Fermentative Production, Kinetic Analysis, Characterization, and Biological Evaluation of Phenazine‐1‐Carboxylic Acid from Pseudomonas chlororaphis subsp. Aureofaciens

This study investigates phenazine‐1‐carboxylic acid (PCA) production by Pseudomonas chlororaphis subsp. aureofaciens under submerged batch fermentation. This nonpathogenic, flexible carbon‐utilizing strain was selected as a safer and industrially adaptable PCA producer. Chemical and physical parameters were optimized, and phenazines were isolated by chloroform extraction followed by adsorption chromatography and characterized using UV–vis, FTIR, and 1 H‐NMR analyses. PCA production was identified as growth‐associated, yielding 129.45 µg/mL—comparable to wild‐type Pseudomonas strains despite the use of low‐cost glycerol medium‐highlighting the process's industrial relevance. Key kinetic parameters were determined as Y XS = 0.64 mg/mg, Y PS = 0.12 mg/mg, Y PX = 0.35 mg/mg, and μ = 0.39 h − 1 . PCA exhibited antimicrobial activity against Klebsiella pneumoniae (10 mm) and Micrococcus luteus (22 mm) at 150 µg/mL. Cytotoxicity was evaluated via MTT assay on A549 and SKOV3 cells, yielding IC 50 values of 17.30 ± 1.72, 13.04 ± 0.76, and 5.69 ± 0.35 µg/mL at 24, 48, and 72 h, respectively, with notable effects also observed in SKOV3 cells. Overall, the findings demonstrate that PCA possesses significant antimicrobial and anticancer potential, supporting its further development as a natural bioactive compound.