Table 2.
Forty-four virulence-defective mutants: 23 conditional prototrophs & 21 auxotrophs. These metabolic mutants can be grouped in metabolic pathways based on the KEGG database25. They are unable to grow in glucose minimal medium (M9), but 23 of them can grow “conditionally” like the wild type strain in M9 supplemented with 5% fly extract and/or in flies. The remaining 21 do not grow like the wild type in any condition tested.
| # | Major Pathways (KEGG database entry) | Metabolic PA14 mutants | In vitro growth | In vivo growth (colonization) | ||
|---|---|---|---|---|---|---|
| Glucose minimal medium (M9) with 5% Fly Extract | Intestinal | Wound | ||||
| 1 | Histidine metabolism (pau00340) | PA14_57770 (hisC1) | + | + | NT | Group (A) (# 6 mutants) Efficient growth in glucose minimal medium with 5% fly extract and in vivo |
| 2 | PA14_57780 (hisD) | + | + | NT | ||
| 3 | PA14_66950 (hisE) | + | + | NT | ||
| 4 | PA14_65250 (hisX) | + | + | NT | ||
| 5 | One carbon pool by folate (pau00670) | PA14_05590 (metF) | + | + | NT | |
| 6 | Pantothenate and CoA biosynthesis (pau00770) | PA14_62580 (panB) | + | + | NT | |
| 7 | PA14_62590 (panC) | + | − | NT | Group (B) (# 6 mutants) Efficient growth only in glucose minimal medium with 5% fly extract | |
| 8 | Purine metabolism (pau00230) | PA14_51240 (purC) | + | − | NT | |
| 9* | PA14_64220 (purD) | + | − | − | ||
| 10 | PA14_23920 (purF) | + | − | NT | ||
| 11* | PA14_64200 (purH) | + | − | NT | ||
| 12 | PA14_15740 (purL) | + | − | NT | ||
| 13* | Pyrimidine metabolism (pau00240) | PA14_05260 (pyrB) | − | + | − | Group (C) (# 11 mutants) Efficient growth only in vivo |
| 14* | PA14_05250 (pyrC) | − | − | + | ||
| 15* | Glycine, serine and threonine metabolism (pau00260) | PA14_16070 (hom) | − | + | + | |
| 16* | PA14_16090 (thrC) | − | − | + | ||
| 17 | Arginine biosynthesis (pau00220) | PA14_08480 (argC) | − | + | NT | |
| 18* | PA14_18740 (argG) | − | + | NT | ||
| 19 | PA14_18610 (argF) | − | NT | + | ||
| 20 | Glycolysis/Gluconeogenesis (pau00010) | PA14_66310 (aceF) | − | + | NT | |
| 21* | PA14_66290 (aceE) | − | + | NT | ||
| 22 | Cysteine and methionine metabolism (pau00270) | PA14_05620 (sahH) | − | − | + | |
| 23 | Lysine biosynthesis (pau00300) | PA14_69670 (lysA) | − | + | NT | |
| 24 | Phenylalanine, tyrosine and tryptophan biosynthesis (pau00400) | PA14_00450 (trpB) | Group (D) (# 21 mutants) Slow growth compared to the wild type PA14 in all the above assays | |||
| 25 | PA14_08360 (trpC) | |||||
| 26 | PA14_08350 (trpD) | |||||
| 27 | PA14_23850 (trpF) | |||||
| 28 | PA14_66600 (aroB) | |||||
| 29 | Valine, leucine and isoleucine biosynthesis (pau00290) | PA14_62130 (ilvC) | ||||
| 30 | PA14_04630 (ilvD) | |||||
| 31 | PA14_62150 (ilvH) | |||||
| 32 | PA14_62160 (ilvI) | |||||
| 33 | PA14_23790 (leuB) | |||||
| 34 | PA14_23750 (leuC) | |||||
| 35* | Pyrimidine metabolism (pau00240) | PA14_18710 (pyrC) | ||||
| 36* | PA14_24640 (pyrD) | |||||
| 37 | PA14_26890 (pyrF) | |||||
| 38* | PA14_62930 (carA) | |||||
| 39 | Pentose phosphate pathway (pau00030) | PA14_22910 (edd) | ||||
| 40 | PA14_23090 (edaA) | |||||
| 41 | Arginine biosynthesis (pau00220) | PA14_70280 (argB) | ||||
| 42 | PA14_69500 (argH) | |||||
| 43 | Glycine, serine and threonine metabolism (pau00260) | PA14_65560 (serB) | ||||
| 44 | Glycolysis/Gluconeogenesis (pau00010) | PA14_62830 (tpiA) | ||||
*: Found attenuated in both assays (pricking and feeding); NT: Not Tested.