Reaction equation
[O-]P(OP([O-])([O-])=O)(=O)[O-].C1C(=O)NC(=O)N([C@@H]2O[C@H](COP([O:28][P:29]([O:32][C@H:33]3[O:38][C@H:37]([CH2:39][OH:40])[C@@H:36]([OH:41])[C@H:35]([OH:42])[C@H:34]3[OH:43])([OH:31])=[O:30])(O)=O)[C@@H](O)[C@H]2O)C=1.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>>[CH2:39]([OH:40])[C@H:37]1[O:38][CH:33]([O:32][P:29]([OH:31])([OH:30])=[O:28])[C@H:34]([OH:43])[C@@H:35]([OH:42])[C@@H:36]1[OH:41]Participants
Procedure
The multi-enzyme system (Scheme 15) started with 1-13C-Gal, [99 Atom Percent, purchased from Isotec Inc., Miamisburg, Ohio), GlcNAcβOallyl (Compound 40), (Lee et al., Carbohydr. Res., 37:193 (1974)] phosphoenolpyruvate (PEP), and catalytic amounts of Glc-1-P, ATP and UDP. UDP was converted into UTP with pyruvate kinase (PK; EC 2.7.1.40) and PEP, and UTP reacted with Glc-1-P catalyzed by UDPGP to produce UDP-Glc. The byproduct inorganic pyrophosphate (PPi) was decomposed by inorganic pyrophosphatase (PPase; EC 3.6.1.1). With Gal-1-P UT, UDP-Glc reacted with 13C-Gal-1-P, generated from 13C-Gal and ATP in the presence of GK, to give UDP-13C-Gal and Glc-1-P. The 13C-Gal of UDP-13C-Gal was transferred onto the acceptor (GlcNAcβOallyl) by GalT to give [Gal-1-13C]-containing LacNAcβOallyl (Compound 41). The produced UDP was again converted to UTP by a reaction of PK and PEP, which reacted with the released Glc-1-P to regenerate UDP-Glc. Using this multienzyme system, [Gal-1-13C]-LacNAcβOallyl was obtained in 54 percent yield. The same procedure was also used in the preparation of unlabelled LacNAc and analogs. Exemplary analogs 41a-c are illustrated in the scheme.
Sources & evidence
- ORD record
- ord-b4975cd3a5a54d8d90565cd62b25ed00
- Source dataset
- uspto-grants-2001_11
- Patent number
- US06319695B1
Linked notes
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