p-nitrophenyl-β-D-arabinopyranoside | 78679-14-8

• 分子结构分类: 有机化合物 - 有机氧化合物
• 英文名称: p-nitrophenyl-β-D-arabinopyranoside
• 英文别名: pNPbA；(4-nitro-phenyl)-β-D-arabinopyranoside；(4-Nitro-phenyl)-β-D-arabinopyranosid；p-Nitrophenyl beta-D-arabinopyranoside；(2S,3S,4R,5R)-2-(4-nitrophenoxy)oxane-3,4,5-triol
• CAS: 78679-14-8
• 化学式: C₁₁H₁₃NO₇
• 分子量: 271.227
• InChiKey: MLJYKRYCCUGBBV-ZNSHCXBVSA-N

物化性质

• 熔点：
  183 °C
• 沸点：
  523.2±50.0 °C(Predicted)
• 密度：
  1.597±0.06 g/cm3(Predicted)

计算性质

• 辛醇/水分配系数(LogP)：
  -0.3
• 重原子数：
  19
• 可旋转键数：
  2
• 环数：
  2.0
• sp3杂化的碳原子比例：
  0.45
• 拓扑面积：
  125
• 氢给体数：
  3
• 氢受体数：
  7

反应信息

• 作为反应物：
  描述：

  p-nitrophenyl-β-D-arabinopyranoside 在 human α-L-fucosidase 、 水 作用下， 生成 对硝基苯酚
  参考文献：
  名称：

  Expression, Purification and Characterization of Human α-l-Fucosidase

  摘要：

  Abstractα‐l‐Fucosidases (EC 3.2.1.51) are exo‐glycosidases. On the basis of the multi‐alignment of amino acid sequence, α‐l‐fucosidases were classified into two families of glycoside hydrolases, GH‐29 and GH‐95. They are responsible for the removal of l‐fucosyl residues from the non‐reducing end of glycoconjugates. Deficiency of α‐l‐fucosidase results in Fucosidosis due to the accumulation of fucose‐containing glycolipids, glycoproteins and oligosaccharides in various tissues. Recent studies discovered that the fucosylation levels are increased on the membrane surfaces of many carcinomas, indicating the biological function of α‐l‐fucosidases may relate to this abnormal cell physiology. Although the gene of human α‐l‐fucosidase (h‐fuc) was cloned, the recombinant enzyme has rarely been overexpressed as a soluble and active from. We report herein that, with carefully control on the growing condition, an active human α‐l‐fucosidases (h‐Fuc) was successfully expressed in Escherichia coli for the first time. After a series steps of ion‐exchange and gel‐filtration chromatographic purification, the recombinant h‐Fuc with 95% homogeneity was obtained. The molecular weight of the enzyme was analyzed by SDS‐PAGE (∼50 kDa) and confirmed by ESI mass (50895 Da). The recombinant h‐Fuc was stable up to 55 °C with incubation at pH 6.8 for 2 h; the optimum temperature for h‐Fuc is approximately 55 °C. The enzyme was stable at pH 2.5–7.0 for 2 h; the enzyme activity decreased greatly for pH greater than 8.0 or less than 2.0. The Km and kcat values of the recombinant h‐Fuc (at pH 6.8) were determined to be 0.28 mM and 17.1 s−1, respectively. The study of pH‐dependent activity showed that the recombinant enzyme exhibited optimum activity at two regions near at pH 4.5 and pH 6.5. These features of the recombinant h‐Fuc are comparable to the native enzyme purified directly from human liver. Studies on the transfucosylation and common intermediate of the enzymatic reaction by NMR support that h‐Fuc functions as a retaining enzyme catalyzing the hydrolysis of substrate via a two‐step, double displacement mechanism.

  DOI：

  10.1002/jccs.200900126
• 作为产物：
  描述：

  对硝基苯酚 在 甲醇 、 sodium methylate 、 zinc(II) chloride 作用下， 生成 p-nitrophenyl-β-D-arabinopyranoside
  参考文献：
  名称：

  Feier; Westphal, Chemische Berichte, 1956, vol. 89, p. 589,593

  摘要：

  DOI：

文献信息

• Use of enzymes of Aureobasidium pullulans in pulp bleaching
  申请人：SANDOZ AG

  公开号：EP0373107B1

  公开（公告）日：1994-03-23
• COLD-ACTIVE BETA-GALACTOSIDASE, A METHOD OF PRODUCING SAME AND USE OF SUCH ENZYME
  申请人：Kobenhavns Universitet

  公开号：EP2396403B1

  公开（公告）日：2015-09-23
• US8288143B2
  申请人：——

  公开号：US8288143B2

  公开（公告）日：2012-10-16
• [EN] COLD-ACTIVE BETA-GALACTOSIDASE, A METHOD OF PRODUCING SAME AND USE OF SUCH ENZYME<br/>[FR] BÊTA-GALACTOSIDASE ACTIVE À FROID, SON PROCÉDÉ DE FABRICATION ET UTILISATION D'UNE TELLE ENZYME
  申请人：UINV KOBENHAVNS

  公开号：WO2010092057A1

  公开（公告）日：2010-08-19

  There is provided a novel cold-active beta-galactosidase enzyme specific for lactose. The enzyme is thus useful in e.g. the food industry for catalyzing at low temperatures the hydrolysis of lactose disaccharide into its constituent monosaccharides, glucose and galactose. The present invention further provides a method of producing the cold-active beta-galactosidase by recombinant DNA technology.
• Expression, Purification and Characterization of Human α-<scp>l</scp>-Fucosidase
  作者：Sheng-Wen Liu、Yaw-Kuen Li

  DOI：10.1002/jccs.200900126

  日期：2009.8

  Abstractα‐l‐Fucosidases (EC 3.2.1.51) are exo‐glycosidases. On the basis of the multi‐alignment of amino acid sequence, α‐l‐fucosidases were classified into two families of glycoside hydrolases, GH‐29 and GH‐95. They are responsible for the removal of l‐fucosyl residues from the non‐reducing end of glycoconjugates. Deficiency of α‐l‐fucosidase results in Fucosidosis due to the accumulation of fucose‐containing glycolipids, glycoproteins and oligosaccharides in various tissues. Recent studies discovered that the fucosylation levels are increased on the membrane surfaces of many carcinomas, indicating the biological function of α‐l‐fucosidases may relate to this abnormal cell physiology. Although the gene of human α‐l‐fucosidase (h‐fuc) was cloned, the recombinant enzyme has rarely been overexpressed as a soluble and active from. We report herein that, with carefully control on the growing condition, an active human α‐l‐fucosidases (h‐Fuc) was successfully expressed in Escherichia coli for the first time. After a series steps of ion‐exchange and gel‐filtration chromatographic purification, the recombinant h‐Fuc with 95% homogeneity was obtained. The molecular weight of the enzyme was analyzed by SDS‐PAGE (∼50 kDa) and confirmed by ESI mass (50895 Da). The recombinant h‐Fuc was stable up to 55 °C with incubation at pH 6.8 for 2 h; the optimum temperature for h‐Fuc is approximately 55 °C. The enzyme was stable at pH 2.5–7.0 for 2 h; the enzyme activity decreased greatly for pH greater than 8.0 or less than 2.0. The Km and kcat values of the recombinant h‐Fuc (at pH 6.8) were determined to be 0.28 mM and 17.1 s−1, respectively. The study of pH‐dependent activity showed that the recombinant enzyme exhibited optimum activity at two regions near at pH 4.5 and pH 6.5. These features of the recombinant h‐Fuc are comparable to the native enzyme purified directly from human liver. Studies on the transfucosylation and common intermediate of the enzymatic reaction by NMR support that h‐Fuc functions as a retaining enzyme catalyzing the hydrolysis of substrate via a two‐step, double displacement mechanism.