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Bücher Englische Bücher Sachbücher Natur & Technik Metabolic engineering of Corynebacterium glutamicum for production of L-leucine and 2-ketoisocaproate
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Metabolic engineering of Corynebacterium glutamicum for production of L-leucine and 2-ketoisocaproate
Michael Vogt (Broschiert, Englisch)
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Beschreibung
Due to the depletion of fossil energy sources, there is an increasing demand for alternative
sustainable production processes utilizing renewable resources. Biotechnological
approaches using microorganisms such as Corynebacterium glutamicum as biocatalysts play
an important role for the production of beneficial substances. These include the three
branched-chain amino acids L-valine, L-isoleucine,… and L-leucine, as well as their respective
keto acid precursors, which have diverse commercial applications in food, feed, and
pharmaceutical industry. In this work, metabolic engineering of the C. glutamicum wild type
was employed to develop efficient strains for the production of L-leucine and
2-ketoisocaproate.
The key-player enzyme in L-leucine biosynthesis is the leuA-encoded 2-isopropylmalate
synthase which is feedback-inhibited by low L-leucine concentrations with a Ki of 0.4 mM. A
feedback-resistant variant of the 2-isopropylmalate synthase was identified and
characterized biochemically in the available weak L-leucine producer B018, which had been
obtained by random mutagenesis and screening. The respective gene leuA_B018, devoid of
the attenuator region and under control of a strong promoter, was integrated in up to three
copies into the genome of C. glutamicum wild type and combined with additional genomic
modifications aimed at increasing L-leucine production. These modifications involved
I) deletion of the gene ltbR encoding the repressor LtbR to increase expression of genes
leuBCD, II) deletion of the gene iolR encoding the transcriptional regulator IolR to increase
glucose uptake, III) reduction of citrate synthase activity to increase precursor supply, and
IV) introduction of a modified ilvN gene encoding a feedback-resistant acetohydroxyacid
synthase. The production performance of the resulting strains was characterized in shake
flask and bioreactor cultivations. Under fed-batch conditions, the best producer strain
accumulated L-leucine to levels exceeding the solubility limit of approximately 24 g l-1. The
maximal molar product yield and volumetric productivity were 0.30 mol per mol glucose and
4.3 mmol l-1 h-1, respectively. Moreover, the achieved values were obtained in a defined
minimal medium with a prototrophic and plasmid-free strain, making this process highly
interesting for industrial application. Dieses Produkt haben wir gerade leider nicht auf Lager.
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Technische Daten
Erscheinungsdatum
30.08.2014
Sprache
Englisch
EAN
9783893369683
Herausgeber
Forschungszentrum Jülich
Sonderedition
Nein
Autor
Michael Vogt
Seitenanzahl
92
Einbandart
Broschiert
Schlagwörter
L-leucine, Corynebacterium glutamicum, -ketoisocaproate
Thema-Inhalt
T - Technologie, Ingenieurswissenschaft, Landwirtschaft
PS - Biologie, Biowissenschaften
Höhe
240 mm
Breite
17 cm
-.-
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