6-chloro-4-methylumbelliferyl 2,2',3,3',4',6,6'-hepta-O-acetyl-β-cellobioside | 1417989-45-7

• 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: 6-chloro-4-methylumbelliferyl 2,2',3,3',4',6,6'-hepta-O-acetyl-β-cellobioside
• CAS: 1417989-45-7
• 化学式: C₃₆H₄₁ClO₂₀
• 分子量: 829.163
• InChiKey: BJALGFGFXNCIAS-UEGIBPQHSA-N

计算性质

• 辛醇/水分配系数(LogP)：
  1.75
• 重原子数：
  57.0
• 可旋转键数：
  13.0
• 环数：
  4.0
• sp3杂化的碳原子比例：
  0.56
• 拓扑面积：
  251.23
• 氢给体数：
  0.0
• 氢受体数：
  20.0

反应信息

• 作为反应物：
  描述：

  6-chloro-4-methylumbelliferyl 2,2',3,3',4',6,6'-hepta-O-acetyl-β-cellobioside 在 sodium methylate 作用下， 以 甲醇 、 二氯甲烷 为溶剂， 以96%的产率得到6-chloro-4-methylumbelliferyl β-cellobioside
  参考文献：
  名称：

  Synthesis and evaluation of a series of 6-chloro-4-methylumbelliferyl glycosides as fluorogenic reagents for screening metagenomic libraries for glycosidase activity

  摘要：

  Screening of large enzyme libraries such as those derived from metagenomic sources requires sensitive substrates. Fluorogenic glycosides typically offer the best sensitivity but typically must be used in a stopped format to generate good signal. Use of fluorescent phenols of pKa < 7, such as halogenated coumarins, allows direct screening at neutral pH. The synthesis and characterisation of a set of nine different glycosides of 6-chloro-4-methylumbelliferone are described. The use of these substrates in a pooled format for screening of expressed metagenomic libraries yielded a "hit rate" of 1 in 60. Hits were then readily deconvoluted with the individual substrates in a single plate to identify specific activities within each clone. The use of such a collection of substrates greatly accelerates the screening process. (C) 2015 Elsevier Ltd. All rights reserved.

  DOI：

  10.1016/j.carres.2015.12.010
• 作为产物：
  描述：

  Cellobiose 在 吡啶 、 氢溴酸 、 四丁基硫酸氢铵 、 乙酸酐 、 溶剂黄146 、 sodium hydroxide 作用下， 以 二氯甲烷 为溶剂， 生成 6-chloro-4-methylumbelliferyl 2,2',3,3',4',6,6'-hepta-O-acetyl-β-cellobioside
  参考文献：
  名称：

  Synthesis and evaluation of a series of 6-chloro-4-methylumbelliferyl glycosides as fluorogenic reagents for screening metagenomic libraries for glycosidase activity

  摘要：

  Screening of large enzyme libraries such as those derived from metagenomic sources requires sensitive substrates. Fluorogenic glycosides typically offer the best sensitivity but typically must be used in a stopped format to generate good signal. Use of fluorescent phenols of pKa < 7, such as halogenated coumarins, allows direct screening at neutral pH. The synthesis and characterisation of a set of nine different glycosides of 6-chloro-4-methylumbelliferone are described. The use of these substrates in a pooled format for screening of expressed metagenomic libraries yielded a "hit rate" of 1 in 60. Hits were then readily deconvoluted with the individual substrates in a single plate to identify specific activities within each clone. The use of such a collection of substrates greatly accelerates the screening process. (C) 2015 Elsevier Ltd. All rights reserved.

  DOI：

  10.1016/j.carres.2015.12.010

文献信息

• Rational design, synthesis, evaluation and enzyme<b>-</b>substrate structures of improved fluorogenic substrates for family 6 glycoside hydrolases
  作者：Miao Wu、Wim Nerinckx、Kathleen Piens、Takuya Ishida、Henrik Hansson、Mats Sandgren、Jerry Ståhlberg

  DOI：10.1111/febs.12060

  日期：2013.1

  Methylumbelliferyl‐β–cellobioside (MUF–G2) is a convenient fluorogenic substrate for certain β–glycoside hydrolases (GH). However, hydrolysis of the aglycone is poor with GH family 6 enzymes (GH6), despite strong binding. Prediction of the orientation of the aglycone of MUF–G2 in the +1 subsite of Hypocrea jecorina Cel6A by automated docking suggested umbelliferyl modifications at C4 and C6 for improved recognition. Four modified umbelliferyl‐β–cellobiosides [6–chloro‐4–methyl‐ (ClMUF); 6–chloro‐4‐trifluoromethyl‐ (ClF3MUF); 4–phenyl‐ (PhUF); 6–chloro‐4–phenyl‐ (ClPhUF)] were synthesized and tested with GH6, GH7, GH9, GH5 and GH45 cellulases. Indeed the rate of aglycone release by H. jecorina Cel6A was 10–150 times higher than with MUF–G2, although it was still three orders of magnitude lower than with H. jecorina Cel7B. The 4–phenyl substitution drastically reduced the fluorescence intensity of the free aglycone, while ClMUF–G2 could be used for determination of kcat and KM for H. jecorina Cel6A and Thermobifida fusca Cel6A. Crystal structures of H. jecorina Cel6A D221A mutant soaked with the MUF‐, ClMUF‐ and ClPhUF‐β–cellobioside substrates show that the modifications turned the umbelliferyl group ‘upside down’, with the glycosidic bond better positioned for protonation than with MUF–G2.DatabaseStructural data have been submitted to the Protein Data Bank under accession numbers pdb 4AU0, 4AX7, 4AX6Structured digital abstract• http://mint.bio.uniroma2.it/mint/search/interaction.do?interactionAc=MINT-7260296• Cel6A and Cel6A bind by x-ray crystallography (View Interaction: 1, 2)