Chenodeoxycholate anion

• 分子结构分类: 有机化合物 - 脂质和类脂质分子 - 类固醇和类固醇衍生物
• 英文名称: Chenodeoxycholate anion
• 英文别名: (4R)-4-[(3R,5S,7R,8R,9S,10S,13R,14S,17R)-3,7-dihydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]pentanoate
• 化学式: C24H39O4-
• 分子量: 391.6
• InChiKey: RUDATBOHQWOJDD-BSWAIDMHSA-M

计算性质

• 辛醇/水分配系数(LogP)：
  5.6
• 重原子数：
  28
• 可旋转键数：
  3
• 环数：
  4.0
• sp3杂化的碳原子比例：
  0.96
• 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：

  Chenodeoxycholate anion 、 聚甘氨酸 生成 Glycochenodeoxycholate (1-) 、 水
  参考文献：
  名称：

  长双歧杆菌胆汁盐水解酶的生化和遗传学表征。

  摘要：

  从长双歧杆菌SBT2928分离出胆汁盐水解酶（BSH），纯化并鉴定。此外，我们首次描述了双歧杆菌属成员中编码BSH（bsh）的基因的克隆和分析。根据推导的氨基酸序列测定，该酶的天然分子量为125,000-130,000，亚单位分子量为35,024，表明该酶是四聚体。长双歧杆菌BSH的最适pH为5至7，最适温度为40℃。该酶被硫醇酶抑制剂强烈抑制，表明Cys残基可能参与催化反应。长双歧杆菌的BSH可以水解所有六种主要的人胆汁盐和至少两种动物胆汁盐。基于特异性和K（m）值，检测到对甘氨酸缀合的胆汁酸略有偏爱。确定了bsh的核苷酸序列，并将其用于同源性研究，转录本分析以及各种突变体的构建和分析。与其他细菌的BSH和球形芽孢杆菌的青霉素V酰基转移酶（PVA）的同源性很高。基于已经阐明晶体结构的BSH和PVA的相似性，可以将BSH分类为N末端亲核水解酶，而将Cys作为N末端氨基酸。通过定点诱变进行的

  DOI：

  10.1128/aem.66.6.2502-2512.2000
• 作为产物：
  描述：

  (3alpha,5beta,7alpha)-3-(beta-D-glucopyranosyloxy)-7-hydroxycholan-24-oate 、 水 生成 Chenodeoxycholate anion 、 D-葡萄糖
  参考文献：
  名称：

  Purification and Characterization of a Microsomal Bile Acid β-Glucosidase from Human Liver

  摘要：

  A human liver microsomal beta-glucosidase has been purified to apparent homogeneity in sodium dodecyl sulfate-polyacrylamide gel electrophoresis where a single protein band of Mr 100,000 was obtained under reducing conditions. The enzyme was enriched about 73, 000-fold over starting microsomal membranes by polyethylene glycol fractionation, anion exchange chromatographies on DEAE-Trisacryl, and Mono Q followed by affinity chromatography on N-(9-carboxynonyl)-1-deoxynojirimycin-AH-Sepharose 4B. The purified enzyme had a pH optimum between 5.0 and 6.4, was activated by divalent metal ions, and required phospholipids for exhibition of activity. The enzyme catalyzed the hydrolysis of 3beta-D-glucosido-lithocholic and 3beta-D-glucosido-chenodeoxycholic acids with high affinity (Km, 1.7 and 6.2 microM, respectively) and of the beta-D-glucoside (Km, 210 microM) and the beta-D-galactoside of 4-methylumbelliferone. The ratio of relative reaction rates for these substrates was about 6:3:11:1. No activity was detectable toward 6beta-D-glucosido-hyodeoxycholic acid, glucocerebroside, and the following glycosides of 4-methylumbelliferone: alpha-D-glucoside, alpha-L-arabinoside, beta-D-fucoside or beta-D-xyloside. Immunoinhibition and immunoprecipitation studies using antibodies prepared against lysosomal glucocerebrosidase showed no cross-reactivity with microsomal beta-glucosidase suggesting that these two enzymes are antigenically unrelated.

  DOI：

  10.1074/jbc.272.17.11261

文献信息

• The Human Bile Acid-CoA:Amino Acid N-Acyltransferase Functions in the Conjugation of Fatty Acids to Glycine
  作者：James O'Byrne、Mary C. Hunt、Dilip K. Rai、Masayumi Saeki、Stefan E.H. Alexson

  DOI：10.1074/jbc.m300987200

  日期：2003.9

  Bile acid-CoA:amino acid N-acyltransferase (BACAT) catalyzes the conjugation of bile acids to glycine and taurine for excretion into bile. By use of site-directed mutagenesis and sequence comparisons, we have identified Cys-235, Asp-328, and His-362 as constituting a catalytic triad in human BACAT (hBACAT) and identifying BACAT as a member of the type I acyl-CoA thioesterase gene family. We therefore

  胆汁酸-CoA：氨基酸N-酰基转移酶（BACAT）催化胆汁酸与甘氨酸和牛磺酸的结合，从而排泄到胆汁中。通过使用定点诱变和序列比较，我们已鉴定出Cys-235，Asp-328和His-362构成人BACAT（hBACAT）中的催化三联体，并将BACAT鉴定为I型酰基辅酶A硫酯酶基因家族。因此，我们假设hBACAT也可能将脂肪酰基辅酶A和/或共轭脂肪酸水解为甘氨酸。我们在这里显示重组hBACAT也可以水解长链和非常长链的饱和酰基CoAs（主要是C16：0-C26：0），并且通过质谱法验证了hBACAT还可以将脂肪酸缀合到甘氨酸上。组织表达研究表明，BACAT在肝，胆囊以及近端和远端肠中都有强表达。然而，BACAT还可以在与胆汁酸形成和运输无关的多种组织中表达，这表明在调节长链脂肪酸的细胞内水平方面也起着重要的作用。在人皮肤成纤维细胞中的绿色荧光蛋白定位实验表明，hBACAT酶主要是胞质的。因此
• The bile acid-inducible baiB gene from Eubacterium sp. strain VPI 12708 encodes a bile acid-coenzyme A ligase
  作者：D H Mallonee、J L Adams、P B Hylemon

  DOI：10.1128/jb.174.7.2065-2071.1992

  日期：1992.4

  baiB gene from Eubacterium sp. strain VPI 12708 was previously cloned, sequenced, and shown to be part of a large bile acid-inducible operon encoding polypeptides believed to be involved in bile acid 7 alpha-dehydroxylation. In the present study, the baiB gene was subcloned and expressed in Escherichia coli and shown to encode a bile acid-coenzyme A (CoA) ligase. This ligase required a C-24 bile acid

  来自Eubacterium sp。的baiB基因。菌株VPI 12708先前已被克隆，测序，并显示出是大胆汁酸诱导的操纵子编码多肽的一部分，该多肽被认为与胆汁酸7α-脱羟基有关。在本研究中，baiB基因被亚克隆并在大肠杆菌中表达，并显示其编码胆汁酸辅酶A（CoA）连接酶。该连接酶需要具有游离羧基，ATP，Mg2 +和CoA的C-24胆汁酸来合成最终的胆汁酸-CoA共轭物。通过反相高效液相色谱法进行的产物分析显示，最终反应产物与胆甾醇CoA和AMP竞争。当反应混合物中未加入CoA时，检测到推测的胆汁酸-AMP中间体。胆汁酸-CoA连接酶与涉及AMP或CoA的ATP依赖性连接的其他几种多肽的氨基酸序列相似，与环状羧化化合物。胆汁酸-CoA的连接被认为是真细菌中胆汁酸7α-脱羟基途径的第一步。VPI 12708株。
• Actinobacterial Acyl Coenzyme A Synthetases Involved in Steroid Side-Chain Catabolism
  作者：Israël Casabon、Kendra Swain、Adam M. Crowe、Lindsay D. Eltis、William W. Mohn

  DOI：10.1128/jb.01012-13

  日期：2014.2

  activate the C8 side chain of cholesterol. Both CasGRHA1 and FadD17Mtb are steroid-24-oyl-CoA synthetases. CasG and its orthologs activate the C5 side chain of cholate, while FadD17 and its orthologs appear to activate the C5 side chain of one or more cholesterol metabolites. CasIRHA1 is a steroid-22-oyl-CoA synthetase, representing orthologs that activate metabolites with a C3 side chain, which accumulate

  细菌类固醇分解代谢是全球碳循环的重要组成部分，在药物合成中有应用。这种分解代谢的途径涉及多种酰基辅酶 A (CoA) 合成酶，它们激活链烷酸酯取代基进行 β-氧化。这些合成酶的功能知之甚少。我们对来自约氏红球菌 RHA1 的胆酸盐分解代谢途径和结核分枝杆菌的胆固醇分解代谢途径的四种不同的酰基辅酶 A 合成酶进行了酶促表征。对预测参与类固醇代谢的 70 种酰基辅酶 A 合成酶进行的系统发育分析表明，所表征的合成酶各自代表在类固醇侧链降解中具有独特功能的直系同源类。合成酶对链烷酸酯取代基的长度具有特异性。FadD19 来自 M。结核分枝杆菌 H37Rv (FadD19Mtb) 转化 3-oxo-4-cholesten-26-oate (kcat/Km = 0.33 × 10(5) ± 0.03 × 10(5) M(-1) s(-1)) 并代表直向同源物激活胆固醇的 C8 侧链。CasGRHA1
• Expanded substrate screenings of human and Drosophila type 10 17β-hydroxysteroid dehydrogenases (HSDs) reveal multiple specificities in bile acid and steroid hormone metabolism: characterization of multifunctional 3α/7α/7β/17β/20β/21-HSD
  作者：Naeem SHAFQAT、Hanns-Ulrich MARSCHALL、Charlotta FILLING、Erik NORDLING、Xiao-Qiu WU、Lars BJÖRK、Johan THYBERG、Eva MÅRTENSSON、Samina SALIM、Hans JÖRNVALL、Udo OPPERMANN

  DOI：10.1042/bj20030877

  日期：2003.11.15

  17β-Hydroxysteroid dehydrogenases (17β-HSDs) catalyse the conversion of 17β-OH (-hydroxy)/17-oxo groups of steroids, and are essential in mammalian hormone physiology. At present, eleven 17β-HSD isoforms have been defined in mammals, with different tissue-expression and substrate-conversion patterns. We analysed 17β-HSD type 10 (17β-HSD10) from humans and Drosophila, the latter known to be essential in development. In addition to the known hydroxyacyl-CoA dehydrogenase, and 3α-OH and 17β-OH activities with sex steroids, we here demonstrate novel activities of 17β-HSD10. Both species variants oxidize the 20β-OH and 21-OH groups in C21 steroids, and act as 7β-OH dehydrogenases of ursodeoxycholic or isoursodeoxycholic acid (also known as 7β-hydroxylithocholic acid or 7β-hydroxyisolithocholic acid respectively). Additionally, the human orthologue oxidizes the 7α-OH of chenodeoxycholic acid (5β-cholanic acid, 3α,7α-diol) and cholic acid (5β-cholanic acid). These novel substrate specificities are explained by homology models based on the orthologous rat crystal structure, showing a wide hydrophobic cleft, capable of accommodating steroids in different orientations. These properties suggest that the human enzyme is involved in glucocorticoid and gestagen catabolism, and participates in bile acid isomerization. Confocal microscopy and electron microscopy studies reveal that the human form is localized to mitochondria, whereas Drosophila 17β-HSD10 shows a cytosolic localization pattern, possibly due to an N-terminal sequence difference that in human 17β-HSD10 constitutes a mitochondrial targeting signal, extending into the Rossmann-fold motif.

  17β-羟类固醇脱氢酶（17β-HSDs）催化类固醇中 17β-OH （-羟基）/17-氧代基团的转化，是哺乳动物激素生理过程中必不可少的物质。目前，哺乳动物中已确定有 11 种 17β-HSD 同工酶，它们的组织表达和底物转换模式各不相同。我们分析了人类和果蝇的 17β-HSD 10 型（17β-HSD10），已知后者在发育过程中至关重要。除了已知的羟基乙酰-CoA 脱氢酶、3α-OH 和 17β-OH 与性类固醇的活性之外，我们在此还证明了 17β-HSD10 的新活性。这两种变体都能氧化 C21 类固醇中的 20β-OH 和 21-OH 基团，并作为熊去氧胆酸或异熊去氧胆酸（也分别称为 7β-hydroxylithocholic acid 或 7β-hydroxyisolithocholic acid）的 7β-OH 脱氢酶发挥作用。此外，人类同源物还能氧化去氧胆酸（5β-胆酸，3α，7α-二醇）和胆酸（5β-胆酸）的 7α-OH 。这些新的底物特异性可以用基于同源大鼠晶体结构的同源模型来解释，该结构显示了一个宽阔的疏水裂隙，能够容纳不同方向的类固醇。这些特性表明，人类酶参与了糖皮质激素和孕激素的分解代谢，并参与了胆汁酸异构化。共聚焦显微镜和电子显微镜研究显示，人类的 17β-HSD10 定位于线粒体，而果蝇的 17β-HSD10 则显示出细胞质定位模式，这可能是由于人类 17β-HSD10 的 N 端序列差异造成的。
• Cloning and sequencing of the 7 alpha-hydroxysteroid dehydrogenase gene from Escherichia coli HB101 and characterization of the expressed enzyme
  作者：T Yoshimoto、H Higashi、A Kanatani、X S Lin、H Nagai、H Oyama、K Kurazono、D Tsuru

  DOI：10.1128/jb.173.7.2173-2179.1991

  日期：1991.4

  The 7 alpha-hydroxysteroid dehydrogenase (EC 1.1.1.159) gene from Escherichia coli HB101 was cloned and expressed in E. coli DH1. The hybrid plasmid pSD1, with a 2.8-kbp insert of chromosomal DNA at the BamHI site of pBR322, was subcloned into pUC19 to construct plasmid pSD3. The entire nucleotide sequence of an inserted PstI-BamHI fragment of plasmid pSD3 was determined by the dideoxy chain-termination method. Within this sequence, the mature enzyme protein-encoding sequence was found to start at a GTG initiation codon and to comprise 765 bp, as judged by comparison with the protein sequence. The deduced amino acid sequence of the enzyme indicated that the molecular weight is 26,778. The transformant of E. coli DH1 harboring pSD3 with a 1.8-kbp fragment showed about 200-fold-higher enzyme activity than the host. The enzyme was purified by a single chromatography step on DEAE-Toyopearl and obtained as crystals, with an activity yield of 39%. The purified enzyme was homogeneous, as judged by sodium dodecyl sulfate gel electrophoresis. The enzyme was most active at pH 8.5 and stable between pH 8 and 9. The enzyme was NAD+ dependent and had a pI of 4.3. The molecular mass was estimated to be 120 kDa by the gel filtration method and 28 kDa by electrophoresis, indicating that the enzyme exists in a tetrameric form.

  大肠杆菌HB101的7α-羟类固醇脱氢酶（EC 1.1.1.159）基因被克隆并在大肠杆菌DH1中表达。将含有2.8-kbp染色体DNA插入到pBR322的BamHI位点的杂交质粒pSD1亚克隆到pUC19中构建出质粒pSD3。通过dideoxy链终止法确定了质粒pSD3的插入PstI-BamHI片段的整个核苷酸序列。在该序列中，成熟酶蛋白编码序列从GTG启动密码子开始，由765个碱基组成，与蛋白质序列比较得出。酶的推导氨基酸序列表明分子量为26,778。携带1.8-kbp片段的大肠杆菌DH1转化体，其酶活性比宿主高约200倍。酶经DEAE-Toyopearl单一色谱步骤纯化，并以晶体形式获得，活性收率为39％。纯化的酶通过十二烷基硫酸钠凝胶电泳判断为均一。该酶在pH 8.5时最活跃，在pH 8和9之间稳定。该酶依赖于NAD+，其等电点为4.3。通过凝胶过滤法估计分子量为120 kDa，通过电泳法估计为28 kDa，表明该酶以四聚体形式存在。