(S)-(+)-ketoprofen ethyl ester

• 分子结构分类: 有机化合物 - 苯类化合物 - 苯和取代衍生物
• 英文名称: (S)-(+)-ketoprofen ethyl ester
• 英文别名: Dexketoprofen Ethyl Ester；ethyl (2S)-2-(3-benzoylphenyl)propanoate
• 化学式: C₁₈H₁₈O₃
• 分子量: 282.339
• InChiKey: CQSMNXCTDMLMLM-ZDUSSCGKSA-N

计算性质

• 辛醇/水分配系数(LogP)：
  3.8
• 重原子数：
  21
• 可旋转键数：
  6
• 环数：
  2.0
• sp3杂化的碳原子比例：
  0.22
• 拓扑面积：
  43.4
• 氢给体数：
  0
• 氢受体数：
  3

反应信息

• 作为产物：
  描述：

  酮洛芬乙酯 在 盐酸 、 水 作用下， 反应 1.0h， 生成 (S)-(+)-ketoprofen ethyl ester
  参考文献：
  名称：

  Improving the enantioselectivity of an esterase toward (S)-ketoprofen ethyl ester through protein engineering

  摘要：

  Esterases, enzymes that hydrolyze ester bonds, have been utilized to synthesize optically pure compounds like (S)-ketoprofen, which can be produced by the esterase-catalyzed hydrolysis of rac-ketoprofen ethyl ester. We previously reported a novel esterase, Est25, which was expressed in active form in Escherichia coli, and showed high hydrolyzing activities for various substrates. However, the enzyme did not show any preference for either of rac-ketoprofen ethyl ester. In this study, to improve the enantioselectivity of Est25, we used a protein engineering approach that involves building a homology model of Est25, library generation based on the model, screening for esterase activity, and screening for specificity toward (S)-ketoprofen ethyl ester. One variant, L255W was isolated showing significantly higher enantioselectivity than the wild-type enzyme. Enantioselectivity was further improved by adding one of the polar organic solvents, namely, ethanol and methanol. However, the mutant enzyme showed lower thermostability, higher sensitivity to pH, and lower resistance to polar solvents than Est25. The L255W mutation improves substrate specificity, but, at the same time, sacrifices structural stability, The approach used in this study is precise enough to engineer the substrate specificity of esterase, and general enough to be adapted to engineer other kinds of enzymes with a minimum amount of modifications. (C) 2013 Elsevier B.V. All rights reserved.

  DOI：

  10.1016/j.molcatb.2013.11.008

文献信息

• Binding of a chiral drug to a protein: an investigation of the 2-(3-benzoylphenyl)propionic acid/bovine serum albumin system by circular dichroism and fluorescence
  作者：Sandra Monti、Francesco Manoli、Salvatore Sortino、Raffaele Morrone、Giovanni Nicolosi

  DOI：10.1039/b509911k

  日期：——

  constants and the absolute circular dichroism spectra of the diastereomeric complexes were determined. The spectra of the 1 : 1 conjugates are opposite in sign, those of the 2 : 1 complexes are both negative, but different in shape from each other (peaks at 358 and 342 nm for S-(+)- and R-(-)-ketoprofen, respectively). A tryptophan residue was shown to be involved in the binding of the drug, in the primary

  采用圆形二色性多波长数据全局分析和时间分辨荧光的组合方法，研究了中性pH缓冲液中R-（-）-和S-（+）-酮洛芬与牛血清白蛋白的相互作用。获得了最稳定的药物：化学计量比为1：1和2：1的蛋白质加合物的特征，作为单独的化学物种。确定了非对映体复合物的稳定性常数和绝对圆二色性光谱。1：1共轭物的光谱符号相反，而2：1配合物的光谱均为负值，但形状互不相同（S-（+）-和R-（- ）-酮洛芬。色氨酸残基被证明与药物结合有关，
• Improving the enantioselectivity of an esterase toward (S)-ketoprofen ethyl ester through protein engineering
  作者：Sangyoung Yoon、Seungbum Kim、Songyi Park、Eunsoo Hong、Jinyeong Kim、Sangchul Kim、Tae Hyeon Yoo、Yeonwoo Ryu

  DOI：10.1016/j.molcatb.2013.11.008

  日期：2014.2

  Esterases, enzymes that hydrolyze ester bonds, have been utilized to synthesize optically pure compounds like (S)-ketoprofen, which can be produced by the esterase-catalyzed hydrolysis of rac-ketoprofen ethyl ester. We previously reported a novel esterase, Est25, which was expressed in active form in Escherichia coli, and showed high hydrolyzing activities for various substrates. However, the enzyme did not show any preference for either of rac-ketoprofen ethyl ester. In this study, to improve the enantioselectivity of Est25, we used a protein engineering approach that involves building a homology model of Est25, library generation based on the model, screening for esterase activity, and screening for specificity toward (S)-ketoprofen ethyl ester. One variant, L255W was isolated showing significantly higher enantioselectivity than the wild-type enzyme. Enantioselectivity was further improved by adding one of the polar organic solvents, namely, ethanol and methanol. However, the mutant enzyme showed lower thermostability, higher sensitivity to pH, and lower resistance to polar solvents than Est25. The L255W mutation improves substrate specificity, but, at the same time, sacrifices structural stability, The approach used in this study is precise enough to engineer the substrate specificity of esterase, and general enough to be adapted to engineer other kinds of enzymes with a minimum amount of modifications. (C) 2013 Elsevier B.V. All rights reserved.