4.5 Article

Production of L-lysine on different silage juices using genetically engineered Corynebacterium glutamicum

Journal

JOURNAL OF BIOTECHNOLOGY
Volume 163, Issue 2, Pages 217-224

Publisher

ELSEVIER SCIENCE BV
DOI: 10.1016/j.jbiotec.2012.07.190

Keywords

Grass silage; Corn silage; Amino acid production; Overexpression; Fermentation; Oxygen limitation

Funding

  1. Fachagentur fur Nachwachsende Rohstoffe (FNR) [FKZ 22004908]
  2. BMBF (Federal Ministry of Education and Research Germany) [FKZ 0315800B]

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Corynebacterium glutamicum, the best established industrial producer organism for lysine was genetically modified to allow the production of lysine on grass and corn silages. The resulting strain C. glutamicum lysC(fbr) dld(Psod) pyc(Psod) malE(Psod) fbp(Psod) gapX(Psod) was based on earlier work (Neuner and Heinzle, 2011). That mutant carries a point mutation in the aspartokinase (lysC) regulatory subunit gene as well as overexpression of d-lactate dehydrogenase (dld), pyruvate carboxylase (pyc) and malic enzyme (malE) using the strong Psod promoter. Here, we additionally overexpressed fructose 1,6-bisphosphatase (fbp) and glyceraldehyde 3-phosphate dehydrogenase (gapX) using the same promoter. The resulting strain grew readily on grass and corn silages with a specific growth rate of 0.35 h(-1) and lysine carbon yields of approximately 90 C-mmol (C-mol)(-1). Lysine yields were hardly affected by oxygen limitation whereas linear growth was observed under oxygen limiting conditions. Overall, this strain seems very robust with respect to the composition of silage utilizing all quantified low molecular weight substrates, e. g. lactate, glucose, fructose, maltose, quinate, fumarate, glutamate, leucine, isoleucine and alanine. (C) 2012 Elsevier B.V. All rights reserved.

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