Improvement of 1,3-propanediol production from crude glycerol by co-cultivation of anaerobic and facultative microbes under non-strictly anaerobic conditions

BACKGROUND: Natural microbial consortia could efficiently produce 1,3-propanediol (1,3-PDO), a most promising bulk biochemical derived from glycerol that can be used as a monomer in the synthesis of polytrimethylene terephthalate (PTT). While natural microbial communities are made up of a diverse range of microbes with frequently unknown functions, the construction of synthetic microbial consortia allows for the creation of more defined systems with lower complexity. RESULTS: In this study, the synthetic microbial consortia were constructed by combining facultative microbes of Klebsiella pneumoniae DUT2 (KP) and/or Escherichia coli DUT3 (EC) cultures with the strictly anaerobic microbe of Clostridium butyricum DUT1 (CB) cultures under micro-aerobic conditions. The function of EC and KP during the fermentation process was to deplete oxygen and create an anaerobic environment for CB. Furthermore, KP competes with CB for the consumption of crude glycerol and the production of 1,3-PDO. The interaction of commensalism and competition resulted in the construction of synthetic microbial consortia capable of efficiently converting crude glycerol to 1,3-PDO even under micro-aerobic conditions. In a batch fermentation, the synthetic CB:KP co-culture at an initial abundance ratio of 92.5:7.5, yielded a maximum 1,3-PDO concentration of 52.08 g/L, with a yield of 0.49 g/g and a productivity of 1.80 g/(L.h), which increased by 10%, 9%, and 12%, respectively, when compared to the CB mono-culture under strictly anaerobic conditions. The final 1,3-PDO concentration, yield, and productivity by the synthetic CB:KP consortia increased by 16%, 19%, and 84%, respectively, when compared to the KP mono-culture. At an initial abundance ratio of 85:7.5:7.5, the synthetic CB:KP:EC co-culture achieved the highest 1,3-PDO flux of 49.17%, while 7.43%, 5.77%, 3.15% 4.24%, and 2.13% of flux was distributed to butyric acid, acetic acid, lactic acid, ethanol, and succinic acid pathways. In a fed-batch fermentation, the synthetic CB:KP:EC co-culture demonstrated a maximum 1,3-PDO concentration of 77.68 g/L with a yield of 0.51 g/g which is 30% and 13% higher than the production by the CB mono-culture at 0.02 vvm (nitrogen volume/culture volume/min) N(2) supply. The initial abundance of CB, which is guaranteed to be at least 85%, enables efficient 1,3-PDO production from crude glycerol via the development of synthetic microbial consortia. CONCLUSION: The synthetic microbial consortia demonstrated excellent performance on 1,3-propanediol production under micro-aerobic conditions through the interaction of commensalism and competition. The experimental results demonstrated the potential benefit of using synthetic microbial consortia to produce 1,3-propanediol from crude glycerol.