trans-o-hydroxybenzylidenepyruvate

• 分子结构分类: 有机化合物 - 苯类化合物 - 苯和取代衍生物
• 英文名称: trans-o-hydroxybenzylidenepyruvate
• 英文别名: 4-hydroxy-4-(2-hydroxyphenyl)-2-oxobutanoate；(3E)-4-(2-hydroxyphenyl)-2-oxobut-3-enoate；2-[(E)-3-carboxy-3-oxoprop-1-enyl]phenolate
• 化学式: C₁₀H₇O₄
• 分子量: 191.163
• InChiKey: HMXOGGUFCBUALL-AATRIKPKSA-M

计算性质

• 辛醇/水分配系数(LogP)：
  2
• 重原子数：
  14
• 可旋转键数：
  2
• 环数：
  1.0
• sp3杂化的碳原子比例：
  0.0
• 拓扑面积：
  77.4
• 氢给体数：
  1
• 氢受体数：
  4

反应信息

• 作为反应物：
  描述：

  trans-o-hydroxybenzylidenepyruvate 在 disodium hydrogenphosphate 、 trans-o-hydroxybenzylidenepyruvate hydratase-aldolase 作用下， 以 乙醇 为溶剂， 生成 piruvate 、 水杨醛
  参考文献：
  名称：

  萘降解途径中的水合酶-醛缩酶 NahE 的诱变分析

  摘要：

  NahE是一种水合酶-醛缩酶，其将邻位取代的反式苯亚甲基丙酮酸盐（H、OH或CO 2 - ）分别转化为苯甲醛、水杨醛或2-羧基苯甲醛和丙酮酸盐。该酶位于萘的细菌降解途径中，萘是一种有毒且持久的环境污染物。序列、晶体学和诱变分析将该酶鉴定为醛缩酶超家族中N-乙酰神经氨酸裂解酶 (NAL) 亚组的成员。因此，它具有用于形成希夫碱的保守赖氨酸 (Lys183) 和酪氨酸 (Tyr155)，以及用于结合丙酮酰羧酸基团的 GXXGE 基序。 NahE 的硒代蛋氨酸衍生物的晶体结构显示了这些活性位点元素以及可能参与机制和/或特异性的附近残基。构建了五个活性位点氨基酸（Thr65、Trp128、Tyr155、Asn157 和 Asn281）的突变并测量了动力学参数，以评估对催化的影响。结果表明，两个Trp128突变体（Phe和Tyr）对催化的影响最小，而Thr65（Val）和Asn281（Leu）侧链庞大的氨基酸的影响最大。将

  DOI：

  10.1016/j.abb.2022.109471
• 作为产物：
  描述：

  2-Hydroxychromene-2-carboxylate 生成 trans-o-hydroxybenzylidenepyruvate
  参考文献：
  名称：

  Identification and functional analysis of the genes for naphthalenesulfonate catabolism by Sphingomonas xenophaga BN6

  摘要：

  BN6能够降解各种（取代）萘磺酸盐为相应的（取代）水杨酸盐。在质粒pBN6上鉴定出一个基因簇，编码了参与萘磺酸盐降解途径的多个酶。测序了含有17个ORF的16,915 bp的DNA片段。在该DNA片段上鉴定出编码萘磺酸盐途径的1,2-二羟基萘二氧化酶、2-羟基咔啉-2-羧酸异构酶和2'-羟基苯丙酮醛缩酮酶的基因，并在大肠杆菌中异源表达编码的蛋白。此外，通过插入失活法鉴定出了多组分环氧化萘磺酸盐二氧化酶的ferredoxin和ferredoxin还原酶的编码基因。鉴定出的基因通常与菲的降解菌 sp. P2或萘和联苯降解菌 F199的megaplasmid pNL1编码的酶具有最高的同源性。参与萘磺酸盐降解的 BN6基因与Sphingomonas sp. P2和S. aromaticivorans F199中先前发现的参与萘、联苯和菲降解的基因具有相同的组织形式，分布在三个不同的转录单元中。这些基因在 BN6中被ORF包围，这些ORF指定的蛋白质与移动基因元件的蛋白质具有最高的同源性。

  DOI：

  10.1099/mic.0.28783-0

文献信息

• Purification and Some Properties of 2-Hydroxychromene-2-Carboxylate Isomerase from Naphthalenesulfonate-Assimilating Pseudomonas sp. TA-2
  作者：T. Ohmoto、T. Kinoshita、K. Moriyoshi、K. Sakai、N. Hamada、T. Ohe

  DOI：10.1093/oxfordjournals.jbchem.a022152

  日期：1998.9.1

  30 amino acid sequence had high homology with the deduced amino acid sequences of the 2HC2CA isomerase of nahD (a gene of naphthalene metabolism), pahD (a gene of naphthalene and phenanthrene metabolism), and doxJ (a gene of dibenzothiophene metabolism). The enzymatic product was a trans isomer. The isomerase activity was inhibited in the presence of monoiodoacetate or Hg2+, but not by preincubation

  从萘磺酸同化的假单胞菌sp。的无细胞提取物中纯化2-羟基亚甲基-2-羧酸异构酶。通过在DEAE-纤维素，DEAE-Toyopearl 650M，Sephadex G-75，羟基磷灰石和Mono Q上的连续柱色谱法将TA-2变为电泳均质状态。通过SDS-PAGE估算，该酶的分子量为25和27 kDa。和Superdex 200。它的N端30个氨基酸序列与nahD（萘代谢基因），pahD（萘和菲代谢基因）和doxJ（二苯并噻吩代谢基因）的2HC2CA异构酶的推导氨基酸序列具有高度同源性。 ）。酶促产物是反式异构体。在单碘乙酸盐或Hg2 +存在下，异构酶活性受到抑制，但不可与碘代乙酸盐或N-乙基马来酰亚胺预孵育。GSH充当辅因子，并在0.15 mM以上激活酶。
• <i>trans-o</i> -Hydroxybenzylidenepyruvate Hydratase-Aldolase as a Biocatalyst
  作者：Richard W. Eaton

  DOI：10.1128/aem.66.6.2668-2672.2000

  日期：2000.6

  The hydratase-aldolase-catalyzed conversion of trans-o -hydroxybenzylidenepyruvate to salicylaldehyde and pyruvate is an intermediate reaction in the conversion of naphthalene to salicylate by bacteria. Here, a variety of aromatic aldehydes and some nonaromatic aldehydes together with pyruvate have been shown to be substrates for aldol condensations catalyzed by this enzyme in extracts of the recombinant strain Escherichia coli JM109(pRE701). Some of the products of these reactions were also compared as substrates in the opposite (hydration-aldol cleavage) reaction.

  摘要 肼基转化酶-醇基转化酶催化的 反式-O -羟基亚苄基丙酮酸转化为水杨醛和丙酮酸是细菌将萘转化为水杨酸的中间反应。研究表明，在重组菌株的提取物中，多种芳香醛和一些非芳香醛与丙酮酸一起成为该酶催化的醛缩合反应的底物。 大肠杆菌 JM109(pRE701)的提取物中，该酶催化的醛醇缩合反应的底物。这些反应的一些产物也被比作相反反应（水合-醛醇裂解）的底物。
• Characterization of the aldol condensation activity of the trans-o-hydroxybenzylidenepyruvate hydratase-aldolase (tHBP-HA) cloned from Pseudomonas fluorescens N3
  作者：Silvia Ferrara、Erika Mapelli、Guido Sello、Patrizia Di Gennaro

  DOI：10.1016/j.bbapap.2011.03.013

  日期：2011.5

  The gene encoding trans-o-hydroxybenzylidenepyruvate hydratase-aldolase (tHBP-HA) was isolated from Pseudomonas fluorescens N3, an environmental strain able to degrade naphthalene. This enzyme is an aldolase of class I that reversibly catalyzes the transformation of the trans-o-hydroxybenzylidenepyruvate (t-HBP), releasing pyruvate and salicylaldehyde. The enzyme was expressed in Escherichia coli as a recombinant protein of 38 kDa with a His6-Tag at its N-terminus. The recombinant protein His-tHBP-HA was purified by affinity chromatography and we present here the biochemical characterization of its activity in the aldol condensation reaction. The aldol condensation reaction parameters were determined using as acceptors both salicylaldehyde, which is the natural substrate taking part to the naphthalene degradative pathway, and benzaldehyde. In both cases, His-tHBP-HA shows similar apparent K(m) and apparent V(max) values. Further analyses showed that the optimal pH and temperature of His-tHBP-HA activity are 7.0 and 30 degrees C, respectively. The tHBP-HA catalytic rates and the availability of an efficient system to produce large amounts of purified protein are relevant from a biotechnological point of view. (C) 2011 Elsevier B.V. All rights reserved.
• 2-Hydroxychromene-2-carboxylic Acid Isomerase:  A Kappa Class Glutathione Transferase from <i>Pseudomonas putida</i>^,^,
  作者：Lawrence C. Thompson、Jane E. Ladner、Simona G. Codreanu、Joel Harp、Gary L. Gilliland、Richard N. Armstrong

  DOI：10.1021/bi700356u

  日期：2007.6.1

  The enzyme 2-hydroxychromene-2-carboxylic acid (HCCA) isomerase catalyzes the glutathione (GSH)-dependent interconversion (Keq = 1.5) of HCCA and trans-o-hydroxybenzylidene pyruvic acid (tHBPA) in the naphthalene catabolic pathway of Pseudomonas putida. The dimeric protein binds one molecule of GSH very tightly (Kd approximately 5 nM) and a second molecule of GSH with much lower affinity (Kd approximately 2 to 11 microM). The enzyme is unstable in the absence of GSH. The turnover number in the forward direction (47 s(-1) at 25 degrees C) greatly exceeds off rates for GSH (koff approximately 10(-3) to 10(-2) s(-1) at 10 degrees C), suggesting that GSH acts as a tightly bound cofactor in the reaction. The crystal structure of the enzyme at 1.7 A resolution reveals that the isomerase is closely related to class kappa GSH transferases. Diffraction quality crystals could only be obtained in the presence of GSH and HCCA/tHBPA. Clear electron density is seen for GSH. Electron density for the organic substrates is located near the GSH and is best modeled to include both HCCA and tHBPA at occupancies of 0.5 for each. Although there is no electron density connecting the sulfur of GSH to the organic substrates, the sulfur is located very close (2.78 A) to C7 of HCCA. Taken together, the results suggest that the isomerization reaction involves a short-lived covalent adduct between the sulfur of GSH and C7 of the substrate.
• Kuhm A.E.; Knackmuss H.J.; Stolz A., J Biol Chem, 1993, 0021-9258, 9484-9
  作者：Kuhm A.E.、Knackmuss H.J.、Stolz A.

  DOI：——

  日期：——