Purification and optimization of pink pigment produced by newly isolated bacterial strain Enterobacter sp. PWN1

Abstract

Pigment-producing bacteria were isolated from kitchen wastewaters of the National Institute of Technology, Rourkela. A pink non-virulent bacterial strain PWN1 was selected based on the India Ink Broth and Coomassie Brilliant Blue (R-250) dye assay. According to morphological and biochemical characterization, the strain PWN1was a Gram-negative, rod-shaped, motile, non-coliform bacterium and could utilize only glucose and adonitol as sole carbon source. The pigment was found to be a growth-associated product, and the pigment production was accelerated after 40 h of bacterial culture. Further, 16S rRNA gene-based molecular identification showed its similarity with Enterobacter sp. The pigments were extracted by the solvent extraction method using chloroform and ethanol (3:1). The extracted pigments were then purified through thin-layer chromatography and column chromatography. To maximize pigment production, the culture condition was optimized for maximum biomass production using statistical software Design Expert v13. A quadratic model was structured describing the process efficiently and it suggested a moderate temperature, pH, and a high inoculum concentration which generated biomass of 3.81 ± 0.02 g/L. At optimized condition, 1 L of cell culture produced 3.77 g of biomass which produced a crude pigment of 0.234 g after solvent extraction and 0.131 g after column chromatography, implying a yield of 6.2% for crude pigment and 3.47% for purified pigment from biomass. The yield of the obtained pigment was high enough to draw interest for industrial production, although the application of the pigment is considerable for further study.