Screening Bacterial Strains Capable of Producing 2,3-Butanediol: Process Optimization and High Diol Production by Klebsiella oxytoca FMCC-197

Abstract

In the present investigation, the potential of various newly isolated strains which belong to the Enterobacteriaceae family to produce 2,3-butanediol (BDO), an important bio-based compound, was studied. The most interesting strain, namely Klebsiella oxytoca FMCC-197, was selected for further investigation. Commercial (raw) sucrose or molasses, which are important agro-industrial surpluses, were employed as carbon sources for most of the trials performed. Different fermentation parameters (viz. incubation te4mperature, utilization of different carbon sources, substrate inhibition, aeration) were tested to optimize the process. Fermentations under non-aseptic conditions were also conducted to investigate the potential of growth of the strain K. oxytoca FMCC-197 to surpass the growth of other microorganisms in the culture medium and produce BDO. Besides BDO production, in trials in which molasses was employed as the sole carbon source, significant color removal was observed simultaneously with the production of microbial metabolites. The very high BDO concentration ≈115 g L−1 was reported in approximately 64 h during a fed-batch bioreactor experiment, using sucrose and molasses as carbon sources at 30 °C, reaching a conversion yield (YBDO) of 0.40 g g−1 and a productivity rate (PBDO) of 1.80 g L−1 h−1, while similar results were also obtained at 37 °C. The strain demonstrated remarkable results in non-previously sterilized media, as it produced 58.0 g L−1 in 62 h during a fed-batch bioreactor experiment, while the potential to decolorize molasses-based substrates over 40% was also recorded. From the results obtained it is shown that this wild-type strain can be used in large-scale microbial BDO production using various raw materials as fermentative substrates. The wastewater derived after BDO fermentation by K. oxytoca FMCC-197 can be disposed relatively safely into the environment.