Cost effective media optimization for PHB production by Bacillus badius MTCC 13004 using the statistical approach

Polyhydroxybutyrate (PHB) has significant potential for replacing non-biodegradable traditional plastic, which is responsible for several global environmental issues. The main problem with switching to bio-based alternatives for petrochemical plastics is the large price gap on the market. To overcome this problem, the present research was focused on the utilization of inexpensive substrates i.e. agricultural residues for cost-effective PHB production by endospore-forming bacteria Bacillus badius MTCC 13004. For efficient PHB production, Box-Behnken Design (BBD) was selected for media optimization and to observe the interactive effects of four variables i.e. pH, Na acetate, Banana peel, and mustard cake. PHB yield of 2.11 g/L was attained under optimized conditions compared to non-optimized conditions (0.72 g/L). FTIR spectra analysis of PHB extracted from Bacillus badius was found to be similar to commercial PHB. NMR data was also matched with the chemical shift signals CH, CH₂, and CH₃ of PHB. The melting temperature (Tm) and glass transition temperature (Tg) of PHB from Bacillus badius was found to be 165.14 and 2.68 °C, respectively. Further, PCR protocol was also designed to amplify key enzymes of the PHB synthesis pathway i.e. PHB synthase (phb C gene).