TABLE 5.
Growth rates, enzyme activities and L-lysine, cadaverine, and 5AVA final titers accumulated in growth media of recombinant MGA3 strains.
| Strain | Growth rate [h–1] | Coupled activity of PatAD or Puo-PatD [mU mg–1] | Lysine [mg l–1] | Cadaverine [mg l–1] | 5AVA [mg l–1] |
| MGA3_Cad | 0.37 ± 0.01 | 0 ± 0 | Not detected | 123.0 ± 5.3 | 0.0 ± 0.0 |
| MGA3_SpuI | 0.33 ± 0.01 | N.A. | Not detected | 118.82 ± 5.1 | 0.0 ± 0.0 |
| MGA3_PatAEc | 0.12 ± 0.02 | 7 ± 4 | Not detected | 1.47 ± 0.17 | 23.7 ± 2.7 |
| MGA3_PatABm | 0.15 ± 0.03 | 170 ± 37 | Not detected | 0.71 ± 0.11 | 8.3 ± 4.1 |
| MGA3_Cad | 0.35 ± 0.01 | 0 ± 0 | Not detected | Supplemented (500 mg L–1) | 0.0 ± 0.0 |
| MGA3_SpuI | 0.32 ± 0.00 | N.A. | Not detected | Supplemented (500 mg L–1) | 0.0 ± 0.0 |
| MGA3_PatAEc | 0.14 ± 0.02 | 7 ± 4 | Not detected | Supplemented (500 mg L–1) | 31.8 ± 2.3 |
| MGA3_PatABm | 0.17 ± 0.04 | 170 ± 37 | Not detected | Supplemented (500 mg L–1) | 77.7 ± 5.5 |
| MGA3_Kat | 0.32 ± 0.00 | 0 ± 0 | 3.1 ± 0.5 | Supplemented (500 mg L–1) | 0.0 ± 0.0 |
| MGA3_PuoEc | 0.28 ± 0.01 | 0 ± 0 | 5.0 ± 0.7 | Supplemented (500 mg L–1) | 0.0 ± 0.0 |
| MGA3_PuoPa | 0.29 ± 0.01 | 0 ± 0 | 4.9 ± 0.9 | Supplemented (500 mg L–1) | 0.0 ± 0.0 |
| MGA3_PuoRq | 0.29 ± 0.00 | 0 ± 0 | 3.7 ± 0.2 | Supplemented (500 mg L–1) | 0.0 ± 0.0 |
The B. methanolicus strains expressing pathways that use cadaverine as an intermediate (SpuI, PatA, or Puo pathways) were cultivated for 24 h, and supernatants were obtained by centrifugation for HPLC analysis. Catalytic activities of PatA and PatD or Puo and PatD were measured by using a coupled reaction, and cadaverine was used as substrate (see Section “Enzyme Assays”). The standard deviation of technical triplicates is shown. NB: RaiP activity and 5AVA production for the RaiP pathway is shown Figure 2.