(3S)-3-氨基-4-(4-硝基苯胺基)-4-氧代丁酸

• 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
• 中文名称: (3S)-3-氨基-4-(4-硝基苯胺基)-4-氧代丁酸
• 英文名称: H-Asp-pNA
• 英文别名: (3S)-3-azaniumyl-4-(4-nitroanilino)-4-oxobutanoate
• 化学式: C₁₀H₁₁N₃O₅
• 分子量: 253.214
• InChiKey: JPSVAMXHSUXSEY-QMMMGPOBSA-N

计算性质

• 辛醇/水分配系数(LogP)：
  -2.7
• 重原子数：
  18
• 可旋转键数：
  4
• 环数：
  1.0
• sp3杂化的碳原子比例：
  0.2
• 拓扑面积：
  138
• 氢给体数：
  3
• 氢受体数：
  6

反应信息

• 作为反应物：
  描述：

  (3S)-3-氨基-4-(4-硝基苯胺基)-4-氧代丁酸 在 soybean cotyledon N-terminal acidic amino acid-specific aminopeptidase 作用下， 生成 L-天门冬氨酸 、 4-硝基苯胺
  参考文献：
  名称：

  Purification and Characterization of an N-Terminal Acidic Amino Acid-Specific Aminopeptidase from Soybean Cotyledons (Glycine max)

  摘要：

  通过硫酸铵分级和 Q-sepharose、Phenyl sepharose 和 Superdex 200 连续柱层析，从大豆子叶中分离出一种催化 Glu-Glu 高效水解的新型酶。该酶的表观分子量为 56 kDa分别通过 SDS-聚丙烯酰胺凝胶电泳和 Superdex 200 HR 10/30 柱层析得出 510 kDa 和 510 kDa。该酶对 Glu-p-硝基苯胺 (pNA) 和 Asp-pNA 具有高活性，而 Leu-pNA、Phe-pNA、Ala-pNA 和 Pro-pNA 不被水解。合成的二肽 Glu-Xxx 和 Asp-Xxx 被水解，但 Xxx-Glu 没有被水解。使用这种纯化酶消化富含 Glu 的寡肽嗜铬粒蛋白 A (Glu-Glu-Glu-Glu-Glu-Met-Ala-Val-Val-Pro-Gln-Gly-Leu-Phe-Arg-Gly-NH2)也被调查。谷氨酸残基从N末端一一被切割。这些观察结果表明该酶从 N 末端含酸性氨基酸的肽中去除谷氨酰或天冬氨酰残基。人们认为它是一种来自植物的 N 末端酸性氨基酸特异性氨肽酶。

  DOI：

  10.1271/bbb.90617

文献信息

• Peptide Synthesis in Organic Media with the Use of Subtilisin 72 Immobilized on a Poly(Vinyl Alcohol) Cryogel
  作者：A. V. Belyaeva、A. V. Bacheva、E. S. Oksenoit、E. N. Lysogorskaya、V. I. Lozinskii、I. Yu. Filippova

  DOI：10.1007/s11171-005-0072-y

  日期：2005.11

  Subtilisin 72 serine protease (EC 3.4.21.14) immobilized on a poly(vinyl alcohol) cryogel was used as a catalyst in the syntheses of N-protected peptide p-nitroanilides of the general formulas Z(or Boc)-Xaa-Phe-pNA (Xaa = Leu or Ala), Z-Ala-Xaa-Yaa-pNA (Xaa = Leu or Ala; Yaa = Leu or Phe), and Z-Ala-Ala-Xaa-Yaa-pNA (Xaa = Leu, Arg, or Gly; Yaa = Phe, Leu, Gly, Asp, or Glu). The syntheses were carried out in DMF-acetonitrile mixtures. A number of protected di-, tri-, and tetrapeptides were prepared in yields up to 99%. The syntheses were found to retain stereoselectivity under the conditions studied. The activation of carboxyl group of the acylating component was shown to have a positive effect upon the coupling rate.

  将固定于聚乙烯醇冻胶上的72丝氨酸蛋白酶（EC 3.4.21.14）用作合成N-保护肽对硝基苯胺的催化剂，其通式为Z（或Boc）-Xaa-Phe-pNA（Xaa=亮氨酸或丙氨酸）、Z-Ala-Xaa-Yaa-pNA（Xaa=亮氨酸或丙氨酸；Yaa=亮氨酸或苯丙氨酸）和Z-Ala-Ala-Xaa-Yaa-pNA（Xaa=亮氨酸、精氨酸或甘氨酸；Yaa=苯丙氨酸、亮氨酸、甘氨酸、天冬氨酸或谷氨酸）。合成在DMF-乙腈混合物中进行。制备了一系列二肽、三肽和四肽保护物，收率高达99%。研究发现，在所研究的条件下，合成保留了立体选择性。酰化成分羧基的活化对偶联率有积极影响。
• ——
  作者：A. V. Bacheva、O. V. Baibak、A. V. Belyaeva、E. N. Lysogorskaya、E. S. Oksenoit、V. I. Lozinskii、I. Yu. Filippova

  DOI：10.1023/a:1026013912147

  日期：——

  that of the native subtilisin. The use of N-acylpeptides with a free carboxyl group was found to be possible in organic solvents during the enzymatic synthesis catalyzed by both native and immobilized subtilisin. A series of tetrapeptide p-nitroanilides of the general formula Z-Ala-Ala-Xaa-Yaa-pNA (where Xaa is Leu, or Glu and Yaa is Phe or Asp) was obtained in the presence of immobilized enzyme in yields

  在低水含量的有机溶剂混合物中，取决于介质的组成，研究了肽偶联反应中天然枯草杆菌蛋白酶，其与聚丙烯酸酯的非共价复合物以及共价固定在聚乙烯醇冷冻凝胶中的枯草杆菌蛋白酶的催化效率，反应时间和生物催化剂浓度。已经确定，在DMF含量＞ 80％的培养基中，修饰的枯草杆菌蛋白酶的合酶活性高于天然枯草杆菌蛋白酶的合酶活性。发现在天然和固定的枯草杆菌蛋白酶催化的酶促合成过程中，可以在有机溶剂中使用具有游离羧基的N-酰基肽。一系列通式为Z-Ala-Ala-Xaa-Yaa-pNA的四肽对硝基苯胺（其中Xaa是Leu，
• ——
  作者：I. Yu. Filippova、E. N. Lysogorskaya

  DOI：10.1023/a:1026061828077

  日期：——

  proteases could catalyze synthesis of a wide variety of peptides of various lengths and structures both in solution and on solid phase in organic solvents. The following modified proteases were studied as catalysts for enzymatic peptide synthesis in polar organic solvents (acetonitrile, dimethylformamide, and ethanol): pepsin sorbed on celite, a noncovalent complex of subtilisin with sodium dodecylsulfate

  我们表明，修饰的蛋白酶可以在有机溶剂中的溶液和固相上催化各种长度和结构各异的肽的合成。研究了以下修饰的蛋白酶，作为在极性有机溶剂（乙腈，二甲基甲酰胺和乙醇）中酶促肽合成的催化剂：吸附在硅藻土上的胃蛋白酶，枯草杆菌蛋白酶与十二烷基硫酸钠的非共价复合物，以及枯草杆菌蛋白酶或嗜热菌蛋白酶共价固定在聚乙烯基冷冻凝胶上醇。枯草杆菌蛋白酶与十二烷基硫酸钠和固定的枯草杆菌蛋白酶的非共价复合物的使用对于多肽片段的片段缩合特别有希望，该片段包含侧链中带有未保护离子源基团的三官能氨基酸残基，例如Lys，Arg，His，Glu和Asp 。
• ——
  作者：I. Yu. Filippova、A. V. Bacheva、O. V. Baibak、F. M. Plieva、E. N. Lysogorskaya、E. S. Oksenoit、V. I. Lozinsky

  DOI：10.1023/a:1014350600760

  日期：——

  Covalent immobilization of subtilisin and thermolysin on cryogel of poly(vinyl alcohol) was carried out. The biocatalysts obtained are characterized by high stability in water and in DMF-MeCN mixtures of various compositions. The synthetic efficiency of immobilized subtilisin in the multiple iterative synthesis of the peptide Z-Ala-Ala-Leu-Phe-pNA was examined in organic mixtures of different solvent compositions, Immobilized subtilisin exhibits high synthetic activity in organic media. A series of N-acylated p-nitroanilides of tetrapeptides of the general formula Z-Ala-Ala-Xaa-Yaa-pNA (Z is benzyloxycarbonyl, Xaa = Leu, Lys, or Glu, Yaa = Phe or Asp; pNA = 4-NO2-C6H4NH-) were synthesized in 70-98% yields using immobilized subtilisin as a biocatalyst without activation and protection of the ionogenic groups of polyfunctional amino acids. Immobilized thermolysin in a DMF-MeCN mixture catalyzed the formation of the peptide Z-Ala-Ala-Leu-pNA, which was obtained in 90% yield (during 1 h). It was demonstrated that the biocatalyst can be used repeatedly and that it retained activity after storage in an aqueous buffer during 6 months.
• Native subtilisin Karlsberg and modified subtilisin 72 as effective catalysts of peptide bond formation in organic media
  作者：O. M. Anikina、T. A. Semashko、E. S. Oksenoit、E. N. Lysogorskaya、I. Yu. Filippova

  DOI：10.1134/s1068162006020026

  日期：2006.3

  The activity and stability of native subtilisin Karlsberg and subtilisin 72 and their complexes with sodium dodecyl sulfate (SDS) in organic solvents were studied. The kinetic constants of the hydrolysis of specific chromogenic peptide substrates Z-Ala-Ala-Leu-pNA and Glp-Ala-Ala-Leu-pNA by the subtilisins were determined. It was found that the subtilisin Karlsberg complex with SDS in anhydrous organic solvents is an effective catalyst of peptide synthesis with multifunctional amino acids in positions P-1 and P-1', (Glu, At-, and Asp) containing unprotected side ionogenic groups.