The contest for precursors: channelling l -isoleucine synthesis in Corynebacterium glutamicum without byproduct formation
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| 008 | 210128e20150101xx s 000 0 eng | ||
| 024 | 7 | 0 | |a 10.1007/s00253-014-6109-5 |2 doi |
| 035 | |a (NATIONALLICENCE)springer-10.1007/s00253-014-6109-5 | ||
| 245 | 0 | 4 | |a The contest for precursors: channelling l -isoleucine synthesis in Corynebacterium glutamicum without byproduct formation |h [Elektronische Daten] |c [Michael Vogt, Karin Krumbach, Won-Gi Bang, Jan van Ooyen, Stephan Noack, Bianca Klein, Michael Bott, Lothar Eggeling] |
| 520 | 3 | |a l-Isoleucine is an essential amino acid, which is required as a pharma product and feed additive. Its synthesis shares initial steps with that of l-lysine and l-threonine, and four enzymes of l-isoleucine synthesis have an enlarged substrate specificity involved also in l-valine and l-leucine synthesis. As a consequence, constructing a strain specifically overproducing l-isoleucine without byproduct formation is a challenge. Here, we analyze for consequences of plasmid-encoded genes in Corynebacterium glutamicum MH20-22B on l-isoleucine formation, but still obtain substantial accumulation of byproducts. In a different approach, we introduce point mutations into the genome of MH20-22B to remove the feedback control of homoserine dehydrogenase, hom, and threonine dehydratase, ilvA, and we assay sets of genomic promoter mutations to increase hom and ilvA expression as well as to reduce dapA expression, the latter gene encoding the dihydrodipicolinate synthase. The promoter mutations are mirrored in the resulting differential protein levels determined by a targeted LC-MS/MS approach for the three key enzymes. The best combination of genomic mutations was found in strain K2P55, where 53mM l-isoleucine could be obtained. Whereas in fed-batch fermentations with the plasmid-based strain, 94mM l-isoleucine with l-lysine as byproduct was formed; with the plasmid-less strain K2P55, 109mM l-isoleucine accumulated with no substantial byproduct formation. The specific molar yield with the latter strain was 0.188mol l-isoleucine (mol glucose)−1 which characterizes it as one of the best l-isoleucine producers available and which does not contain plasmids. | |
| 540 | |a Springer-Verlag Berlin Heidelberg, 2014 | ||
| 690 | 7 | |a Genomic mutations |2 nationallicence | |
| 690 | 7 | |a Activity control |2 nationallicence | |
| 690 | 7 | |a Promoter mutations |2 nationallicence | |
| 690 | 7 | |a Branched-chain amino acids |2 nationallicence | |
| 690 | 7 | |a Flux directioning |2 nationallicence | |
| 690 | 7 | |a Byproduct reduction |2 nationallicence | |
| 690 | 7 | |a Enzyme quantification |2 nationallicence | |
| 700 | 1 | |a Vogt |D Michael |u Institut für Bio- und Geowissenschaften, IBG-1: Biotechnologie, Forschungszentrum Jülich GmbH, Jülich, Germany |4 aut | |
| 700 | 1 | |a Krumbach |D Karin |u Institut für Bio- und Geowissenschaften, IBG-1: Biotechnologie, Forschungszentrum Jülich GmbH, Jülich, Germany |4 aut | |
| 700 | 1 | |a Bang |D Won-Gi |u Department of Agricultural Chemistry, College of Life and Environmental Sciences, Korea University, Seoul, Sungbook-gu, South Korea |4 aut | |
| 700 | 1 | |a van Ooyen |D Jan |u Institut für Bio- und Geowissenschaften, IBG-1: Biotechnologie, Forschungszentrum Jülich GmbH, Jülich, Germany |4 aut | |
| 700 | 1 | |a Noack |D Stephan |u Institut für Bio- und Geowissenschaften, IBG-1: Biotechnologie, Forschungszentrum Jülich GmbH, Jülich, Germany |4 aut | |
| 700 | 1 | |a Klein |D Bianca |u Institut für Bio- und Geowissenschaften, IBG-1: Biotechnologie, Forschungszentrum Jülich GmbH, Jülich, Germany |4 aut | |
| 700 | 1 | |a Bott |D Michael |u Institut für Bio- und Geowissenschaften, IBG-1: Biotechnologie, Forschungszentrum Jülich GmbH, Jülich, Germany |4 aut | |
| 700 | 1 | |a Eggeling |D Lothar |u Institut für Bio- und Geowissenschaften, IBG-1: Biotechnologie, Forschungszentrum Jülich GmbH, Jülich, Germany |4 aut | |
| 773 | 0 | |t Applied Microbiology and Biotechnology |d Springer Berlin Heidelberg |g 99/2(2015-01-01), 791-800 |x 0175-7598 |q 99:2<791 |1 2015 |2 99 |o 253 | |
| 856 | 4 | 0 | |u https://doi.org/10.1007/s00253-014-6109-5 |q text/html |z Onlinezugriff via DOI |
| 898 | |a BK010053 |b XK010053 |c XK010000 | ||
| 900 | 7 | |a Metadata rights reserved |b Springer special CC-BY-NC licence |2 nationallicence | |
| 908 | |D 1 |a research-article |2 jats | ||
| 949 | |B NATIONALLICENCE |F NATIONALLICENCE |b NL-springer | ||
| 950 | |B NATIONALLICENCE |P 856 |E 40 |u https://doi.org/10.1007/s00253-014-6109-5 |q text/html |z Onlinezugriff via DOI | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Vogt |D Michael |u Institut für Bio- und Geowissenschaften, IBG-1: Biotechnologie, Forschungszentrum Jülich GmbH, Jülich, Germany |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Krumbach |D Karin |u Institut für Bio- und Geowissenschaften, IBG-1: Biotechnologie, Forschungszentrum Jülich GmbH, Jülich, Germany |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Bang |D Won-Gi |u Department of Agricultural Chemistry, College of Life and Environmental Sciences, Korea University, Seoul, Sungbook-gu, South Korea |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a van Ooyen |D Jan |u Institut für Bio- und Geowissenschaften, IBG-1: Biotechnologie, Forschungszentrum Jülich GmbH, Jülich, Germany |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Noack |D Stephan |u Institut für Bio- und Geowissenschaften, IBG-1: Biotechnologie, Forschungszentrum Jülich GmbH, Jülich, Germany |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Klein |D Bianca |u Institut für Bio- und Geowissenschaften, IBG-1: Biotechnologie, Forschungszentrum Jülich GmbH, Jülich, Germany |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Bott |D Michael |u Institut für Bio- und Geowissenschaften, IBG-1: Biotechnologie, Forschungszentrum Jülich GmbH, Jülich, Germany |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Eggeling |D Lothar |u Institut für Bio- und Geowissenschaften, IBG-1: Biotechnologie, Forschungszentrum Jülich GmbH, Jülich, Germany |4 aut | ||
| 950 | |B NATIONALLICENCE |P 773 |E 0- |t Applied Microbiology and Biotechnology |d Springer Berlin Heidelberg |g 99/2(2015-01-01), 791-800 |x 0175-7598 |q 99:2<791 |1 2015 |2 99 |o 253 | ||