Table 3.
Acceptor | Percentage conversion (%) | |||
---|---|---|---|---|
CMP-Sialic acid | Transferase reaction | |||
10 μg/mL, 10 min | 3.3 mg/mL, 10 h | |||
1 | G4 | aCMP-Neu5Ac (Quant.) | 35±0.1 | 97±1 |
2 | 7N3-G4 | 13±1 | 83±1 | |
3 | 9N3-G4 | 14±0.2 | 89±3 | |
4 | 7NAc-G4 | 0 | 85±2 | |
5 | 9NAc-G4 | 0 | 97±1 | |
6 | G4 | bCMP-Neu5Ac7N3 (Quant.) | 0 | 90±1 |
7 | 7N3-G4 | 0 | 82±2 | |
8 | 9N3-G4 | 0 | 36±1 | |
9 | 7NAc-G4 | 0 | 0 | |
10 | 9NAc-G4 | 0 | 0 | |
11 | G4 | bCMP-Neu5Ac9N3 (Quant.) | 0 | 97±1 |
12 | 7N3-G4 | 0 | 83±2 | |
13 | 9N3-G4 | 0 | 31±1 | |
14 | 7NAc-G4 | 0 | 0 | |
15 | 9NAc-G4 | 0 | 0 |
Step 1 of the reaction was carried out with aNeu5Ac, bManNAc4N3, or bManNAc6N3 (1.2 equiv.) in the presence of NmCSS (1.5 mg/mL), CTP (1 equiv.) and Tris-HCl (pH 8.5) awithout or bwith PmAldolase (4 mg/mL) and sodium pyruvate (5 equiv.).