cis-2-Hydroxypenta-2,4-dienoate

• 分子结构分类: 有机化合物 - 脂质和类脂质分子 - 脂肪酰基
• 英文名称: cis-2-Hydroxypenta-2,4-dienoate
• 英文别名: (2E)-1-hydroxy-1-oxopenta-2,4-dien-2-olate
• 化学式: C5H5O3-
• 分子量: 113.09
• InChiKey: VHTQQDXPNUTMNB-ONEGZZNKSA-M

计算性质

• 辛醇/水分配系数(LogP)：
  1.5
• 重原子数：
  8
• 可旋转键数：
  1
• 环数：
  0.0
• sp3杂化的碳原子比例：
  0.0
• 拓扑面积：
  60.4
• 氢给体数：
  1
• 氢受体数：
  3

反应信息

• 作为反应物：
  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
• 作为产物：
  描述：

  (2E,4Z,7E)-2-hydroxy-6-oxonona-2,4,7-trienedioate 、 水 生成 cis-2-Hydroxypenta-2,4-dienoate 、 富马酸酯(2-) 、 氢(+1)阳离子
  参考文献：
  名称：

  Characterization of the hca Cluster Encoding the Dioxygenolytic Pathway for Initial Catabolism of 3-Phenylpropionic Acid in Escherichia coli K-12

  摘要：

  摘要 我们鉴定、克隆并测序了 hca 簇，该簇编码大肠杆菌中 3-苯基丙酸（PP）初始分解的二氧分解途径。 大肠杆菌 K-12.该基因簇位于染色体的第 57.5 小节，由五个分解基因组成，这些基因组成了一个假定操作子（hcaA1A2CBD）。 hcaA1A2CBD ) 和另外两个反向转录的基因，这两个基因编码一种潜在的渗透酶 ( hcaT )和一个调节器( hcaR ).序列比较显示 hcaA1A2CD 基因编码 3-苯基丙酸二氧合酶的四个亚基，而 hcaB 基因编码相应的 顺式 -二氢二醇脱氢酶。这种分解模块与编码 IIB 类二加氧酶的模块同源，是大肠杆菌中第一个此类分解簇的实例。 大肠杆菌 .诱导表达 hca 基因的诱导表达需要 hcaR 基因产物，它是一种转录激活因子，与 LysR 家族的调节因子序列非常相似。有趣的是，HcaA1A2CD 和 HcaB 酶不仅能将 PP 氧化成 3-（2,3-二羟基苯基）丙酸（DHPP），还能将肉桂酸（CI）氧化成其相应的 2,3- 二羟基衍生物。DHPP 的进一步分解需要 mhp -编码的 元 Ferrández, J. L. Garcı́a, and E. Dı́az, J. Bacteriol.179:2573-2581, 1997).在 鼠伤寒沙门氏菌 的 mhp 基因单独或与 hca 簇的重组细菌可分别在 3-羟基肉桂酸（3HCI）和 CI 中生长。因此，趋同的 mhp - 和 hca -编码的途径在 鼠伤寒杆菌 它们负责分解自然界中广泛存在的不同苯丙类化合物（3HPP、3HCI、PP 和 CI）。

  DOI：

  10.1128/jb.180.11.2915-2923.1998

文献信息

• Purification and Characterization of<i>meta</i>-Cleavage Compound Hydrolase from a Carbazole Degrader<i>Pseudomonas resinovorans</i>Strain CA10
  作者：Hideaki NOJIRI、Hiroko TAIRA、Kenichi IWATA、Kenichi MORII、Jeong-Won NAM、Takako YOSHIDA、Hiroshi HABE、Shugo NAKAMURA、Kentaro SHIMIZU、Hisakazu YAMANE、Toshio OMORI

  DOI：10.1271/bbb.67.36

  日期：2003.1

  2-Hydroxy-6-oxo-6-(2′-aminophenyl)-hexa-2,4- dienoic acid [6-(2′-aminophenyl)-HODA] hydrolase, involved in carbazole degradation by Pseudomonas resinovorans strain CA10, was purified to near homogeneity from an overexpressing Escherichia coli strain. The enzyme was dimeric, and its optimum pH was 7.0-7.5. Phylogenetic analysis showed the close relationship of this enzyme to other hydrolases involved in the degradation of monocyclic aromatic compounds, and this enzyme was specific for 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid (6-phenyl-HODA), having little activity toward 2-hydroxy-6-oxohepta-2,4-dienoic acid and 2-hydroxymuconic semialdehyde. The enzyme had a Km of 2.51 μM and kcat of 2.14 (s−1) for 6-phenyl-HODA (50 mM sodium phosphate, pH 7.5, 25°C). The effect of the presence of an amino group or hydroxyl group at the 2′-position of phenyl moiety of 6-phenyl-HODA on the enzyme activity was found to be small; the activity decreased only in the order of 6-(2′-aminophenyl)-HODA (2.44 U/mg)>6-phenyl-HODA (1.99 U/mg)>2-hydroxy-6-oxo-6-(2′-hydroxyphenyl)-hexa-2,4-dienoic acid (1.05 U/mg). The effects of 2′-substitution on the activity were in accordance with the predicted reactivity based on the calculated lowest unoccupied molecular orbital energy for these substrates.

  2-羟基-6-氧代-6-(2'-氨基苯基)-六-2,4-二烯酸[6-(2'-氨基苯基)-HODA]水解酶参与食树脂假单胞菌菌株 CA10 的咔唑降解，从过度表达的大肠杆菌菌株中纯化至接近同质。该酶为二聚体，其最适pH为7.0-7.5。系统发育分析表明该酶与参与单环芳香族化合物降解的其他水解酶有密切关系，并且该酶对2-羟基-6-氧代-6-苯基六-2,4-二烯酸（6-苯基- HODA)，对 2-羟基-6-氧七-2,4-二烯酸和 2-羟基粘康半醛几乎没有活性。对于 6-苯基-HODA（50 mM 磷酸钠，pH 7.5，25°C），该酶的 Km 为 2.51 μM，kcat 为 2.14 (s−1)。发现6-苯基-HODA的苯基部分2'位上的氨基或羟基的存在对酶活性的影响很小；活性降低的顺序为：6-(2'-氨基苯基)-HODA (2.44 U/mg)>6-苯基-HODA (1.99 U/mg)>2-羟基-6-氧代-6-(2' -羟基苯基）-六-2,4-二烯酸（1.05 U/mg）。 2'-取代对活性的影响与基于计算的这些底物的最低未占据分子轨道能量所预测的反应性一致。
• Genetic characterization and expression in heterologous hosts of the 3-(3-hydroxyphenyl)propionate catabolic pathway of Escherichia coli K-12
  作者：A Ferrández、J L Garciá、E Díaz

  DOI：10.1128/jb.179.8.2573-2581.1997

  日期：1997.4

  We report the complete nucleotide sequence of the gene cluster encoding the 3-(3-hydroxyphenyl)propionate (3-HPP) catabolic pathway of Escherichia coli K-12. Sequence analysis revealed the existence of eight genes that map at min 8 of the chromosome, between the lac and hemB regions. Six enzyme-encoding genes account for a flavin-type monooxygenase (mhpA), the extradiol dioxygenase (mhpB), and the meta-cleavage pathway (mhpCDFE). The order of these catabolic genes, with the sole exception of mhpF, parallels that of the enzymatic steps of the pathway. The mhpF gene may encode the terminal acetaldehyde dehydrogenase (acylating) not reported previously in the proposed pathway. Enzymes that catalyze the early reactions of the pathway, MhpA and MhpB, showed the lowest level of sequence similarity to analogous enzymes of other aromatic catabolic pathways. However, the genes mhpCDFE present the same organization and appear to be homologous to the Pseudomonas xyl, dmp, and nah meta-pathway genes, supporting the hypothesis of the modular evolution of catabolic pathways and becoming the first example of this type of catabolic module outside the genus Pseudomonas. Two bacterial interspersed mosaic elements were found downstream of the mhpABCDFE locus and flank a gene, orfT, which encodes a protein related to the superfamily of transmembrane facilitators that might be associated with transport. All of the genes of the 3-HPP cluster are transcribed in the same direction, with the sole exception of mhpR. Inducible expression of the mhp catabolic genes depends upon the presence, in the cis or trans position, of a functional mhpR gene, which suggests that the mhpR gene product is the activator of the 3-HPP biodegradative pathway. The primary structure of MhpR revealed significant similarities to that of members of the IclR subfamily of transcriptional regulators. A 3-HPP catabolic DNA cassette was engineered and shown to be functional not only in enteric bacteria (E. coli and Salmonella typhimurium) but also in Pseudomonas putida and Rhizobium meliloti, thus facilitating its potential application to improve the catabolic abilities of bacterial strains for degradation of aromatic compounds.

  我们报告了大肠杆菌K-12的3-(3-羟基苯基)丙酸(3-HPP)分解途径的基因簇的完整核苷酸序列。序列分析揭示了存在8个基因，这些基因位于染色体的min 8区域，介于lac和hemB区域之间。六个编码酶的基因解释了黄酮型单加氧酶(mhpA)、外二元酸双加氧酶(mhpB)和间位裂解途径(mhpCDFE)。这些分解代谢基因的顺序，除了mhpF之外，与途径的酶步骤的顺序相似。mhpF基因可能编码末端乙醛脱氢酶(酰化)，这在提出的途径中以前没有报道过。催化途径早期反应的酶MhpA和MhpB，与其他芳香族分解途径中类似的酶相比，显示出最低的序列相似性。然而，mhpCDFE基因的组织方式相同，并且似乎与假单胞菌xyl、dmp和nah的间位裂解途径基因同源，支持分解代谢途径模块化进化的假说，并成为该类分解代谢模块在假单胞菌属以外的第一个例子。在mhpABCDFE位点下游发现了两个细菌间杂合嵌合元件，并夹挟一个orfT基因，该基因编码与跨膜促进因子超家族相关的蛋白质，可能与转运有关。3-HPP簇的所有基因都以相同的方向转录，唯一的例外是mhpR。mhp分解代谢基因的诱导表达取决于功能性mhpR基因的存在，无论是在顺式还是反式位置，这表明mhpR基因产物是3-HPP生物降解途径的激活剂。MhpR的主要结构与转录调节因子IclR亚家族的成员显著相似。3-HPP分解代谢DNA盒被工程化，显示其不仅在肠道细菌(E. coli和沙门氏菌伤寒杆菌)中功能正常，而且在假单胞菌putida和苜蓿根瘤菌中也是如此，因此有助于提高细菌菌株降解芳香族化合物的分解代谢能力。
• Purification and properties of 2-hydroxy-6-oxo-6-(2′-aminophenyl)hexa-2,4-dienoic acid hydrolase involved in microbial degradation of carbazole
  作者：Robert R Riddle、Phillip R Gibbs、Richard C Willson、Michael J Benedik

  DOI：10.1016/s1046-5928(02)00676-9

  日期：2003.3

  Hydrolysis following meta-ring cleavage by a dioxygenase is a well-known step in aromatic compound metabolism. The 2-hydroxy-6-oxo-6-(2'-aminophenyl)hexa-2,4-dienoic acid hydrolase from Pseudomonas LD2 is a new member of the small group of characterized aromatic hydrolases that catalyze the cleavage of C-C bonds. In this study, the His(6)-tagged 2-hydroxy-6-oxo-6-(2'aminophenyl)hexa-2,4-dienoic acid (HOPDA) hydrolase was purified from a recombinant Escherichia coli strain utilizing immobilized metal affinity chromatography. 2-Hydroxy-6-oxo-6-(2'-aminophenyl)hexa-2,4-dienoic acid hydrolase is a colorless homodimer with no cofactor requirement. The enzyme actively converted HOPDA into benzoic acid and 2-hydroxypenta-2,4-dienoic acid. The enzyme exhibited activity between pH 6.5 and 10.5 with a maximum activity at pH 7.0. The optimum temperature at pH 7.0 was 60degreesC. The calculated K-m' for HOPDA was 4.6 muM, the V-max, was 3.3 mumol min(-1), and the K-s was 70.0 muM. This corresponds to a maximum specific turnover rate of 1300 HOPDA s(-1) dimer(-1). The deduced amino acid sequence of CarC showed 30.3, 31.3, and 31.8% identity with TodF (P. putida F1), XylF (P. putida), and DmpD (Pseudomonas sp. CF600), respectively, which are meta-cleavage compound hydrolases from other Pseudomonads. The amino acid sequence Gly-X-Ser-X-Gly, which is highly conserved in these hydrolases, is also found in CarC. Lysates from a strain expressing enzyme in which the putative active site serine is mutated to alanine showed a significant reduction in activity. (C) 2002 Elsevier Science (USA). All rights reserved.
• Characterization of the <i>hca</i> Cluster Encoding the Dioxygenolytic Pathway for Initial Catabolism of 3-Phenylpropionic Acid in <i>Escherichia coli</i> K-12
  作者：Eduardo Díaz、Abel Ferrández、José L. García

  DOI：10.1128/jb.180.11.2915-2923.1998

  日期：1998.6

  We have identified, cloned, and sequenced the hca cluster encoding the dioxygenolytic pathway for initial catabolism of 3-phenylpropionic acid (PP) in Escherichia coli K-12. This cluster maps at min 57.5 of the chromosome and is composed of five catabolic genes arranged as a putative operon ( hcaA1A2CBD ) and two additional genes transcribed in the opposite direction that encode a potential permease ( hcaT ) and a regulator ( hcaR ). Sequence comparisons revealed that while hcaA1A2CD genes encode the four subunits of the 3-phenylpropionate dioxygenase, the hcaB gene codes for the corresponding cis -dihydrodiol dehydrogenase. This type of catabolic module is homologous to those encoding class IIB dioxygenases and becomes the first example of such a catabolic cluster in E. coli . The inducible expression of the hca genes requires the presence of the hcaR gene product, which acts as a transcriptional activator and shows significant sequence similarity to members of the LysR family of regulators. Interestingly, the HcaA1A2CD and HcaB enzymes are able to oxidize not only PP to 3-(2,3-dihydroxyphenyl)propionate (DHPP) but also cinnamic acid (CI) to its corresponding 2,3-dihydroxy derivative. Further catabolism of DHPP requires the mhp -encoded meta fission pathway for the mineralization of 3-hydroxyphenylpropionate (3HPP) (A. Ferrández, J. L. Garcı́a, and E. Dı́az, J. Bacteriol. 179:2573–2581, 1997). Expression in Salmonella typhimurium of the mhp genes alone or in combination with the hca cluster allowed the growth of the recombinant bacteria in 3-hydroxycinnamic acid (3HCI) and CI, respectively. Thus, the convergent mhp - and hca -encoded pathways are also functional in S. typhimurium , and they are responsible for the catabolism of different phenylpropanoid compounds (3HPP, 3HCI, PP, and CI) widely available in nature.

  摘要 我们鉴定、克隆并测序了 hca 簇，该簇编码大肠杆菌中 3-苯基丙酸（PP）初始分解的二氧分解途径。 大肠杆菌 K-12.该基因簇位于染色体的第 57.5 小节，由五个分解基因组成，这些基因组成了一个假定操作子（hcaA1A2CBD）。 hcaA1A2CBD ) 和另外两个反向转录的基因，这两个基因编码一种潜在的渗透酶 ( hcaT )和一个调节器( hcaR ).序列比较显示 hcaA1A2CD 基因编码 3-苯基丙酸二氧合酶的四个亚基，而 hcaB 基因编码相应的 顺式 -二氢二醇脱氢酶。这种分解模块与编码 IIB 类二加氧酶的模块同源，是大肠杆菌中第一个此类分解簇的实例。 大肠杆菌 .诱导表达 hca 基因的诱导表达需要 hcaR 基因产物，它是一种转录激活因子，与 LysR 家族的调节因子序列非常相似。有趣的是，HcaA1A2CD 和 HcaB 酶不仅能将 PP 氧化成 3-（2,3-二羟基苯基）丙酸（DHPP），还能将肉桂酸（CI）氧化成其相应的 2,3- 二羟基衍生物。DHPP 的进一步分解需要 mhp -编码的 元 Ferrández, J. L. Garcı́a, and E. Dı́az, J. Bacteriol.179:2573-2581, 1997).在 鼠伤寒沙门氏菌 的 mhp 基因单独或与 hca 簇的重组细菌可分别在 3-羟基肉桂酸（3HCI）和 CI 中生长。因此，趋同的 mhp - 和 hca -编码的途径在 鼠伤寒杆菌 它们负责分解自然界中广泛存在的不同苯丙类化合物（3HPP、3HCI、PP 和 CI）。
• 4-Oxalocrotonate Tautomerase, Its Homologue YwhB, and Active Vinylpyruvate Hydratase:  Synthesis and Evaluation of 2-Fluoro Substrate Analogues
  作者：William H. Johnson,、Susan C. Wang、Thanuja M. Stanley、Robert M. Czerwinski、Jeffrey J. Almrud、Gerrit J. Poelarends、Alexey G. Murzin、Christian P. Whitman

  DOI：10.1021/bi049489p

  日期：2004.8.1

  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.