Table 2.
Reaction interventions needed to create mutant M3 and enable 1-octanol-coupled growth in iJN678_OctFA.
| Reaction name in iJN678 | Enzyme (s) | Reaction* | Locus to target** |
|---|---|---|---|
| NDH1_1u | NAD(P)H dehydrogenase NDH-1 (thylakoid) | 4h[c]+nadph[c]+pq[p]→nadp[c]+3h[p]+pqh2[p] | slr0331 (ndhD1) and slr1291 (ndhD2) |
| NDH1_2u | NAD(P)H dehydrogenase NDH-1 (thylakoid) | 4h[c]+nadh[c]+pq[u]→nad[c]+3h[u]+pqh2[u] | slr0331 (ndhD1) and slr1291 (ndhD2) |
| NDH2_syn | NdbA, NdbB, NdbC (thylakoid) | h[c]+nadh[c]+pq[u]→nad[c]+pqh2[u] | slr0851, slr1743, and sll1484 |
| NDH2_2p | NdbA, NdbB, NdbC (periplasm) | h[c]+nadh[c]+pq[p]→nad[c]+pqh2[p] | slr0851, slr1743, and sll1484 |
| NDH1_3u | Active CO2 transporter facilitator (thylakoid) | 3h[c]+h2o[c]+nadph[c]+pq[u]+co2[p]→nadp[c]+hco3[c]+3h[u]+pqh2[u] | sll1733 (ndhD3) and sll0027 (ndhD4) |
| Mehler | Flavodiiron proteins Flv1 and Flv3 | h[c]+0.5 o2[c]+nadph[c]→h2o[c]+nadp[c] | sll1521 (flv1) |
| sll0550 (flv3) | |||
| Cyo1b_syn | Cytochrome c oxidase | 4h[c]+2 focytc6[u]+0.5 o2[u]→2h[u]+2 ficytc6[u]+ h2o[u] | slr1137 |
| GLYCTO1 | Glycolate oxidase | o2[c]+glyclt[c]→h2o2[c]+glx[c] | sll0404 (glcD2) |
| GLUSx | Glutamate synthase GOGAT (NADH-dependent) | h[c]+nadh[c]+akg[c]+gln-L[c]→nad[c]+2 glu-L[c] | sll1502 |
| ACKr | Acetate kinase | atp[c]+ac[c]⇔adp[c]+actp[c] | sll1299 |
| H2ase_syn | [NiFe] Hydrogenase | h[c]+ nadph[c]⇔nadp[c]+h2[c] | sll1224 (hoxY) |
| ATPS4rpp | ATP synthase (periplasmic) | 3 adp[c]+3 pi[c]+14h[p]→3 atp[c]+11h[c]+3 h2o[c] | slr1330 (atpE) |
| FNOR*** | Ferredoxin: NADP+reductase | h[c]+nadp[c]+2 fdxr-2:2[c]⇔nadph[c]+2 fdxo-2:2[c] | slr1643 |
*
[c] cytoplasmic, [u] thylakoid, [p] periplasmic compartments.
**
Locus to target is suggestion for gene deletion to eliminate enzyme activity. For multi-domain proteins a core subunit is given. NDH-1 (Battchikova et al., 2011), GlcD2 (Eisenhut et al., 2008), Hox (Eckert et al., 2012), AtpE (Imashimizu et al., 2011).
***
Overexpression required.