Carminomycin(1+)

• 分子结构分类: 有机化合物 - 苯丙烷和聚酮 - 蒽环类药物
• 英文名称: Carminomycin(1+)
• 英文别名: [(2S,3S,4S,6R)-6-[[(1S,3S)-3-acetyl-3,5,10,12-tetrahydroxy-6,11-dioxo-2,4-dihydro-1H-tetracen-1-yl]oxy]-3-hydroxy-2-methyloxan-4-yl]azanium
• 化学式: C26H28NO10+
• 分子量: 514.5
• InChiKey: XREUEWVEMYWFFA-CSKJXFQVSA-O

计算性质

• 辛醇/水分配系数(LogP)：
  1.5
• 重原子数：
  37
• 可旋转键数：
  3
• 环数：
  5.0
• sp3杂化的碳原子比例：
  0.42
• 拓扑面积：
  199
• 氢给体数：
  6
• 氢受体数：
  10

反应信息

• 作为反应物：
  描述：

  Carminomycin(1+) 、 S-腺苷蛋氨酸 生成 (7S,9S)-9-acetyl-7-[(2R,4S,5S,6S)-4-amino-5-hydroxy-6-methyloxan-2-yl]oxy-6,9,11-trihydroxy-4-methoxy-8,10-dihydro-7H-tetracene-5,12-dione;hydron 、 氢(+1)阳离子 、 S-(5'-腺苷)-L-高半胱氨酸
  参考文献：
  名称：

  Partial purification and properties of carminomycin 4-O-methyltransferase from Streptomyces sp. strain C5

  摘要：

  一种甲基转移酶作用于卡米霉素和13-二氢卡米霉素，被认为是柔红霉素生物合成途径中的末端酶，从产生柔红霉素和鲍麦霉素的链霉菌C5菌株中纯化得到接近纯的酶。该酶的纯化得率约为5%，比用30-50%硫酸铵沉淀的样品的特定活性高114倍。聚丙烯酰胺凝胶电泳在变性条件下显示亚基分子量为41000左右。凝胶过滤色谱显示该酶的分子量约为166000，表明它是一种同四聚体。动力学分析表明它对S-腺苷-L-蛋氨酸的亲和力与典型的抗生素甲基转移酶相似；该酶对卡米霉素的亲和力略高于13-二氢卡米霉素。卡米霉素甲基化的反应产物经色谱和质谱分析证实为柔红霉素。纯化的酶不催化无糖苷卡米霉素酮或13-二氢卡米霉素酮的甲基化。S-腺苷-L-高半胱氨酸抑制甲基转移酶，而高半胱氨酸、腺苷、腺嘌呤、ε-罗霉素酮、柔红霉素和柔红霉素酮几乎没有抑制活性。

  DOI：

  10.1099/00221287-139-6-1353
• 作为产物：
  描述：

  (1S,3S)-3,5,10,12-tetrahydroxy-3-[(1R)-1-hydroxyethyl]-6,11-dioxo-1,2,3,4,6,11-hexahydrotetracen-1-yl 3-azaniumyl-2,3,6-trideoxy-alpha-L-lyxo-hexopyranoside 、 氢(+1)阳离子 、 NADPH(4-) 、 氧气 生成 Carminomycin(1+) 、 水 、 NADP⁺
  参考文献：
  名称：

  In vivo and in vitro bioconversion of epsilon-rhodomycinone glycoside to doxorubicin: functions of DauP, DauK, and DoxA

  摘要：

  我们最近确定了Streptomyces sp.菌株C5 doxA基因产物的功能，这是一种类似于细胞色素P-450的蛋白质，被确定为多柔比星C-14羟化酶(M. L. Dickens和W. R. Strohl，J. Bacteriol. 178: 3389-3395，1996)。在本研究中，我们展示了DoxA也可以催化13-去氧卡尔米霉素和13-去氧多柔比星的羟基化反应，分别形成13-二氢卡尔米霉素和13-二氢多柔比星，同时氧化13-二氢蒽环类化合物为它们相应的13-酮形式。Streptomyces sp.菌株C5 dauP基因产物也被明确地证明可以去除epsilon-洛多霉素酮-糖苷（洛多霉素D）的羧甲氧基，形成10-羧基-13-去氧卡尔米霉素。此外，发现Streptomyces sp.菌株C5 DauK可以在4-羟基位甲基化蒽环类化合物洛多霉素D，10-羧基-13-去氧卡尔米霉素和13-去氧卡尔米霉素，表明它具有比以前已知的更广泛的底物特异性。Streptomyces sp.菌株C5 doxA、dauK和dauP的产物足以并且必要地赋予Streptomyces lividans TK24将洛多霉素和多柔比星生物合成中的第一个糖苷洛多霉素D转化为多柔比星的能力。

  DOI：

  10.1128/jb.179.8.2641-2650.1997

文献信息

• Purification, Properties, and Characterization of Recombinant <i>Streptomyces</i> sp. Strain C5 DoxA, a Cytochrome P-450 Catalyzing Multiple Steps in Doxorubicin Biosynthesis
  作者：Robbie J. Walczak、Michael L. Dickens、Nigel D. Priestley、William R. Strohl

  DOI：10.1128/jb.181.1.298-304.1999

  日期：1999.1

  DoxA is a cytochrome P-450 monooxygenase involved in the late stages of daunorubicin and doxorubicin biosynthesis that has a broad substrate specificity for anthracycline glycone substrates. Recombinant DoxA was purified to homogeneity from Streptomyces lividans transformed with a plasmid containing the Streptomyces sp. strain C5 doxA gene under the control of the strong SnpR-activated snpA promoter. The purified enzyme was a monomeric, soluble protein with an apparent M _r of 47,000. Purified DoxA catalyzed the 13-hydroxylation of 13-deoxydaunorubicin, the 13-oxidation of 13-dihydrocarminomycin and 13-dihydrodaunorubicin, and the 14-hydroxylation of daunorubicin. The pH optimum for heme activation was pH 7.5, and the temperature optimum was 30°C. The k _cat / K _m values for the oxidation of anthracycline substrates by purified DoxA, incubated with appropriate electron-donating components, were as follows: for 13-deoxydaunorubicin, 22,000 M ⁻¹ · s ⁻¹ ; for 13-dihydrodaunorubicin, 14,000 M ⁻¹ · s ⁻¹ ; for 13-dihydrocarminomycin, 280 M ⁻¹ · s ⁻¹ ; and for daunorubicin, 130 M ⁻¹ · s ⁻¹ . Our results indicate that the conversion of daunorubicin to doxorubicin by this enzyme is not a favored reaction and that the main anthracycline flux through the late steps of the daunorubicin biosynthetic pathway catalyzed by DoxA is likely directed through the 4- O -methyl series of anthracyclines.

  摘要 DoxA 是一种细胞色素 P-450 单加氧酶，参与达柔比星和多柔比星生物合成的后期阶段，对蒽环类糖酮底物具有广泛的底物特异性。重组的 DoxA 可从铁锈色链霉菌（Streptomyces lividans）中纯化至均一。 重组 DoxA 转化了含有 Streptomyces 菌株 C5 doxA 基因的质粒进行转化。 snpA 启动子的控制下。纯化的酶是一种单体可溶性蛋白，其表观分子量为 M r 为 47,000。纯化的 DoxA 催化了 13-脱氧大柔比星的 13-羟基化、13-二氢金霉素和 13-二氢大柔比星的 13-氧化以及大柔比星的 14-羟基化。血红素活化的最佳 pH 值为 7.5，最佳温度为 30°C。在 k cat / K m 纯化的 DoxA 与适当的电子供能成分孵育后氧化蒽环类底物的值如下：13-脱氧腺苷酸，22,000 M -1 - s -1 ；13-二氢杜冷丁为 14,000 M -1 - s -1 ；对于 13-二氢金霉素，280 M -1 - s -1 ；而对于 daunorubicin，则为 130 M -1 - s -1 .我们的研究结果表明，该酶将 daunorubicin 转化为多柔比星并不是一个有利的反应，由 DoxA 催化的通过 daunorubicin 生物合成途径后期步骤的主要蒽环类通量很可能是通过 4- O. O -甲基系列蒽环类化合物。
• Bioconversion of the Anthracycline Analogue Desacetyladriamycin by Recombinant DoxA, a P450-Monooxygenase from <i>Streptomyces</i> sp. Strain C5
  作者：Robbie J. Walczak、Jennifer V. Hines、William R. Strohl、Nigel D. Priestley

  DOI：10.1021/ol015998x

  日期：2001.7.1

  [GRAPHICS]A recombinant P450-monooxygenase, DoxA, obtained from Streptomyces sp, strain C5, the producer of the anticancer compound daunorubicin, was expressed in S. lividans TK24 and therein used to catalyze the conversion of the anthracycline analogue desacetyladriamycin into the new anthracycline, 10-hydroxydesacetyladriamycin. This work establishes a new function for DoxA and demonstrates the use of a recombinant enzyme to prepare a new anthracycline analogue.
• Crystal Structure of a Ternary Complex of DnrK, a Methyltransferase in Daunorubicin Biosynthesis, with Bound Products
  作者：Anna Jansson、Hanna Koskiniemi、Pekka Mäntsälä、Jarmo Niemi、Gunter Schneider

  DOI：10.1074/jbc.m407081200

  日期：2004.9

  One of the final steps in the biosynthesis of the widely used anti-tumor drug daunorubicin in Streptomyces peucetius is the methylation of the 4-hydroxyl group of the tetracyclic ring system. This reaction is catalyzed by the S-adenosyl-L-methionine-dependent carminomycin 4-O-methyltransferase DnrK. The crystal structure of the ternary complex of this enzyme with the bound products S-adenosyl-L-homocysteine and 4-methoxy-epsilon-rhodomycin T has been determined to a 2.35-Angstrom resolution. DnrK is a homodimer, and the subunit displays the typical fold of small molecule O-methyltransferases. The structure provides insights into the recognition of the anthracycline substrate and also suggests conformational changes as part of the catalytic cycle of the enzyme. The position and orientation of the bound ligands are consistent with an S(N)2 mechanism of methyl transfer. Mutagenesis experiments on a putative catalytic base confirm that DnrK most likely acts as an entropic enzyme in that rate enhancement is mainly due to orientational and proximity effects. This contrasts the mechanism of DnrK with that of other O-methyltransferases where acid/base catalysis has been demonstrated to be an essential contribution to rate enhancement.
• In vivo and in vitro bioconversion of epsilon-rhodomycinone glycoside to doxorubicin: functions of DauP, DauK, and DoxA
  作者：M L Dickens、N D Priestley、W R Strohl

  DOI：10.1128/jb.179.8.2641-2650.1997

  日期：1997.4

  We recently determined the function of the gene product of Streptomyces sp. strain C5 doxA, a cytochrome P-450-like protein, to be daunorubicin C-14 hydroxylase (M. L. Dickens and W. R. Strohl, J. Bacteriol. 178: 3389-3395, 1996). In the present study, we show that DoxA also catalyzes the hydroxylation of 13-deoxycarminomycin and 13-deoxydaunorubicin to 13-dihydrocarminomycin and 13-dihydrodaunorubicin, respectively, as well as oxidizing the 13-dihydro-anthracyclines to their respective 13-keto forms. The Streptomyces sp. strain C5 dauP gene product also was shown unequivocally to remove the carbomethoxy group of the epsilon-rhodomycinone-glycoside (rhodomycin D) to form 10-carboxy-13-deoxycarminomycin. Additionally, Streptomyces sp. strain C5 DauK was found to methylate the anthracyclines rhodomycin D, 10-carboxy-13-deoxycarminomycin, and 13-deoxy-carminomycin, at the 4-hydroxyl position, indicating a broader substrate specificity than was previously known. The products of Streptomyces sp. strain C5 doxA, dauK, and dauP were sufficient and necessary to confer on Streptomyces lividans TK24 the ability to convert rhodomycin D, the first glycoside in daunorubicin and doxorubicin biosynthesis, to doxorubicin.

  我们最近确定了Streptomyces sp.菌株C5 doxA基因产物的功能，这是一种类似于细胞色素P-450的蛋白质，被确定为多柔比星C-14羟化酶(M. L. Dickens和W. R. Strohl，J. Bacteriol. 178: 3389-3395，1996)。在本研究中，我们展示了DoxA也可以催化13-去氧卡尔米霉素和13-去氧多柔比星的羟基化反应，分别形成13-二氢卡尔米霉素和13-二氢多柔比星，同时氧化13-二氢蒽环类化合物为它们相应的13-酮形式。Streptomyces sp.菌株C5 dauP基因产物也被明确地证明可以去除epsilon-洛多霉素酮-糖苷（洛多霉素D）的羧甲氧基，形成10-羧基-13-去氧卡尔米霉素。此外，发现Streptomyces sp.菌株C5 DauK可以在4-羟基位甲基化蒽环类化合物洛多霉素D，10-羧基-13-去氧卡尔米霉素和13-去氧卡尔米霉素，表明它具有比以前已知的更广泛的底物特异性。Streptomyces sp.菌株C5 doxA、dauK和dauP的产物足以并且必要地赋予Streptomyces lividans TK24将洛多霉素和多柔比星生物合成中的第一个糖苷洛多霉素D转化为多柔比星的能力。
• Partial purification and properties of carminomycin 4-O-methyltransferase from Streptomyces sp. strain C5
  作者：N. C. Connors、W. R. Strohl

  DOI：10.1099/00221287-139-6-1353

  日期：1993.6.1

  A methyltransferase that acts on carminomycin and 13-dihydrocarminomycin, and that is postulated to be the terminal enzyme in the daunomycin biosynthesis pathway, was purified to near-homogeneity from the daunomycin-and baumycin-producing Streptomyces sp. strain C5. The enzyme was obtained in approximately 5% yield with a purification of 114-fold in specific activity over the sample precipitated with 30–50% ammonium sulphate. Polyacrylamide gel electrophoresis under denaturing conditions indicated a subunit M r of about 41000. The enzyme was shown by gel filtration chromatography to have an M r of approximately 166000, suggesting that it is a homotetramer. Kinetic analysis indicated an affinity for S-adenosyl-l-methionine typical of antibiotic methyltransferases; the enzyme had a slightly higher affinity for carminomycin than for 13-dihydrocarminomycin. The reaction product from methylation of carminomycin was confirmed by chromatography and mass spectral analysis to be daunomycin. The purified enzyme did not catalyse methylation of the aglycones carminomycinone or 13-dihydrocarminomycinone. S-Adenosyl-l-homocysteine inhibited the methyltransferase, whereas homocysteine, adenosine, adenine, ε-rhodomycinone, daunomycin, and daunomycinone showed little or no inhibitory activity.

  一种甲基转移酶作用于卡米霉素和13-二氢卡米霉素，被认为是柔红霉素生物合成途径中的末端酶，从产生柔红霉素和鲍麦霉素的链霉菌C5菌株中纯化得到接近纯的酶。该酶的纯化得率约为5%，比用30-50%硫酸铵沉淀的样品的特定活性高114倍。聚丙烯酰胺凝胶电泳在变性条件下显示亚基分子量为41000左右。凝胶过滤色谱显示该酶的分子量约为166000，表明它是一种同四聚体。动力学分析表明它对S-腺苷-L-蛋氨酸的亲和力与典型的抗生素甲基转移酶相似；该酶对卡米霉素的亲和力略高于13-二氢卡米霉素。卡米霉素甲基化的反应产物经色谱和质谱分析证实为柔红霉素。纯化的酶不催化无糖苷卡米霉素酮或13-二氢卡米霉素酮的甲基化。S-腺苷-L-高半胱氨酸抑制甲基转移酶，而高半胱氨酸、腺苷、腺嘌呤、ε-罗霉素酮、柔红霉素和柔红霉素酮几乎没有抑制活性。