Evaluation of yeast alcohol acetyltransferases for ethyl acetate production in Clostridium ljungdahlii.

Boto ST, Gerges K, Bardl B, Rosenbaum MA (2025) Evaluation of yeast alcohol acetyltransferases for ethyl acetate production in Clostridium ljungdahlii. Eng Life Sci 25(1), e202400076.

Abstract

Sustainable chemical production from C1 gaseous substrates, such as syngas or CO2 /H2 , can be achieved through gas fermentation. In gas fermentation, acetogenic bacteria are able to utilize oxidized inorganic carbon sources as the sole carbon source and electron acceptor, while reduced inorganic species are used as the electron donor. Clostridium ljungdahlii, a model acetogen, is only capable of reducing CO2 to acetate and ethanol, with H 2 as electron donor. In order to expand the product profile of this bacterium, five alcohol acetyltransferases (AATs) from yeast were heterologously expressed in C. ljungdahlii to evaluate its potential to produce ethyl acetate. When growing on CO2 and H 2 , up to 7.38 ± 0.43 mg/L of ethyl acetate were produced. Using fructose as the main carbon and energy source, up to 23.15 ± 1.28 mg/L of ethyl acetate were produced. Ethanol and fumarate supplementation were
able to boost ethyl acetate titers (up to 37.51 ± 9.44 mg/L). Hence, ethyl acetate production was enabled in C. ljungdahlii at low titers, which could be explained by the high energetic cost of operation of AATs, and their shown promiscuity. However, we also show that this opens the door to more complex esterification reactions of higher added value biomolecules.

Leibniz-HKI-Authors

Miriam Agler-Rosenbaum
Bettina Bardl
Santiago Boto
Kareem Gerges

Identifier

doi: 10.1002/elsc.202400076

PMID: 39850489