Table 3. Product distribution determined after incubation of wild-type GTFO-∆N and GTFA-∆N enzymes, and of GTFO-derived mutants, with 100 mM sucrose and 100 mM maltose.
| Enzyme | Oligosaccharide yield (%) | Panose (%) | Maltotriose | Glucosyl-(α1 → 4)-panose | Glucosyl-(α1 → 6)- (α1 → 4)-panose | Maltotetraose |
|---|---|---|---|---|---|---|
| GTFA-ΔN | 69.9 ± 3.4 | 51.1 ± 3.5 | 3.3 ± 0.4 | 8.9 ± 0.9 | 6.5 ± 0.7 | ND |
| GTFO-ΔN | 60.7 ± 2.8 | 29.5 ± 2.7 | 12.1 ± 1.1 | 14.2 ± 0.8 | ND | 4.9 ± 0.8 |
| GTFO-ΔN A977Q | 69.1 ± 3.7 | 44.3 ± 3.1 | 7.0 ± 0.9 | 14.8 ± 1.1 | 0.5 ± 0.2 | 2.5 ± 0.5 |
| GTFO-ΔN lp977 | 68.0 ± 3.8 | 43.2 ± 2.9 | 6.1 ± 1.0 | 15.1 ± 1.6 | 0.5 ± 0.2 | 3.1 ± 0.5 |
| GTFO-ΔN SS-HA | 59.7 ± 2.1 | 29.5 ± 2.3 | 11.9 ± 1.2 | 14.0 ± 1.1 | ND | 4.2 ± 0.6 |
| GTFO-ΔN α4 | 55.6 ± 3.0 | 25.8 ± 2.0 | 10.7 ± 1.4 | 9.9 ± 1.2 | 5.7 ± 0.8 | 3.6 ± 0.5 |
| GTFO-ΔN A977Q/α4 | 65.7 ± 2.1 | 42.4 ± 2.4 | 5.8 ± 0.6 | 9.2 ± 1.1 | 6.2 ± 1.2 | 2.0 ± 0.5 |
| GTFO-ΔN lp977/α4 | 67.8 ± 6.3 | 43.8 ± 3.8 | 5.5 ± 0.8 | 9.4 ± 1.2 | 6.7 ± 1.0 | 2.4 ± 0.4 |
| GTFO-ΔN SS-HA/α4 | 58.0 ± 1.5 | 26.6 ± 1.7 | 11.4 ± 1.5 | 12.6 ± 1.5 | 2.1 ± 0.4 | 5.2 ± 0.5 |
| GTFO-ΔN lp977/SS-HA | 69.0 ± 3.7 | 43.0 ± 4.3 | 6.4 ± 1.0 | 16.1 ± 1.6 | ND | 3.4 ± 0.6 |
| GTFO-ΔN lp977/SS-HA/α4 | 68.7 ± 3.5 | 40.2 ± 2.1 | 7.2 ± 1.1 | 14.2 ± 0.9 | 3.8 ± 1.1 | 3.3 ± 1.2 |
| GTFO-ΔN A977Q/SS-HA/α4 | 65.8 ± 2.7 | 39.2 ± 2.7 | 7.1 ± 1.3 | 13.3 ± 1.3 | 3.2 ± 0.6 | 3.0 ± 0.4 |
| GTFO-ΔN A977Q/V1083D | 69.6 ± 4.5 | 44.8 ± 2.9 | 5.7 ± 0.9 | 13.2 ± 0.9 | 3.5 ± 0.4 | 2.3 ± 0.5 |
| GTFO-ΔN A977Q/G1087N | 70.7 ± 4.9 | 41.4 ± 3.2 | 7.0 ± 0.7 | 17.5 ± 1.4 | 0.5 ± 0.2 | 4.2 ± 0.5 |
| GTFO-ΔN A977Q/V1083D/G1087N | 68.4 ± 5.4 | 43.4 ± 2.8 | 5.7 ± 0.8 | 10.1 ± 0.9 | 6.7 ± 0.8 | 2.4 ± 0.4 |
aAfter depletion of sucrose, the amounts of individual oligosaccharides formed were determined by HPAEC-PAD and expressed as the percentage of maltose initially present in the incubation. ND: not detected.