| Popis: |
Background: Ethyl acetate and hydrogen are industrially relevant compounds that preferably are produced via sustainable, non-petrochemical production processes. Both compounds are volatile and can be produced by Escherichia coli before. However, relatively low yields for hydrogen are obtained and a mix of by-products render the sole production of hydrogen by micro-organisms unfeasible. High yields for ethyl acetate have been achieved but accumulation of formate remained an undesired but inevitable obstacle. Coupling ethyl acetate production to the conversion of formate into H2 may offer an interesting solution to both drawbacks. Ethyl acetate production requires equimolar amounts of ethanol and acetyl-CoA, which enables a redox neutral fermentation, without the need for production of by-products, other than hydrogen and CO2. Results: We engineered Escherichia coli, towards improved conversion of formate into hydrogen and CO2 by inactivating the formate hydrogen lyase repressor (hycA), both uptake hydrogenases (hyaAB, hybBC) and/or overexpressing the hydrogen formate lyase activator (fhlA). Initially 10 strains were evaluated in anaerobic serum bottles with respect to growth, after which four strains were further analyzed. Anaerobic co-production of hydrogen and ethyl acetate via heterologous ethanol acyltransferase (Eat1) was achieved in 1.5-L pH controlled bioreactors. Conclusions: We showed that the engineered strains co-produced ethyl acetate and hydrogen to yields exceeding 70 % of the pathway maximum for ethyl acetate and hydrogen, and propose in-situ product removal via gas stripping as efficient technique to isolate the products of interest. |
