High production of 2,3-butanediol from glycerol by Klebsiella pneumoniae G31.

The microbial production of high amounts of 2,3-butanediol (2,3-BD) from glycerol as a sole carbon source by the Bulgarian isolate Klebsiella pneumoniae G31 was studied in a series of fed-batch processes. The following conditions were evaluated as optimal: micro-aerobic cultivation in modified media, without pH control. Beginning at pH 8, 49.2 g/l of 2,3-BD was produced as negligible concentrations of by-products were received. The pH is the most important factor ruling the 2,3-BD production. Spontaneous pH changes and products formation in time were investigated, performing fermentations with non-controlled pH starting at different initial pH. In lack of external maintenance, the microorganism attempted to control the pH using acetate/2,3-BD alternations of the oxidative pathway of glycerol catabolism, which resulted in pH fluctuations. Thus, the culture secreted 2,3-BD at unequal portions, either allowing or detaining the acetate synthesis. More alkaline initial pH led to enhanced 2,3-BD accumulation as a response to the increased amplitudes of the pH variations. When the pH was maintained constant, the yield of 2,3-BD was very poor. These cultures remained viable only 72 h; whereas, the pH self-controlling cells lived and produced 2,3-BD up to 280 h. In conclusion, the formation of 2,3-BD is a result of an adaptive mechanism of pH self-control, responding to spontaneous pH drops during glycerol fermentation.