Journal
METABOLIC ENGINEERING
Volume 30, Issue -, Pages 156-165Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymben.2015.06.003
Keywords
Cotynebacterium glutamicum; Cis-aconitate decarboxylase; Aconitase; Isocitrate clehydrogenase; MalE fusion protein; Nitrogen-limitation
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Funding
- Federal Ministry of Food, Agriculture, and Consumer Protection (BMELV) within the ERA-IB project Bio-based production of chemical building blocks (BioProChemBB) [220-095-08D]
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The capability of Corynebacterium glutamicum for glucose-based synthesis of itaconate was explored, which can serve as building block for production of polymers, chemicals, and fuels. C glutamicum was highly tolerant to itaconate and did not metabolize it. Expression of the Aspergillus terreus CAD1 gene encoding cis-aconitate decarboxylase (CAD) in strain ATCC13032 led to the production of 1.4 mM itaconate in the stationary growth phase. Fusion of CAD with the Escherichia colt maltose-binding protein increased its activity and the itaconate titer more than two-fold. Nitrogen-limited growth conditions boosted CAD activity and itaconate titer about 10-fold to values of 1440 mU mg(-1) and 30 mM. Reduction of isocitrate dehydrogenase activity via exchange of the ATG start codon to GTG or TUG resulted in maximal itaconate titers of 60 mM (7.8 gl(-1)), a molar yield of 0.4 mol mol(-1), and a volumetric productivity of 2.1 mmol 1(-1) h(-1). (C) 2015 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.
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