Journal
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
Volume 99, Issue 12, Pages 5033-5044Publisher
SPRINGER
DOI: 10.1007/s00253-015-6556-7
Keywords
Fumaric acid; Escherichia coli; Urea cycle; Metabolic analysis
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Funding
- National Basic Research Program of China [2013CB733600, 2012CB725200]
- Amoy Industrial Biotechnology RAMP
- D and Pilot Conversion Platform [3502Z20121009 and 3502Z20141054]
- National Nature Science Foundation of China [21390202, 21436002]
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Fumaric acid (FA) is an important raw material in the chemical and pharmaceutical industries. In this work, Escherichia coli was metabolically engineered for the production of FA. The fumA, fumB, fumC, and frdABCD genes were deleted to cut off the downstream pathway of FA. In addition, the iclR and arcA genes were also deleted to activate the glyoxylate shunt and to reinforce the oxidative Krebs cycle. To increase the FA yield, this base strain was further engineered to be pulled in the urea cycle by overexpressing the native carAB, argI, and heterologous rocF genes. The metabolites and the proteins of the Krebs cycle and the urea cycle were analyzed to confirm that the induced urea cycle improved the FA accumulation. With the induced urea cycle, the resulting strain ABCDIA-RAC was able to produce 11.38 mmol/L of FA from 83.33 mmol/L of glucose in a flask culture during 24 h of incubation.
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