Table 1.
Kinetic properties of Adh1, Adh2 and candidate ancestral AdhA
| KM (µM) | ||||
|---|---|---|---|---|
| Samplea | Ethanol | NAD+ | Acetaldehyde | NADH |
| Adh1 | 20,060 | 218 | 1,492 | 164 |
| MKD | 17,280 | 511 | 1,019 | 144 |
| MKN | 13,750 | 814 | 1,067 | 1,106 |
| MRD | 11,590 | 734 | 1,265 | 287 |
| MRN | 10,960 | 554 | 1,163 | 894 |
| MTD | 10,740 | 467 | 959 | 190 |
| MTN | NA | NA | NA | NA |
| RKD | 8,497 | 449 | 1,066 | 142 |
| RKN | 7,238 | 407 | 1,085 | 735 |
| RRD | 7,784 | 400 | 1,074 | 203 |
| RRN | 8,403 | 172 | 1,156 | 1,142 |
| RTD | 6,639 | 254 | 1,083 | 316 |
| RTN | 7,757 | 564 | 1,158 | 477 |
| Adh1b | 24,000 | 240 | 3,400 | 140 |
| Adh1c | 17,000 | 170 | 1,100 | 110 |
| Adh2b | 2,700 | 140 | 45 | 28 |
| Adh2c | 810 | 110 | 90 | 50 |
| Adh3c | 12,000 | 240 | 440 | 70 |
| Adh1c (S. pombe) | 14,000 | 160 | 1,600 | 100 |
| Adh1 (M270L)c | 19,000 | 630 | 1,000 | 80 |
| KlP20369d | 27,000 | 2,800 | 1,200 | 110 |
| KlX64397d | 23,000 | 2,200 | 1,700 | 180 |
| KlX62766d | 2,570 | 310 | 100 | 20 |
| KlX62767d | 1,560 | 200 | 3,100 | 30 |
a
The three letters designate the amino acids at positions 168, 211 and 236 (e.g., MKD = Met168 Lys211 Asp236). The remaining residues were the same as in Adh1, except for the following changes (using sequence numbering of Adh1 from S. cerevisiae): Asn15 Pro30 Thr58 Ala74 Glu147 Leu213 Ile232 Cys259 Val265 Leu270 Ser277 Asn324.
b
From ref. 8.
c
From ref. 16.
d
From ref. 30.
Kl, Kluyveromyces lactis.