This study involves the isolation of succinic acid (SA)‐producing microorganisms from different samples, including the rumen, sludge, soil, and wastewater. For primary screening, 29 isolates exhibited a zone of clearance around the colony, indicating acid production. For secondary screening using thin‐layer chromatography, only two isolates symbolized SA production according to their Rf values. These two isolates were further identified as Bacillus velezensis and Enterococcus gallinarum by phylogenetic analysis using the neighbor‐joining method. The high SA concentrations of 50.2 and 66.9 g/L were produced by B. velezensis and E. gallinarum with an SA yield of 0.836 and 1.12 g/g glucose, respectively. The high SA concentration from these newly isolated strains was achieved with a low formation of unwanted acids compared with those from Actinobacillus succinogenes ATCC 55618. Moreover, E. gallinarum was cultured in palm oil mill wastewater (POMW) and molasses, which were cheap substrates. The high SA production of 73.9 g/L with low other acids (the ratio of SA to total acids = 0.917) was achieved using POMW and molasses (80:20) as substrates.
Bio-succinic acid production using microorganisms has been interesting as an environmentally friendly process. Palm oil mill effluent (POME) was considered as a cheap substrate to lower the cost of production. It was revealed that 2-fold diluted POME produced more succinic acid than undiluted and 5-fold diluted POME. In addition, the effects of various neutralizing agents on succinic acid production utilized to manage pH and CO2 supply indicated that the utilization of MgCO3 as a neutralizing agent produced succinic acid of 11.5 g/L with a small amount of by-product synthesis. Plackett–Burman Design (PBD) was used to screen the most significant nutrients for bio-succinic acid production from 2-fold diluted POME using E. gallinarum. From the Pareto chart, MgCO3 and peptone presented the highest positive effect on the production of succinic acid. In addition, Box–Behnken Design (BBD) was conducted to increase bio-succinic acid production. Experiments showed the highest production of succinic acid of 23.7 g/L with the addition of 22.5 g/L MgCO3 and 12.0 g/L peptone in 2-fold diluted POME. Moreover, the experiment of replacing MgCO3 with CO2 from biogas resulted in 19.1 g/L of succinic acid, simultaneously creating the high purity of biogas and a higher CH4 content.
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