Influence of nutritional and operational parameters on the production of butanol or 1,3-propanediol from glycerol by a mutant Clostridium pasteurianum

- Random chemical mutagenesis of Clostridium pasteurianum in solid medium was conducted.
- A moderate butanol-resistant strain (M2) was isolated.
- The strain M2 produced higher amounts of butanol under optimal iron concentration and inoculum age.
- N2 sparging is a suitable strategy to increase 1,3-propanediol titer, yield and productivity.
- A low operational pH favours the production of butanol.

Butanol and 1,3-propanediol (1,3-PDO) are simultaneously produced by Clostridium pasteurianum from glycerol. In this study, random chemical mutagenesis of C. pasteurianum DSM 525 was conducted to improve its tolerance to butanol. Selected nutritional and operational parameters were evaluated to identify strategies that favour the production of each metabolite. From those experiments, it was possible to isolate cells able to produce 22% more butanol than the parent strain in serum bottles. The supplementation of the culture medium with 2 mg l−1 of iron increased the production of butanol by 163%, and the optimum inoculum age was found to be 12 hours. Overall, the experiments conducted in bioreactor led to lower butanol titers than in serum bottles, which is attributed to the higher pressure present in the bottles. At pH 6.0, N2 sparging notoriously favoured the production of biomass and 1,3-PDO, while a lower pH (5.0) led to a higher butanol yield, although growth was negatively affected. The results herein gathered allowed the identification of specific conditions that favour the production of either butanol or 1,3-PDO. Furthermore, it was found that N2 sparging is a suitable strategy to maximize the titer, yield and productivity of 1,3-PDO using C. pasteurianum.