Butyl butyrate is widely used as a fragrance additive for foods and beverages. The first step in the currently used process is the production of precursors, including butanol and butyrate, from petroleum using chemical catalysts, followed by the conversion of precursors to butyl butyrate by immobilized lipase. In this work, we engineered Clostridium acetobutylicum for the selective, one-step production of butyl butyrate from glucose. C. acetobutylicum ATCC 824, possessing a strong carbon flux that yields butanol and butyryl-CoA, was selected as a host and was engineered by introducing alcohol acyltransferases (AATs) from Fragaria x ananassa (strawberry) or Malus sp. (apple). Batch culture of the engineered C. acetobutylicum strain CaSAAT expressing the strawberry SAAT gene produced 50.07 mg/L of butyl butyrate with a selectivity of 84.8% of total esters produced. Also, the engineered C. acetobutylicum strain CaAAAT expressing the apple AAAT gene produced 40.60 mg/L of butyl butyrate with a selectivity of 87.4%. This study demonstrated the feasibility of the one-step fermentation of butyl butyrate from glucose in the engineered C. acetobutylicum, as a proof of concept.
Clostridium is a genus of Gram-positive, rod shape, spore-forming obligate anaerobe. Recently, Clostridium has been attracted as a host for bio-based chemical production, due to its diversity of substrate utilization and the production ability for metabolites which can be used as a building block for chemical production. Especially, butanol produced from Clostridium has been considered as an alternative fuel. As a transportation fuel, butanol has a higher energy density and lower hygroscopicity compared to ethanol, the first generation biofuel. Recently, metabolic engineering of Clostridium has been massively conducted for butanol production. In this study, the metabolic engineering strategy of Clostridium for butanol production has been reviewed with a brief perspective.
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