N-acetyl-L-phenylalanine

• 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: N-acetyl-L-phenylalanine
• 英文别名: N-acetyl-L-phenylalaninate；N-[(1S)-1-carboxy-2-phenylethyl]ethanimidate
• 化学式: C₁₁H₁₂NO₃
• 分子量: 206.221
• InChiKey: CBQJSKKFNMDLON-JTQLQIEISA-M

计算性质

• 辛醇/水分配系数(LogP)：
  1.3
• 重原子数：
  15
• 可旋转键数：
  3
• 环数：
  1.0
• sp3杂化的碳原子比例：
  0.27
• 拓扑面积：
  69.2
• 氢给体数：
  1
• 氢受体数：
  3

反应信息

• 作为反应物：
  描述：

  N-acetyl-L-phenylalanine 、 (+)-tubocurarine 在 phosphate buffer 、 borate buffer 、 acetate buffer 、 氯化筒箭毒碱 作用下， 以 乙腈 为溶剂， 生成
  参考文献：
  名称：

  Godoy-Alcantar, Carolina; Nelen, Marina I.; Eliseev, Alexey V., Journal of the Chemical Society. Perkin Transactions 2 (2001), 1999, # 2, p. 353 - 361

  摘要：

  DOI：
• 作为产物：
  描述：

  N-acetyl-D-phenylalanine 生成 N-acetyl-L-phenylalanine
  参考文献：
  名称：

  Structural Basis for Catalytic Racemization and Substrate Specificity of an N-Acylamino Acid Racemase Homologue from Deinococcus radiodurans

  摘要：

  N-acylamino acid racemase (NAAAR) catalyzes the racemization of N-acylamino acids and can be used in concert with an aminoacylase to produce enantiopure alpha-amino acids, a process that has potential industrial applications. Here we have cloned and characterized an NAAAR homologue from a radiation-resistant ancient bacterium, Deinococcus radiodurans. The expressed NAAAR racemized various substrates at an optimal temperature of 60 degrees C and had Km values of 24.8 mM and 12.3 mM for N-acetyl-D-methionine and N-acetyl-L-methionine, respectively. The crystal structure of NAAAR was solved to 1.3 A resolution using multiwavelength anomalous dispersion (MAD) methods. The structure consists of a homooctamer in which each subunit has an architecture characteristic of enolases with a capping domain and a (beta/alpha)7 beta barrel domain. The NAAAR.Mg2+ and NAAAR.N-acetyl-L-glutamine.Mg2+ structures were also determined, allowing us to define the Lys170-Asp195-Glu220-Asp245-Lys269 framework for catalyzing 1,1-proton exchange of N-acylamino acids. Four subsites enclosing the substrate are identified: catalytic site, metal-binding site, side-chain-binding region, and a flexible lid region. The high conservation of catalytic and metal-binding sites in different enolases reflects the essentiality of a common catalytic platform, allowing these enzymes to robustly abstract alpha-protons of various carboxylate substrates efficiently. The other subsites involved in substrate recognition are less conserved, suggesting that divergent evolution has led to functionally distinct enzymes.

  DOI：

  10.1016/j.jmb.2004.07.023

文献信息

• [EN] CARBONYL SULFIDE-MEDIATED SYNTHESIS OF PEPTIDES WITH AMINO ACID IONIC LIQUIDS<br/>[FR] SYNTHÈSE DE PEPTIDES À MÉDIATION PAR SULFURE DE CARBONYLE AVEC DES LIQUIDES IONIQUES D'ACIDES AMINÉS
  申请人：CHEN MING

  公开号：WO2020034878A1

  公开（公告）日：2020-02-20

  The present invention provides an OCS (carbonyl sulfide)-mediated approach for solid-phase peptide synthesis using amino acid ionic liquids as both recyclable reactants and reaction media. The coupling reactions required no base and solvent and was completed in minutes at room temperature.

  本发明提供了一种以OCS（硫酰基硫醇）为介质的固相肽合成方法，使用氨基酸离子液体作为可回收反应物和反应介质。偶联反应不需要碱和溶剂，并且可以在室温下在几分钟内完成。
• Nontargeted in vitro metabolomics for high-throughput identification of novel enzymes in Escherichia coli
  作者：Daniel C Sévin、Tobias Fuhrer、Nicola Zamboni、Uwe Sauer

  DOI：10.1038/nmeth.4103

  日期：2017.2

  A method to screen proteins for enzymatic activity by incubating purified or overexpressed proteins with a metabolite extract and measuring changes in metabolite abundance using mass spectrometry enables high-throughput characterization of functionally uncharacterized proteins in Escherichia coli. Our understanding of metabolism is limited by a lack of knowledge about the functions of many enzymes. Here, we develop a high-throughput mass spectrometry approach to comprehensively profile proteins for in vitro enzymatic activity. Overexpressed or purified proteins are incubated in a supplemented metabolome extract containing hundreds of biologically relevant candidate substrates, and accumulating and depleting metabolites are determined by nontargeted mass spectrometry. By combining chemometrics and database approaches, we established an automated pipeline for unbiased annotation of the functions of novel enzymes. In screening all 1,275 functionally uncharacterized Escherichia coli proteins, we discovered 241 potential novel enzymes, 12 of which we experimentally validated. Our high-throughput in vitro metabolomics method is generally applicable to any purified protein or crude cell lysate of its overexpression host and enables performing up to 1,200 nontargeted enzyme assays per working day.

  通过将纯化或过表达的蛋白质与代谢物提取物孵育，并使用质谱法测量代谢物丰度的变化，筛选蛋白质的酶活性，这种方法能够对大肠杆菌中功能未定的蛋白质进行高通量表征。我们对代谢的理解受到许多酶功能缺乏了解的限制。在这里，我们开发了一种高通量质谱法，用于全面分析蛋白质的体外酶活性。将过表达或纯化的蛋白质与含有数百种生物相关候选底物的补充代谢物提取物孵育，并通过非靶向质谱法确定累积和耗竭的代谢物。通过结合化学计量学和数据库方法，我们建立了一个自动化的管道，用于对新型酶的功能进行无偏注解。在筛选所有1275种功能未定的大肠杆菌蛋白质时，我们发现了241种潜在的新型酶，其中12种通过实验验证。我们的高通量体外代谢组学方法通常适用于任何纯化的蛋白质或过表达宿主的粗细胞裂解液，每个工作日最多可以进行1200次非靶向酶检测。
• Structural characterization, tissue distribution, and functional expression of murine aminoacylase III
  作者：Alexander Pushkin、Gerardo Carpenito、Natalia Abuladze、Debra Newman、Vladimir Tsuprun、Sergey Ryazantsev、Srilakshmi Motemoturu、Pakan Sassani、Nadezhda Solovieva、Ramnath Dukkipati、Ira Kurtz

  DOI：10.1152/ajpcell.00192.2003

  日期：2004.4

  Many xenobiotics are detoxified through the mercapturate metabolic pathway. The final product of the pathway, mercapturic acids ( N-acetylcysteine S-conjugates), are secreted predominantly by renal proximal tubules. Mercapturic acids may undergo a transformation mediated by aminoacylases and cysteine S-conjugate β-lyases that leads to nephrotoxic reactive thiol formation. The deacetylation of cysteine S-conjugates of N-acyl aromatic amino acids is thought to be mediated by an aminoacylase whose molecular identity has not been determined. In the present study, we cloned aminoacylase III, which likely mediates this process in vivo, and characterized its function and structure. The enzyme consists of 318 amino acids and has a molecular mass (determined by SDS-PAGE) of ∼35 kDa. Under nondenaturing conditions, the molecular mass of the enzyme is ∼140 kDa as determined by size-exclusion chromatography, which suggests that it is a tetramer. In agreement with this hypothesis, transmission electron microscopy and image analysis of aminoacylase III showed that the monomers of the enzyme are arranged with a fourfold rotational symmetry. Northern analysis demonstrated an ∼1.4-kb transcript that was expressed predominantly in kidney and showed less expression in liver, heart, small intestine, brain, lung, testis, and stomach. In kidney, aminoacylase III was immunolocalized predominantly to the apical domain of S1 proximal tubules and the cytoplasm of S2 and S3 proximal tubules. The data suggest that in kidney proximal tubules, aminoacylase III plays an important role in deacetylating mercapturic acids. The predominant cytoplasmic localization of aminoacylase III may explain the greater sensitivity of the proximal straight tubule to the nephrotoxicity of mercapturic acids.

  许多异物质通过巯基酸代谢途径进行解毒。途径的最终产物，巯基酸（N-乙酰半胱氨酸S-结合物），主要由肾近曲小管分泌。巯基酸可能通过由氨酰化酶和半胱氨酸S-结合物β-裂解酶介导的转化而发生反应性硫醇形成的转化。认为N-酰基芳香族氨基酸的半胱氨酸S-结合物的脱乙酰化是由一种分子尚未确定的氨酰化酶介导的。在本研究中，我们克隆了氨酰化酶III，该酶可能在体内介导这一过程，并对其功能和结构进行了表征。该酶由318个氨基酸组成，其分子量（通过SDS-PAGE确定）约为35 kDa。在非变性条件下，该酶的分子量通过分子排阻色谱法确定为约140 kDa，这表明它是四聚体。与此假设一致，氨酰化酶III的透射电子显微镜和图像分析显示，该酶的单体以四倍旋转对称排列。北方分析显示了一个约1.4 kb的转录本，主要在肾脏中表达，在肝脏、心脏、小肠、大脑、肺、睾丸和胃中表达较少。在肾脏中，氨酰化酶III主要免疫定位于S1近曲小管的顶端区域和S2和S3近曲小管的细胞质中。数据表明，在肾近曲小管中，氨酰化酶III在去乙酰化巯基酸中发挥重要作用。氨酰化酶III的主要细胞质定位可能解释了近曲直小管对巯基酸的肾毒性反应更敏感的原因。
• Godoy-Alcantar, Carolina; Nelen, Marina I.; Eliseev, Alexey V., Journal of the Chemical Society. Perkin Transactions 2 (2001), 1999, # 2, p. 353 - 361
  作者：Godoy-Alcantar, Carolina、Nelen, Marina I.、Eliseev, Alexey V.、Yatsimirsky, Anatoly K.

  DOI：——

  日期：——
• Structural Basis for Catalytic Racemization and Substrate Specificity of an N-Acylamino Acid Racemase Homologue from Deinococcus radiodurans
  作者：Wen-Ching Wang、Wei-Chun Chiu、Shih-Kuang Hsu、Chun-Li Wu、Cheng-Yu Chen、Jai-Shin Liu、Wen-Hwei Hsu

  DOI：10.1016/j.jmb.2004.07.023

  日期：2004.9

  N-acylamino acid racemase (NAAAR) catalyzes the racemization of N-acylamino acids and can be used in concert with an aminoacylase to produce enantiopure alpha-amino acids, a process that has potential industrial applications. Here we have cloned and characterized an NAAAR homologue from a radiation-resistant ancient bacterium, Deinococcus radiodurans. The expressed NAAAR racemized various substrates at an optimal temperature of 60 degrees C and had Km values of 24.8 mM and 12.3 mM for N-acetyl-D-methionine and N-acetyl-L-methionine, respectively. The crystal structure of NAAAR was solved to 1.3 A resolution using multiwavelength anomalous dispersion (MAD) methods. The structure consists of a homooctamer in which each subunit has an architecture characteristic of enolases with a capping domain and a (beta/alpha)7 beta barrel domain. The NAAAR.Mg2+ and NAAAR.N-acetyl-L-glutamine.Mg2+ structures were also determined, allowing us to define the Lys170-Asp195-Glu220-Asp245-Lys269 framework for catalyzing 1,1-proton exchange of N-acylamino acids. Four subsites enclosing the substrate are identified: catalytic site, metal-binding site, side-chain-binding region, and a flexible lid region. The high conservation of catalytic and metal-binding sites in different enolases reflects the essentiality of a common catalytic platform, allowing these enzymes to robustly abstract alpha-protons of various carboxylate substrates efficiently. The other subsites involved in substrate recognition are less conserved, suggesting that divergent evolution has led to functionally distinct enzymes.