Fig. 7: Synergistic multi-target engineering for improvement of thaxtomin and surfactin production. | Nature Communications

Fig. 7: Synergistic multi-target engineering for improvement of thaxtomin and surfactin production.

From: A rational multi-target combination strategy for synergistic improvement of non-ribosomal peptide production

Fig. 7

a Schematic diagram of increased thaxtomin and surfactin production by the analog co-expression and co-biosynthesis reporter system. The promoterless reporter genes idgS-sfp are fused downstream of txtB, resulting in an overlap between the TGA stop codon of txtB and the ATG start codon of idgS. The codon-optimized reporter genes idgS-sfp are fused downstream of srfAD, resulting in an overlap between the TGA stop codon of srfAD and the ATG start codon of idgS. b Indigoidine level and thaxtomin A titer when inhibiting the identified targets in Scotxti. [p = 4.65485E−05, 5.35998E−06, 5.04019E−06, 6.08803E−07, 1.08422E−05, 5.02854E−06, 2.08711E−06, 1.08422E−05, 4.80512E−06, 4.43093E−06 (indigoidine: left to right); p = 1.4452E−05, 4.57848E−05, 1.64923E−05, 5.12443E−06, 4.2004E−05, 8.81769E−07, 1.55787E−05, 2.38202E−06, 6.58047E−07, 5.21529E−06 (thaxtomin A: left to right)]. c Interactions network between targets based on synergy coefficient. Red dots indicate four targets that have a synergistic relationship with each other. Other targets are indicated by blue dots. d Thaxtomin titer of CRISPRi-mediated strains. “+“ indicates the use of CRISPRi to inhibit the corresponding gene, while “−“ indicates no inhibition. e Indigoidine level and surfactin titer when inhibiting the identified targets in 168S1R. [p = 0.016500953, 0.000413121, 0.000963969, 0.00063344, 0.001814368, 0.000499214, 0.0008904, 0.004535417, 0.000745307, 0.000522386, 0.000235722 (indigoidine: left to right, control:168S1R); p = 0.953066195, 0.010555248, 0.00110127, 0.002547204, 0.000626228, 0.00415879, 0.000214603, 0.003250768, 0.016483722, 0.006590528, 0.00105454, 0.001113834 (surfactin: left to right, control:168S1)]. f Interactions network between targets based on synergy coefficient. Red dots indicate four targets that have a synergistic relationship with each other. Other targets are indicated by blue dots. g Surfactin titer of CRISPRi-mediated strains. “+“ indicates the use of CRISPRi to inhibit the corresponding gene, while “−“ indicates no inhibition. [p = 0.722658162, 0.000105794, 0.000105702, 9.6318E−06, 6.99555E−05, 1.3685E−05, 0.000100527, 1.62203E−06, 4.15269E−06, 1.64117E−06, 8.92386E−07, 6.09771E−07 (left to right)]. Data in (b, d, e, g) are shown as the mean ± SD (n = 3 biological replicates). Two-tailed Student’s t test was used in (b, e, g) to analyze the statistical significance (n.s. not significant (p >  0.05); **p  <  0.01; ***p  <  0.001; ****p  <  0.0001).

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