Table 1.
Enzyme | Organism | Native/Mutant | Activity (units)b | Half-life in 8% isobutanol (hours) | |
---|---|---|---|---|---|
TmHex | Hexokinasea | T. maritima | Native | 14.5 ± 0.3 | >>96 |
TmPgi | Glucose-6-phosphate isomerasea | T. maritima | Native | 29.6 ± 1.1 | >>113 |
EcPfkB | Phosphofructokinase B | E. coli | Native | 8.3 ± 0.1 | 55/123d |
TtFba | Fructose-1,6-bisphosphate aldolasea | T. thermophilus | Codon-optimized | 4.7 ± 0.10 | >>117 |
TmTpi | Triosephosphate isomerasea | T. maritima | Native | 210 ± 0.7 | >>100 |
TkGapN’ | Glyceraldehyde-3-phosphate dehydrogenase (non-phosphorylating)a | T. kodakarensis | Designed Codon-optimized | 7.6 ± 0.8 | >>117 |
AfGapDH | Glyceraldehyde-3-phosphate dehydrogenase (phosphorylating)a | A. fulgidus | Native | 10.9 ± 0.6 | >>100 |
TmPgk | Phosphoglycerate kinasea | T. maritima | Native | 8.4 ± 0.5 | >>100 |
Gs iPgm | Phosphoglycerate mutase (2,3-bisphosphoglycerate independent)a | G. stearothermophilus | Native | 49.4 ± 2.2 | >>117 |
TtEno | Phosphoenolpyruvate hydratasea | T. thermophilus | Codon-optimized | 117 ± 5 | >>117 |
TtPyk | Pyruvate kinasea | T. thermophilus | Codon-optimized | 23.8 ± 0.8 | >>139 |
BsAlsS-P | Acetolactate synthase | B. subtilis | PROSSc Codon-optimized | 4.9 ± 0.2 | >>115 |
GsIlvC | Ketol-acid reductoisomerasea | G. stearothermophilus | Native | 0.9 ± 0.1 | >>123 |
SmIlvD | Dihydroxyacid dehydratase | S. mutans | Native | 2.7 ± 0.2 | 125 |
KivD-S | Alpha-ketoisovalerate decarboxylase | L. lactis | PROSSc/DE Codon-optimized | 18.0 ± 1.6 | 123 |
EcYahK | Aldehyde reductase | E. coli | Native | 4.5 ± 0.4 | 138 |
DE directed evolution.
aEnzyme from hyperthermophilic organism.
bUnit defined as µmole product min−1 mg−1 enzyme.
cPROSS design algorithm33.
dBiphasic inactivation in 8% isobutanol.