Journal: NPJ Systems Biology and Applications
Article Title: Addressing genome scale design tradeoffs in Pseudomonas putida for bioconversion of an aromatic carbon source
doi: 10.1038/s41540-024-00480-z
Figure Lengend Snippet: A Serial dilution plating for growth on solid agar medium supplemented with rich (LB) or p -CA minimal medium. Abbreviated genotypes are indicated to the left of each image. B The native PP_0897 promoter was replaced with two different low abundance promoters and the impact on viability in rich medium or M9 p- CA minimal medium was quantified. C P. putida strains assayed for indigoidine production in M9 60 mM p -CA medium at the indicated time points following induction with 1.5% arabinose (w/v). Control is WT P. putida harboring genome integrated indigoidine pathway cassette and ∆PP_0897 is the single gene deletion strain. Strain D1b_gf ∆PP_0897 did not grow nor produced in M9 p- CA medium. D P. putida strains with promoter-varied PP_0897 expression assayed for indigoidine production. Specific indigoidine yield (mg/CFU) was calculated by normalizing the indigoidine with the number of total viable cells at 24 h. E P. putida strains with promoter-varied PP_0897 expression with or without induction of the indigoidine pathway in M9 60 mM p- CA, sampled at 24 h and 48 h for glutamate and indigoidine (Supplementary Fig. ). F , G Strains D1b_gf with or without ∆PP_0897 were transformed with an empty vector plasmid or a plasmid containing a P BAD -PP_0897 construct. F Growth in M9 60 mM p- CA medium with basal BAD promoter induction without inducer ( F ) or with 1.5% (w/v) arabinose ( G ) in 24 well microtiter dishes. Strain genotypes are indicated above each panel. Measurements are average of at least three independent replicates. Error bars represent mean ± S.D. ( n = 4) in ( C – E ). The shaded area represents mean ± S.D. ( n = 3) in ( F , G ).
Article Snippet: A BIOLOG TM phenotype microarray assay (Cat. No. 12111, & Cat. No. 12112, Hayward CA) was conducted using strain D1b_gf ΔPP_0897.
Techniques: Serial Dilution, Control, Produced, Expressing, Transformation Assay, Plasmid Preparation, Construct