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

HRID 429738

Reaktionsgleichung

Reaktionsgleichung von HRID 429738
[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]

Komponenten

7

Edukt 5

HCID 5235Sodium FluorideHCID 5884NADPHHCID 440994Lacto-N-bioseHCID 178434963HCID 178434963, VerbindungHCID 178508932HCID 178508932, Verbindung

Produkt 1

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

Reagenzien, Katalysatoren, Lösungsmittel 1

HCID 3776IsopropanolLösungsmittel

Versuchsvorschrift

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).]

Aufarbeitung

  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

Quellen und Nachweise

ORD-Eintrag
ord-65c9469dadb846bebc350df6d831b48b
Quelldatensatz
uspto-grants-2001_11
Patentnummer
US06319695B1

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