Lebrun、Maeva Subileau、Estelle Grousseau、Eric Dubreucq、Jullien Drone

  DOI：10.1021/acscatal.6b01332

  日期：2016.8.5

  On the basis of bioinformatic analysis of 137 putative and characterized bacterial terpene synthases (TS), we applied ancestral reconstruction strategy to design a biosynthetic protein sequence (AncCL1). We biochemically confirmed its catalytic competence and product profile. This enzyme catalyzes not only epi-isozizaene (minor product) but also epi-zizaene (major product) formation. We compared AncCL1 with two related TS to allow a more detailed insight into the product specificity determinants. In the future, the presented method will be useful for engineering biosynthetic TS and to elucidate the function of unknown enzymes.
• Biosynthesis of the Sesquiterpene Antibiotic Albaflavenone in Streptomyces coelicolor A3(2)
  作者：Bin Zhao、Xin Lin、Li Lei、David C. Lamb、Steven L. Kelly、Michael R. Waterman、David E. Cane

  DOI：10.1074/jbc.m710421200

  日期：2008.3

  Cytochrome P450 170A1 (CYP170A1) is encoded by the sco5223 gene of the Gram-positive, soil-dwelling bacterium Streptomyces coelicolor A3(2) as part of a two-gene cluster with the sco5222 gene. The SCO5222 protein is a sesquiterpene synthase that catalyzes the cyclization of farnesyl diphosphate to the novel tricyclic hydrocarbon, epi-isozizaene (Lin, X., Hopson, R., and Cane, D. E. (2006) J. Am. Chem. Soc. 128, 6022 - 6023). The presence of CYP170A1 (sco5223) suggested that epiisozizaene might be further oxidized by the transcriptionally coupled P450. We have now established that purified CYP170A1 carries out two sequential allylic oxidations to convert epi-isozizaene to an epimeric mixture of albaflavenols and thence to the sesquiterpene antibiotic albaflavenone. Gas chromatography/ mass spectrometry analysis of S. coelicolor culture extracts established the presence of albaflavenone in the wildtype strain, along with its precursors epi-isozizaene and the albaflavenols. Disruption of the CYP170A1 gene abolished biosynthesis of both albaflavenone and the albaflavenols, but not epi-isozizaene. The combined results establish for the first time the presence of albaflavenone in S. coelicolor and clearly demonstrate that the biosynthesis of this antibiotic involves the coupled action of epi-isozizaene synthase and CYP170A1.
• Lin, Xin; Cane, David E., Journal of the American Chemical Society, 2009, vol. 131, p. 6332 - 6333
  作者：Lin, Xin、Cane, David E.

  DOI：——

  日期：——
• Genome Mining in <i>Streptomyces </i><i>c</i><i>oelicolor</i>:  Molecular Cloning and Characterization of a New Sesquiterpene Synthase
  作者：Xin Lin、Russell Hopson、David E. Cane

  DOI：10.1021/ja061292s

  日期：2006.5.1

  The terpene synthase encoded by the SCO5222 (SC7E4.19) gene of Streptomyces coelicolor was cloned by PCR and expressed in Escherichia coli as an N-terminal-His6-tag protein. Incubation of the recombinant protein, SCO5222p, with farnesyl diphosphate (1, FPP) in the presence of Mg(II) gave a new sesquiterpene, (+)-epi-isozizaene (2), whose structure and stereochemistry were determined by a combination of 1H, 13C, COSY, HMQC, HMBC, and NOESY NMR. The steady-state kinetic parameters were kcat 0.049 +/- 0.001 s-1 and a Km (FPP) of 147 +/- 14 nM. Individual incubations of recombinant epi-isozizaene synthase with [1,1-2H2]FPP (1a), (1R)-[1-2H]-FPP (1b), and (1S)-[1-2H]-FPP (1c) and NMR analysis of the resulting deuterated epi-isozizaenes supported an isomerization-cyclization-rearrangement mechanism involving the intermediacy of (3R)-nerolidyl diphosphate (3).