Abstract

In this study, the ability of a novel strain of Clostridium tyrobutyricum NRRL 67062 to produce butyric acid during glucose fermentation was evaluated. The strain was evaluated for substrate and product inhibition in batch experiments using anaerobic tubes. To characterize glucose inhibition, initial glucose concentrations ranging from 60 to 250 g L−1 were used, and it was demonstrated that a glucose concentration of 250 g L−1 exerted strong inhibition on cell growth and fermentation. To evaluate butyric acid inhibition, the culture was challenged with 5–50 g L−1 of butyric acid at an initial pH of 6.5. These experiments were performed without pH control. When challenged with a butyric acid concentration of 50 g L−1, cell growth was slow; however, it produced 8.25 g L−1 of butyric acid. This suggested that the butyric acid tolerance of the culture was 58 g L−1. In a scaled-up batch experiment, which was performed in a 2.5 L fermentor with an initial glucose concentration of 100 g L−1, the pH was controlled at 6.5. In this experiment, the strain produced 57.86 g L−1 of butyric acid and 12.88 g L−1 of acetic acid, thus producing 70.74 g L−1 of total acids with a productivity of 0.69 g·L−1·h−1. A concentration of 70.74 g L−1 of acids equates to a yield of 0.71 g of acid per g consumed glucose. The maximum cell concentration was 3.80 g L−1, which may have been the reason for high productivity in the batch culture. Finally, corn steep liquor (CSL; a commercial nutrient solution) provided greater growth and acid production than the refined medium.

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