Fed-batch two-phase production of alanine by a metabolically engineered Escherichia coli |
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Authors: | Geoffrey M. Smith Sarah A. Lee Kevin C. Reilly Mark A. Eiteman Elliot Altman |
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Affiliation: | (1) Center for Molecular BioEngineering, Driftmier Engineering, University of Georgia, Athens, GA 30602, USA |
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Abstract: | dl-Alanine was produced from glucose in an Escherichia coli pfl pps poxB ldhA aceEF pTrc99A-alaD strain which lacked pyruvate-formate lyase, phosphoenolpyruvate (PEP) synthase, pyruvate oxidase, lactate dehydogenase, components of the pyruvate dehydogenase complex and over-produced alanine dehydrogenase (ALD). A two-phase process was developed with cell growth under aerobic conditions followed by alanine production under anaerobic conditions. Using the batch mode, cells grew to 5.3 g/l in 9 h with the accumulation of 6–10 g acetate/l, and under subsequent anaerobic conditions achieved 34 g alanine/l in 13 h with a yield of 0.86 g/g glucose. Using the fed-batch mode at μ = 0.15 h−1, only about 1 g acetate/l formed in the 25 h required for the cells to reach 5.6 g/l, and 88 g alanine/l accumulated during the subsequent 23 h. This fed-batch process attained an alanine volumetric productivity of 4 g/lh during the production phase, and a yield that was essentially 1 g/g. |
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Keywords: | Acetate Alanine Alanine dehydrogenase Fed-batch process Overflow metabolism |
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