Sodium Fluoride → beta-D-Galp-(1->3)-(alpha-L-Fucp-(1->4))-D-GlcpNAc

HRID 429738

反応式

HRID 429738 の反応式
[O:1]([C@H:13]1[C@H:19]([OH:20])[C@@H:18]([CH2:21][OH:22])[O:17][CH:15]([OH:16])[C@@H:14]1[NH:23][C:24]([CH3:26])=[O:25])[C@@H:2]1[O:10][C@H:9]([CH2:11][OH:12])[C@H:7]([OH:8])[C@H:5]([OH:6])[C@H:3]1[OH:4].[F-].[Na+].C1N=C(N)C2N=CN([C@@H]3O[C@H](COP(OP(OC[C@H]4O[C@@H](N5C=C(C(N)=O)CC=C5)[C@H](O)[C@@H]4O)(O)=O)(O)=O)[C@@H](O)[C@H]3OP(O)(O)=O)C=2N=1.C1N([C@@H]2O[C@H](COP(OP(O[C@H:97]3[O:102][C@H:101]([CH2:103]O)[C@@H:100]([OH:105])[C@H:99]([OH:106])[C@@H:98]3[OH:107])(O)=O)(O)=O)[C@@H](O)[C@H]2O)C2NC(N)=NC(=O)C=2N=1.CC1OC(OP(OP(OCC2OC(N3C4NC(N)=NC(=O)C=4N=C3)C(O)C2O)(O)=O)(O)=O)C(O)C(O)C1O>CC(O)C>[O:1]([C@H:13]1[C@H:19]([O:20][C@@H:97]2[O:102][C@@H:101]([CH3:103])[C@@H:100]([OH:105])[C@@H:99]([OH:106])[C@@H:98]2[OH:107])[C@@H:18]([CH2:21][OH:22])[O:17][CH:15]([OH:16])[C@@H:14]1[NH:23][C:24]([CH3:26])=[O:25])[C@@H:2]1[O:10][C@H:9]([CH2:11][OH:12])[C@H:7]([OH:8])[C@H:5]([OH:6])[C@H:3]1[OH:4]

参加物

7

反応物 5

HCID 5235Sodium FluorideHCID 5884NADPHHCID 440994Lacto-N-bioseHCID 178434963HCID 178434963、化合物HCID 178508932HCID 178508932、化合物

生成物 1

HCID 22806751beta-D-Galp-(1->3)-(alpha-L-Fucp-(1->4))-D-GlcpNAc

試薬・触媒・溶媒 1

HCID 3776Isopropanol溶媒

実験手順

A mixture of GTP Na salt (6.0 mg, 10 μmol), Man-1-P K salt (3.5 mg, 10 μmol), Galβ1,3GlcNAc (3.8 mg, 10 μmol), NaF (0.42 mg, 10 μmol), NADPH (9.4 mg, 10 μmol), PEP K salt (4.1 mg, 20 μmol), MgCl2—6H2O (2.6 mg, 10 μmol), MnCl2—4H2O (2 mg, 10 μmol), 2-propanol, (50 μL), ADH (12 U), PK (200 U), PPase (100 U), crude enzyme preparation of GDP-mannose pyrophosphorylase (1.0 mL), and crude enzyme preparation of GDP-Fuc producing enzyme (1.0 mL) in 100 mM tris buffer (pH 7.5) and diluted to 3 mL. α1,3/4-Fucosyltransferase (0.01 U) was added to the mixture and the resulting mixture was stirred under Ar for three days at room temperature. The mixture was filtered and the filtrate was applied to a column of Dowex 1-X8 [OH−) form followed with a column of Dowex 50W-X8 [H+] with water. The fractions were collected and lyophilized. The residual material was purified with a column of Sephadex G-25 (superfine) with water. The appropriate fractions were pooled and lyophilized to give Galβ1,3(Fucα1,4)GlcNAc. Its 1H-NMR spectrum was in good agreement with that reported. [Dumas et al., Biomed. Chem. Lett., 1:425 (1991).]

後処理

  1. additiondiluted to 3 mL
  2. additionα1,3/4-Fucosyltransferase (0.01 U) was added to the mixture
  3. filtrationThe mixture was filtered
  4. customThe fractions were collected
  5. customThe residual material was purified with a column of Sephadex

出典と根拠

ORDレコード
ord-65c9469dadb846bebc350df6d831b48b
出典データセット
uspto-grants-2001_11
特許番号
US06319695B1

関連ノート

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