对硝基苯基-b-D-昆布二糖苷 | 26255-70-9

• 物质功能分类: 生物化工 - 糖类化合物 - 寡糖
• 分子结构分类: 有机化合物 - 有机氧化合物
• 中文名称: 对硝基苯基-b-D-昆布二糖苷
• 中文别名: 4-硝基苯基-BETA-昆布二糖苷
• 英文名称: 4-nitrophenyl β-D-glucopyranosyl-(1->3)-β-D-glucopyranoside
• 英文别名: p-nitrophenyl β-D-laminaribioside；4-nitrophenyl β-D-glucopyranosyl-(1→3)-β-D-glucopyranoside；4-nitrophenyl β-laminaribioside；p-nitrophenyl3-O-(β-D-glucopyranosyl)-β-D-glucopyranoside；4-nitrophenyl β-cellobiose；4-Nitrophenyl 3-O-(b-D-glucopyranosyl)-b-D-glucopyranoside；(2S,3R,4S,5S,6R)-2-[(2R,3R,4S,5R,6S)-3,5-dihydroxy-2-(hydroxymethyl)-6-(4-nitrophenoxy)oxan-4-yl]oxy-6-(hydroxymethyl)oxane-3,4,5-triol
• CAS: 26255-70-9
• 化学式: C₁₈H₂₅NO₁₃
• 分子量: 463.395
• InChiKey: LBTDRWMZFQVCAR-GZQMJUEHSA-N

物化性质

• 熔点：
  232-234°C dec.
• 溶解度：
  可溶于水

计算性质

• 辛醇/水分配系数(LogP)：
  -2
• 重原子数：
  32
• 可旋转键数：
  6
• 环数：
  3.0
• sp3杂化的碳原子比例：
  0.67
• 拓扑面积：
  224
• 氢给体数：
  7
• 氢受体数：
  13

反应信息

• 作为反应物：
  描述：

  对硝基苯基-b-D-昆布二糖苷 、 (3-azido-3-deoxy-β-D-glucopyranosyl)-(1->3)-α-D-glucopyranosyl fluoride 在 mutated (1,3)-β-D-glucan endohydrolase Glu231Gly 作用下， 以 phosphate buffer 为溶剂， 以95%的产率得到p-nitrophenyl (3-azido-3-deoxy-β-D-glucopyranosyl)-(1->3)-(β-D-glucopyranosyl)-(1->3)-(β-D-glucopyranosyl)-(1->3)-β-D-glucopyranoside
  参考文献：
  名称：

  通过使用来自大麦的突变的（1,3）-β-D-葡聚糖内切酶合成复杂的寡糖。

  摘要：

  当取代或未取代的α-laminaribiosyl氟化物，作为供体，通过缩合生成单糖和二糖β-D-己基吡喃糖苷受体时，可以良好的产率获得具有新形成的（1,3）-β-糖苷键的复杂寡糖。 （1,3）-β-D-糖基合成酶。这些线性和分支的（1,3）-β-连接的寡糖在医疗，制药和农业应用领域中可能被证明是重要的。此外，观察到（1,3）-β-D-葡聚糖葡糖合酶在其受体亚位点中容纳了（1,3）-，（1,4），-和（1,6）-β-寡糖，但来自高等植物的野生型（1,3）-β-D-葡聚糖内切酶的生理作用却出乎意料。

  DOI：

  10.1002/chem.200304733
• 作为产物：
  描述：

  2,3,4,6-四-O-乙酰基-α-D-吡喃葡萄糖酰氟 在 Thermotoga neapolitana β-glucosidase 1A E349G mutant 、 sodium methylate 作用下， 以 甲醇 为溶剂， 反应 15.0h， 生成 对硝基苯基-b-D-昆布二糖苷
  参考文献：
  名称：

  嗜热栖热菌β-葡萄糖苷酶1A中的糖合酶：α-葡萄糖基氟化物与原位生成的α-糖基甲酸供体的比较

  摘要：

  来自嗜热嗜热菌Theromotoga neapolitana的Tn Bgl1A是一种二聚体β-葡萄糖苷酶，属于糖苷水解酶家族1（GH1），具有通过保留机制的水解活性，并且对β-1,4-，β-1具有广泛的底物特异性，一系列糖寡糖上的3-和β-1,6-键。该酶的三种变体（Tn Bgl1A_E349G，Tn Bgl1A_E349A和TnBgl1A_E349S），在催化亲核试剂上发生突变，用于评估其对寡糖合成的糖合酶活性。有两种方法用于合成反应，两种方法均使用4-硝基苯基β-d-吡喃葡萄糖苷（4NPGlc）作为受体分子：第一种方法是在经典糖合酶的低温（35°C）下使用α-葡萄糖基氟化物供体反应，第二步是用（4NPGlc）原位生成糖基供体，其中甲酸在较高温度（70°C）下充当外源亲核试剂。使用第一种方法Tn Bgl1A_E349G和TnBgl1A_E349A经过长时间孵育（15小时）后，以良好的产率（高达61％）合成了具有β-1

  DOI：

  10.1016/j.molcatb.2014.05.021

文献信息

• Glycosynthase with Broad Substrate Specificity - an Efficient Biocatalyst for the Construction of Oligosaccharide Library
  作者：Jinhua Wei、Xun Lv、Yang Lü、Gangzhu Yang、Lifeng Fu、Liu Yang、Jianjun Wang、Jianhui Gao、Shuihong Cheng、Qian Duan、Cheng Jin、Xuebing Li

  DOI：10.1002/ejoc.201201507

  日期：2013.4

  A versatile glycosynthase (TnG-E338A) with strikingly broad substrate scope has been developed from Thermus nonproteolyticus β-glycosidase (TnG) by using site-directed mutagenesis. The practical utility of this biocatalyst has been demonstrated by the facile generation of a small library containing various oligosaccharides and a steroidal glycoside (total 25 compounds) in up to 100 % isolated yield

  通过使用定点诱变从非蛋白水解栖热菌 β-糖苷酶 (TnG) 中开发出一种多功能糖合酶 (TnG-E338A)，其底物范围非常广泛。这种生物催化剂的实际效用已通过以高达 100% 的分离产率轻松生成包含各种寡糖和甾体糖苷（总共 25 种化合物）的小型文库得到证明。此外，该酶在一锅平行反应中很容易合成了一系列八种低聚糖，这突出了其在碳水化合物库的组合构建中的潜力，这将有助于糖组学和糖治疗研究。值得注意的是，该酶提供了一种可以将糖合酶技术扩展到组合化学的方法。
• Rational design of a thermostable glycoside hydrolase from family 3 introduces β-glycosynthase activity
  作者：Tania Pozzo、Javier Romero-García、Magda Faijes、Antoni Planas、Eva Nordberg Karlsson

  DOI：10.1093/glycob/cww081

  日期：2017.1

  were successfully converted into beta-glucosynthases that were assayed using two separate biosynthetic methods. The first reaction used an alpha-glucosyl fluoride donor with a 4-nitrophenyl beta-D-glucopyranoside (4NPGlc) acceptor, and the second used 4NPGlc as both the donor and acceptor in the presence of the exogenous nucleophile formate. The primary specificity observed was a beta-1,3-linked disaccharide

  来自Thermatoga neapolitana的热稳定β-葡萄糖苷酶TnBgl3B是糖苷水解酶家族3（GH3）的单体三结构域代表。在先前的TnBg13B研究中，通过使用外源亲核试剂进行化学活化，可以确定催化亲核试剂（D242）和相应的酸/碱残基（E458）。鉴定这些残基导致尝试将TnBgl3B转化为β-葡萄糖合成酶，在其中创建了三个亲核变异体（TnBgl3B_D242G，TnBgl3B_D242A，TnBgl3B_D242S），并且所有这些均未显示出葡萄糖合酶活性。对TnBgl3B活性位点的更深入分析导致生成了三个其他变体，每个变体都接受一个单点突变。其中的两个变体在-1子位点（Y210F，W243F）和第三个在结合位点的糖苷配基区域（N248R）附近接受取代。对这三个变体的动力学评估表明，W243F取代减少了水解，同时又保持了KM。此关键的W243F突变随后被引入到原始亲核体变体中，并
• Acceptor-dependent regioselectivity of glycosynthase reactions by Streptomyces E383A β-glucosidase
  作者：Magda Faijes、Marc Saura-Valls、Xavi Pérez、Marta Conti、Antoni Planas

  DOI：10.1016/j.carres.2006.04.049

  日期：2006.9

  The normucleophilic mutant E383A beta-glucosidase from Streptomyces sp. has proven to be an efficient glycosynthase enzyme, catalyzing the condensation of alpha-glucosyl and alpha-galactosyl fluoride donors to a variety of acceptors. The enzyme has maximal activity at 45 degrees C, and a pH-dependence reflecting general base catalysis with an apparent kinetic pK(a) of 7.2. The regioselectivity of the new glycosidic linkage depends unexpectedly on the acceptor substrate. With aryl monosaccharide acceptors, beta-(1 -> 3) disaccharides are obtained in good to excellent yields, thus expanding the synthetic products available with current exo-glycosynthases. With xylopyranosyl acceptor, regioselectivity is poorer and results in the formation of a mixture of beta-(1 -> 3) and beta-(1 -> 4) linkages. In contrast, disaccharide acceptors produce exclusively beta-(1 -> 4) linkages. Therefore, the presence of a glycosyl unit in subsite +II redirects regioselectivity from beta-(1 -> 3) to beta-(1 -> 4). To improve operational performance, the E383A mutant was immobilized on a Ni(2+)-chelating Sepharose resin. Immobilization did not increase stability to pH and organic solvents, but the operational stability and storage stability were clearly enhanced for recycling and scaling-up. (c) 2006 Elsevier Ltd. All rights reserved.
• Major Change in Regiospecificity for the Exo-1,3-β-glucanase from <i>Candida albicans</i> following Its Conversion to a Glycosynthase
  作者：Y. Nakatani、D. S. Larsen、S. M. Cutfield、J. F. Cutfield

  DOI：10.1021/bi500239m

  日期：2014.5.27

  The exo-1,3-beta-glucanase (Exg) from Candida albicans is involved in cell wall beta-D-glucan metabolism and morphogenesis through its hydrolase and transglycosidase activities. Previous work has shown that both these activities strongly favor beta-1,3-linkages. The E292S Exg variant displayed modest glycosynthase activity using alpha-D-glucopyranosyl fluoride (alpha-GlcF) as the donor and pNP-beta-D-glucopyranoside (pNPGlc) as the acceptor but surprisingly showed a marked preference for synthesizing beta-1,6-linked over beta-1,3- and beta-1,4-linked disaccharide products. With pNPXyl as the acceptor, the preference became beta-1,4 over beta-1,3. The crystal structure of the glycosynthase bound to both of its substrates, alpha-GlcF and pNPGlc, is the first such ternary complex structure to be determined. The results revealed that the donor bound in the -1 subsite, as expected, while the acceptor was oriented in the +1 subsite to facilitate beta-1,6-linkage, thereby supporting the results from solution studies. A second crystal structure containing the major product of glycosynthesis, pNP-gentiobiose, showed that the -1 subsite allows another docking position for the terminal sugar; i.e., one position is set up for catalysis, whereas the other is an intermediate stage prior to the displacement of water from the active site by the incoming sugar hydroxyls. The +1 subsite, an aromatic "clamp", permits several different sugar positions and orientations, including a 180 degrees flip that explains the observed variable regiospecificity. The p-nitrophenyl group on the acceptor most likely influences the unexpectedly observed beta-1,6-specificity through its interaction with F229. These results demonstrate that tailoring the specificity of a particular glycosynthase depends not only on the chemical structure of the acceptor but also on understanding the structural basis of the promiscuity of the native enzyme.
• Rare Keto-Aldoses from Enzymatic Oxidation: Substrates and Oxidation Products of Pyranose 2-Oxidase
  作者：Stefan Freimund、Alexander Huwig、Friedrich Giffhorn、Sabine Köpper

  DOI：10.1002/(sici)1521-3765(19981204)4:12<2442::aid-chem2442>3.0.co;2-a

  日期：1998.12.4