2,5-Didehydro-D-gluconate

• 分子结构分类: 有机化合物 - 有机酸及其衍生物 - 酮酸及其衍生物
• 英文名称: 2,5-Didehydro-D-gluconate
• 英文别名: (3S,4S)-3,4,6-trihydroxy-2,5-dioxohexanoate
• 化学式: C6H7O7-
• 分子量: 191.12
• InChiKey: RXMWXENJQAINCC-DMTCNVIQSA-M

计算性质

• 辛醇/水分配系数(LogP)：
  -1.8
• 重原子数：
  13
• 可旋转键数：
  4
• 环数：
  0.0
• sp3杂化的碳原子比例：
  0.5
• 拓扑面积：
  135
• 氢给体数：
  3
• 氢受体数：
  7

反应信息

• 作为反应物：
  描述：

  2,5-Didehydro-D-gluconate 、 氢(+1)阳离子 、 NADH 生成 5-dehydro-D-gluconate 、 nicotinamide adenine dinucleotide
  参考文献：
  名称：

  The yiaE Gene, Located at 80.1 Minutes on the Escherichia coli Chromosome, Encodes a 2-Ketoaldonate Reductase

  摘要：

  摘要 位于 bisC-cspA 基因间区域的开放阅读框，或位于 大肠杆菌 编码一个假定的 2-羟基酸脱氢酶。 大肠杆菌 基因组测序项目的结果。我们在此报告，该基因的产物 ( yiaE ) 是一种 2-Ketoaldonate 还原酶（2KR）。该基因被克隆并以 C 端 His 标记表达于 大肠杆菌 用金属螯合物亲和层析法纯化了该蛋白。NH 2 -末端氨基酸序列，确定了该基因的翻译起始位点。研究发现，该酶是一种 2KR 催化剂，可催化 2,5-二酮还原为 d -葡萄糖酸还原为 5-酮- d d -葡萄糖酸、2-酮 d -葡萄糖酸（2KDG）转化为 d -葡萄糖酸 2-酮 l -转化为 l -还原酶在 pH 值为 7.5 时活性最佳，NADPH 是首选的电子供体。该还原酶在 pH 值为 7.5 时活性最佳，NADPH 是首选的电子供体。推导出的氨基酸序列与来自 的 2KR .该基因在染色体上的断裂导致 2KR 在大肠杆菌中失去活性。 大肠杆菌。大肠杆菌 W3110 可以在 2KDG 上生长，而缺乏 2KR 活性的突变体则无法在 2KDG 作为碳源的条件下生长。 大肠杆菌 产生的 2KDG 减少。 d -葡萄糖酸产生的 2KDG 减少。 大肠杆菌 葡萄糖酸脱氢酶基因。

  DOI：

  10.1128/jb.180.22.5984-5988.1998
• 作为产物：
  描述：

  2-ketogluconate 、 A 生成 2,5-Didehydro-D-gluconate 、 AH₂
  参考文献：
  名称：

  2-酮葡萄糖酸被黑醋杆菌的部分纯化的酶氧化。

  摘要：

  DOI：

  10.1016/0003-9861(56)90218-1

文献信息

• Identification of the <i>yqhE</i> and <i>yafB</i> Genes Encoding Two 2,5-Diketo- <scp>d</scp> -Gluconate Reductases in <i>Escherichia coli</i>
  作者：Do-Young Yum、Bong-Yong Lee、Jae-Gu Pan

  DOI：10.1128/aem.65.8.3341-3346.1999

  日期：1999.8

  The identification of a gene ( yiaE ) encoding 2-ketoaldonate reductase (2KR) in our previous work led to the hypothesis that Escherichia coli has other ketogluconate reductases including 2,5-diketo- d -gluconate reductase (25DKGR) and to study of the related ketogluconate metabolism. By using the deduced amino acid sequences of 5-diketo- d -gluconate reductase (5KDGR) of Gluconobacter oxydans and 25DKGR of Corynebacterium sp., protein databases were screened to detect homologous proteins. Among the proteins of E. coli , an oxidoreductase encoded by yjgU and having 56% similarity to 5KDGR of G. oxydans and two hypothetical oxidoreductases encoded by yqhE and yafB and having 49.8 and 42% similarity, respectively, to 25DKGR of Corynebacterium sp. were detected. Recently, the yjgU gene was identified as encoding 5KDGR and renamed idnO (C. Bausch, N. Peekhaus, C. Utz, T. Blais, E. Murray, T. Lowary, and T. Conway, J. Bacteriol. 180:3704–3710, 1998). The pathways involved in the metabolism of ketogluconate by E. coli have been predicted by biochemical analysis of purified enzymes and chemical analysis of the pathway intermediates. The gene products of yqhE and yafB were identified as 25DKGR-A, and 25DKGR-B, respectively, catalyzing the reduction of 25KDG to 2-keto- l -gulonate (2KLG). The native 25DKGR-A, 25DKGR-B, and 5KDGR had apparent molecular weights of about 30,000, 30,000, and 54,000, respectively. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels, all three enzymes showed protein bands with a molecular weight of about 29,000, which indicated that 25DKGR-A, 25DKGR-B, and 5KDGR may exist as monomeric, monomeric, and dimeric proteins, respectively. The optimum pHs for reduction were 7.5, 7.0, and 8.0, respectively. The 5KDGR was active with NADH, whereas 25DKGR-A and 25DKGR-B were active with NADPH as a preferred electron donor. 25DKG can be converted to 5KDG by 2KR, which is then reduced to d -gluconate by 5KDGR. The pathways were compared with those of Erwinia sp. and Corynebacterium sp. A BLAST search of published and incomplete microbial genome sequences revealed that the ketogluconate reductases and their related metabolism may be widespread in many species.

  摘要 一种基因( yiaE 编码 2-酮醛酸还原酶（2KR）的基因，从而提出了这样的假设 大肠杆菌 有其他酮基葡萄糖酸还原酶，包括 2,5-二酮-2KR。 d -葡萄糖酸还原酶（25DKGR），并对相关的酮基葡萄糖酸代谢进行了研究。通过推导 5-二酮-d-葡萄糖酸还原酶（25DKGR）的氨基酸序列，研究了相关的酮基葡萄糖酸代谢。 d -葡萄糖酸还原酶（5KDGR）的氨基酸序列，对其相关的酮基葡萄糖酸代谢进行研究。 和 25DKGR 和 25DKGR 的 25DKGR 的氨基酸序列。 蛋白数据库来检测同源蛋白。在这些蛋白质中 大肠杆菌 编码的氧化还原酶 yjgU 编码的氧化还原酶与大肠杆菌的 5KDGR 有 56% 的相似性。 氧化还原酶 编码的氧化还原酶，以及由 yqhE 和 yafB 的 25DKGR 的相似度分别为 49.8% 和 42%。 的 25DKGR 相似度分别为 49.8%和 42%。 的 25DKGR 相似度分别为 49.8 和 42。最近 yjgU 基因被确定为编码 5KDGR 的基因，并被重新命名为 idnO (最近，yjgU 基因被确定为编码 5KDGR 并更名为 idnO（C. Bausch、N. Peekhaus、C. Utz、T. Blais、E. Murray、T. Lowary 和 T. Conway，J. Bacteriol.180:3704-3710, 1998).大肠杆菌代谢酮基葡萄糖酸的途径 大肠杆菌 通过对纯化酶的生化分析和途径中间产物的化学分析，预测了大肠杆菌代谢酮基葡萄糖酸盐的途径。其中 yqhE 和 yafB 的基因产物分别被鉴定为 25DKGR-A 和 25DKGR-B，它们催化 25KDG 还原为 2-keto- l -丙酮酸（2KLG）。原生 25DKGR-A、25DKGR-B 和 5KDGR 的表观分子量分别约为 30,000、30,000 和 54,000。在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳凝胶中，三种酶都显示出分子量约为 29,000 的蛋白质条带，这表明 25DKGR-A、25DKGR-B 和 5KDGR 可能分别以单体、单体和二聚体蛋白质的形式存在。还原的最佳 pH 值分别为 7.5、7.0 和 8.0。5KDGR 在 NADH 的作用下具有活性，而 25DKGR-A 和 25DKGR-B 在 NADPH 作为首选电子供体时具有活性。25DKG 可通过 2KR 转化为 5KDG，然后被还原为 d -葡萄糖酸。将这些途径与 Erwinia 和 和 Corynebacterium 通过对已发表的和不完整的微生物基因组序列进行 BLAST 搜索，发现酮基葡萄糖酸还原酶及其相关代谢可能广泛存在于许多物种中。
• Crystal structure of 2,5-diketo- <scp>d</scp> -gluconic acid reductase A complexed with NADPH at 2.1-Å resolution
  作者：Sumit Khurana、David B. Powers、Stephen Anderson、Michael Blaber

  DOI：10.1073/pnas.95.12.6768

  日期：1998.6.9

  The three-dimensional structure of Corynebacterium 2,5-diketo- d -gluconic acid reductase A (2,5-DKGR A; EC 1.1.1.-), in complex with cofactor NADPH, has been solved by using x-ray crystallographic data to 2.1-Å resolution. This enzyme catalyzes stereospecific reduction of 2,5-diketo- d -gluconate (2,5-DKG) to 2-keto- l -gulonate. Thus the three-dimensional structure has now been solved for a prokaryotic example of the aldo–keto reductase superfamily. The details of the binding of the NADPH cofactor help to explain why 2,5-DKGR exhibits lower binding affinity for cofactor than the related human aldose reductase does. Furthermore, changes in the local loop structure near the cofactor suggest that 2,5-DKGR will not exhibit the biphasic cofactor binding characteristics observed in aldose reductase. Although the crystal structure does not include substrate, the two ordered water molecules present within the substrate-binding pocket are postulated to provide positional landmarks for the substrate 5-keto and 4-hydroxyl groups. The structural basis for several previously described active-site mutants of 2,5-DKGR A is also proposed. Recent research efforts have described a novel approach to the synthesis of l -ascorbate (vitamin C) by using a genetically engineered microorganism that is capable of synthesizing 2,5-DKG from glucose and subsequently is transformed with the gene for 2,5-DKGR. These modifications create a microorganism capable of direct production of 2-keto- l -gulonate from d -glucose, and the gulonate can subsequently be converted into vitamin C. In economic terms, vitamin C is the single most important specialty chemical manufactured in the world. Understanding the structural determinants of specificity, catalysis, and stability for 2,5-DKGR A is of substantial commercial interest.

  使用X射线晶体学数据解析了与辅因子NADPH结合的Corynebacterium 2,5-二酮-D-葡萄糖酸还原酶A（2,5-DKGR A; EC 1.1.1.-）的三维结构，分辨率为2.1埃。该酶催化2,5-二酮-D-葡萄糖酸（2,5-DKG）的立体特异性还原为2-酮-L-古龙酸。因此，现在已经解决了一种原核生物的醛酮还原酶超家族的三维结构。NADPH辅因子结合的细节有助于解释为什么2,5-DKGR的结合亲和力比相关的人类醛糖还原酶低。此外，辅因子附近的局部环结构的变化表明，2,5-DKGR不会表现出醛糖还原酶观察到的双相辅因子结合特性。虽然晶体结构不包括底物，但位于底物结合口袋内的两个有序水分子被推测为底物5-酮基和4-羟基的位置标志。还提出了2,5-DKGR A的几个先前描述的活性位点突变的结构基础。最近的研究努力描述了一种新的方法，即使用能够从葡萄糖合成2,5-DKG的基因工程微生物，随后转化为2,5-DKGR基因，从而创建一种能够直接从D-葡萄糖生产2-酮-L-古龙酸的微生物，然后将古龙酸转化为维生素C。在经济方面，维生素C是全球最重要的特种化学品。了解2,5-DKGR A的特异性、催化和稳定性的结构决定因素具有重要的商业利益。
• The oxidation of 2-ketogluconate by a partially purified enzyme from Acetobactor melanogenum
  作者：Asoke G. Datta、H. Katznelson

  DOI：10.1016/0003-9861(56)90218-1

  日期：1956.12
• The <i>yiaE</i> Gene, Located at 80.1 Minutes on the <i>Escherichia coli</i> Chromosome, Encodes a 2-Ketoaldonate Reductase
  作者：Do-Young Yum、Bong-Yong Lee、Dae-Hyum Hahm、Jae-Gu Pan

  DOI：10.1128/jb.180.22.5984-5988.1998

  日期：1998.11.15

  An open reading frame located in the bisC-cspA intergenic region, or at 80.1 min on the Escherichia coli chromosome, encodes a hypothetical 2-hydroxyacid dehydrogenase, which was identified as a result of the E. coli Genome Sequencing Project. We report here that the product of the gene ( yiaE ) is a 2-ketoaldonate reductase (2KR). The gene was cloned and expressed with a C-terminal His tag in E. coli , and the protein was purified by metal-chelate affinity chromatography. The determination of the NH ₂ -terminal amino acid sequence of the protein defined the translational start site of this gene. The enzyme was found to be a 2KR catalyzing the reduction of 2,5-diketo- d -gluconate to 5-keto- d -gluconate, 2-keto- d -gluconate (2KDG) to d -gluconate, 2-keto- l -gulonate to l -idonate. The reductase was optimally active at pH 7.5, with NADPH as a preferred electron donor. The deduced amino acid sequence showed 69.4% identity with that of 2KR from Erwinia herbicola . Disruption of this gene on the chromosome resulted in the loss of 2KR activity in E. coli. E. coli W3110 was found to grow on 2KDG, whereas the mutant deficient in 2KR activity was unable to grow on 2KDG as the carbon source, suggesting that 2KR is responsible for the catabolism of 2KDG in E. coli and the diminishment of produced 2KDG from d -gluconate in the cultivation of E. coli harboring a cloned gluconate dehydrogenase gene.

  摘要 位于 bisC-cspA 基因间区域的开放阅读框，或位于 大肠杆菌 编码一个假定的 2-羟基酸脱氢酶。 大肠杆菌 基因组测序项目的结果。我们在此报告，该基因的产物 ( yiaE ) 是一种 2-Ketoaldonate 还原酶（2KR）。该基因被克隆并以 C 端 His 标记表达于 大肠杆菌 用金属螯合物亲和层析法纯化了该蛋白。NH 2 -末端氨基酸序列，确定了该基因的翻译起始位点。研究发现，该酶是一种 2KR 催化剂，可催化 2,5-二酮还原为 d -葡萄糖酸还原为 5-酮- d d -葡萄糖酸、2-酮 d -葡萄糖酸（2KDG）转化为 d -葡萄糖酸 2-酮 l -转化为 l -还原酶在 pH 值为 7.5 时活性最佳，NADPH 是首选的电子供体。该还原酶在 pH 值为 7.5 时活性最佳，NADPH 是首选的电子供体。推导出的氨基酸序列与来自 的 2KR .该基因在染色体上的断裂导致 2KR 在大肠杆菌中失去活性。 大肠杆菌。大肠杆菌 W3110 可以在 2KDG 上生长，而缺乏 2KR 活性的突变体则无法在 2KDG 作为碳源的条件下生长。 大肠杆菌 产生的 2KDG 减少。 d -葡萄糖酸产生的 2KDG 减少。 大肠杆菌 葡萄糖酸脱氢酶基因。