Butanol Production by a Novel Efficient Method Using Mixed Cultures of Clostridium beijerinckii and Arthrobacter sp. in Stirred-Tank and Gas-Lift Bioreactors

Abstract

Arthrobacter sp. BCC 72131, an obligate aerobic bacterium, was used to create anaerobic conditions, and Clostridium beijerinckii TISTR 1461 was used as a butanol producer in an acetone-butanol–ethanol (ABE) fermentation. Sweet sorghum juice (SSJ) medium containing 60 g/L of total sugar supplemented with 1.27 g/L of (NH4)2SO4 was used as a butanol production (BP) medium. Arthrobacter sp. was inoculated into the BP medium in 1-L screw-capped bottles. After 2, 4, 6 and 12 h of Arthrobacter sp. cultivation at 30 °C, C. beijerinckii was transferred into the BP medium to start butanol production at 37 °C. The results showed that C. beijerinckii inoculation after 6 h of Arthrobacter sp. cultivation gave the highest butanol titer (PB) at 12.56 g/L, with a butanol yield (YB/S) and volumetric butanol productivity (QB) of 0.34 g/g and 0.23 g/L·h, respectively. These values are approximately 10–27% higher than those of the control experiment using a single culture of C. beijerinckii TISTR 1461 and oxygen-free nitrogen (OFN) gas flushing to create anaerobic conditions. Field emission scanning electron microscopic (FE-SEM) images of Clostridium cells, as well as protein and free amino nitrogen concentrations in the broth during butanol fermentation were also studied to confirm the results. The butanol fermentation was then carried out in a 5.6-L stirred-tank and a 1.2-L low-cost gas-lift bioreactor by the mixed cultures using the optimal time of Clostridium inoculation. The PB, YB/S and QB values obtained were not significantly different from those in the 1-L screw-capped bottles. Hence, Arthrobacter sp. can be used as a novel method to create anaerobic conditions instead of a traditional method employing OFN gas flushing. Using mixed cultures of Arthrobacter sp. BCC 72131 and C. beijerinckii TISTR 1461 is a practical method to produce butanol on a large-scale, both in complex and low-cost bioreactors.