(2S)-2-{[(2S)-2-azaniumylpropanoyl]amino}-3-[(1R,2S,6R)-5-oxo-7-oxabicyclo[4.1.0]hept-2-yl]propanoate | 29393-20-2

• 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-{[(2S)-2-azaniumylpropanoyl]amino}-3-[(1R,2S,6R)-5-oxo-7-oxabicyclo[4.1.0]hept-2-yl]propanoate
• 英文别名: (2S)-2-[[(2S)-2-azaniumylpropanoyl]amino]-3-[(1R,2S,6R)-5-oxo-7-oxabicyclo[4.1.0]heptan-2-yl]propanoate
• CAS: 29393-20-2
• 化学式: C12H18N2O5
• 分子量: 270.28
• InChiKey: XFOUAXMJRHNTOP-PFQXTLEHSA-N

物化性质

• 沸点：
  413.4°C (rough estimate)
• 密度：
  1.2232 (rough estimate)

计算性质

• 辛醇/水分配系数(LogP)：
  -3.4
• 重原子数：
  19
• 可旋转键数：
  5
• 环数：
  2.0
• sp3杂化的碳原子比例：
  0.75
• 拓扑面积：
  122
• 氢给体数：
  3
• 氢受体数：
  6

安全信息

• 安全说明：
  Sx.

制备方法与用途

类别：有毒物品

毒性分级：中毒

急性毒性：静脉-小鼠 LD50：450 毫克/公斤

可燃性危险特性：可燃；燃烧时产生有毒氮氧化物烟雾

储运特性：通风、低温、干燥

灭火剂：干粉、泡沫、沙土、二氧化碳、雾状水

反应信息

• 作为产物：
  描述：

  adenosine 5'-triphosphate 、 L-丙氨酸 、 抗荚膜菌素 生成 adenosine 5'-diphosphate 、 (2S)-2-{[(2S)-2-azaniumylpropanoyl]amino}-3-[(1R,2S,6R)-5-oxo-7-oxabicyclo[4.1.0]hept-2-yl]propanoate 、 氢(+1)阳离子 、 H3PO4
  参考文献：
  名称：

  枯草芽孢杆菌中的ywfE编码一种新型酶L-氨基酸连接酶。

  摘要：

  已知ATP依赖的羧酸胺/硫醇连接酶超家族包含催化各种类型的肽（例如d-丙氨酰-d-丙氨酸，聚谷氨酸和γ-肽）形成的酶，但奇怪的是，没有可合成α的酶1-氨基酸的β-二肽是已知的。我们试图找到这种酶。通过基于超家族的共有序列进行计算机筛选，然后使用纯化的酶进行体外测定以避免合成肽的降解，发现枯草芽孢杆菌的ywfE编码形成L-丙氨酰-1的活性-来自谷氨酰胺和谷氨酰胺的谷氨酰胺，ATP水解成ADP。没有形成AMP，支持了该酶属于超家族的观点。出人意料的是，该酶接受了各种各样的1-氨基酸。在测试的231种L-氨基酸组合中，获得了111种组合的反应产物，并通过高效液相色谱分析确定了44种α-二肽，而未检测到三肽或更长的肽，并且d-氨基酸呈惰性。根据这些结果，我们提出ywfE编码超家族的一个新成员，即I-氨基酸连接酶。

  DOI：

  10.1128/jb.187.15.5195-5202.2005

文献信息

• Structural and enzymatic characterization of BacD, an<scp>L</scp>-amino acid dipeptide ligase from<i>Bacillus subtilis</i>
  作者：Yasuhito Shomura、Emi Hinokuchi、Hajime Ikeda、Akihiro Senoo、Yuichi Takahashi、Jun-ichi Saito、Hirofumi Komori、Naoki Shibata、Yoshiyuki Yonetani、Yoshiki Higuchi

  DOI：10.1002/pro.2058

  日期：2012.5

  BacD is an ATP‐dependent dipeptide ligase responsible for the biosynthesis of L‐alanyl‐L‐anticapsin, a precursor of an antibiotic produced by Bacillus spp. In contrast to the well‐studied and phylogenetically related D‐alanine: D‐alanine ligase (Ddl), BacD synthesizes dipeptides using L‐amino acids as substrates and has a low substrate specificity in vitro. The enzyme is of great interest because of

  BACD是一种依赖ATP的二肽连接酶，负责L-丙氨酰-L-抗溶衣蛋白酶的生物合成，这是一种由芽孢杆菌属产生的抗生素的前体。与经过深入研究和系统发育相关的D-丙氨酸：D-丙氨酸连接酶（Ddl）相比，BACD使用L-氨基酸作为底物来合成二肽，并且在体外具有较低的底物特异性。该酶备受关注，因为它在工业蛋白质工程中可能用于环境友好地生物学生产有用的肽化合物（例如生理活性肽，人造甜味剂和抗生素），但其底物特异性及其催化机制的决定因素尚无定论。由于缺乏结构信息而尚未建立。在这项研究中，我们报告BACD的晶体结构在复杂与ADP和中间模拟，磷酸化次膦大号-alanyl-大号-苯丙氨酸，精制至2.5Å分辨率。复杂的结构表明，ADP和两个镁离子以类似于Ddl的方式结合。但是，二肽的方向相反，因此，氨基酸结合腔的入口位置不同。对两个突变体Y265F和S185A的酶促表征表明，至少在以L-苯丙氨酸为底物的反应中，
• Action and Timing of BacC and BacD in the Late Stages of Biosynthesis of the Dipeptide Antibiotic Bacilysin
  作者：Jared B. Parker、Christopher T. Walsh

  DOI：10.1021/bi3016229

  日期：2013.2.5

  Biosynthesis of the dipeptide antibiotic bacilysin, encoded by the seven Bacillus subtilis genes bacA-G, involves diversion of flux from prephenate to the noncognate amino acid anticapsin. The anticapsin warhead is then ligated to the C-terminus of L-alanine to produce mature bacilysin. We have previously noted the formation of two diastereomers of tetrahydrotyrosine (4S- and 4R-H(4)Tyr) by tandem action of the four purified enzymes BacABGF. BacC (oxidase) and BacD (ligase) have been hypothesized to be remaining late stage enzymes in bacilysin biosynthesis. Using a combination of BacCD in vitro studies, B. subtilis deletion mutants, and isotopic feeding studies, we were able to determine that the H(4)Tyr diastereomers are actually shunt products that are not on-pathway to bacilysin biosynthesis. Dihydroanticapsin and dihydrobacilysin accumulate in extracts of a Delta bacC strain and are processed to anticapsin and then bacilysin upon addition of BacC and BacD, respectively. These results suggest the epoxide group in bacilysin is installed in an earlier step of bacilysin biosynthesis, while BacC oxidation of the C-7-hydroxyl and the subsequent BacD ligation of anticapsin to L-Ala are the penultimate and ultimate steps of bacilysin biosynthesis, respectively.
• Single Mutation Alters the Substrate Specificity of <scp>l</scp>-Amino Acid Ligase
  作者：Takeo Tsuda、Mana Asami、Yoshiaki Koguchi、Shuichi Kojima

  DOI：10.1021/bi500292b

  日期：2014.4.29

  L-Amino acid ligase (Lal) catalyzes the formation of a dipeptide from two L-amino acids in an ATP-dependent manner and belongs to the ATP-grasp superfamily. Bacillus subtilis YwfE, the first identified Lal, produces the dipeptide antibiotic bacilysin, which consists of L-Ala and L-anticapsin. Its substrate specificity is restricted to smaller amino acids such as L-Ala for the N-terminal end of the dipeptide, whereas a wide range of hydrophobic amino acids including L-Phe and L-Met are recognized for the C-terminal end in vitro. We determined the crystal structures of YwfE with bound ADP-Mg2+-Pi and ADP-Mg2+-L-Ala at 1.9 and 2.0 angstrom resolutions, respectively. On the basis of these structures, we generated point mutants of residues that are considered to participate in the recognition of L-Ala and measured their ATPase activity. The conserved Arg328 is suggested to be a crucial residue for L-Ala recognition and catalysis. The mutation of Trp332 to Ala caused the enzyme to hydrolyze ATP, even in the absence of L-Ala, and the structure of this mutant protein appeared to show a cavity in the N-terminal substrate-binding pocket. These results suggest that Trp332 plays a key role in restricting the substrate specificity to smaller amino acids such as L-Ala. Moreover, Trp332 mutants can alter the substrate specificity and activity depending on the size and shape of substituted amino acids. These observations provide sufficient scope for the rational design of Lal to produce desirable dipeptides. We propose that the positioning of the conserved Arg residue in Lal is important for enantioselective recognition of L-amino acids.