Reducing cofactors contribute to the increase of butanol production by a wild-type Clostridium sp. strain BOH3.

Availability of reducing factors (e.g., NADH and NADPH) plays an important role in improving the efficacy of products conversion in cofactor-dependent production systems. In this study, nicotinic acid (NA), the precursor of NADH and NADPH, was supplemented to the growth medium of a wild-type Clostridium sp. strain BOH3. Results showed that the addition of precursor NA to the medium led to a significant increase in the levels of NADH and NADPH. Meanwhile, a maximal cell growth rate and butanol generation rate were reached by applying a two-stage pH-shift strategy, achieving 18.7g/L butanol with a yield of 24.6% and a productivity of 0.26g/Lh. The metabolic patterns were shifted towards more reduced metabolites as reflected by higher butanol-to-acetone ratio (11%) and butanol-to-acid ratio (292%). Redistributing metabolic flux to butanol via manipulations of reducing cofactor and pH shift could become an alternative tool to realize metabolic engineering goals.