4-oxalomesaconate

• 分子结构分类: 有机化合物 - 有机酸及其衍生物 - 羧酸及其衍生物
• 英文名称: 4-oxalomesaconate
• 英文别名: (1E)-4-oxobut-1-ene-1,2,4-tricarboxylate；(E)-4-oxobut-1-ene-1,2,4-tricarboxylate
• 化学式: C₇H₃O₇
• 分子量: 199.097
• InChiKey: ODTDYYZJDQGKQT-NSCUHMNNSA-K

计算性质

• 辛醇/水分配系数(LogP)：
  0.9
• 重原子数：
  14
• 可旋转键数：
  2
• 环数：
  0.0
• sp3杂化的碳原子比例：
  0.14
• 拓扑面积：
  138
• 氢给体数：
  0
• 氢受体数：
  7

反应信息

• 作为反应物：
  描述：

  4-oxalomesaconate 生成 (1E,3E)-4-hydroxybuta-1,3-diene-1,2,4-tricarboxylate
  参考文献：
  名称：

  Unravelling the gallic acid degradation pathway in bacteria: the gal cluster from Pseudomonas putida

  摘要：

  SummaryGallic acid (3,4,5‐trihydroxybenzoic acid, GA) is widely distributed in nature, being a major phenolic pollutant and a commonly used antioxidant and building‐block for drug development. We have characterized the first complete cluster (gal genes) responsible for growth in GA in a derivative of the model bacterium Pseudomonas putida KT2440. GalT mediates specific GA uptake and chemotaxis, and highlights the critical role of GA transport in bacterial adaptation to GA consumption. The proposed GA degradation via the central intermediate 4‐oxalomesaconic acid (OMA) was revisited and all enzymes involved have been identified. Thus, GalD is the prototype of a new subfamily of isomerases that catalyses a biochemical step that remained unknown, i.e. the tautomerization of the OMAketo generated by the GalA dioxygenase to OMAenol. GalB is the founding member of a new family of zinc‐containing hydratases that converts OMAenol into 4‐carboxy‐4‐hydroxy‐2‐oxoadipic acid (CHA). galC encodes the aldolase catalysing CHA cleavage to pyruvic and oxaloacetic acids. The presence of homologous gal clusters outside the Pseudomonas genus sheds light on the evolution and ecology of the gal genes in GA degraders. The gal genes were used for expanding the metabolic abilities of heterologous hosts towards GA degradation, and for engineering a GA cellular biosensor.

  DOI：

  10.1111/j.1365-2958.2010.07448.x
• 作为产物：
  描述：

  6-氧代吡喃-2,4-二甲酸 在 sodium hydroxide 作用下， 以 水 为溶剂， 反应 1.0h， 生成 4-oxalomesaconate
  参考文献：
  名称：

  半胱氨酸残基在och草假单胞菌NGJ1的4-草酸酯中的水合酶的作用。

  摘要：

  在没有还原剂（如二硫苏糖醇）的情况下，来自草假单胞菌NGJ1的4-氧杂草酸水合酶不稳定，并被5,5'-二硫代双（2-硝基苯甲酸）（DTNB）强烈抑制。为了研究半胱氨酸残基在酶催化中的作用，通过定点诱变将酶的八个单独的半胱氨酸残基替换为丝氨酸残基。DTNB对野生型和突变型酶的催化性质和化学修饰表明：（i）8个半胱氨酸残基都不是酶催化所必需的；（ii）DTNB的抑制作用主要是由于Cys-186的修饰；（iii）Cys-96可能是与DTNB发生反应的另一个残基，其修饰导致4-草酸酯的K（m）值增加；（iv）DTNB无法接近其他六个半胱氨酸残基，但易受HgCl（2）影响；（v）在没有还原剂的情况下，仅置换Cys-186可显着提高酶的稳定性。

  DOI：

  10.1271/bbb.60503

文献信息

• Purification and Properties of 2-Pyrone-4,6-Dicarboxylate Hydrolase
  作者：Kiyofumi MARUYAMA

  DOI：10.1093/oxfordjournals.jbchem.a134210

  日期：1983.2

  A hydrolase which catalyzes specifically the interconversion between 2-pyrone-4,6-dicarboxylate and 4-oxalmesaconate was purified about 410-fold with a 16% yield from cell-free extracts of Pseudomonas ochraceae grown with phthalate. Upon disc gel electrophoresis, the enzyme preparation gave a single band which was coincident with the enzyme activity. The molecular weight of the enzyme was estimated

  从邻苯二甲酸酯生长的无假单胞菌提取物中，水解催化特定水解2-吡喃酮4,6-二羧酸酯和4-草酸酯的水解酶约410倍，收率16％。经圆盘凝胶电泳后，酶制剂产生一条与酶活性一致的条带。通过在Sephadex G-75上进行凝胶过滤，估计该酶的分子量为31,000，通过十二烷基硫酸钠凝胶电泳估计该酶的分子量为33,000。通过等电聚焦测定该酶的等电点为pH 5.49。该酶对2-pyrone-4,6-dicarboxylate具有特异性，而其他各种内酯均不能用作底物。2-吡喃酮4-，6-二羧酸酯水解，4-草酸酯的形成和质子产生的化学计量约为1：1：1。水解和合成2-pyrone-4,6-dicarboxylate的最佳pH分别为8.5和6.0。2-pyrone-4,6-dicarboxylate和4-oxalmesaconate的Km值分别为87和26 microM。在pH 8.5下，处于平衡状态的
• 2-pyrone-4,6-dicarboxylic acid, a catabolite of gallic acids in Pseudomonas species
  作者：P J Kersten、S Dagley、J W Whittaker、D M Arciero、J D Lipscomb

  DOI：10.1128/jb.152.3.1154-1162.1982

  日期：1982.12

  2-Pyrone-4,6-dicarboxylate hydrolase was purified from 4-hydroxybenzoate-grown Pseudomonas testosteroni. Gel filtration and electrophoretic measurements indicated that the preparation was homogeneous and gave a molecular weight of 37,200 for the single subunit of the enzyme. Hydrolytic activity was dependent upon a functioning sulfhydryl group(s) and was freely reversible; the equilibrium position was dependent upon pH, with equimolar amounts of pyrone and open-chain form present at pH 7.9. Since the hydrolase was strongly induced when the nonfluorescent organisms P. testosteroni and P. acidovorans grew with 4-hydroxybenzoate, it is suggested that 2-pyrone-4,6-dicarboxylate is a normal intermediate in the meta fission degradative pathway of protocatechuate. Laboratory strains of fluorescent pseudomonads did not metabolize 2-pyrone-4,6-dicarboxylate, but a strain of P. putida was isolated from soil that utilized this compound for growth; the hydrolase was then induced, but it was absent from extracts of 4-hydroxybenzoate-grown cells that readily catabolized protocatechuate by ortho fission reactions. 2-Pyrone-4,6-dicarboxylic acid was the major product formed when gallic acid was oxidized by purified protocatechuate 3,4-dioxygenase. Protocatechuate 4,5-dioxygenase gave only the open-chain ring fission product when gallic acid was oxidized, but the enzyme attacked 3-O-methylgallic acid, giving 2-pyrone-4,6-dicarboxylic acid as the major product. Cell suspensions of 4-hydroxybenzoate-grown P. testosteroni readily oxidized 3-O-methylgallate with accumulation of methanol.

  2-吡喃-4,6-二羧酸水解酶从生长在4-羟基苯甲酸上的睾丸酮假单胞菌中纯化。凝胶过滤和电泳测量表明该制备是均一的，并给出了该酶单个亚基的分子量为37,200。水解活性取决于功能性巯基，并且是自由可逆的。平衡位置取决于pH，在pH 7.9时呈现出吡喃和开链形式的等摩尔量。由于当非荧光菌P. testosteroni和P. acidovorans在4-羟基苯甲酸上生长时，水解酶被强烈诱导，因此建议2-吡喃-4,6-二羧酸是原儿茶酸的代谢途径中间体。荧光假单胞菌的实验室菌株不代谢2-吡喃-4,6-二羧酸，但是从土壤中分离出一株利用该化合物生长的P. putida菌株，然后诱导水解酶，但在容易通过邻位裂解反应降解原儿茶酸的4-羟基苯甲酸生长的细胞提取物中缺少该酶。当用纯化的原儿茶酸3,4-双加氧酶氧化没食子酸时，2-吡喃-4,6-二羧酸是主要产物。当氧化没食子酸时，原儿茶酸4,5-双加氧酶只产生开链环裂解产物，但是该酶攻击3-O-甲基没食子酸，将2-吡喃-4,6-二羧酸作为主要产物。4-羟基苯甲酸生长的P. testosteroni细胞悬液容易氧化3-O-甲基没食子酸，并积累甲醇。
• Molecular Characterization of the Gallate Dioxygenase from Pseudomonas putida KT2440
  作者：Juan Nogales、Ángeles Canales、Jesús Jiménez-Barbero、José Luis García、Eduardo Díaz

  DOI：10.1074/jbc.m502585200

  日期：2005.10

  mutant strain was unable to use gallate as the sole carbon source and it did not show gallate dioxygenase activity, suggesting that the GalA protein is the only dioxygenase involved in gallate cleavage in this bacterium. This work points to the existence of a new pathway that is devoted to the catabolism of gallic acid and that remained unknown in the paradigmatic P. putida KT2440 strain.

  在这项工作中，我们已经鉴定了恶臭假单胞菌KT2440的galA基因产物，该产物是一种环裂解双加氧酶，专门作用于没食子酸酯以产生4-草酸酯。该蛋白质是三聚体，由三个相同的47.6 kDa（419个氨基酸）亚基组成，并使用Fe2 +作为主要辅因子。没食子酸酯双加氧酶在pH 7.0下显示最大活性，没食子酸酯的Km和Vmax值分别为蛋白质的144 microM和53.2 micromol / min / mg。系统发育研究表明，恶臭假单胞菌KT2440的没食子酸双加氧酶是II型外二醇双加氧酶新亚组的原型，该亚组与原儿茶酸4,5-双加氧酶有共同的祖先，并且其两个结构域的结构可能是由于后者的大和小亚基。与恶臭假单胞菌KT2440的没食子酸双加氧酶的N末端结构域（残基1-281）和C末端结构域（残基294-420）的三维模型是通过与大型（LigB）晶体结构进行比较而生成的）和来自小孢子单胞菌（Sphingomonas
• Functional Annotation of LigU as a 1,3-Allylic Isomerase during the Degradation of Lignin in the Protocatechuate 4,5-Cleavage Pathway from the Soil Bacterium <i>Sphingobium</i> sp. SYK-6
  作者：Tessily N. Hogancamp、Frank M. Raushel

  DOI：10.1021/acs.biochem.8b00295

  日期：2018.5.15

  Sphingobium sp. SYK-6 is a Gram-negative soil bacterium that contributes to the degradation of lignin. Lignin provides structural support and protection to plants as a complex aromatic heteropolymer. The lignin degradation pathway of guaiacyl moieties leads to the intermediate, protocatechuate (PCA), which is further degraded via the 4,5-cleavage pathway in which PCA is ultimately metabolized to pyruvate and oxaloacetate. In this pathway, LigI has been shown to catalyze the hydrolysis of 2-pyrone-4,6-dicarboxylate to (4E)-oxalomesaconate (OMA). Here we have demonstrated, using 1H and 13C nuclear magnetic resonance spectroscopy, that LigU catalyzes the isomerization of the double bond between C4 and C5 in (4E)-OMA to (3Z)-2-keto-4-carboxy-3-hexenedioate (KCH), where the double bond has migrated to be between C3 and C4 via a 1,3-allylic isomerization. LigU is most closely related in amino acid sequence to methylaconitate isomerase (PrpF) from Shewanella oneidensis and methylitaconate-Δ-isomerase (Mii) from Eubacterium barkeri. The kinetic constants for the isomerization of OMA to KCH by LigU at pH 8.0 were determined to be 1300 ± 120 s–1 and (7.7 ± 1.5) × 106 M–1 s–1 for kcat and kcat/Km, respectively. We have also shown that the product of the LigU-catalyzed reaction is the preferred substrate for the LigJ hydratase. In this reaction, LigJ catalyzes the hydration of KCH to 4-carboxy-4-hydroxy-2-oxoadipate.

  鞘氨醇单胞菌（Sphingobium sp. SYK-6）是一种革兰氏阴性土壤细菌，有助于木质素的降解。木质素是一种复杂的芳香杂多聚物，可为植物提供结构支撑和保护。木质素降解途径中的愈创木酚部分会形成中间产物原儿茶酸（PCA），后者会通过4,5-裂解途径进一步降解，最终代谢为丙酮酸和草酰乙酸。在此途径中，LigI已被证明可催化2-吡喃酮-4,6-二羧酸的水解反应，生成（4E）-草酰草酸（OMA）。在此，我们使用1H和13C核磁共振光谱法证明，LigU可催化（4E）-OMA中C4和C5之间的双键异构化为（3Z）-2-酮-4-羧基-3-己烯二酸（KCH），其中双键通过1,3-烯丙基异构化移动到C3和C4之间。在氨基酸序列上，LigU与单胞菌（Shewanella oneidensis）的甲基乙二酸异构酶（PrpF）和巴克氏真杆菌（Eubacterium barkeri）的甲基异丁酸-Δ-异构酶（Mii）最为接近。在pH 8.
• Role of Cysteine Residues in 4-Oxalomesaconate Hydratase from<i>Pseudomonas ochraceae</i>NGJ1
  作者：Suhong LI、Maho KIMURA、Teruo TAKASHIMA、Kunihiko HAYASHI、Kazunori INOUE、Ryou ISHIGURO、Hiroyuki SUGISAKI、Kiyofumi MARUYAMA

  DOI：10.1271/bbb.60503

  日期：2007.2.23

  4-Oxalomesaconate hydratase from Pseudomonas ochraceae NGJ1 is unstable in the absence of reducing reagents such as dithiothreitol, and strongly inhibited by 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB). To study the role of cysteine residues in enzyme catalysis, the eight individual cysteine residues of the enzyme were replaced with serine residues by site-directed mutagenesis. The catalytic properties and chemical

  在没有还原剂（如二硫苏糖醇）的情况下，来自草假单胞菌NGJ1的4-氧杂草酸水合酶不稳定，并被5,5'-二硫代双（2-硝基苯甲酸）（DTNB）强烈抑制。为了研究半胱氨酸残基在酶催化中的作用，通过定点诱变将酶的八个单独的半胱氨酸残基替换为丝氨酸残基。DTNB对野生型和突变型酶的催化性质和化学修饰表明：（i）8个半胱氨酸残基都不是酶催化所必需的；（ii）DTNB的抑制作用主要是由于Cys-186的修饰；（iii）Cys-96可能是与DTNB发生反应的另一个残基，其修饰导致4-草酸酯的K（m）值增加；（iv）DTNB无法接近其他六个半胱氨酸残基，但易受HgCl（2）影响；（v）在没有还原剂的情况下，仅置换Cys-186可显着提高酶的稳定性。