Aromatics valorization to polyhydroxyalkanoate by the ligninolytic bacteria isolated from soil sample

Polyhydroxyalkanoates (PHA) are ecofriendly alternatives to conventional plastics due to their biodegradable nature. However, the high production cost limits their applications. Exploring novel bacteria with ligninolytic potential would be crucial to advance cost-effective PHA synthesis. The current study aims to unveil soil bacteria capable of aromatics valorization to PHA. Considering this, six aromatics resistance bacteria from a soil sample were isolated through culture acclimatization strategy and their growth was analyzed in various lignin model compounds. Ralstonia sp. BPSS-1 and Arthrobacter sp. BPSS-3 presented high-cell-densities in 4-hydroxybenzoic acid (4-HBA) and benzoate, respectively. Fluorescence microscopy confirmed the strains to be PHA positive and were subsequently evaluated for PHA synthesis from 4-HBA and benzoate at a concentration of 2 g L^-1 in a nitrogen-limited M9 medium. However, applying a co-feeding strategy by the integration of 4-HBA and benzoate further increased the substrates consumption efficiency, biomass and PHA titer compared to single carbon sources. The maximum dry cell weight (DCW) and PHA yield by Ralstonia sp. BPSS-1 through the substrate co-feeding under optimized fermentation conditions was 0.69 ± 0.03, and 0.4 ± 0.02 g L^-1, respectively. The draft genome analysis confirmed the genes involved in aromatic degradation. Besides, the proposed metabolic pathway was validated by studying the expression level of key genes, analyzing key intermediates and associated enzymes activities. The FTIR, ¹H NMR and GC-MS determined the PHA functional group, chemical structure and monomers analysis, respectively. Overall, the current study highlighted the aromatic valorization potential of newly isolated PHA producing bacteria for sustainable biomanufacturing.