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. 2020 Aug 27;11:4292. doi: 10.1038/s41467-020-18124-1

Table 1.

Stable enzyme set.

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.