DIMBOA-Glc | 113565-32-5

• 分子结构分类: 有机化合物 - 有机氧化合物
• 英文名称: DIMBOA-Glc
• 英文别名: (2R)-4-hydroxy-7-methoxy-2-[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-1,4-benzoxazin-3-one
• CAS: 113565-32-5
• 化学式: C₁₅H₁₉NO₁₀
• 分子量: 373.317
• InChiKey: WTGXAWKVZMQEDA-XFWGRBSCSA-N

计算性质

• 辛醇/水分配系数(LogP)：
  -1.6
• 重原子数：
  26
• 可旋转键数：
  4
• 环数：
  3.0
• sp3杂化的碳原子比例：
  0.53
• 拓扑面积：
  158
• 氢给体数：
  5
• 氢受体数：
  10

反应信息

• 作为反应物：
  描述：

  DIMBOA-Glc 、 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-O-β-D-glucopyranosyloxy-4,7-dimethoxy-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 DIMBOA-Glc
  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

• Reglucosylation of the Benzoxazinoid DIMBOA with Inversion of Stereochemical Configuration is a Detoxification Strategy in Lepidopteran Herbivores
  作者：Felipe C. Wouters、Michael Reichelt、Gaétan Glauser、Eugen Bauer、Matthias Erb、Jonathan Gershenzon、Daniel G. Vassão

  DOI：10.1002/anie.201406643

  日期：2014.10.13

  Benzoxazinoids are chemical defenses against herbivores and are produced by many members of the grass family. These compounds are stored as stable glucosides in plant cells and require the activity of glucosidases to release the corresponding toxic aglucones. In maize leaves, the most abundant benzoxazinoid is (2R)‐DIMBOA‐Glc, which is converted into the toxic DIMBOA upon herbivory. The ways in which

  苯并恶嗪类化合物是抗草食动物的化学防御剂，由草科的许多成员产生。这些化合物以稳定的糖苷形式储存在植物细胞中，需要糖苷酶的活性才能释放出相应的有毒糖苷。在玉米叶片中，最丰富的苯并恶嗪类化合物是（2 R）-DIMBOA-Glc，在食草时会转化为有毒的DIMBOA。研究了三种斜纹夜蛾代谢这种毒素的方式。（2秒）-在昆虫的茎中观察到了最初植物化合物的差向异构体-DIMBOA-Glc，并且在昆虫的肠道组织中检测到了相关的葡萄糖基转移酶活性。发现由昆虫产生的差向异构糖苷不再对植物葡糖苷酶具有反应性，因此不能转化为毒素。因此，立体选择性再糖基化代表了斜纹夜蛾物种的一种排毒策略，这可能有助于解释其作为农业害虫在含苯并恶嗪类作物上的成功之处。
• 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。