2-oxo-4-pentenoate

• 分子结构分类: 有机化合物 - 有机酸及其衍生物 - 酮酸及其衍生物
• 英文名称: 2-oxo-4-pentenoate
• 英文别名: 2-Oxopent-4-enoate
• 化学式: C₅H₅O₃
• 分子量: 113.093
• InChiKey: NOXRYJAWRSNUJD-UHFFFAOYSA-M

计算性质

• 辛醇/水分配系数(LogP)：
  1.1
• 重原子数：
  8
• 可旋转键数：
  2
• 环数：
  0.0
• sp3杂化的碳原子比例：
  0.2
• 拓扑面积：
  57.2
• 氢给体数：
  0
• 氢受体数：
  3

反应信息

• 作为产物：
  描述：

  cis-2-Hydroxypenta-2,4-dienoate 生成 2-oxo-4-pentenoate
  参考文献：
  名称：

  4-Oxalocrotonate Tautomerase, Its Homologue YwhB, and Active Vinylpyruvate Hydratase:  Synthesis and Evaluation of 2-Fluoro Substrate Analogues

  摘要：

  A series of 2-fluoro-4-alkene and 2-fluoro-4-alkyne substrate analogues were synthesized and examined as potential inhibitors of three enzymes: 4-oxalocrotonate tautomerase (4-OT) and vinylpyruvate hydratase (VPH) from the catechol meta-fission pathway and a closely related 4-OT homologue found in Bacillus subtilis designated YwhB. All of the compounds were potent competitive inhibitors of 4-OT with the monocarboxylated 2E-fluoro-2,4-pentadienoate and the dicarboxylated 2E-fluoro-2-en-4-ynoate being the most potent. Despite the close mechanistic and structural similarities between 4-OT and YwhB, these compounds were significantly less potent inhibitors of YwhB with K-i values ranging from 5- to 633-fold lower than those determined for 4-OT. The study of VPH is complicated by the fact that the enzyme is only active as a complex with the metal-dependent 4-oxalocrotonate decarboxylase (4-OD), the enzyme following 4-OT in the catechol meta-fission pathway. A structure-based sequence analysis identified 4-OD as a member of the fumarylacetoacetate hydrolase (FAH) superfamily and implicated Glu-109 and Glu-111 as potential metal-binding ligands. Changing these residues to a glutamine verified their importance for enzymatic activity and enabled the production of soluble E109Q4-OD/VPH or E111/Q4OD/VPH complexes, which retained full hydratase activity but had little decarboxylase activity. Subsequent incubation of the E109Q4-OD/VPH complex with the substrate analogues identified the 2E and 2Z isomers of the monocarboxylated 2-fluoropent-2-en-4-ynoate as competitive inhibitors. The combined results set the stage for crystallographic studies of 4-OT, YwhB, and VPH using these inhibitors as ligands.

  DOI：

  10.1021/bi049489p

文献信息

• The meta Cleavage of Catechol by Azotobacter Species. 4-Oxalocrotonate Pathway
  作者：Jose M. Sala-trepat、W. Charles Evans

  DOI：10.1111/j.1432-1033.1971.tb01406.x

  日期：1971.6

  Catechol was metabolized through 2‐hydroxymuconic semialdehyde by cell‐free extracts of benzoate‐grown Azotobacter Strains. Some properties of catechol 2,3 oxygenase preparations from Azotobacter vinelandii 206 are described. Two different enzymatic activities able to attack 2‐hydroxymuconic semialdehyde have been found in crude extracts from benzoate‐grown cells; one catalyses a hydrolytic release of formate from the semialdehyde and the other a dehydrogenation of this compound to 4‐oxalocrotonate. However, the low, non‐inducible levels of 2‐hydroxymuconic semialdehyde hydrolase activity appear negligible for metabolic purposes and the semialdehyde seems to be dissimilated almost exclusively via 4‐oxalocrotonate, by the action of a NAD+‐dependent dehydrogenase, in Azotobacter strains. A tautomerase activity responsible for the interconversion of the enol and keto forms of 4‐oxalocrotonic acid was found in extracts from benzoate‐grown cells. 4‐Oxalocrotonate was stoicheiometrically converted to CO2 and 4‐hydroxy‐2‐oxovalerate by a partially purified extract, with the transient formation of a compound that appears to be 2‐oxopent‐4‐enoic acid. The 4‐oxalocrotonate decarboxylase activity was stimulated by Mg2+ or Mn2+ ions and was inhibited by EDTA. Cell‐free extracts from Azotobacter strains converted synthetic 4‐hydroxy‐2‐oxovalerate to acetaldehyde and pyruvate. A reaction sequence, termed the 4‐oxalocrotonate pathway, for the dissimiation of catechol to acetaldehyde and pyruvate by Azotobacter species is presented. All the enzymes operative in this pathway were inducible, except the 4‐hydroxy‐2‐oxovalerate aldolase. The findings described here are discussed in connection with the two previously reported meta cleavage pathways for the oxidation of catechol in Pseudomonas strains.

  以下是上述文本的中文翻译： 1. 邻苯二酚通过2-羟基邻苯二甲酰半醛被苯甲酸培养的斜生盐单胞菌（*Azotobacter*）菌株的游离细胞提取物代谢。文中描述了来自*Azotobacter vinelandii* 206菌株的邻苯二酚2,3-双加氧酶制剂的一些特性。 2. 在苯甲酸培养细胞的粗提取物中发现了两种能够攻击2-羟基邻苯二甲酰半醛的不同酶活性：一种催化半醛的水解，释放出甲酸；另一种使该化合物脱氢生成4-氧代海湾氨酸。然而，2-羟基邻苯二甲酰半醛水解酶活性的水平较低且不可诱导，似乎对代谢目的无足轻重。在*Azotobacter*菌株中，半醛似乎几乎完全通过4-氧代海湾氨酸（通过一种NAD⁺依赖的脱氢酶的作用）进行异化。 3. 在苯甲酸培养细胞的提取物中发现了一种tautomerase（tautomer酶）活性，可介导4-氧代海湾氨酸烯醇和酮形式之间的相互转换。 4. 通过部分纯化的提取物，4-氧代海湾氨酸被化学计量地转化为二氧化碳和4-羟基-2-氧代戊酸，同时形成了一种暂时性的化合物，似乎为2-氧代戊-4-烯酸。4-氧代海湾氨酸脱羧酶活性被Mg²⁺或Mn²⁺离子刺激，并被EDTA抑制。 5. 苯甲酸培养的*Azotobacter*菌株的游离细胞提取物将合成的4-羟基-2-氧代戊酸转化为乙醛和丙酮酸。 6. 本文提出了一条称为“4-氧代海湾氨酸途径”的反应序列，用于*Azotobacter*物种将邻苯二酚异化为乙醛和丙酮酸。此途径中所涉及的所有酶均诱导表达，除4-羟基-2-氧代戊酸醛缩酶外。 7. 在文中讨论了先前报道的关于苯甲酸菌株中邻苯二酚氧化的两种*meta*裂解途径的发现。
• Identification of Functional Residues in a 2-Hydroxymuconic Semialdehyde Hydrolase
  作者：Eduardo Díaz、Kenneth N. Timmis

  DOI：10.1074/jbc.270.11.6403

  日期：1995.3

  hydrolase, XylF, of the Pseudomonas putida TOL plasmid-encoded pathway for the catabolism of toluene and xylenes, catalyzes one of the rarest types of enzyme reaction (EC 3.7.1.9), the hydrolysis of a carbon-carbon bond in its substrate, the ring-fission product of 3-alkyl-substituted catechols. In this study, amino acid sequence comparisons between XylF and other hydrolases, and analysis of the similarity

  恶臭假单胞菌TOL质粒编码途径中的2-羟基粘康半醛水解酶XylF用于甲苯和二甲苯的分解代谢，催化一种最罕见的酶反应类型（EC 3.7.1.9），即碳-碳键的水解在其底物中是3-烷基取代的邻苯二酚的环裂变产物。在这项研究中，XylF与其他水解酶之间的氨基酸序列比较，以及预测的XylF二级结构与来自自养黄单胞菌GJ10的卤代烷脱卤酶的已知二级结构之间的相似性分析，使我们鉴定了几个保守残基，这些残基可能在XylF的催化中心具有功能性作用。Ser107，Asp228和His256这三个氨基酸，发现α-β水解酶的顺序与α/β水解酶折叠酶的顺序相似。通过氨基酸修饰和体外定点诱变实验对这些残基和其他残基的潜在功能性作用进行的研究提供了证据支持XylF是α/β水解酶折叠酶家族的丝氨酸水解酶这一假说，并指出残基被鉴定为XylF的催化三联体。这些研究还提供了有关XylF相关酶中其他保守残基的信息。有趣的是，在
• Uncovering the Protocatechuate 2,3-Cleavage Pathway Genes
  作者：Daisuke Kasai、Toshihiro Fujinami、Tomokuni Abe、Kohei Mase、Yoshihiro Katayama、Masao Fukuda、Eiji Masai

  DOI：10.1128/jb.00840-09

  日期：2009.11

  Paenibacillus sp. (formerly Bacillus macerans ) strain JJ-1b is able to grow on 4-hydroxybenzoate (4HB) as a sole source of carbon and energy and is known to degrade 4HB via the protocatechuate (PCA) 2,3-cleavage pathway. However, none of the genes involved in this pathway have been identified. In this study, we identified and characterized the JJ-1b genes for the 4HB catabolic pathway via the PCA 2,3-cleavage pathway, which consisted of praR and praABEGFDCHI . Based on the enzyme activities of cell extracts of Escherichia coli carrying praI , praA , praH , praB , praC , and praD , these genes were found to code for 4HB 3-hydroxylase, PCA 2,3-dioxygenase, 5-carboxy-2-hydroxymuconate-6-semialdehyde decarboxylase, 2-hydroxymuconate-6-semialdehyde dehydrogenase, 4-oxalocrotonate (OCA) tautomerase, and OCA decarboxylase, respectively, which are involved in the conversion of 4HB into 2-hydroxypenta-2,4-dienoate (HPD). The praE , praF , and praG gene products exhibited 45 to 61% amino acid sequence identity to the corresponding enzymes responsible for the catabolism of HPD to pyruvate and acetyl coenzyme A. The deduced amino acid sequence of praR showed similarity with those of IclR-type transcriptional regulators. Reverse transcription-PCR analysis revealed that praABEGFDCHI constitute an operon, and these genes were expressed during the growth of JJ-1b on 4HB and PCA. praR-praABEGFDCHI conferred the ability to grow on 4HB to E . coli , suggesting that praEGF were functional for the conversion of HPD to pyruvate and acetyl coenzyme A. A promoter analysis suggested that praR encodes a repressor of the pra operon.

  摘要 Paenibacillus sp. Bacillus macerans ）菌株 JJ-1b 能够以 4-hydroxybenzoate (4HB) 作为唯一的碳和能量来源进行生长，而且已知它能通过原儿茶酸盐 (PCA) 2,3 裂解途径降解 4HB。然而，参与这一途径的基因尚未被确定。在这项研究中，我们鉴定并描述了通过 PCA 2,3 裂解途径分解 4HB 的 JJ-1b 基因，这些基因包括 praR 和 praABEGFDCHI .根据大肠杆菌细胞提取物的酶活性 大肠杆菌 携带 praI , praA , praH , praB , praC 和 praD 发现这些基因编码 4HB 3-羟化酶、PCA 2,3-二氧 化酶、5-羧基-2-羟基琥珀酸-6-半乳糖醛脱羧酶、2-羟基琥珀酸-6-半乳糖醛脱氢酶、4-oxalocrotonate (OCA) tautomerase 和 OCA decarboxylase，它们分别参与将 4HB 转化为 2-hydroxypenta-2,4-dienoate (HPD)。这些 肽 , praF 和 和 基因产物的氨基酸序列与负责将HPD分解为丙酮酸和乙酰辅酶A的相应酶具有45%至61%的相同性。 praR 的氨基酸序列与 IclR 型转录调节因子的氨基酸序列相似。反转录-PCR分析表明 praABEGFDCHI 构成一个操作子，这些基因在 JJ-1b 在 4HB 和 PCA 上的生长过程中表达。 praR-praABEGFDCHI 将在 4HB 上生长的能力赋予了 E . 大肠杆菌 表明 启动子分析表明 启动子分析表明，praEGF praR 的抑制因子。 启动子 操作子的抑制因子。
• Expression and Stereochemical and Isotope Effect Studies of Active 4-Oxalocrotonate Decarboxylase
  作者：Thanuja M. Stanley、William H. Johnson、Elizabeth A. Burks、Christian P. Whitman、Chi-Ching Hwang、Paul F. Cook

  DOI：10.1021/bi9918902

  日期：2000.2.1

  the conversion of 2-oxo-3-hexenedioate to 2-oxo-4-hydroxypentanoate has been revised [Lian, H., and Whitman, C. P. (1994) J. Am. Chem. Soc. 116, 10403-10411]. Finally, the observed (13)C kinetic isotope effect on the decarboxylation of 2-oxo-3-hexenedioate by the 4-OD/VPH complex suggests that the decarboxylation step is nearly rate-limiting. Because the value is not sensitive to either magnesium or

  来自恶臭假单胞菌mt-2的4-氧代巴豆酸脱羧酶（4-OD）和乙烯基丙酮酸水合酶（VPH）形成一个复合物，该复合物在邻苯二酚间裂变途径中将2-氧代-3-己烯二酸酯转化为2-氧代-4-羟基戊酸酯。为了促进复合物的机理和结构研究，两种酶已被共表达，并且复合物已被纯化至均质。另外，Vlu中潜在的催化残基Glu-106已变为谷氨酰胺，所得的E106QVPH突变体已与4-OD共表达并纯化至均一。4-OD / E106QVPH复合物保留了完整的脱羧酶活性，其动力学参数与在野生型复合物中观察到的4-OD相当，但没有任何可检测的水合酶活性。（5S）-2-oxo-3- [5-D]己烯二酸酯通过4-OD / VPH络合物或突变体络合物的脱羧反应在D（2）中生成2-羟基-2,4E- [5-D]戊二烯酸酯O. 野生型配合物对2-羟基-2,4-戊二烯酸酯的酮化反应具有高度立体选择性，并导致2-氧代-（3S）-[3-D]
• Nucleotide sequence and functional analysis of the complete phenol/3,4-dimethylphenol catabolic pathway of Pseudomonas sp. strain CF600
  作者：V Shingler、J Powlowski、U Marklund

  DOI：10.1128/jb.174.3.711-724.1992

  日期：1992.2

  The meta-cleavage pathway for catechol is one of the major routes for the microbial degradation of aromatic compounds. Pseudomonas sp. strain CF600 grows efficiently on phenol, cresols, and 3,4-dimethylphenol via a plasmid-encoded multicomponent phenol hydroxylase and a subsequent meta-cleavage pathway. The genes for the entire pathway were previously found to be clustered, and the nucleotide sequences of dmpKLMNOPBC and D, which encode the first four biochemical steps of the pathway, were determined. By using a combination of deletion mapping, nucleotide sequence determinations, and polypeptide analysis, we identified the remaining six genes of the pathway. The fifteen genes, encoded in the order dmpKLMNOPQBCDEFGHI, lie in a single operon structure with intergenic spacing that varies between 0 to 70 nucleotides. Homologies found between the newly determined gene sequences and known genes are reported. Enzyme activity assays of deletion derivatives of the operon expressed in Escherichia coli were used to correlate dmpE, G, H, and I with known meta-cleavage enzymes. Although the function of the dmpQ gene product remains unknown, dmpF was found to encode acetaldehyde dehydrogenase (acylating) activity (acetaldehyde:NAD+ oxidoreductase [coenzyme A acylating]; E.C.1.2.1.10). The role of this previously unknown meta-cleavage pathway enzyme is discussed.

  邻苯二酚的元解聚途径是微生物降解芳香化合物的主要途径之一。假单胞菌属CF600菌株通过质粒编码的多组分酚羟化酶和随后的元解聚途径，有效地生长于苯酚、甲酚和3,4-二甲基苯酚上。先前已发现整个途径的基因被聚集在一起，并确定了编码途径前四个生化步骤的dmpKLMNOPBC和D的核苷酸序列。通过使用删除映射、核苷酸序列确定和多肽分析的组合方法，我们确定了途径中其余的六个基因。这十五个基因按顺序编码为dmpKLMNOPQBCDEFGHI，以介基间隔在0到70个核苷酸之间的单个操纵子结构中。报道了新确定的基因序列与已知基因之间的同源性。在大肠杆菌中表达的操纵子的删除衍生物的酶活性测定被用来将dmpE、G、H和I与已知的元解聚酶相关联。虽然dmpQ基因产物的功能仍未知，但发现dmpF编码醛脱氢酶（酰化）活性（乙醛：NAD+氧化还原酶[辅酶A酰化]；E.C.1.2.1.10）。讨论了这个先前未知的元解聚途径酶的作用。