L-dihydroorotate

• 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: L-dihydroorotate
• 英文别名: (S)-dihydro-orotate；(4S)-4-carboxy-6-oxo-4,5-dihydro-1H-pyrimidin-2-olate
• 化学式: C₅H₅N₂O₄
• 分子量: 157.106
• InChiKey: UFIVEPVSAGBUSI-REOHCLBHSA-M

计算性质

• 辛醇/水分配系数(LogP)：
  -0.9
• 重原子数：
  11
• 可旋转键数：
  0
• 环数：
  1.0
• sp3杂化的碳原子比例：
  0.4
• 拓扑面积：
  98.3
• 氢给体数：
  2
• 氢受体数：
  4

反应信息

• 作为反应物：
  描述：

  L-dihydroorotate 在 Mycobacterium tuberculosis dihydroorotate dehydrogenase, recombinant 作用下， 以 二甲基亚砜 为溶剂， 生成 2,6-二氧代-3H-嘧啶-4-羧酸酯
  参考文献：
  名称：

  结核分枝杆菌二氢乳清酸脱氢酶的动力学和结构研究揭示了对 2 类 DHODH 抑制的新见解

  摘要：

  结核病 (TB) 是全世界死亡的主要原因。结核病是一种严重的公共卫生威胁，其特点是传播率高、流行于贫困地区、合并感染HIV的比例高。此外，降低患者依从性的长期治疗的严重副作用，以及结核分枝杆菌（结核病的致病因子）多重耐药菌株的出现，对其根除提出了若干挑战。寻找新的结核病治疗方法是必要且紧迫的。二氢乳清酸脱氢酶 (DHODH) 负责 ( S )-二氢乳清酸 (DHO) 在从头合成的第四步也是唯一的氧化还原步骤中立体特异性氧化成乳清酸嘧啶核苷酸生物合成途径。DHODH 被认为是对抗传染病的有吸引力的目标。作为利用 DHODH 作为抗结核药物靶标的第一步，我们使用底物辅酶 Q0 (Q0) 和维生素 K3 (K3) 对细菌 MtDHODH 及其人类直系同源物 (HsDHDOH) 进行了完整的动力学表征。MtDHODH 遵循乒乓催化机制，与人类酶具有相似的催化参数。偶然发现 Q0 抑制 MtDHODH

  DOI：

  10.1016/j.bbagen.2023.130378

文献信息

• Stereospecificity of the Dihydroorotate-Dehydrogenase Reaction
  作者：Paul Blattmann、Janos Retey

  DOI：10.1111/j.1432-1033.1972.tb02079.x

  日期：1972.10

  Dihydroorotate dehydrogenase from Zymobacterium oroticum is shown to catalyse the anti‐elimination of hydrogen from the substrate. In the presence of a catalytic amount of NAD+ the enzyme catalyses the exchange between the abstractable hydrogen atoms of dihydroorotate and solvent protons, the exchange of (5S)‐H atom being twice as fast as that of the 4‐H atom. Sodium‐ethoxide‐catalysed exchange affects both diastereotopic protons in the methylene group of dihydroorotate, the (5S)‐H atom being exchanged somewhat faster than the (5R)‐H atom. These findings are discussed in terms of stereospecificity and mechanism of flavin dependent dehydrogenase reactions.

  Zymobacterium oroticum的二氢乳清酸脱氢酶被发现催化底物的anti型氢消除反应。 在NAD+的催化下，该酶催化二氢乳清酸中可.Abstractable氢原子与溶剂质子之间的交换，其中(5S)-H的交换速度是4-H的两倍。 在钠-乙酸盐催化下，交换作用影响二氢乳清酸中甲基上的两个对映异构体质子，其中(5S)-H的交换稍快于(5R)-H。 这些发现从立体特异性和黄素依赖性脱氢酶反应机制的角度进行了讨论。
• Two Functionally Different Dihydroorotic Dehydrogenases in Bacteria
  作者：W. Herman Taylor、Mary L. Taylor、Donald F. Eames

  DOI：10.1128/jb.91.6.2251-2256.1966

  日期：1966.6

  Taylor , W. H. (Portland State College, Portland, Ore.), M. L. Taylor, and D. F. Eames . Two functionally different dihydroorotic dehydrogenases in bacteria. J. Bacteriol. 91: 2251–2256. 1966.—We have investigated the relationship between the two kinds of dihydroorotic dehydrogenases produced by bacteria. A pseudomonad, capable of growth on a salts medium with glucose, aspartate, glycerol, or orotate as the carbon source, was isolated from lake bank mud. A particle-bound dihydroorotic dehydrogenase, similar to the biosynthetic enzyme in Escherichia coli , was formed by the pseudomonad when the carbon source was orotate, glucose, glycerol, or aspartate. A soluble, degradative nicotinamide adenine dinucleotide phosphate-linked dihydroorotic dehydrogenase, as well as the particle-bound biosynthetic enzyme, was formed when the pseudomonad was cultivated on orotate. The biosynthetic enzyme links to oxygen or ferricyanide, but not to pyridine nucleotides. Zymobacterium oroticum , when cultivated on glucose, contained only the biosynthetic type of dihydroorotic dehydrogenase. The presence of two functionally different dihydroorotic dehydrogenases in the pseudomonad was suggested on the basis of the following observations: (i) the two enzyme activities were separated by centrifugation; (ii) the pyridine nucleotide-linked activity was formed only when orotate was present in the growth medium; and (iii) the biosynthetic enzyme was stable to storage at −20 C for 4 months, whereas the degradative enzyme activity was destroyed by storage under these conditions.

  泰勒（Taylor），W.H.（俄勒冈州波特兰州州立学院），M.L.泰勒和D.F.伊姆斯（Eames）。细菌中两种功能不同的双氢乳酸脱氢酶。《细菌学杂志》91：2251-2256。1966年。——我们研究了细菌所产生的两种双氢乳酸脱氢酶之间的关系。从湖岸泥土中分离出一种假单胞菌，能够在含有葡萄糖、天冬氨酸、甘油或乳酸盐作为碳源的盐基培养基上生长。当碳源为乳酸盐、葡萄糖、甘油或天冬氨酸时，该假单胞菌形成了一种类似于大肠杆菌中生物合成酶的粒子结合型双氢乳酸脱氢酶。当该假单胞菌在乳酸盐上培养时，形成可溶性的、降解性的烟酰胺腺嘌呤二核苷酸磷酸型双氢乳酸脱氢酶，以及粒子结合型的生物合成酶。生物合成酶与氧或铁氰化物结合，但不与吡啶核苷酸结合。当以葡萄糖培养的嗜酸性乳酸杆菌中，只含有生物合成型的双氢乳酸脱氢酶。基于以下观察结果，建议在假单胞菌中存在两种功能不同的双氢乳酸脱氢酶：（i）通过离心分离了两种酶活性；（ii）只有在生长基质中存在乳酸盐时，才形成吡啶核苷酸连接的酶活性；（iii）生物合成酶在-20℃下储存4个月后仍稳定，而降解酶活性在这些条件下被破坏。
• Biochemical Characterization of the Heteromeric Bacillus subtilis Dihydroorotate Dehydrogenase and Its Isolated Subunits
  作者：Andrea E Kahler、Finn S Nielsen、Robert L Switzer

  DOI：10.1006/abbi.1999.1455

  日期：1999.11

  Bacillus subtilis dihydroorotate dehydrogenase (DHOD) consists of two subunits, PyrDI (M(r) = 33,094) and PyrDII (M(r) = 28,099). The two subunits were overexpressed jointly and individually and purified. PyrDI was an FMN-containing flavoprotein with an apparent native molecular mass of 85,000. Overexpressed PyrDII formed inclusion bodies and was purified by refolding and reconstitution. Refolded PyrDII

  枯草芽孢杆菌二氢乳清酸脱氢酶（DHOD）由两个亚基组成，PyrDI（M（r）= 33,094）和PyrDII（M（r）= 28,099）。这两个亚基共同和单独过表达并纯化。PyrDI是一种含FMN的黄素蛋白，其表观天然分子量为85,000。过表达的PyrDII形成包涵体，并通过重折叠和重组纯化。重新折叠的PyrDII每摩尔PyrDII结合1摩尔FAD和1摩尔[2Fe-2S]。PyrDI和PyrDII的共表达和纯化产生了天然全酶复合物，其表观天然分子量为114,000，表明存在异四聚体（PyrDI（2）PyrDII（2））。全酶具有二氢乳清酸酯：NAD（+）氧化还原酶活性，还可以还原甲萘醌和人造染料。纯化的PyrDI也具有DHOD活性，但不能降低NAD（+）。与PyrDI相比，全酶对二氢乳清酸酯的K（m）值小20倍以上，对乳清酸酯的K（i）值小约50倍，催化效率高约500倍。通过混合纯化的亚
• Thermodynamic Basis of Electron Transfer in Dihydroorotate Dehydrogenase B from <i>Lactococcus lactis</i>:  Analysis by Potentiometry, EPR Spectroscopy, and ENDOR Spectroscopy
  作者：Al-Walid A. Mohsen、Stephen E. J. Rigby、Kaj Frank Jensen、Andrew W. Munro、Nigel S. Scrutton

  DOI：10.1021/bi036179i

  日期：2004.6.1

  formation of a spin interacting state between the FMN semiquinone species and the reduced 2Fe-2S center. Reduction of DHODB using an excess of NADH or dihydroorotate produces EPR spectra that are distinct from those produced by dithionite. From potentiometric studies, the reduction of the 2Fe-2S center and the reduction of the FMN occur concomitantly. The study provides a detailed thermodynamic framework for

  二氢乳清酸脱氢酶B（DHODB）是一种复杂的铁硫黄素蛋白，可催化二氢乳清酸转化为乳清酸和NAD（+）还原。该酶是含有FMN，FAD和2Fe-2S中心的异二聚体的二聚体。紫外可见，EPR和ENDOR光谱已用于确定黄素和2Fe-2S中心的还原电势，并表征自由基及其相互作用。使用连二亚硫酸盐的还原滴定表明DHODB的五电子容量。2Fe-2S中心的中点还原电位（-212 +/- 3 mV）是通过分析540 nm处的吸收数据确定的，其中两个黄素的吸收贡献很小。FMN的氧化/半醌（E（1））和半醌/对苯二酚（E（2））对的中点还原电位（E（1）= -301 +/- 6 mV; E（2）= -252 +/- 8 mV）和FAD（E（1）= -312 +/- 6 mV; E（2）= -297 +/- 5 mV）通过分析光谱变化确定630纳米 分离的催化亚基（亚基D，缺少2Fe-）的FMN中点还原电位的相应值（E（1）=
• Two different dihydroorotate dehydrogenases in Lactococcus lactis
  作者：P S Andersen、P J Jansen、K Hammer

  DOI：10.1128/jb.176.13.3975-3982.1994

  日期：1994.7

  enzymatic steps. In the characterization of the pyrimidine pathway of Lactococcus lactis, two different pyrD genes encoding dihydroorotate dehydrogenase were isolated. The nucleotide sequences of the two genes, pyrDa and pyrDb, have been determined. One of the deduced amino acid sequences has a high degree of homology to the Saccharomyces cerevisiae dihydroorotate dehydrogenase, and the other resembles

  嘧啶从头开始的生物合成途径已经针对许多生物进行了表征。一般途径包括六个酶促步骤。在表征乳酸乳球菌的嘧啶途径中，分离了两个不同的编码二氢乳清酸脱氢酶的pyrD基因。已经确定了两个基因pyrDa和​​pyrDb的核苷酸序列。推导的氨基酸序列之一与酿酒酵母二氢乳清酸脱氢酶具有高度同源性，另一个类似于来自枯草芽孢杆菌的二氢乳清酸脱氢酶。通过使用不同的电子受体，可以区分粗提物中的两种酶。我们构建了一个突变体，其中含有一个或两个或两个pyrD基因的突变形式。只有双重突变体是嘧啶营养缺陷的。