R-3-(4-hydroxyphenyl)lactate

• 分子结构分类: 有机化合物 - 苯丙烷和聚酮 - 苯丙酸
• 英文名称: R-3-(4-hydroxyphenyl)lactate
• 英文别名: (R)-4-hydroxyphenyllactate；(R)-3-(4-Hydroxyphenyl)lactate；(2R)-2-hydroxy-3-(4-hydroxyphenyl)propanoate
• 化学式: C₉H₉O₄
• 分子量: 181.168
• InChiKey: JVGVDSSUAVXRDY-MRVPVSSYSA-M

计算性质

• 辛醇/水分配系数(LogP)：
  1.4
• 重原子数：
  13
• 可旋转键数：
  2
• 环数：
  1.0
• sp3杂化的碳原子比例：
  0.22
• 拓扑面积：
  80.6
• 氢给体数：
  2
• 氢受体数：
  4

反应信息

• 作为反应物：
  描述：

  R-3-(4-hydroxyphenyl)lactate 、 (E)-4-coumaroyl-CoA 生成 3-(4'-hydroxyphenyl)-trans-2-propenoate 、 (R)-3-(4-hydroxyphenyl)lactoyl-CoA
  参考文献：
  名称：

  辅酶A酯参与了产气荚膜梭菌（R.-phenyllactate）脱水生成（E）-肉桂酸酯的过程。

  摘要：

  在L-苯丙氨酸上厌氧生长的产孢梭状芽孢杆菌的苯乳酸脱水酶催化（R）-苯乳酸可逆脱水合成为（E）-肉桂酸酯。纯化产生由FldA（46kDa），F1dB（43kDa）和FldC（40kDa）组成的异三聚酶复合物（130 +/- 15kDa）。通过对Q-琼脂糖进行重层析，可以分离出FldA的大部分，并将其鉴定为对氧不敏感的肉桂酰基-CoA：苯基乳酸CoA-转移酶，而消耗转移酶的三聚体保留了对氧敏感的苯基乳酸脱水酶的活性，并含有约一个[4Fe- 4S]群集。脱水酶活性需要10 microM FAD，0.4 mM ATP，2.5 mM MgCl2、0.1 mM NADH，5 microM肉桂酰-CoA和少量无细胞提取物（每毫升10 microg蛋白）类似于酸发酵球菌中的2-羟基戊二酰-CoA脱水酶。均质的FldA的N-末端（39个氨基酸）与大肠杆菌中的肉碱代谢有关的CaiB的N-末端（39％序列同一

  DOI：

  10.1046/j.1432-1327.2000.01427.x
• 作为产物：
  描述：

  在 citrate synthase 、 acetyl-CoA synthetase 、 Agrobacterium tumefaciens C₅₈ recombinant Atu₃₂₆₆ protein 、 L-malate dehydrogenase 、 β-烟酰胺腺嘌呤二核苷酸 、 水 、 5’-三磷酸腺苷 、 辅酶 A 、 magnesium chloride 作用下， 生成 R-3-(4-hydroxyphenyl)lactate
  参考文献：
  名称：

  Functional Annotation and Three-Dimensional Structure of an Incorrectly Annotated Dihydroorotase from cog3964 in the Amidohydrolase Superfamily

  摘要：

  The substrate specificities of two incorrectly annotated enzymes belonging to cog3964 from the amidohydrolase superfamily were determined. This group of enzymes are currently misannotated as either dihydroorotases or adenine deaminases. Atu3266 from Agrobacterium tumefaciens C58 and Oant2987 from Ochrobactrum anthropi ATCC 49188 were found to catalyze the hydrolysis of acetyl-(R)-mandelate and similar esters with values of K-cat/K-m that exceed 10(5) M-1 s(-1). These enzymes do not catalyze the deamination of adenine or the hydrolysis of dihydroorotate. Atu3266 was crystallized and the structure determined to a resolution of 2.62 angstrom. The protein folds as a distorted (beta/alpha)(8) barrel and binds two zincs in the active site. The substrate profile was determined via a combination of computational docking to the three-dimensional structure of Atu3266 and screening of a highly focused library of potential substrates. The initial weak hit was the hydrolysis of N-acetyl-D-serine (k(cat)/K-m = 4 M-1 s(-1)). This was followed by the progressive identification of acetyl-(R)-glycerate (k(cat)/K-m = 4 x 10(2) M-1 s(-1)), acetyl glycolate (k(cat)/K-m = 1.3 x 10(4) M-1 s(-1)), and ultimately acetyl-(R)-mandelate (k(cat)/K-m = 2.8 x 10(2) M-1 s(-1)).

  DOI：

  10.1021/bi301483z

文献信息

• 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。
• The involvement of coenzyme A esters in the dehydration of (R)-phenyllactate to (E)-cinnamate by Clostridium sporogenes
  作者：Sandra Dickert、Antonio J. Pierik、Dietmar Linder、Wolfgang Buckel

  DOI：10.1046/j.1432-1327.2000.01427.x

  日期：2000.6

  retained oxygen sensitive phenyllactate dehydratase activity and contained about one [4Fe-4S] cluster. The dehydratase activity required 10 microM FAD, 0.4 mM ATP, 2.5 mM MgCl2, 0.1 mM NADH, 5 microM cinnamoyl-CoA and small amounts of cell-free extract (10 microg protein per mL) similar to that known for 2-hydroxyglutaryl-CoA dehydratase from Acidaminococcus fermentans. The N-terminus of the homogenous FldA

  在L-苯丙氨酸上厌氧生长的产孢梭状芽孢杆菌的苯乳酸脱水酶催化（R）-苯乳酸可逆脱水合成为（E）-肉桂酸酯。纯化产生由FldA（46kDa），F1dB（43kDa）和FldC（40kDa）组成的异三聚酶复合物（130 +/- 15kDa）。通过对Q-琼脂糖进行重层析，可以分离出FldA的大部分，并将其鉴定为对氧不敏感的肉桂酰基-CoA：苯基乳酸CoA-转移酶，而消耗转移酶的三聚体保留了对氧敏感的苯基乳酸脱水酶的活性，并含有约一个[4Fe- 4S]群集。脱水酶活性需要10 microM FAD，0.4 mM ATP，2.5 mM MgCl2、0.1 mM NADH，5 microM肉桂酰-CoA和少量无细胞提取物（每毫升10 microg蛋白）类似于酸发酵球菌中的2-羟基戊二酰-CoA脱水酶。均质的FldA的N-末端（39个氨基酸）与大肠杆菌中的肉碱代谢有关的CaiB的N-末端（39％序列同一
• Purification, Cloning and Functional Expression of hydroxyphenylpyruvate Reductase Involved in Rosmarinic Acid Biosynthesis in Cell Cultures of Coleus Blumei
  作者：Kyung Hee Kim、Verena Janiak、Maike Petersen

  DOI：10.1023/b:plan.0000036367.03056.b2

  日期：2004.2

  Hydroxyphenylpyruvate reductase ( HPPR) is an enzyme involved in the biosynthesis of rosmarinic acid in Lamiaceae reducing hydroxyphenylpyruvates in dependence of NAD( P) H to the corresponding hydroxyphenyllactates. The HPPR protein was purified from suspension cells of Coleus blumei accumulating high levels of rosmarinic acid by ammonium sulfate precipitation, anion exchange chromatography, hydroxylapatite chromatography, chromatography on 2', 5'- ADP-Sepharose 4B and SDS-polyacrylamide gel electrophoresis. The protein was tryptically digested and the peptides sequenced. Sequence information was used to isolate a full-length cDNA-clone for HPPR ( EMBL accession number AJ507733) by RT-PCR, screening of a C. blumei cDNA-library and 5'-RACE-PCR. The open reading frame of the HPPR-cDNA consists of 939 nucleotides encoding a protein of 313 amino acid residues. The sequence showed that HPPR belongs to the family of D-isomer-specific 2-hydroxyacid dehydrogenases. The HPPR-cDNA was heterologously expressed in Escherichia coli and the protein was shown to catalyse the NAD( P) H-dependent reduction of 4-hydroxyphenylpyruvate to 4-hydroxyphenyllactate and 3,4-dihydroxyphenylpyruvate to 3,4-dihydroxyphenyllactate.
• Kim Y.B.; Uddina M.R.; Kim Y., Nat Prod Commun, 2014, 1934-578X, 1311-4
  作者：Kim Y.B.、Uddina M.R.、Kim Y.、Park C.G.、Park S.U.

  DOI：——

  日期：——
• HPPR encodes the hydroxyphenylpyruvate reductase required for the biosynthesis of hydrophilic phenolic acids in Salvia miltiorrhiza
  作者：Guo-Quan WANG、Jun-Feng CHEN、Bo YI、He-Xin TAN、Lei ZHANG、Wan-Sheng CHEN

  DOI：10.1016/s1875-5364(18)30008-6

  日期：2017.12

  Salvia miltiorrhiza is a medicinal plant widely used in the treatment of cardiovascular and cerebrovascular diseases. Hydrophilic phenolic acids, including rosmarinic acid (RA) and lithospermic acid B (LAB), are its primary medicinal ingredients. However, the biosynthetic pathway of RA and LAB in S. miltiorrhiza is still poorly understood. In the present study, we accomplished the isolation and characterization of a novel S. miltiorrhiza Hydroxyphenylpyruvate reductase (HPPR) gene, SmHPPR, which plays an important role in the biosynthesis of RA. SmHPPR contained a putative catalytic domain and a NAD(P)H-binding motif. The recombinant SmHPPR enzyme exhibited high HPPR activity, converting 4-hydroxyphenylpyruvic acid (pHPP) to 4-hydroxyphenyllactic acid (pHPL), and exhibited the highest affinity for substrate 4-hydroxyphenylpyruvate. SmHPPR expression could be induced by various treatments, including SA, GA3, MeJA and Ag+, and the changes in SmHPPR activity were correlated well with hydrophilic phenolic acid accumulation. SmHPPR was localized in cytoplasm, most likely close to the cytosolic NADPH-dependent hydroxypyruvate reductase active in photorespiration. In addition, the transgenic S. miltiorrhiza hairy roots overexpressing SmHPPR exhibited up to 10-fold increases in the products of hydrophilic phenolic acid pathway. In conclusion, our findings provide a new insight into the synthesis of active pharmaceutical compounds at molecular level.