Fig. 4: Screening targets that inhibit daptomycin production. | Nature Communications

Fig. 4: Screening targets that inhibit daptomycin production.

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

Fig. 4

a Workflow for screening targets that inhibit daptomycin production. b Coverage of sgRNAs in CRISPRi plasmid library. c Coverage of sgRNAs in Srdi transformants. Since three target sites were designed for each gene, “sgRNA=3” means that all three target sites were detected. Similarly, “sgRNA=2”, “sgRNA=1”, and “sgRNA=0” mean that only 2, 1, and 0 of the three target sites were detected, respectively. d Indigoidine level and daptomycin titer when inhibiting the identified targets in Srdi. The blue and brown lines indicate the indigoidine levels and daptomycin in the control strain Srdi-, respectively. [p = 0.003282553, 0.007464338, 0.001446173, 0.00034476, 0.006491144, 0.010929629, 0.002683903, 0.0044472, 0.000722168, 0.00359535, 0.031076087, 0.13338661, 0.009880467, 0.001124236, 0.000753792, 0.00158046 (indigoidine: left to right); p = 0.036945733, 0.0432256, 0.015491013, 0.008536545, 0.03695489, 0.036148854, 0.032078525, 0.037558817, 0.022461226, 0.034004881, 0.041198351, 0.043306871, 0.035889288, 0.030190684, 0.024581079, 0.016597474 (daptomycin: left to right)]. e Daptomycin titers of WT and double mutant strains Sr4-Q57*, Sr5-Q22*, Sr6-Q71*, Sr7-Q6*. [p = 0.004047725, 0.011340892, 0.013594525, 0.015901674 (left to right)]. f Transcription levels of daptomycin BGC in WT, Sr4-Q57*, Sr5-Q22*, Sr6-Q71*, and Sr7-Q6*. Samples were collected after 2 days culture in seed medium. [p = 0.000182357, 0.000231765, 0.000228029, 0.000764954, 0.000249225, 0.000577618, 0.002118873, 0.000747447, 0.000276501, 0.000261965, 0.000660066, 0.000529196, 6.63681E−05, 0.000301623, 0.001158026, 0.000150468, 0.000139481, 3.71049E−05, 0.000910931, 0.000764598 (left to right)]. g Daptomycin titer in WT and double mutant strains Sr45, Sr46, Sr47, Sr56, Sr57, Sr67. [p = 0.021964853, 0.03252863, 0.739909874, 0.003015842, 0.182164021, 0.003489912 (left to right)]. Data in d–g are shown as the mean ± SD (n = 3 biological replicates). Two-tailed Student’s t test was used in d–g to analyze the statistical significance (n.s., not significant (p >  0.05); *p  <  0.05; **p  <  0.01; ***p  <  0.001).

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