5'-Atp → UDP-N-acetylgalactosamine

HRID 928871

反応式

HRID 928871 の反応式
[OH:1][CH:2]1[O:9][C@H:8]([CH2:10][OH:11])[C@H:6]([OH:7])[C@H:4]([OH:5])[C@H:3]1[NH:12][C:13]([CH3:15])=[O:14].P(OC[C@H]1O[C@@H](N2C3N=CN=C(N)C=3N=C2)[C@H](O)[C@@H]1O)(OP(OP(O)(O)=O)(O)=O)(=O)O.[CH:47]1[C:53](=[O:54])[NH:52][C:50](=[O:51])[N:49]([C@@H:55]2[O:59][C@H:58]([CH2:60][O:61][P:62]([O:65][P:66](OP(O)(O)=O)([OH:68])=[O:67])([OH:64])=[O:63])[C@@H:57]([OH:74])[C@H:56]2[OH:75])[CH:48]=1.C(O)C(N)(CO)CO.Cl.[Mg+2].[Cl-].[Cl-]>O.CO.CCOC(C)=O>[CH3:15][C:13]([NH:12][C@H:3]1[C@@H:2]([O:1][P:66]([O:65][P:62]([O:61][CH2:60][C@H:58]2[O:59][C@@H:55]([N:49]3[C:50](=[O:51])[NH:52][C:53](=[O:54])[CH:47]=[CH:48]3)[C@H:56]([OH:75])[C@@H:57]2[OH:74])([OH:64])=[O:63])([OH:68])=[O:67])[O:9][C@H:8]([CH2:10][OH:11])[C@H:6]([OH:7])[C@@H:4]1[OH:5])=[O:14]

参加物

9

反応物 5

HCID 59575'-AtpHCID 6133Uridine 5'-triphosphateHCID 35717N-Acetyl-D-GalactosamineHCID 93573Tromethamine hydrochlorideHCID 5360315Magnesium Chloride

生成物 1

HCID 23700UDP-N-acetylgalactosamine

試薬・触媒・溶媒 3

HCID 887Methanol溶媒HCID 962Water溶媒3 回HCID 8857Ethyl Acetate溶媒

実験手順

One-Pot Three-Enzyme Synthesis of uridine 5′-diphospho-2-acetamido-2-deoxy-α-D-glacopyranoside (UDP-GalNAc). GalNAc (100 mg, 1.0 eq), ATP (1.2 eq.), and UTP (1.2 eq.) were dissolved in water in a 50 mL centrifuge tube containing Tris-HCl buffer (100 mM, pH 8.0) and MgCl2 (10 mM). After the addition of NanK_ATCC55813 (3.5 mg), PmGlmU (5 mg), and PmPpA (2.5 mg), water was added to bring the volume of the reaction mixture to 20 mL. The reaction was carried out by incubating the solution in an isotherm incubator at 37° C. for 24 h with gentle shaking. Product formation was monitored by TLC (EtOAc:MeOH:H2O=3:2:1 by volume) with p-anisaldehyde sugar staining. The reaction was stopped by adding the same volume of ice-cold ethanol and incubating at 4° C. for 30 min. The mixture was concentrated and passed through a BioGel P-2 gel filtration column to obtain the desired product. Silica gel column purification (EtOAc:MeOH:H2O=4:2:1) was applied for further purification to give pure target compound. Yield, 83% (228 mg); white foam. 1H NMR (600 MHz, D2O) δ 7.93 (d, J=8.4 Hz, 1H), 5.94-5.96 (m, 2H), 5.55 (dd, J=6.6, 3.0 Hz, 1H), 4.27-4.36 (m, 2H), 4.22-4.27 (m, 3H), 4.16-4.18 (m, 2H), 4.02 (d, J=3.0 Hz, 1H), 3.95 (dd, J=10.8, 3.0 Hz, 1H), 3.71-3.78 (m, 2H), 2.06 (s, 3H). 13C NMR (150 MHz, D2O) δ 175.05, 166.31, 151.85, 141.89, 102.82, 94.75, 88.70, 83.03 (d, J=8.6 Hz), 73.97, 72.32, 69.79, 68.50, 67.64, 65.14, 61.17, 49.95 (d, J=7.8 Hz), 22.24. HRMS (ESI) m/z calcd for C17H28N3O17P2 (M+H) 608.0894, found 608.0906.

後処理

  1. additionwas added
  2. customat 37° C.
  3. customfor 24 h
  4. additionby adding the same volume of ice-cold ethanol
  5. concentrationThe mixture was concentrated
  6. filtrationpassed through a BioGel P-2 gel filtration column

出典と根拠

ORDレコード
ord-7dc54a020d76451896b20307fb5581fb
出典データセット
uspto-grants-2016_03
特許番号
US09290530B2

関連ノート

読み込み中…