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Purpose The purpose of this paper was to identify Serratia marcescens to extract and purify prodigiosin pigment to evaluate the antibacterial potential of the pigment. Design/methodology/approach Samples were collected from shrimp aquaculture ponds. Species identification was conducted using morphological, biochemical and molecular tests. Pigment extraction and purification were carried out using column chromatography. The antibacterial effect of crude and purified prodigiosin pigment was evaluated on Escherichia coli, Bacillus subtilis, Pseudomonas aeruginosa and Staphylococcus aureus as biofouling bacteria. In addition, the interaction between prodigiosin and proteins involved in biofilm formation was evaluated using molecular docking. Findings The results of prodigiosin extraction with solvents showed the highest percentage of pigment presence with methanol solvent in the second day of culture. The chemical structure of pure prodigiosin obtained from the column chromatography was confirmed by Fourier-transform infrared spectroscopy. Both crude and purified pigments exhibited antibacterial effects against selected bacterial strains. The antibacterial effect of the purified pigment was higher, and the highest antibacterial effect was observed on B. subtilis. Prodigiosin docking was carried out with all target proteins, and the docked energy in all of them was at an acceptable level. Originality/value Prodigiosin extracted from S. marcescens can be used as a bioactive compound to design and manufacture of anti-biofouling and anti-biofilm formation products to use extensively for industrial applications as a natural color in marine industries, food industry, cosmetics and textile productions.