3-Hydroxy-5-(hydroxymethyl)-2-methylpyridine-4-carboxylate

• 分子结构分类: 有机化合物 - 有机杂环化合物 - 吡啶及其衍生物
• 英文名称: 3-Hydroxy-5-(hydroxymethyl)-2-methylpyridine-4-carboxylate
• 英文别名: 4-carboxy-5-(hydroxymethyl)-2-methylpyridin-3-olate
• 化学式: C8H8NO4-
• 分子量: 182.15
• InChiKey: HXACOUQIXZGNBF-UHFFFAOYSA-M

计算性质

• 辛醇/水分配系数(LogP)：
  0.7
• 重原子数：
  13
• 可旋转键数：
  1
• 环数：
  1.0
• sp3杂化的碳原子比例：
  0.25
• 拓扑面积：
  93.5
• 氢给体数：
  2
• 氢受体数：
  5

反应信息

• 作为反应物：
  描述：

  3-Hydroxy-5-(hydroxymethyl)-2-methylpyridine-4-carboxylate 、 A 生成 5-Formyl-3-hydroxy-2-methylpyridine-4-carboxylate 、 AH₂
  参考文献：
  名称：

  Gene Identification and Characterization of the Pyridoxine Degradative Enzyme 4-Pyridoxic Acid Dehydrogenase from the Nitrogen-fixing Symbiotic Bacterium Mesorhizobium loti MAFF303099

  摘要：

  在固氮共生细菌Mesorhizobium loti MAFF303099的染色体中，编码4-吡哆酸脱氢酶的基因被确定为mlr6792。该酶是维生素B6（吡哆醇）降解途径I中的第四种酶。在大肠杆菌细胞中过量表达的重组酶带有组氨酸标签，是一种膜结合蛋白，纯化后具有同质性。该酶是一种单体蛋白，分子量为59,000，是一种黄素蛋白，每摩尔亚基含一摩尔FAD。4-吡哆酸的最佳pH值和温度以及Km分别为pH 8.5和30°C，以及29 μM。该酶是一种葡萄糖-甲醇-胆碱（GMC）家族蛋白，具有两种特征模式，即FAD结合残基、假定的活性位点组氨酸残基和可能的跨膜片段。

  DOI：

  10.1093/jb/mvn010
• 作为产物：
  描述：

  4-吡哆酸内酯 、 水 生成 3-Hydroxy-5-(hydroxymethyl)-2-methylpyridine-4-carboxylate 、 氢(+1)阳离子
  参考文献：
  名称：

  Structure of 4-pyridoxolactonase fromMesorhizobium loti

  摘要：

  在维生素 B6 降解途径 I 中，荷包菌（Mesorhizobium loticatalyzes）的 4-吡哆醇内酯酶催化锌依赖性内酯环水解 4-吡哆醇内酯（4PAL）为 4-吡哆酸（4PA）。测定了 4-吡哆醇内酯酶及其与 5-吡哆醇内酯（5PAL，竞争性抑制剂）的复合物的晶体结构。整体结构为αβ/βα夹层折叠，有两个锌离子配位。这有力地表明该酶属于 B 类 β-内酰胺酶的 B3 亚类。在复合物结构中，5PAL 的羰基远离活性位点，这揭示了其作为竞争性抑制剂的原因。根据与 4PAL、4PA 和反应中间体的对接模拟，4-吡哆醇内酰胺酶可能是通过类似 B2 亚类的机制而不是 B3 亚类的机制来催化反应的。

  DOI：

  10.1107/s2053230x14003926

文献信息

• Gene Identification and Characterization of 5-Formyl-3-Hydroxy-2-Methylpyridine 4-Carboxylic Acid 5-Dehydrogenase, an NAD+-Dependent Dismutase
  作者：N. Yokochi、Y. Yoshikane、S. Matsumoto、M. Fujisawa、K. Ohnishi、T. Yagi

  DOI：10.1093/jb/mvp007

  日期：——

  A chromosomal gene, mlr6793, in Mesorhizobium loti was identified as the gene encoding 5-formyl-3-hydroxy-2-methylpyridine 4-carboxylic acid (FHMPC) dehydrogenase (dismutase) involved in the degradation pathway for pyridoxine (vitamin B6). The homogenously purified recombinant enzyme has a molecular mass of 59.1 kDa and is a homodimeric protein. FHMPC dehydrogenase catalyses practically irreversible oxidation (kcat = 204 s–1) of FHMPC (Km = 48.2 μM) by NAD+ (Km = 34.3 μM) to 3-hydroxy-2-methyl-pyridine 4, 5-dicarboxylic acid (HMPDC), and practically irreversible reduction (kcat = 217 s–1) of FHMPC (Km = 24.9 μM) by NADH (Km = 12.4 μM) to 4-pyridoxic acid. When the enzyme reaction was started with the combination of FHMPC and NAD+ or that of FHMPC and NADH, HMPDC and 4-pyridoxic acid were produced in an almost equimolar ratio throughout the reaction. FHMPC dehydrogenase belongs to the 3-hydroxyacyl-CoA dehydrogenase family with 31% identity with the human enzyme: it has probable catalytic diad residues, i.e. His137 and Glu149. The H137L mutant enzyme showed no measurable activity. The E149Q one was stable in contrast to the corresponding human 3-hydroxyacyl-CoA dehydrogenase mutant, and showed unique pH optima depending on the co-substrates used for the reaction.

  Mesorhizobium loti中的染色体基因mlr6793被确定为编码5-甲酰基-3-羟基-2-甲基吡啶4-羧酸（FHMPC）脱氢酶（歧化酶）的基因，该酶参与吡哆醇（维生素B6）的降解途径。均相纯化的重组酶分子量为59.1 kDa，是一种同二聚体蛋白。FHMPC脱氢酶催化NAD+（Km = 34.3 μM）对FHMPC（Km = 48.2 μM）的几乎不可逆氧化（kcat = 204 s–1）反应，生成3-羟基-2-甲基吡啶4,5-二羧酸（HMPDC），以及催化NADH（Km = 12.4 μM）对FHMPC（Km = 24.9 μM）的几乎不可逆还原（kcat = 217 s–1）反应，生成4-吡哆酸。当FHMPC和NAD+或FHMPC和NADH的组合开始酶反应时，在整个反应过程中，HMPDC和4-吡哆酸以几乎等摩尔比生成。FHMPC脱氢酶属于3
• Crystal structure of 5-formyl-3-hydroxy-2-methylpyridine 4-carboxylic acid 5-dehydrogenase, an NAD+-dependent dismutase from Mesorhizobium loti
  作者：Andrew Njagi Mugo、Jun Kobayashi、Bunzo Mikami、Yu Yoshikane、Toshiharu Yagi、Kouhei Ohnishi

  DOI：10.1016/j.bbrc.2014.11.028

  日期：2015.1

  5-Formyl-3-hydroxy-2-methylpyridine 4-carboxylic acid 5-dehydrogenase (FHMPCDH) from Mesorhizobium loti is the fifth enzyme in degradation pathway I for pyridoxine. The enzyme catalyzes a dismutation reaction: the oxidation of 5-formyl-3-hydroxy-2-methylpyridine 4-carboxylic acid (FHMPC) to 3-hydroxy-2-methylpyridine 4,5-dicarboxylic acid with NAD(+) and reduction of FHMPC to 4-pyridoxic acid with NADH. FHMPCDH belongs to the (L)-3-hydroxyacyl-CoA dehydrogenase (HAD) family. The crystal structure was determined by molecular replacement and refined to a resolution of 1.55 angstrom (R-factor of 16.4%, R-free = 19.4%). There were two monomers in the asymmetric unit. The overall structure of the monomer consisted of N- and C-terminal domains connected by a short linker loop. The monomer was similar to members of the HAD family (RMSD = 1.9 angstrom). The active site was located between the domains and highly conserved to that of human heart (L)-3-hydroxyacyl-CoA dehydrogenase (HhHAD). His-Glu catalytic dyad, a serine and two asparagine residues of HhHAD were conserved. Ser116, His137 and Glu149 in FHMPCDH are connected by a hydrogen bonding network forming a catalytic triad. The functions of the active site residues in the reaction mechanism are discussed. (C) 2014 Elsevier Inc. All rights reserved.
• Yagi T.; Kishore G.M.; Snell E.E., J Biol Chem, 1983, 0021-9258, 9419-25
  作者：Yagi T.、Kishore G.M.、Snell E.E.

  DOI：——

  日期：——
• 4-Pyridoxolactonase from a symbiotic nitrogen-fixing bacterium Mesorhizobium loti: Cloning, expression, and characterization
  作者：Junichi Funami、Yu Yoshikane、Hitoshi Kobayashi、Nana Yokochi、Baiqiang Yuan、Kozo Iwasaki、Kouhei Ohnishi、Toshiharu Yagi

  DOI：10.1016/j.bbapap.2005.08.026

  日期：2005.12

  4-Pyridoxolactonase is involved in the degradation pathway for pyridoxine, a free form of vitamin B-6. The gene (mlr6805) encoding the putative 4-pyridoxolactonase of nitrogen fixing symbiotic microorganism Mesorhizobium loti MAFF303099 has been identified based on the genome database. The gene was cloned and overexpressed in a cotransformant Escherichia coli cell. The recombinant enzyme was dimeric protein and contained one mole of Zn2+ per mole of subunit. The enzyme showed about 30% identity with various N-acylhomoserine lactone lactonases and metallo-beta-lactamases. The phylogram made with ClustalW shows that 4-pyridoxolactonase makes a cluster with Agrobacterium tumefaciens acyl-homoserine lactone lactonase. The alignment of amino acid sequences suggests that 4-pyridoxolactonase has three histidine residues probably involved in binding of Zn2+. (c) 2005 Elsevier B.V. All rights reserved.
• Gene Identification and Characterization of the Pyridoxine Degradative Enzyme 4-Pyridoxic Acid Dehydrogenase from the Nitrogen-fixing Symbiotic Bacterium Mesorhizobium loti MAFF303099
  作者：F. Ge、N. Yokochi、Y. Yoshikane、K. Ohnishi、T. Yagi

  DOI：10.1093/jb/mvn010

  日期：——

  The gene encoding 4-pyridoxic acid dehydrogenase was identified as mlr6792 in a chromosome of a nitrogen-fixing symbiotic bacterium Mesorhizobium loti MAFF303099. The enzyme is the fourth enzyme in the vitamin B6 (pyridoxine)-degradation pathway I. The recombinant enzyme with a his-tag over-expressed in Escherichia coli cells was a membrane-bound protein, and purified to homogeneity. The enzyme was a monomeric protein with a molecular weight of 59,000, and a flavoprotein containing one mole of FAD per mole of subunit. The optimum pH and temperature, and Km for 4-pyridoxic acid were pH 8.5 and 30°C, and 29 μM, respectively. The enzyme was a glucose-methanol-choline (GMC) family protein with two signature patterns, FAD-binding residues, a putative active site histidine residue and a probable transmembrane segment.

  在固氮共生细菌Mesorhizobium loti MAFF303099的染色体中，编码4-吡哆酸脱氢酶的基因被确定为mlr6792。该酶是维生素B6（吡哆醇）降解途径I中的第四种酶。在大肠杆菌细胞中过量表达的重组酶带有组氨酸标签，是一种膜结合蛋白，纯化后具有同质性。该酶是一种单体蛋白，分子量为59,000，是一种黄素蛋白，每摩尔亚基含一摩尔FAD。4-吡哆酸的最佳pH值和温度以及Km分别为pH 8.5和30°C，以及29 μM。该酶是一种葡萄糖-甲醇-胆碱（GMC）家族蛋白，具有两种特征模式，即FAD结合残基、假定的活性位点组氨酸残基和可能的跨膜片段。