2-hydroxyethylphosphonate

• 分子结构分类: 有机化合物 - 有机酸及其衍生物 - 有机膦酸及其衍生物
• 英文名称: 2-hydroxyethylphosphonate
• 英文别名: 2-HEP；2-Phosphonatoethanol
• 化学式: C₂H₅O₄P
• 分子量: 124.033
• InChiKey: SEHJHHHUIGULEI-UHFFFAOYSA-L

计算性质

• 辛醇/水分配系数(LogP)：
  -2.4
• 重原子数：
  7
• 可旋转键数：
  1
• 环数：
  0.0
• sp3杂化的碳原子比例：
  1.0
• 拓扑面积：
  83.4
• 氢给体数：
  1
• 氢受体数：
  4

反应信息

• 作为反应物：
  描述：

  2-hydroxyethylphosphonate 、 5'-胞苷酸 在 三乙胺 、 N,N'-羰基二咪唑 作用下， 以 N,N-二甲基甲酰胺 为溶剂， 反应 48.0h， 以51%的产率得到Cytidine 5'-{[hydroxy(2-hydroxyethyl)phosphonoyl]phosphate}
  参考文献：
  名称：

  通过改善大肠杆菌中钴胺素的吸收来增强B类自由基S-腺苷甲硫氨酸甲基化酶的增溶作用

  摘要：

  未激活的碳和磷中心的甲基化是生物化学的新兴领域，尤其是考虑到此类反应是众多酶辅因子，抗生素和其他具有临床价值的天然产物生物合成中的关键步骤。这些动力学上具有挑战性的反应仅由自由基S-腺苷甲硫氨酸（SAM）超家族中的酶催化，并且已分为四类（AD）。B类自由基SAM（RS）甲基化酶除了具有RS酶特征的[4Fe-4S]簇外，还需要钴胺素辅助因子。但是，它们在过度表达时的溶解性差，并且其营业额通常较差，这妨碍了详细的体外试验这些酶的研究。已经提出，不适当的折叠可能是由于在大肠杆菌中过量生产时钴胺素不足引起的，导致包涵体的形成。在本文中，我们报告了通过改造大肠杆菌菌株以吸收更多钴胺素来改善可溶性B类RS甲基化酶过量生产的努力。我们克隆了五个基因（btuC，btuE，btuD，btuF和btuB），这些基因编码负责钴胺素在大肠杆菌中摄取和运输的蛋白质。并将这些基因与编码TsrM，Fom3，PhpK和T

  DOI：

  10.1021/acs.biochem.7b01205
• 作为产物：
  描述：

  二乙基[2-(苄氧基)乙基]膦酸酯 在 盐酸 作用下， 以 水 为溶剂， 以87%的产率得到2-hydroxyethylphosphonate
  参考文献：
  名称：

  通过改善大肠杆菌中钴胺素的吸收来增强B类自由基S-腺苷甲硫氨酸甲基化酶的增溶作用

  摘要：

  未激活的碳和磷中心的甲基化是生物化学的新兴领域，尤其是考虑到此类反应是众多酶辅因子，抗生素和其他具有临床价值的天然产物生物合成中的关键步骤。这些动力学上具有挑战性的反应仅由自由基S-腺苷甲硫氨酸（SAM）超家族中的酶催化，并且已分为四类（AD）。B类自由基SAM（RS）甲基化酶除了具有RS酶特征的[4Fe-4S]簇外，还需要钴胺素辅助因子。但是，它们在过度表达时的溶解性差，并且其营业额通常较差，这妨碍了详细的体外试验这些酶的研究。已经提出，不适当的折叠可能是由于在大肠杆菌中过量生产时钴胺素不足引起的，导致包涵体的形成。在本文中，我们报告了通过改造大肠杆菌菌株以吸收更多钴胺素来改善可溶性B类RS甲基化酶过量生产的努力。我们克隆了五个基因（btuC，btuE，btuD，btuF和btuB），这些基因编码负责钴胺素在大肠杆菌中摄取和运输的蛋白质。并将这些基因与编码TsrM，Fom3，PhpK和T

  DOI：

  10.1021/acs.biochem.7b01205

文献信息

• Initial characterization of Fom3 from Streptomyces wedmorensis: The methyltransferase in fosfomycin biosynthesis
  作者：Kylie D. Allen、Susan C. Wang

  DOI：10.1016/j.abb.2013.12.004

  日期：2014.2

  Fosfomycin is a broad-spectrum antibiotic that is useful against multi-drug resistant bacteria. Although its biosynthesis was first studied over 40 years ago, characterization of the penultimate methyl transfer reaction has eluded investigators. The enzyme believed to catalyze this reaction, Fom3, has been identified as a radical S-adenosyl-L-methionine (SAM) superfamily member. Radical SAM enzymes use SAM and a four-iron, four-sulfur ([4Fe-4S]) cluster to catalyze complex chemical transformations. Fom3 also belongs to a family of radical SAM enzymes that contain a putative cobalamin-binding motif, suggesting that it uses cobalamin for methylation. Here we describe the first biochemical characterization of Fom3 from Streptomyces wedmorensis. Since recombinant Fom3 is insoluble, we developed a successful refolding and iron-sulfur cluster reconstitution procedure. Spectroscopic analyses demonstrate that Fom3 binds a [4Fe-4S] cluster which undergoes a transition between a +2 "resting" state and a +1 active state characteristic of radical SAM enzymes. Site-directed mutagenesis of the cysteine residues in the radical SAM CxxxCxxC motif indicates that each residue is essential for functional cluster formation. We also provide preliminary evidence that Fom3 adds a methyl group to 2-hydroxyethylphosphonate (2-HEP) to form 2-hydroxypropylphosphonate (2-HPP) in an apparently SAM-, sodium dithionite-, and methylcobalamin-dependent manner. (C) 2013 Elsevier Inc. All rights reserved.
• Spectroscopic characterization and mechanistic investigation of P-methyl transfer by a radical SAM enzyme from the marine bacterium Shewanella denitrificans OS217
  作者：Kylie D. Allen、Susan C. Wang

  DOI：10.1016/j.bbapap.2014.09.009

  日期：2014.12

  Natural products containing carbon-phosphorus bonds elicit important bioactivity in many organisms. L-Phosphinothricin contains the only known naturally-occurring carbon-phosphorus-carbon bond linkage. In actinomycetes, the cobalamin-dependent radical S-adenosyl-L-methionine (SAM) methyltransferase PhpK catalyzes the formation of the second C-P bond to generate the complete C-P-C linkage in phosphinothricin. Here we use electron paramagnetic resonance and nuclear magnetic resonance spectroscopies to characterize and demonstrate the activity of a cobalamin-dependent radical SAM methyltransferase denoted SD_1168 from Shewanella denitrificans OS217, a marine bacterium that has not been reported to synthesize phosphinothricin. Recombinant, refolded, and reconstituted SD_1168 binds a four-iron, four-sulfur cluster that interacts with SAM and cobalamin. In the presence of SAM, a reductant, and methylcobalamin, SD_1168 surprisingly catalyzes the P-methylation of N-acetyl-demethylphosphinothricin and demethylphosphinothricin to produce N-acetyl-phosphinothricin and phosphinothricin, respectively. In addition, this enzyme is active in the absence of methylcobalamin if the strong reductant titanium (III) citrate and hydroxocobalamin are provided. When incubated with [methyl-C-13] cobalamin and titanium citrate, both [methyl-C-13] and unlabeled N-acetylphosphinothricin are produced. Our results suggest that SD_1168 catalyzes P-methylation using radical SAM-dependent chemistry with cobalamin as a coenzyme. In light of recent genomic information, the discovery of this P-methyltransferase suggests that S. denitnficans produces a phosphinate natural product. (C) 2014 Elsevier B.V. All rights reserved.
• Connolly, James A.; Banaszczyk, Mariusz; Hynes, Rosemary C., Inorganic Chemistry, 1994, vol. 33, # 4, p. 665 - 669
  作者：Connolly, James A.、Banaszczyk, Mariusz、Hynes, Rosemary C.、Chin, Jik

  DOI：——

  日期：——
• Enhanced Solubilization of Class B Radical <i>S</i>-Adenosylmethionine Methylases by Improved Cobalamin Uptake in <i>Escherichia coli</i>
  作者：Nicholas D. Lanz、Anthony J. Blaszczyk、Erin L. McCarthy、Bo Wang、Roy X. Wang、Brianne S. Jones、Squire J. Booker

  DOI：10.1021/acs.biochem.7b01205

  日期：2018.3.6

  improve the overproduction of class B RS methylases in a soluble form by engineering a strain of E. coli to take in more cobalamin. We cloned five genes (btuC, btuE, btuD, btuF, and btuB) that encode proteins that are responsible for cobalamin uptake and transport in E. coli and co-expressed these genes with those that encode TsrM, Fom3, PhpK, and ThnK, four class B RS methylases that suffer from poor solubility

  未激活的碳和磷中心的甲基化是生物化学的新兴领域，尤其是考虑到此类反应是众多酶辅因子，抗生素和其他具有临床价值的天然产物生物合成中的关键步骤。这些动力学上具有挑战性的反应仅由自由基S-腺苷甲硫氨酸（SAM）超家族中的酶催化，并且已分为四类（AD）。B类自由基SAM（RS）甲基化酶除了具有RS酶特征的[4Fe-4S]簇外，还需要钴胺素辅助因子。但是，它们在过度表达时的溶解性差，并且其营业额通常较差，这妨碍了详细的体外试验这些酶的研究。已经提出，不适当的折叠可能是由于在大肠杆菌中过量生产时钴胺素不足引起的，导致包涵体的形成。在本文中，我们报告了通过改造大肠杆菌菌株以吸收更多钴胺素来改善可溶性B类RS甲基化酶过量生产的努力。我们克隆了五个基因（btuC，btuE，btuD，btuF和btuB），这些基因编码负责钴胺素在大肠杆菌中摄取和运输的蛋白质。并将这些基因与编码TsrM，Fom3，PhpK和T