(2R)-2-O-β-D-glucopyranosyl-4-hydroxy-(2H)-1,4-benzoxazin-3(4H)-one | 155835-54-4

• 分子结构分类: 有机化合物 - 有机氧化合物
• 英文名称: (2R)-2-O-β-D-glucopyranosyl-4-hydroxy-(2H)-1,4-benzoxazin-3(4H)-one
• 英文别名: (2R)-2-β-D-glucopyranosyloxy-4-hydroxy-2H-1,4-benzoxazin-3(4H)-one；(2R)-4-hydroxy-2-[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-1,4-benzoxazin-3-one
• CAS: 155835-54-4
• 化学式: C₁₄H₁₇NO₉
• 分子量: 343.29
• InChiKey: OUSLYTBGQGKTME-TWTZXXGESA-N

计算性质

• 辛醇/水分配系数(LogP)：
  -1.6
• 重原子数：
  24
• 可旋转键数：
  3
• 环数：
  3.0
• sp3杂化的碳原子比例：
  0.5
• 拓扑面积：
  149
• 氢给体数：
  5
• 氢受体数：
  9

反应信息

• 作为反应物：
  描述：

  (2R)-2-O-β-D-glucopyranosyl-4-hydroxy-(2H)-1,4-benzoxazin-3(4H)-one 、 S-adenosylmethionine p-toluenesulfonate 在 APMSF 乙二胺四乙酸 、 Triticum aestivum DIMBOA-Glc 4-O-methyltransferase 、 三羟甲基氨基甲烷盐酸盐 作用下， 以 水 为溶剂， 反应 1.0h， 生成 (2R)-2-β-D-glucopyranosyloxy-4-methoxy-2H-1,4-benzoxazin-3(4H)-one
  参考文献：
  名称：

  Induction of HDMBOA-Glc accumulation and DIMBOA-Glc 4-O-methyltransferase by jasmonic acid in poaceous plants

  摘要：

  Induction of the accumulation of 2-(2-hydroxy-4,7-dimethoxy-1,4-benzoxazin-3-one)-beta-D-glucopyranose (HDMBOA-Glc) by jasmonic acid (JA) was investigated in wheat, Job's tears (Coix lacryma-jobi), and rye. An increase in HDMBOA-Glc and a corresponding decrease in 2-(2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one)-beta-D-glucopyranose (DIMBOA-Glc) were found in wheat and Job's tears, whereas no such changes were observed in rye. The activity of S-adenosyl-L-methionine:DIMBOA-Glc 4-O-methyltransferase which catalyzes the conversion of DIMBOA-Glc to HDMBOA-Glc was detected in wheat leaves treated with 50 muM JA. The activity started to increase 3 h after treatment with JA, reached a maximum after 9 h, and then decreased gradually. This mode of induction was well correlated with that for the accumulation of HDMBOA-Glc, indicating the induction of enzyme activity was responsible for the accumulation of HDMBOA-Glc. The enzyme was purified from JA-treated wheat leaves by three steps of chromatography, resulting in 95-fold purification. The enzyme showed strict substrate specificity for DIMBOA-Glc with a K value of 0.12 mM. DIBOA-Glc was also accepted as substrate, but the K-m value was 10 times larger than that for DIMBOAGlc. The aglycones, DIMBOA and DIBOA, were not methylated by the enzyme. The K-m value for S-adenoSyl-L-methionine was 0.06 mM The optimum pH and temperature were 7.5 and 35 degreesC, respectively. The activity was slightly enhanced by the presence of I mM EDTA, while heavy metal ions at 5 mM completely inhibited the activity. (C) 2002 Elsevier Science Ltd. All rights reserved.

  DOI：

  10.1016/s0031-9422(02)00225-x
• 作为产物：
  描述：

  2,3,4,6-四-O-乙酰基-Β-D-吡喃半乳糖酰基-2,2,2-三氯代亚氨乙酸酯 在 三氟化硼乙醚 、 sodium methylate 作用下， 以 甲醇 、 二氯甲烷 为溶剂， 反应 0.5h， 生成 (2R)-2-O-β-D-glucopyranosyl-4-hydroxy-(2H)-1,4-benzoxazin-3(4H)-one
  参考文献：
  名称：

  环状半缩醛的双非对映选择性糖苷化：从禾本科植物中合成1,4-苯并恶嗪酮缩醛糖苷GDIBOA和GDIMBOA

  摘要：

  在一个步骤中的双非对映选择性glucosidation过程引起自然（2 - [R）-2- β构型的6，并且对于第一次，7，苯并嗪乙缩醛糖苷从禾本科物种，则通过的反应所描述ø - （2- ，3,4,6-四ö乙酰基β -D-吡喃葡萄糖基）三氯乙酰亚胺酯3作为葡糖供体与2,4-二羟基-2- ħ -1,4-苯并恶嗪-3（4 H ^） -酮1或它的7-甲氧基衍生物2 在过量三氟化硼醚化物作为促进剂存在下，作为半缩醛葡糖基受体，不需要对葡糖苷中的环状异羟肟酸单元进行共价保护。

  DOI：

  10.1016/0040-4020(96)00577-7

文献信息

• Hydroxamic Acid Glucosyltransferases from Maize Seedlings
  作者：Bryan A. Bailey、Russell L. Larson

  DOI：10.1104/pp.90.3.1071

  日期：1989.7.1

  Hydroxamic acids occur in several forms in maize (Zea mays L.) with 2,4-dihydroxy-7-methoxy-2H-1,4-benzoxazin-3(4H)-one (DIMBOA) being the predominant form and others including 2,4-dihydroxy-1,4-benzoxazin-3-one (DIBOA) being found at lower concentrations. Two enzymes capable of glucosylating hydroxamic acids were identified in maize protein extracts and partially purified and characterized. The total

  异羟肟酸以多种形式存在于玉米 (Zea mays L.) 中，其中 2,4-dihydroxy-7-methoxy-2H-1,4-benzoxazin-3(4H)-one (DIMBOA) 是主要形式，其他形式包括 2 ,4-dihydroxy-1,4-benzoxazin-3-one (DIBOA) 浓度较低。在玉米蛋白提取物中鉴定出两种能够对异羟肟酸进行葡萄糖基化的酶，并对其进行了部分纯化和表征。在萌发的前 4 天，每株幼苗的总酶活性增加，并且与 DIMBOA 的积累同时发生。通过硫酸铵沉淀纯化酶，随后使用 Sephadex G-200 和 Q-Sepharose 凝胶层析，导致比活性增加 13 倍。酶最初通过 Q-Sepharose 凝胶色谱分离为两个活性峰（峰 1 和峰 2）。当 DIBOA 用作底物时，峰 1 葡糖基转移酶具有 3.6% 的 DIMBOA 糖基化活性，而峰 2 酶的这一百分比增加到
• Expression of soluble and catalytically active plant (monocot) ?-glucosidases inE. coli
  作者：Muzaffer Cicek、Asim Esen

  DOI：10.1002/(sici)1097-0290(19990520)63:4<392::aid-bit2>3.0.co;2-m

  日期：1999.5.20

  and Glu2 of maize were amplified by the polymerase chain reaction (PCR) and cloned into the expression vector pET21a. Both Glu1 and Glu2 isozymes were expressed in high yield ( approximately 3.8% of the total soluble protein and 32% of the total expressed protein) in E. coli. Recombinant enzymes were active on a variety of artificial and natural substrates at levels similar to those of their native

  通过聚合酶链反应（PCR）扩增编码玉米成熟β-葡萄糖苷酶蛋白Glu1和Glu2的互补DNA，并将其克隆到表达载体pET21a中。Glu1和Glu2同工酶均在大肠杆菌中以高收率表达（约占可溶性蛋白总量的3.8％，占表达蛋白总量的32％）。重组酶在多种人工和天然底物上的活性水平与从玉米幼苗中分离出来的天然酶相当。Western印迹分析证实这两种重组同工酶均与玉米抗β-葡萄糖苷酶血清发生免疫反应，其分子大小与天然玉米Glu1和Glu2同工酶相同。
• Purification and characterization of a β-glucosidase from rye (Secale cereale L.) seedlings
  作者：Masayuki Sue、Atsushi Ishihara、Hajime Iwamura

  DOI：10.1016/s0168-9452(00)00204-1

  日期：2000.6

  timing of the increase and decrease was concurrent with that of cyclic hydroxamic acid glucosides. The glucosidase was isolated from 48-h-old rye shoots and purified to apparent homogeneity by using isoelectric precipitation, anion exchange chromatography, and gel filtration. The isoelectric point and the optimum reaction temperature were 4.9-5.1 and 25-30 degrees C, respectively. The N-terminal amino acid

  研究了出现在黑麦幼苗中的环异羟肟酸和葡萄糖苷酶。芽中 2,4-二羟基-1,4-苯并恶嗪-3-one (DIBOA-Glc) 的葡萄糖苷浓度在萌发后很快增加，并随着植物开始自养生长而降至较低的恒定水平。环异羟肟酸糖苷β-葡糖苷酶活性也发生了短暂的变化，其增减时间与环异羟肟酸糖苷酶活性同步。葡萄糖苷酶是从 48 小时的黑麦芽中分离出来的，并通过使用等电沉淀、阴离子交换层析和凝胶过滤纯化到明显的同质性。等电点和最佳反应温度分别为4.9-5.1和25-30℃。N-末端氨基酸序列与小麦葡糖苷酶的序列几乎相同，但与其他植物来源的葡糖苷酶的序列没有任何相似性。SDS 和非变性 PAGE 分析表明，黑麦具有多种葡萄糖苷酶同工酶，每个同工酶都是 60-kDa 单体的寡聚物，分子量约为 300 kDa。该酶不仅对 DIMBOA-Glc 具有高度活性，而且对其 7-脱甲氧基类似物 DIBOA-Glc 也具有高活
• The mechanism of substrate (aglycone) specificity in β-glucosidases is revealed by crystal structures of mutant maize β-glucosidase-DIMBOA, -DIMBOAGlc, and -dhurrin complexes
  作者：Mirjam Czjzek、Muzaffer Cicek、Véronique Zamboni、David R. Bevan、Bernard Henrissat、Asim Esen

  DOI：10.1073/pnas.97.25.13555

  日期：2000.12.5

  The mechanism and the site of substrate (i.e., aglycone) recognition and specificity were investigated in maize β-glucosidase (Glu1) by x-ray crystallography by using crystals of a catalytically inactive mutant (Glu1E191D) in complex with the natural substrate 2- O -β- d -glucopyranosyl-4-hydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOAGlc), the free aglycone DIMBOA, and competitive inhibitor para -hydroxy- S -mandelonitrile β-glucoside (dhurrin). The structures of these complexes and of the free enzyme were solved at 2.1-, 2.1-, 2.0-, and 2.2-Å resolution, respectively. The structural data from the complexes allowed us to visualize an intact substrate, free aglycone, or a competitive inhibitor in the slot-like active site of a β-glucosidase. These data show that the aglycone moiety of the substrate is sandwiched between W378 on one side and F198, F205, and F466 on the other. Thus, specific conformations of these four hydrophobic amino acids and the shape of the aglycone-binding site they form determine aglycone recognition and substrate specificity in Glu1. In addition to these four residues, A467 interacts with the 7-methoxy group of DIMBOA. All residues but W378 are variable among β-glucosidases that differ in substrate specificity, supporting the conclusion that these sites are the basis of aglycone recognition and binding (i.e., substrate specificity) in β-glucosidases. The data also provide a plausible explanation for the competitive binding of dhurrin to maize β-glucosidases with high affinity without being hydrolyzed.

  通过X射线晶体学，使用玉米β-葡萄糖苷酶（Glu1）的催化失活突变体（Glu1E191D）晶体，在天然底物2-O-β-D-葡萄糖吡喃基-4-羟基-7-甲氧基-1,4-苯并噁唑-3-酮（DIMBOAGlc）、游离的底物DIMBOA和竞争性抑制剂对羟基-S-甘氨酸苯丙氰酸酯（dhurrin）的复合物中，研究了底物（即去糖苷基）识别和特异性的机制和位点。这些复合物和游离酶的结构分别在2.1、2.1、2.0和2.2 Å分辨率下解决。从这些复合物的结构数据中，我们可以在β-葡萄糖苷酶的槽状活性位点中可视化完整的底物、游离的去糖苷基或竞争性抑制剂。这些数据表明，底物的去糖苷基部分被夹在一侧的W378和另一侧的F198、F205和F466之间。因此，这四个疏水氨基酸的特定构象和它们形成的去糖苷基结合位点的形状决定了Glu1中去糖苷基识别和底物特异性。除了这四个氨基酸外，A467还与DIMBOA的7-甲氧基团相互作用。除了W378外，所有氨基酸都是β-葡萄糖苷酶中可变的，这些酶在底物特异性上有所不同，支持这些位点是β-葡萄糖苷酶中去糖苷基识别和结合（即底物特异性）的基础的结论。这些数据还提供了一个合理的解释，为什么dhurrin可以与玉米β-葡萄糖苷酶具有高亲和力而不被水解。
• Molecular and Structural Characterization of Hexameric <i>β</i>-<scp>d</scp>-Glucosidases in Wheat and Rye
  作者：Masayuki Sue、Kana Yamazaki、Shunsuke Yajima、Taiji Nomura、Tetsuya Matsukawa、Hajime Iwamura、Toru Miyamoto

  DOI：10.1104/pp.106.077693

  日期：2006.8.1

  The wheat (Triticum aestivum) and rye (Secale cereale) β-d-glucosidases hydrolyze hydroxamic acid-glucose conjugates, exist as different types of isozyme, and function as oligomers. In this study, three cDNAs encoding β-d-glucosidases (TaGlu1a, TaGlu1b, and TaGlu1c) were isolated from young wheat shoots. Although the TaGlu1s share very high sequence homology, the mRNA level of Taglu1c was much lower than the other two genes in 48- and 96-h-old wheat shoots. The expression ratio of each gene was different between two wheat cultivars. Recombinant TaGlu1b expressed in Escherichia coli was electrophoretically distinct fromTaGlu1a and TaGlu1c. Furthermore, coexpression of TaGlu1a and TaGlu1b gave seven bands on a native-PAGE gel, indicating the formation of both homo- and heterohexamers. One distinctive property of the wheat and rye glucosidases is that they function as hexamers but lose activity when dissociated into smaller oligomers or monomers. The crystal structure of hexameric TaGlu1b was determined at a resolution of 1.8 Å. The N-terminal region was located at the dimer-dimer interface and plays a crucial role in hexamer formation. Mutational analyses revealed that the aromatic side chain at position 378, which is located at the entrance to the catalytic center, plays an important role in substrate binding. Additionally, serine-464 and leucine-465 of TaGlu1a were shown to be critical in the relative specificity for DIMBOA-glucose (2-O-β-d-glucopyranosyl-4-hydroxy-7-methoxy-1,4-benzoxazin-3-one) over DIBOA-glucose (7-demethoxy-DIMBOA-glucose).

  小麦（Triticum aestivum）和黑麦（Secale cereale）β-d-葡萄糖苷酶水解羟基酰胺酸-葡萄糖共轭物，存在不同类型的同工酶，并作为寡聚体发挥作用。本研究从年轻的小麦幼苗中分离出三个编码β-d-葡萄糖苷酶（TaGlu1a，TaGlu1b和TaGlu1c）的cDNA。尽管TaGlu1s具有非常高的序列同源性，但48和96小时的小麦幼苗中Taglu1c的mRNA水平远低于其他两个基因。每个基因的表达比例在两个小麦品种之间不同。在大肠杆菌中表达的重组TaGlu1b在电泳上与TaGlu1a和TaGlu1c有所不同。此外，TaGlu1a和TaGlu1b的共表达在天然PAGE凝胶上产生七个条带，表明形成了同型和异型六聚体。小麦和黑麦葡萄糖苷酶的一个独特特性是它们作为六聚体发挥作用，但当解离成较小的寡聚体或单体时失去活性。六聚体TaGlu1b的晶体结构在1.8Å的分辨率下确定。N末端区域位于二聚体-二聚体界面，并在六聚体形成中发挥关键作用。突变分析表明，位于催化中心入口处的378号芳香侧链在底物结合中起重要作用。此外，TaGlu1a的丝氨酸-464和亮氨酸-465被证明在相对特异性DIMBOA-葡萄糖（2-O-β-d-葡萄糖吡喃基-4-羟基-7-甲氧基-1,4-苯并噁唑-3-酮）和DIBOA-葡萄糖（7-去甲氧基-DIMBOA-葡萄糖）中起关键作用。