(2S)-2-formamidopentanedioate

• 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: (2S)-2-formamidopentanedioate
• 化学式: C6H7NO5-2
• 分子量: 173.12
• InChiKey: ADZLWSMFHHHOBV-BYPYZUCNSA-L

计算性质

• 辛醇/水分配系数(LogP)：
  0.3
• 重原子数：
  12
• 可旋转键数：
  3
• 环数：
  0.0
• sp3杂化的碳原子比例：
  0.5
• 拓扑面积：
  109
• 氢给体数：
  1
• 氢受体数：
  5

反应信息

• 作为反应物：
  描述：

  水 、 (2S)-2-formamidopentanedioate 生成 草氨酸 、 2-氨基戊二酸酯
  参考文献：
  名称：

  恶臭假单胞菌甲酰谷氨酸酰胺水解酶的纯化和性质。

  摘要：

  甲酰谷氨酸酰胺水解酶（FGase）催化恶臭假单胞菌利用组氨酸利用的五步途径中的末端反应。通过该作用，N-甲酰基-L-谷氨酸（FG）被水解以产生L-谷氨酸加甲酸。尿酸尿酸盐是该途径的第一个产物，可诱导所有5种酶，但FG能够单独诱导FGase，尽管效率不及尿酸盐。FG的这种诱导导致形成具有与尿烷酸酯诱导的FGase相同的电泳迁移率的FGase。将含有恶臭假单胞菌FGase基因的9.6碱基对HindIII DNA片段克隆到大肠杆菌中维持的质粒pBEU1的相应位点。在载体的任一方向上插入片段都会导致表达，但在一个方向上却发现了更高的水平，这表明除了效率较低的假单胞菌启动子外，FGase基因还可以由效率不同的两个载体启动子或单个载体启动子表达。通过六步纯化，从高表达克隆中纯化FGase 1110倍，收率为10％。二价金属离子刺激了活性，在所测试的那些（钴，铁，锌，钙，镍，镉，锰和镁）中，钴（Ⅱ）是最好的活化剂，其次是铁（Ⅱ）。在5

  DOI：

  10.1128/jb.169.10.4696-4702.1987
• 作为产物：
  描述：

  水 、 (2S)-2-(aminomethylideneazaniumyl)pentanedioate 生成 (2S)-2-formamidopentanedioate 、 铵
  参考文献：
  名称：

  Mechanistic Characterization of N-Formimino-l-glutamate Iminohydrolase from Pseudomonas aeruginosa

  摘要：

  N-Formimino-L-glutamate iminohydrolase (HutF) from Pseudomonas aeruginosa catalyzes the deimination of N-formimino-L-glutamate in the histidine degradation pathway. An amino acid sequence alignment between HutF and members of the amidohydrolase superfamily containing mononuclear metal centers indicated that residues Glu-235, His-269, and Asp-320 are involved in substrate binding and activation of the nucleophilic water molecule. The purified enzyme contained up to one equivalent of zinc. The metal was removed by dialysis against the metal chelator dipicolinate with the complete loss of catalytic activity. Enzymatic activity was restored by incubation of the apoprotein with Zn2+, Cd2+, Ni2+, or Cu2+. The mutation of Glu-235, His-269, or Asp-320 resulted in the diminution of catalytic activity by two to six orders of magnitude. Bell-shaped profiles were observed for k(cat) and k(cat)/K-m as a function of pH. The pK(a) of the group that must be unprotonated for catalytic activity was consistent with the ionization of His-269. This residue is proposed to function as a general base in the abstraction of a proton from the metal-bound water molecule. In the proposed catalytic mechanism, the reaction is initiated by the abstraction of a proton from the metal-bound water molecule by the side chain imidazole of His-269 to generate a tetrahedral intermediate of the substrate. The collapse of the tetrahedral intermediate commences with the abstraction of a second proton via the side chain carboxylate of Asp-320. The C-N bond of the substrate is subsequently cleaved with proton transfer from His-269 to form ammonia and the N-formyl product. The postulated role of the invariant Glu-235 is to ion pair with the positively charged formimino group of the substrate.

  DOI：

  10.1021/bi061673i

文献信息

• Moonlighting Glutamate Formiminotransferases Can Functionally Replace 5-Formyltetrahydrofolate Cycloligase
  作者：Linda Jeanguenin、Aurora Lara-Núñez、Anne Pribat、Melissa Hamner Mageroy、Jesse F. Gregory、Kelly C. Rice、Valérie de Crécy-Lagard、Andrew D. Hanson

  DOI：10.1074/jbc.m110.190504

  日期：2010.12

  5-Formyltetrahydrofolate (5-CHO-THF) is formed by a side reaction of serine hydroxymethyltransferase. Unlike other folates, it is not a one-carbon donor but a potent inhibitor of folate enzymes and must therefore be metabolized. Only 5-CHO-THF cycloligase (5-FCL) is generally considered to do this. However, comparative genomic analysis indicated (i) that certain prokaryotes lack 5-FCL, implying that they have an alternative 5-CHO-THF-metabolizing enzyme, and (ii) that the histidine breakdown enzyme glutamate formiminotransferase (FT) might moonlight in this role. A functional complementation assay for 5-CHO-THF metabolism was developed in Escherichia coli, based on deleting the gene encoding 5-FCL (ygfA). The deletion mutant accumulated 5-CHO-THF and, with glycine as sole nitrogen source, showed a growth defect; both phenotypes were complemented by bacterial or archaeal genes encoding FT. Furthermore, utilization of supplied 5-CHO-THF by Streptococcus pyogenes was shown to require expression of the native FT. Recombinant bacterial and archaeal FTs catalyzed formyl transfer from 5-CHO-THF to glutamate, with k(cat) values of 0.1-1.2 min(-1) and K-m values for 5-CHO-THF and glutamate of 0.4-5 mu M and 0.03-1 mM, respectively. Although the formyltransferase activities of these proteins were far lower than their formiminotransferase activities, the K-m values for both substrates relative to their intracellular levels in prokaryotes are consistent with significant in vivo flux through the formyltransferase reaction. Collectively, these data indicate that FTs functionally replace 5-FCL in certain prokaryotes.
• A β-citryl-L-glutamate-hydrolysing enzyme in rat testes
  作者：Masaharu Miyake、Toshihiko Innami、Yasuo Kakimoto

  DOI：10.1016/0304-4165(83)90165-4

  日期：1983.10

  An enzyme responsible for the deacylation of beta-citryl-L-glutamate to citrate and glutamate has been characterized in rat testis. The enzyme required manganese ion for full activity and was strongly inhibited by nucleotides such as ATP or GTP. The activity was localized in the particulate fractions. The enzyme favored N-formyl-L-glutamate greater than beta-citrly-L-glutamate greater than beta-citryl-L-glutamine in a decreasing order. The amidohydrolyase activity was highest in the testis and lung, a moderate activity was detected in heart, kidney and intestine, and low in brain, thymus, stomach, skeletal muscle, spleen and liver. These findings suggest that the amidohydrolase is different from any of amidohydrolases reported so far, amidohydrolase I (EC 3.5.1.14), II (EC 3.5.1.15), III, N-acetyl-lysine deacylase (EC 3.5.1.17) and N-acetyl-beta-alanine deacetylase (EC 3.5.1.21), and various peptidases.
• Mechanistic Characterization of <i>N</i>-Formimino-<scp>l</scp>-glutamate Iminohydrolase from <i>Pseudomonas aeruginosa</i>
  作者：Ricardo Martí-Arbona、Frank M. Raushel

  DOI：10.1021/bi061673i

  日期：2006.12.1

  N-Formimino-L-glutamate iminohydrolase (HutF) from Pseudomonas aeruginosa catalyzes the deimination of N-formimino-L-glutamate in the histidine degradation pathway. An amino acid sequence alignment between HutF and members of the amidohydrolase superfamily containing mononuclear metal centers indicated that residues Glu-235, His-269, and Asp-320 are involved in substrate binding and activation of the nucleophilic water molecule. The purified enzyme contained up to one equivalent of zinc. The metal was removed by dialysis against the metal chelator dipicolinate with the complete loss of catalytic activity. Enzymatic activity was restored by incubation of the apoprotein with Zn2+, Cd2+, Ni2+, or Cu2+. The mutation of Glu-235, His-269, or Asp-320 resulted in the diminution of catalytic activity by two to six orders of magnitude. Bell-shaped profiles were observed for k(cat) and k(cat)/K-m as a function of pH. The pK(a) of the group that must be unprotonated for catalytic activity was consistent with the ionization of His-269. This residue is proposed to function as a general base in the abstraction of a proton from the metal-bound water molecule. In the proposed catalytic mechanism, the reaction is initiated by the abstraction of a proton from the metal-bound water molecule by the side chain imidazole of His-269 to generate a tetrahedral intermediate of the substrate. The collapse of the tetrahedral intermediate commences with the abstraction of a second proton via the side chain carboxylate of Asp-320. The C-N bond of the substrate is subsequently cleaved with proton transfer from His-269 to form ammonia and the N-formyl product. The postulated role of the invariant Glu-235 is to ion pair with the positively charged formimino group of the substrate.
• Purification and properties of formylglutamate amidohydrolase from Pseudomonas putida
  作者：L Hu、L M Mulfinger、A T Phillips

  DOI：10.1128/jb.169.10.4696-4702.1987

  日期：1987.10

  fragment containing the P. putida FGase gene was cloned into the corresponding site on plasmid pBEU1 maintained in Escherichia coli. Insertion of the fragment in either orientation on the vector resulted in expression, but a higher level was noted in one direction, suggesting that the FGase gene can be expressed from either of two vector promoters with different efficiencies or from a single vector promoter

  甲酰谷氨酸酰胺水解酶（FGase）催化恶臭假单胞菌利用组氨酸利用的五步途径中的末端反应。通过该作用，N-甲酰基-L-谷氨酸（FG）被水解以产生L-谷氨酸加甲酸。尿酸尿酸盐是该途径的第一个产物，可诱导所有5种酶，但FG能够单独诱导FGase，尽管效率不及尿酸盐。FG的这种诱导导致形成具有与尿烷酸酯诱导的FGase相同的电泳迁移率的FGase。将含有恶臭假单胞菌FGase基因的9.6碱基对HindIII DNA片段克隆到大肠杆菌中维持的质粒pBEU1的相应位点。在载体的任一方向上插入片段都会导致表达，但在一个方向上却发现了更高的水平，这表明除了效率较低的假单胞菌启动子外，FGase基因还可以由效率不同的两个载体启动子或单个载体启动子表达。通过六步纯化，从高表达克隆中纯化FGase 1110倍，收率为10％。二价金属离子刺激了活性，在所测试的那些（钴，铁，锌，钙，镍，镉，锰和镁）中，钴（Ⅱ）是最好的活化剂，其次是铁（Ⅱ）。在5