N-(3alpha,7alpha,12alpha-trihydroxy-5beta-cholan-24-oyl)glycinate

• 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: N-(3alpha,7alpha,12alpha-trihydroxy-5beta-cholan-24-oyl)glycinate
• 英文别名: 2-[[(4R)-4-[(3R,5S,7R,8R,9S,10S,12S,13R,14S,17R)-3,7,12-trihydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]pentanoyl]amino]acetate
• 化学式: C26H42NO6-
• 分子量: 464.6
• InChiKey: RFDAIACWWDREDC-FRVQLJSFSA-M

计算性质

• 辛醇/水分配系数(LogP)：
  3.6
• 重原子数：
  33
• 可旋转键数：
  5
• 环数：
  4.0
• sp3杂化的碳原子比例：
  0.92
• 拓扑面积：
  130
• 氢给体数：
  4
• 氢受体数：
  6

反应信息

• 作为反应物：
  描述：

  N-(3alpha,7alpha,12alpha-trihydroxy-5beta-cholan-24-oyl)glycinate 、 水 生成 Cholate 、 聚甘氨酸
  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：

  choloyl-CoA(4-) 、 聚甘氨酸 生成 coenzyme A 、 N-(3alpha,7alpha,12alpha-trihydroxy-5beta-cholan-24-oyl)glycinate 、 氢(+1)阳离子
  参考文献：
  名称：

  大鼠肝胆汁酸CoA连接酶的分子克隆和表达。

  摘要：

  胆汁酸CoA连接酶（BAL）负责催化胆汁酸与氨基酸结合的第一步。推定的大鼠肝脏BAL cDNA的序列确定了cDNA（rBAL-1），其具有51个核苷酸的5'非翻译区，2,070个碱基的开放阅读框，编码分子量为75,960 Da的690 aa蛋白和138个核苷酸3 '-非翻译区，后接poly（A）尾巴。cDNA的同一性是通过以下方法建立的：1）通过对纯化的rBAL蛋白进行化学测序获得的rBAL-1开放阅读框编码的肽。2）在SDS-聚丙烯酰胺凝胶电泳中表达与纯化的rBAL相对应的rBAL-1蛋白。3）在昆虫Sf9细胞中表达的rBAL-1具有与从大鼠肝脏分离的酶相当的酶学性质。顺式不饱和脂肪酸对rBAL-1的特异性抑制及其与人超长链脂肪酸CoA连接酶的高度同源性提示了脂肪酸与胆汁酸代谢之间关系的证据。总之，这些结果表明已经分离出大鼠肝脏BAL的cDNA，并且在昆虫Sf9细胞中rBAL cDNA的

  DOI：

  10.1194/jlr.m200260-jlr200

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
• Cloning and expression of a conjugated bile acid hydrolase gene from Lactobacillus plantarum by using a direct plate assay
  作者：H Christiaens、R J Leer、P H Pouwels、W Verstraete

  DOI：10.1128/aem.58.12.3792-3798.1992

  日期：1992.12

  The conjugated bile acid hydrolase gene from the silage isolate Lactobacillus plantarum 80 was cloned and expressed in Escherichia coli MC1061. For the screening of this hydrolase gene within the gene bank, a direct plate assay developed by Dashkevicz and Feighner (M. P. Dashkevicz and S. D. Feighner, Appl. Environ. Microbiol. 53:331-336, 1989) was adapted to the growth requirements of E. coli. Because of hydrolysis and medium acidification, hydrolase-active colonies were surrounded with big halos of precipitated, free bile acids. This phenomenon was also obtained when the gene was cloned into a multicopy shuttle vector and subsequently reintroduced into the parental Lactobacillus strain. The cbh gene and surrounding regions were characterized by nucleotide sequence analysis. The deduced amino acid sequence was shown to have 52% similarity with a penicillin V amidase from Bacillus sphaericus. Preliminary characterization of the gene product showed that it is a cholylglycine hydrolase (EC 3.5.1.24) with only slight activity against taurine conjugates. The optimum pH was between 4.7 and 5.5. Optimum temperature ranged from 30 to 45 degrees C. Southern blot analysis indicated that the cloned gene has similarity with genomic DNA of bile acid hydrolase-active Lactobacillus spp. of intestinal origin.

  从青贮分离的植物乳杆菌80中克隆和表达了共轭胆汁酸水解酶基因，并在大肠杆菌MC1061中进行了筛选。为了筛选该水解酶基因，采用了Dashkevicz和Feighner（M.P. Dashkevicz和S.D. Feighner，应用环境微生物学，53：331-336，1989）开发的直接平板检测法，并根据大肠杆菌的生长要求进行了改进。由于水解和介质酸化，水解酶活性菌落周围出现了大量的沉淀胆汁酸自由酸环。当将该基因克隆到多拷贝穿梭载体中，并随后重新引入到母体植物乳杆菌菌株中时，也观察到了这种现象。通过核苷酸序列分析对cbh基因及其周围区域进行了表征。推导出的氨基酸序列显示，它与来自球形芽孢杆菌的青霉素V酰胺酶有52％的相似性。基因产物的初步表征表明它是一种胆酰甘氨酸水解酶（EC 3.5.1.24），对牛磺酸共轭物的活性仅略微。最适pH在4.7到5.5之间。最适温度范围为30到45摄氏度。南方杂交分析表明，克隆基因与肠源性胆汁酸水解酶活性的植物乳杆菌属基因组DNA具有相似性。
• Cloning and characterization of a conjugated bile acid hydrolase gene from Clostridium perfringens
  作者：J P Coleman、L L Hudson

  DOI：10.1128/aem.61.7.2514-2520.1995

  日期：1995.7

  The gene encoding a conjugated bile acid hydrolase (CBAH) from Clostridium perfringens 13 has been cloned and expressed in Escherichia coli, and its nucleotide sequence has been determined. Nucleotide and predicted amino acid sequence analyses indicated that the gene product is related to two previously characterized amidases, a CBAH from Lactobacillus plantarum (40% identity) and a penicillin V amidase from Bacillus sphaericus (34% identity). The product is apparently unrelated to a CBAH from C. perfringens for which N-terminal sequence information was determined. The gene product was purified from recombinant E. coli and used to raise antibody in rabbits. The presence of the protein in C. perfringens was then confirmed by immunoblot analysis. The protein was shown to have a native molecular weight of 147,000 and a subunit molecular weight of 36,100, indicating its probable existence as a tetramer. Disruption of the chromosomal C. perfringens CBAH gene with a chloramphenicol resistance cartridge resulted in a mutant strain which retained partial CBAH activity. Polyacrylamide gel electrophoresis followed by enzymatic activity staining and immunoblotting indicated that the mutant strain no longer expressed the cloned CBAH (CBAH-1) but did express at least one additional CBAH (CBAH-2). CBAH-2 was immunologically distinct from CBAH-1, and its mobility on native polyacrylamide gels was different from that of CBAH-1. Furthermore, comparisons of pH optima and substrate specificities of CBAH activities from recombinant E. coli and wild-type and mutant C. perfringens provided further evidence for the presence of multiple CBAH activities in C. perfringens.

  来自Clostridium perfringens 13的编码共轭胆汁酸水解酶（CBAH）基因已在大肠杆菌中克隆和表达，并确定了其核苷酸序列。核苷酸和预测的氨基酸序列分析表明，该基因产物与两种先前表征的酰胺酶有关，即来自LactoBAcillus plantarum的CBAH（40％同源性）和来自BAcillus sphaericus的青霉素V酰胺酶（34％同源性）。该产物显然与确定N末端序列信息的C. perfringens的CBAH不相关。从重组E. coli中纯化了基因产物，并用于兔子中提高抗体。然后通过免疫印迹分析确认了C. perfringens中的蛋白质存在。该蛋白质显示具有147,000的天然分子量和36,100的亚基分子量，表明其可能存在为四聚体。使用氯霉素耐药性卡带破坏染色体C. perfringens CBAH基因导致突变株保留部分CBAH活性。聚丙烯酰胺凝胶电泳随后进行酶活性染色和免疫印迹分析表明，突变株不再表达克隆的CBAH（CBAH-1），但至少表达另外一种CBAH（CBAH-2）。CBAH-2在免疫学上与CBAH-1不同，并且其在天然聚丙烯酰胺凝胶上的迁移率与CBAH-1不同。此外，从重组E. coli和野生型和突变C. perfringens中的CBAH活性的pH最适值和底物特异性的比较，为C. perfringens中存在多种CBAH活性提供了进一步的证据。
• Rat liver bile acid CoA:amino acid N-acyltransferase
  作者：Dongning He、Stephen Barnes、Charles N. Falany

  DOI：10.1194/jlr.m300128-jlr200

  日期：2003.12

  Bile acid CoA:amino acid N-acyltransferase (BAT) is responsible for the amidation of bile acids with the amino acids taurine and glycine. Rat liver BAT (rBAT) cDNA was isolated from a rat liver lambdaZAP cDNA library and expressed in Sf9 insect cells using a baculoviral vector. rBAT displayed 65% amino acid sequence homology with human BAT (hBAT) and 85% homology with mouse BAT (mBAT). Similar to hBAT

  胆汁酸CoA：氨基酸N-酰基转移酶（BAT）负责胆汁酸与牛磺酸和甘氨酸的酰胺化。从鼠肝lambdaZAP cDNA文库中分离出鼠肝BAT（rBAT）cDNA，并使用杆状病毒载体在Sf9昆虫细胞中表达。rBAT与人BAT（hBAT）的氨基酸序列同源性为65％，与小鼠BAT（mBAT）的同源性为85％。与hBAT相似，表达的rBAT能够与胆固醇CoA形成牛磺酸和甘氨酸共轭物。与rBAT高度同源的mBAT仅形成牛磺酸缀合的胆汁酸（Falany，CN，H。Fortinberry，EH Leiter和S.BArnes。1997。小鼠肝胆汁酸CoA的克隆和表达：氨基酸N-酰基转移酶。 J.Lipid Res.38：86-95）。大鼠组织的免疫印迹分析仅在匀浆和超速离心后在大鼠肝细胞溶质中检测到rBAT。rBAT的亚细胞定位检测到了细胞溶质和分离的过氧化物酶体中的活性和免疫反应蛋白。大鼠胆汁酸CoA连接
• The critical role of His48 in mouse cytosolic sulfotransferase SULT2A8 for the 7α-hydroxyl sulfation of bile acids
  作者：Takehiko Shimohira、Katsuhisa Kurogi、Ming-Cheh Liu、Masahito Suiko、Yoichi Sakakibara

  DOI：10.1080/09168451.2018.1464897

  日期：2018.8.3

  involved in the homeostasis of steroids and bile acids. SULT2A8, a 7α-hydroxyl bile acid-preferring mouse SULT, has been identified as the major enzyme responsible for the mouse-specific 7-O-sulfation of bile acids. Interestingly, SULT2A8 lacks a conservative catalytic His residue at position 99th. The catalytic mechanism underlying the SULT2A8-mediated 7-O-sulfation of bile acids thus remained unclear. In

  已知胞质磺基转移酶（SULT）SULT2A亚家族的成员严重参与类固醇和胆汁酸的稳态。SULT2A8是7α-羟基胆汁酸偏爱的小鼠SULT，已被确定为负责小鼠特异性7-O-胆汁酸硫酸化的主要酶。有趣的是，SULT2A8在第99位缺少保守的催化His残基。因此，尚不清楚SULT2A8介导的胆汁酸7-O-硫酸化的催化机理。在这项研究中，我们进行了突变分析，以便深入了解这个尚未解决的问题。获得的结果表明，两个氨基酸残基His48和Leu99对小鼠SULT2A8而言是唯一的，但对其他SULTs而言则不是，对于其对胆汁酸的7-O硫酸化活性至关重要。