Table 1.
Enrichment of apratoxin A by culture of M. bouillonii PNG 5-198 supplemented with [methyl-13C]methionine. Final enrichment values are derived by normalization of enriched 13C-NMR resonances to five carbon atoms. Calculation steps are detailed in Methods section. SAM-derived enrichment values are in bold and italicized in column 21 and 22.
| Column # → | |||||||||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 13 | 14 | 15 | 16 | 17 | 18 | 19 | 20 | 21 | 22 |
|
| |||||||||||||||||||||
| C# | Biosynthetic source |
ppm | Integral of unlabeled resonance |
Normalization factor using: | Integral of enriched resonance |
Normalized integral of enriched sample using: |
Enrichment compared to: | Average % enrichment |
Fold change |
||||||||||||
| C7 | C13 | C20 | C30 | C37 | C7 | C13 | C20 | C30 | C37 | C7 | C13 | C20 | C30 | C37 | |||||||
| 1 | Pro | 172.8 | 42.7 | 0.4 | 0.7 | 0.4 | 0.4 | 0.6 | 45.9 | 37.2 | 40.2 | 30.6 | 36.2 | 40.1 | 23.3 | 14.1 | 50.0 | 26.7 | 14.5 | 25.7 | 1.26 |
| 2 | Pro | 59.9 | 112.6 | 1.1 | 1.8 | 1.1 | 1.0 | 1.6 | 79.2 | 98.1 | 106.1 | 80.7 | 95.5 | 105.7 | −19.3 | −25.3 | −1.9 | −17.1 | −25.1 | −17.7 | 0.82 |
| 3 | Pro | 29.5 | 119.5 | 1.1 | 2.0 | 1.2 | 1.1 | 1.6 | 126.5 | 104.1 | 112.6 | 85.6 | 101.3 | 112.2 | 21.5 | 12.4 | 47.7 | 24.8 | 12.8 | 23.8 | 1.24 |
| 4 | Pro | 25.8 | 91.9 | 0.9 | 1.5 | 0.9 | 0.8 | 1.3 | 81.7 | 80.1 | 86.6 | 65.9 | 78.0 | 86.3 | 1.9 | −5.7 | 24.0 | 4.8 | −5.4 | 3.9 | 1.04 |
| 5 | Pro | 47.8 | 133.9 | 1.3 | 2.2 | 1.3 | 1.2 | 1.8 | 88.1 | 116.7 | 126.2 | 96.0 | 113.6 | 125.7 | −24.5 | −30.2 | −8.2 | −22.5 | −30.0 | −23.1 | 0.77 |
| 6 | Ile | 170.8 | 37.4 | 0.4 | 0.6 | 0.4 | 0.3 | 0.5 | 51.4 | 32.6 | 35.2 | 26.8 | 31.7 | 35.1 | 57.9 | 46.1 | 92.1 | 62.3 | 46.6 | 61.0 | 1.61 |
| 7 | Ile | 56.8 | 104.0 | 1.0 | 1.7 | 1.0 | 0.9 | 1.4 | 90.7 | 90.7 | 98.0 | 74.6 | 88.2 | 97.7 | 0.0 | −7.5 | 21.6 | 2.8 | −7.2 | 1.9 | 1.02 |
| 8 | Ile | 31.9 | 82.9 | 0.8 | 1.4 | 0.8 | 0.7 | 1.1 | 68.2 | 72.3 | 78.1 | 59.4 | 70.3 | 77.9 | −5.7 | −12.7 | 14.7 | −3.1 | −12.4 | −3.8 | 0.96 |
| 9 | Ile | 24.8 | 93.7 | 0.9 | 1.5 | 0.9 | 0.8 | 1.3 | 66.0 | 81.7 | 88.3 | 67.2 | 79.5 | 88.0 | −19.2 | −25.3 | −1.7 | −17.0 | −25.0 | −17.6 | 0.82 |
| 10 | Ile | 9.2 | 124.2 | 1.2 | 2.0 | 1.2 | 1.1 | 1.7 | 83.4 | 108.3 | 117.0 | 89.0 | 105.4 | 116.6 | −23.0 | −28.8 | −6.4 | −20.9 | −28.5 | −21.5 | 0.79 |
| 11 | Ile | 14.2 | 145.1 | 1.4 | 2.4 | 1.4 | 1.3 | 2.0 | 86.1 | 126.5 | 136.7 | 104.0 | 123.1 | 136.3 | −31.9 | −37.0 | −17.2 | −30.1 | −36.8 | −30.6 | 0.69 |
| 12 | SAM | 30.7 | 89.0 | 0.9 | 1.5 | 0.9 | 0.8 | 1.2 | 175.7 | 77.6 | 83.8 | 63.8 | 75.5 | 83.6 | 126.5 | 109.6 | 175.5 | 132.8 | 110.3 | 130.9 | 2.31 |
| 13 | Ala | 170.2 | 61.1 | 0.6 | 1.0 | 0.6 | 0.5 | 0.8 | 57.6 | 53.3 | 57.6 | 43.8 | 51.8 | 57.4 | 8.1 | 0.0 | 31.5 | 11.1 | 0.3 | 10.2 | 1.10 |
| 14 | Ala | 60.9 | 76.9 | 0.7 | 1.3 | 0.8 | 0.7 | 1.1 | 90.6 | 67.0 | 72.5 | 55.1 | 65.2 | 72.2 | 35.1 | 25.0 | 64.3 | 38.9 | 25.5 | 37.8 | 1.38 |
| 15 | Ala | 14.1 | 62.4 | 0.6 | 1.0 | 0.6 | 0.6 | 0.9 | 70.6 | 54.4 | 58.8 | 44.7 | 52.9 | 58.6 | 29.9 | 20.1 | 57.9 | 33.5 | 20.6 | 32.4 | 1.32 |
| 16 | SAM | 36.9 | 38.8 | 0.4 | 0.6 | 0.4 | 0.3 | 0.5 | 109.7 | 33.8 | 36.6 | 27.8 | 32.9 | 36.4 | 224.3 | 200.0 | 294.4 | 233.2 | 201.0 | 230.6 | 3.31 |
| 17 | Tyr | 170.6 | 58.8 | 0.6 | 1.0 | 0.6 | 0.5 | 0.8 | 58.5 | 51.3 | 55.4 | 42.2 | 49.9 | 55.2 | 14.1 | 5.6 | 38.8 | 17.3 | 5.9 | 16.3 | 1.16 |
| 18 | Tyr | 50.6 | 97.8 | 0.9 | 1.6 | 1.0 | 0.9 | 1.3 | 13.8 | 85.2 | 92.1 | 70.1 | 82.9 | 91.8 | −83.8 | −85.0 | −80.3 | −83.4 | −85.0 | −83.5 | 0.17 |
| 19 | Tyr | 37.3 | 116.5 | 1.1 | 1.9 | 1.2 | 1.0 | 1.6 | 95.5 | 101.6 | 109.8 | 83.5 | 98.8 | 109.4 | −6.0 | −13.0 | 14.4 | −3.4 | −12.7 | −4.1 | 0.96 |
| 20 | Tyr | 128.4 | 100.6 | 1.0 | 1.6 | 1.0 | 0.9 | 1.4 | s72.1 | 87.7 | 94.8 | 72.1 | 85.3 | 94.5 | −17.8 | −23.9 | 0.0 | −15.5 | −23.7 | −16.2 | 0.84 |
| 21/25 | Tyr | 130.8 | 293.1 | 2.8 | 4.8 | 2.9 | 2.6 | 4.0 | 237.2 | 255.4 | 276.1 | 210.0 | 248.6 | 275.2 | −7.2 | −14.1 | 12.9 | −4.6 | −13.8 | −5.4 | 0.95 |
| 22/24 | Tyr | 114.0 | 254.6 | 2.4 | 4.2 | 2.5 | 2.3 | 3.5 | 225.7 | 221.9 | 239.9 | 182.5 | 215.9 | 239.0 | 1.7 | −5.9 | 23.7 | 4.5 | −5.6 | 3.7 | 1.04 |
| 23 | Tyr | 158.8 | 43.0 | 0.4 | 0.7 | 0.4 | 0.4 | 0.6 | 46.0 | 37.5 | 40.5 | 30.8 | 36.5 | 40.4 | 22.6 | 13.5 | 49.1 | 26.0 | 13.8 | 25.0 | 1.25 |
| 26 | SAM | 55.4 | 124.2 | 1.2 | 2.0 | 1.2 | 1.1 | 1.7 | 270.1 | 108.2 | 117.0 | 89.0 | 105.3 | 116.6 | 149.6 | 130.9 | 203.5 | 156.5 | 131.7 | 154.5 | 2.55 |
| 27 | Acetate | 169.7 | 44.5 | 0.4 | 0.7 | 0.4 | 0.4 | 0.6 | 50.1 | 38.8 | 41.9 | 31.9 | 37.7 | 41.7 | 29.2 | 19.6 | 57.2 | 32.8 | 20.0 | 31.8 | 1.32 |
| 28 | Acetate | 130.6 | 42.1 | 0.4 | 0.7 | 0.4 | 0.4 | 0.6 | 56.6 | 36.7 | 39.6 | 30.1 | 35.7 | 39.5 | 54.4 | 42.8 | 87.7 | 58.7 | 43.3 | 57.4 | 1.57 |
| 29 | Cys | 136.4 | 90.2 | 0.9 | 1.5 | 0.9 | 0.8 | 1.2 | 98.7 | 78.6 | 85.0 | 64.6 | 76.5 | 84.7 | 25.6 | 16.2 | 52.7 | 29.1 | 16.6 | 28.0 | 1.28 |
| 30 | Cys | 72.6 | 111.7 | 1.1 | 1.8 | 1.1 | 1.0 | 1.5 | 94.8 | 97.4 | 105.3 | 80.1 | 94.8 | 104.9 | −2.7 | −10.0 | 18.3 | 0.0 | −9.7 | −0.8 | 0.99 |
| 31 | Cys | 37.7 | 71.5 | 0.7 | 1.2 | 0.7 | 0.6 | 1.0 | 37.6 | 62.3 | 67.4 | 51.3 | 60.7 | 67.2 | −39.7 | −44.2 | −26.7 | −38.0 | −44.0 | −38.5 | 0.62 |
| 32 | SAM | 13.5 | 81.2 | 0.8 | 1.3 | 0.8 | 0.7 | 1.1 | 167.6 | 70.8 | 76.5 | 58.2 | 68.9 | 76.2 | 136.8 | 119.1 | 188.0 | 143.4 | 119.8 | 141.4 | 2.41 |
| 33 | Acetate | 177.6 | 57.3 | 0.6 | 0.9 | 0.6 | 0.5 | 0.8 | 65.3 | 50.0 | 54.0 | 41.1 | 48.6 | 53.8 | 30.8 | 21.0 | 59.0 | 34.4 | 21.4 | 33.3 | 1.33 |
| 34 | Acetate | 49.3 | 76.3 | 0.7 | 1.2 | 0.8 | 0.7 | 1.1 | 84.9 | 66.5 | 71.9 | 54.7 | 64.7 | 71.7 | 27.6 | 18.0 | 55.2 | 31.1 | 18.5 | 30.1 | 1.30 |
| 35 | Acetate | 71.8 | 116.2 | 1.1 | 1.9 | 1.2 | 1.0 | 1.6 | 122.4 | 101.3 | 109.5 | 83.3 | 98.6 | 109.1 | 20.8 | 11.8 | 46.9 | 24.1 | 12.1 | 23.2 | 1.23 |
| 36 | Acetate | 38.3 | 94.7 | 0.9 | 1.6 | 0.9 | 0.8 | 1.3 | 104.0 | 82.6 | 89.3 | 67.9 | 80.3 | 89.0 | 25.9 | 16.5 | 53.1 | 29.4 | 16.9 | 28.4 | 1.28 |
| 37 | Acetate | 24.4 | 72.6 | 0.7 | 1.2 | 0.7 | 0.6 | 1.0 | 68.4 | 63.3 | 68.4 | 52.0 | 61.6 | 68.2 | 8.0 | −0.1 | 31.4 | 11.0 | 0.3 | 10.1 | 1.10 |
| 38 | Acetate | 37.8 | 111.1 | 1.1 | 1.8 | 1.1 | 1.0 | 1.5 | 129.8 | 96.8 | 104.7 | 79.6 | 94.2 | 104.3 | 34.0 | 24.0 | 63.0 | 37.7 | 24.4 | 36.6 | 1.37 |
| 39 | Acetate | 77.4 | N.P.* | – | – | – | – | – | – | – | – | – | – | – | – | – | – | – | – | – | – |
| 40 | Acetate | 35.0 | 93.4 | 0.9 | 1.5 | 0.9 | 0.8 | 1.3 | 94.2 | 81.4 | 88.0 | 66.9 | 79.2 | 87.7 | 15.8 | 7.1 | 40.8 | 19.0 | 7.5 | 18.0 | 1.18 |
| 41–43 | SAM | 26.2 | 387.6 | 3.7 | 6.3 | 3.9 | 3.5 | 5.3 | 659.6 | 337.8 | 365.2 | 277.8 | 328.7 | 363.9 | 95.2 | 80.6 | 137.4 | 100.6 | 81.2 | 99.0 | 1.99 |
| 44 | SAM | 16.8 | 94.1 | 0.9 | 1.5 | 0.9 | 0.8 | 1.3 | 203.0 | 82.0 | 88.6 | 67.4 | 79.8 | 88.3 | 147.5 | 129.0 | 201.0 | 154.4 | 129.8 | 152.3 | 2.52 |
| 45 | Acetate | 20.0 | 109.2 | 1.0 | 1.8 | 1.1 | 1.0 | 1.5 | 70.7 | 95.2 | 102.9 | 78.3 | 92.6 | 102.5 | −25.7 | −31.2 | −9.6 | −23.6 | −31.0 | −24.2 | 0.76 |
*
N.P. = No quantitation due to CDCl3 peak overlap