12-(4-trifluoromethylcoumarin-7-yloxy)dodecanoic acid | 1610978-43-2

• 分子结构分类: 有机化合物 - 苯丙烷和聚酮 - 香豆素及其衍生物
• 英文名称: 12-(4-trifluoromethylcoumarin-7-yloxy)dodecanoic acid
• 英文别名: 12-[4-(Trifluoromethyl)-2-oxo-2H-1-benzopyran-7-yloxy]dodecanoic acid；12-[2-oxo-4-(trifluoromethyl)chromen-7-yl]oxydodecanoic acid
• CAS: 1610978-43-2
• 化学式: C₂₂H₂₇F₃O₅
• 分子量: 428.449
• InChiKey: ZGDRMWQAOCKNEP-UHFFFAOYSA-N

计算性质

• 辛醇/水分配系数(LogP)：
  6.1
• 重原子数：
  30
• 可旋转键数：
  13
• 环数：
  2.0
• sp3杂化的碳原子比例：
  0.55
• 拓扑面积：
  72.8
• 氢给体数：
  1
• 氢受体数：
  8

反应信息

• 作为反应物：
  描述：

  12-(4-trifluoromethylcoumarin-7-yloxy)dodecanoic acid 在 alcohol dehydrogenase from Lactobacillus kefir 、 cytochrome P₄₅₀ BM₃ monooxygenase A₇₄G/F₈₇V mutant 、 异丙醇 、 还原型辅酶II(NADPH)四钠盐 、 magnesium chloride 作用下， 以 aq. phosphate buffer 为溶剂， 生成 12-oxododecanoic acid 、 7-羟基-4-三氟甲基香豆素
  参考文献：
  名称：

  Evaluation of coumarin-based fluorogenic P450 BM3 substrates and prospects for competitive inhibition screenings

  摘要：

  Fluorescence-based assays for the cytochrome P450 BM3 monooxygenase from Bacillus megaterium address an attractive biotechnological challenge by facilitating enzyme engineering and the identification of potential substrates of this highly promising biocatalyst. In the current study, we used the scarcity of corresponding screening systems as an opportunity to evaluate a novel and continuous high-throughput assay for this unique enzyme. A set of nine catalytically diverse P450 BM3 variants was constructed and tested toward the native substrate-inspired fluorogenic substrate 12-(4-trifluoromethylcoumarin-7-yloxy)dodecanoic acid. Particularly high enzyme-mediated 0-dealkylation yielding the fluorescent product 7-hydroxy-4-trifluoromethylcoumarin was observed with mutants containing the F87V substitution, with A74G/F87V showing the highest catalytic efficiency (0.458 min(-1) mu M-1). To simplify the assay procedure and show its versatility, different modes of application were successfully demonstrated, including (i) the direct use of NADPH or its oxidized form NADP(+) along with diverse NADPH recycling systems for electron supply, (ii) the use of cell-free lysates and whole-cell preparations as the biocatalyst source, and (iii) its use for competitive inhibition screens to identify or characterize substrates and inhibitors. A detailed comparison with known, fluorescence-based P450 BM3 assays finally emphasizes the relevance of our contribution to the ongoing research. (C) 2014 Elsevier Inc. All rights reserved.

  DOI：

  10.1016/j.ab.2014.03.022
• 作为产物：
  描述：

  12-溴十二烷酸 在 硫酸 、 水 、 potassium carbonate 、 potassium hydroxide 作用下， 以 四氢呋喃 、 N,N-二甲基甲酰胺 为溶剂， 反应 39.0h， 生成 12-(4-trifluoromethylcoumarin-7-yloxy)dodecanoic acid
  参考文献：
  名称：

  Evaluation of coumarin-based fluorogenic P450 BM3 substrates and prospects for competitive inhibition screenings

  摘要：

  Fluorescence-based assays for the cytochrome P450 BM3 monooxygenase from Bacillus megaterium address an attractive biotechnological challenge by facilitating enzyme engineering and the identification of potential substrates of this highly promising biocatalyst. In the current study, we used the scarcity of corresponding screening systems as an opportunity to evaluate a novel and continuous high-throughput assay for this unique enzyme. A set of nine catalytically diverse P450 BM3 variants was constructed and tested toward the native substrate-inspired fluorogenic substrate 12-(4-trifluoromethylcoumarin-7-yloxy)dodecanoic acid. Particularly high enzyme-mediated 0-dealkylation yielding the fluorescent product 7-hydroxy-4-trifluoromethylcoumarin was observed with mutants containing the F87V substitution, with A74G/F87V showing the highest catalytic efficiency (0.458 min(-1) mu M-1). To simplify the assay procedure and show its versatility, different modes of application were successfully demonstrated, including (i) the direct use of NADPH or its oxidized form NADP(+) along with diverse NADPH recycling systems for electron supply, (ii) the use of cell-free lysates and whole-cell preparations as the biocatalyst source, and (iii) its use for competitive inhibition screens to identify or characterize substrates and inhibitors. A detailed comparison with known, fluorescence-based P450 BM3 assays finally emphasizes the relevance of our contribution to the ongoing research. (C) 2014 Elsevier Inc. All rights reserved.

  DOI：

  10.1016/j.ab.2014.03.022

文献信息

• VERFAHREN ZUR SELEKTIVEN BINDUNG VON ZELLEN AN EINEN BINDUNGSPARTNER UND ZELLEN UMFASSENDE VORRICHTUNGEN
  申请人：Karlsruher Institut für Technologie

  公开号：EP3336099A1

  公开（公告）日：2018-06-20

  Die vorliegende Erfindung betrifft Verfahren zur selektiven Bindung von Zellen an einen Bindungspartner, etwa ein festes Trägermaterial, andere Zellen oder Proteine, Vorrichtungen, welche entsprechende Trägermaterialien mit darauf immobilisierten Zellen umfassen, Zellaggregate und Bindungskomplexe, sowie deren Verwendung als Biokatalysator, Biosensor und/oder Bioreaktor.

  本发明涉及细胞与结合伙伴（如固体载体材料、其他细胞或蛋白质）选择性结合的方法，包括固定有细胞的相应载体材料、细胞聚集体和结合复合物的装置，以及它们作为生物催化剂、生物传感器和/或生物反应器的用途。
• Evaluation of coumarin-based fluorogenic P450 BM3 substrates and prospects for competitive inhibition screenings
  作者：Katharina Neufeld、Sonja Meyer zu Berstenhorst、Jörg Pietruszka

  DOI：10.1016/j.ab.2014.03.022

  日期：2014.7

  Fluorescence-based assays for the cytochrome P450 BM3 monooxygenase from Bacillus megaterium address an attractive biotechnological challenge by facilitating enzyme engineering and the identification of potential substrates of this highly promising biocatalyst. In the current study, we used the scarcity of corresponding screening systems as an opportunity to evaluate a novel and continuous high-throughput assay for this unique enzyme. A set of nine catalytically diverse P450 BM3 variants was constructed and tested toward the native substrate-inspired fluorogenic substrate 12-(4-trifluoromethylcoumarin-7-yloxy)dodecanoic acid. Particularly high enzyme-mediated 0-dealkylation yielding the fluorescent product 7-hydroxy-4-trifluoromethylcoumarin was observed with mutants containing the F87V substitution, with A74G/F87V showing the highest catalytic efficiency (0.458 min(-1) mu M-1). To simplify the assay procedure and show its versatility, different modes of application were successfully demonstrated, including (i) the direct use of NADPH or its oxidized form NADP(+) along with diverse NADPH recycling systems for electron supply, (ii) the use of cell-free lysates and whole-cell preparations as the biocatalyst source, and (iii) its use for competitive inhibition screens to identify or characterize substrates and inhibitors. A detailed comparison with known, fluorescence-based P450 BM3 assays finally emphasizes the relevance of our contribution to the ongoing research. (C) 2014 Elsevier Inc. All rights reserved.