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

HRID 429738

Reaction equation

HRID 429738 reaction equation
[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]

Participants

7

Reactant 5

HCID 5235Sodium FluorideHCID 5884NADPHHCID 440994Lacto-N-bioseHCID 178434963HCID 178434963, compoundHCID 178508932HCID 178508932, compound

Product 1

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

Reagents, catalysts & solvents 1

HCID 3776IsopropanolSolvent

Procedure

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).]

Workup

  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

Sources & evidence

ORD record
ord-65c9469dadb846bebc350df6d831b48b
Source dataset
uspto-grants-2001_11
Patent number
US06319695B1

Linked notes

Loading…