PUMA publications for /user/bastian/networks%20gene%20andhttps://puma.ub.uni-stuttgart.de/user/bastian/networks%20gene%20andPUMA RSS feed for /user/bastian/networks%20gene%20and2024-03-28T17:19:27+01:00Metabolic engineering of Corynebacterium glutamicum for 2-ketoisovalerate productionhttps://puma.ub.uni-stuttgart.de/bibtex/2bc1670b16ed84beabd8820d13e46b2d2/bastianbastian2018-02-09T13:18:17+01:00Acids, Bacterial, Corynebacterium Deletion, Engineering, Expression, Gene Genes, Genetic Genetically Glucose, Keto Metabolic Modified Networks Organisms, Pathways, and glutamicum, myown <span data-person-type="author" class="authorEditorList "><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Felix S. Krause" itemprop="url" href="/person/13154880de1726a3496ebe8869a0a7468/author/0"><span itemprop="name">F. Krause</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Bastian Blombach" itemprop="url" href="/person/13154880de1726a3496ebe8869a0a7468/author/1"><span itemprop="name">B. Blombach</span></a></span>, </span> and <span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Bernhard J. Eikmanns" itemprop="url" href="/person/13154880de1726a3496ebe8869a0a7468/author/2"><span itemprop="name">B. Eikmanns</span></a></span></span>. </span><span class="additional-entrytype-information"><span itemtype="http://schema.org/PublicationIssue" itemscope="itemscope" itemprop="isPartOf"><em><span itemprop="journal">Appl. Environ. Microbiol.</span>, </em> <em><span itemtype="http://schema.org/PublicationVolume" itemscope="itemscope" itemprop="isPartOf"><span itemprop="volumeNumber">76 </span></span>(<span itemprop="issueNumber">24</span>):
<span itemprop="pagination">8053--8061</span></em> </span>(<em><span>December 2010<meta content="December 2010" itemprop="datePublished"/></span></em>)</span>Fri Feb 09 13:18:17 CET 2018Appl. Environ. Microbiol.dec248053--8061Metabolic engineering of {Corynebacterium} glutamicum for 2-ketoisovalerate production762010Acids, Bacterial, Corynebacterium Deletion, Engineering, Expression, Gene Genes, Genetic Genetically Glucose, Keto Metabolic Modified Networks Organisms, Pathways, and glutamicum, myown 2-Ketoisovalerate is used as a therapeutic agent, and a 2-ketoisovalerate-producing organism may serve as a platform for products deriving from this 2-keto acid. We engineered the wild type of Corynebacterium glutamicum for the growth-decoupled production of 2-ketoisovalerate from glucose by deletion of the aceE gene encoding the E1p subunit of the pyruvate dehydrogenase complex, deletion of the transaminase B gene ilvE, and additional overexpression of the ilvBNCD genes, encoding the l-valine biosynthetic enzymes acetohydroxyacid synthase (AHAS), acetohydroxyacid isomeroreductase, and dihydroxyacid dehydratase. 2-Ketoisovalerate production was further improved by deletion of the pyruvate:quinone oxidoreductase gene pqo. In fed-batch fermentations at high cell densities, the newly constructed strains produced up to 188 ± 28 mM (21.8 ± 3.2 g liter(-1)) 2-ketoisovalerate and showed a product yield of about 0.47 ± 0.05 mol per mol (0.3 ± 0.03 g per g) of glucose and a volumetric productivity of about 4.6 ± 0.6 mM (0.53 ± 0.07 g liter(-1)) 2-ketoisovalerate per h in the overall production phase. In studying the influence of the three branched-chain 2-keto acids 2-ketoisovalerate, 2-ketoisocaproate, and 2-keto-3-methylvalerate on the AHAS activity, we observed a competitive inhibition of the AHAS enzyme by 2-ketoisovalerate.Platform engineering of Corynebacterium glutamicum with reduced pyruvate dehydrogenase complex activity for improved production of L-lysine, L-valine, and 2-ketoisovaleratehttps://puma.ub.uni-stuttgart.de/bibtex/2fc097b674eaff7966119cba97aaacd40/bastianbastian2018-02-09T13:18:17+01:00Acids, Biomass, Complex, Corynebacterium Dehydrogenase Deletion, Down-Regulation, Engineering Expression, Gene Genetic, Glucose, Keto Lysine, Metabolic Networks Pathways, Promoter Pyruvate Recombination, Regions, Valine, and glutamicum, myown <span data-person-type="author" class="authorEditorList "><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Jens Buchholz" itemprop="url" href="/person/1671300cf6c01886cf28889b6f3b9a19e/author/0"><span itemprop="name">J. Buchholz</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Andreas Schwentner" itemprop="url" href="/person/1671300cf6c01886cf28889b6f3b9a19e/author/1"><span itemprop="name">A. Schwentner</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Britta Brunnenkan" itemprop="url" href="/person/1671300cf6c01886cf28889b6f3b9a19e/author/2"><span itemprop="name">B. Brunnenkan</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Christina Gabris" itemprop="url" href="/person/1671300cf6c01886cf28889b6f3b9a19e/author/3"><span itemprop="name">C. Gabris</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Simon Grimm" itemprop="url" href="/person/1671300cf6c01886cf28889b6f3b9a19e/author/4"><span itemprop="name">S. Grimm</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Robert Gerstmeir" itemprop="url" href="/person/1671300cf6c01886cf28889b6f3b9a19e/author/5"><span itemprop="name">R. Gerstmeir</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Ralf Takors" itemprop="url" href="/person/1671300cf6c01886cf28889b6f3b9a19e/author/6"><span itemprop="name">R. Takors</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Bernhard J. Eikmanns" itemprop="url" href="/person/1671300cf6c01886cf28889b6f3b9a19e/author/7"><span itemprop="name">B. Eikmanns</span></a></span>, </span> and <span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Bastian Blombach" itemprop="url" href="/person/1671300cf6c01886cf28889b6f3b9a19e/author/8"><span itemprop="name">B. Blombach</span></a></span></span>. </span><span class="additional-entrytype-information"><span itemtype="http://schema.org/PublicationIssue" itemscope="itemscope" itemprop="isPartOf"><em><span itemprop="journal">Appl. Environ. Microbiol.</span>, </em> <em><span itemtype="http://schema.org/PublicationVolume" itemscope="itemscope" itemprop="isPartOf"><span itemprop="volumeNumber">79 </span></span>(<span itemprop="issueNumber">18</span>):
<span itemprop="pagination">5566--5575</span></em> </span>(<em><span>September 2013<meta content="September 2013" itemprop="datePublished"/></span></em>)</span>Fri Feb 09 13:18:17 CET 2018Appl. Environ. Microbiol.sep185566--5575Platform engineering of {Corynebacterium} glutamicum with reduced pyruvate dehydrogenase complex activity for improved production of {L}-lysine, {L}-valine, and 2-ketoisovalerate792013Acids, Biomass, Complex, Corynebacterium Dehydrogenase Deletion, Down-Regulation, Engineering Expression, Gene Genetic, Glucose, Keto Lysine, Metabolic Networks Pathways, Promoter Pyruvate Recombination, Regions, Valine, and glutamicum, myown Exchange of the native Corynebacterium glutamicum promoter of the aceE gene, encoding the E1p subunit of the pyruvate dehydrogenase complex (PDHC), with mutated dapA promoter variants led to a series of C. glutamicum strains with gradually reduced growth rates and PDHC activities. Upon overexpression of the l-valine biosynthetic genes ilvBNCE, all strains produced l-valine. Among these strains, C. glutamicum aceE A16 (pJC4 ilvBNCE) showed the highest biomass and product yields, and thus it was further improved by additional deletion of the pqo and ppc genes, encoding pyruvate:quinone oxidoreductase and phosphoenolpyruvate carboxylase, respectively. In fed-batch fermentations at high cell densities, C. glutamicum aceE A16 Δpqo Δppc (pJC4 ilvBNCE) produced up to 738 mM (i.e., 86.5 g/liter) l-valine with an overall yield (YP/S) of 0.36 mol per mol of glucose and a volumetric productivity (QP) of 13.6 mM per h [1.6 g/(liter × h)]. Additional inactivation of the transaminase B gene (ilvE) and overexpression of ilvBNCD instead of ilvBNCE transformed the l-valine-producing strain into a 2-ketoisovalerate producer, excreting up to 303 mM (35 g/liter) 2-ketoisovalerate with a YP/S of 0.24 mol per mol of glucose and a QP of 6.9 mM per h [0.8 g/(liter × h)]. The replacement of the aceE promoter by the dapA-A16 promoter in the two C. glutamicum l-lysine producers DM1800 and DM1933 improved the production by 100\% and 44\%, respectively. These results demonstrate that C. glutamicum strains with reduced PDHC activity are an excellent platform for the production of pyruvate-derived products.Engineering Corynebacterium glutamicum for the production of pyruvatehttps://puma.ub.uni-stuttgart.de/bibtex/246bcc08cb2281fd8c7391745cce2ec97/bastianbastian2018-02-09T13:18:17+01:00Acid, Corynebacterium Deletion, Engineering Expression, Gene Glucose, Metabolic Networks Pathways, Pyruvic Valine, and glutamicum, myown <span data-person-type="author" class="authorEditorList "><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Stefan Wieschalka" itemprop="url" href="/person/148b87aa35f4559e7a9ead73d1df42fd1/author/0"><span itemprop="name">S. Wieschalka</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Bastian Blombach" itemprop="url" href="/person/148b87aa35f4559e7a9ead73d1df42fd1/author/1"><span itemprop="name">B. Blombach</span></a></span>, </span> and <span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Bernhard J. Eikmanns" itemprop="url" href="/person/148b87aa35f4559e7a9ead73d1df42fd1/author/2"><span itemprop="name">B. Eikmanns</span></a></span></span>. </span><span class="additional-entrytype-information"><span itemtype="http://schema.org/PublicationIssue" itemscope="itemscope" itemprop="isPartOf"><em><span itemprop="journal">Appl. Microbiol. Biotechnol.</span>, </em> <em><span itemtype="http://schema.org/PublicationVolume" itemscope="itemscope" itemprop="isPartOf"><span itemprop="volumeNumber">94 </span></span>(<span itemprop="issueNumber">2</span>):
<span itemprop="pagination">449--459</span></em> </span>(<em><span>April 2012<meta content="April 2012" itemprop="datePublished"/></span></em>)</span>Fri Feb 09 13:18:17 CET 2018Appl. Microbiol. Biotechnol.apr2449--459Engineering {Corynebacterium} glutamicum for the production of pyruvate942012Acid, Corynebacterium Deletion, Engineering Expression, Gene Glucose, Metabolic Networks Pathways, Pyruvic Valine, and glutamicum, myown A Corynebacterium glutamicum strain with inactivated pyruvate dehydrogenase complex and a deletion of the gene encoding the pyruvate:quinone oxidoreductase produces about 19 mM L: -valine, 28 mM L: -alanine and about 55 mM pyruvate from 150 mM glucose. Based on this double mutant C. glutamicum △aceE △pqo, we engineered C. glutamicum for efficient production of pyruvate from glucose by additional deletion of the ldhA gene encoding NAD(+)-dependent L: -lactate dehydrogenase (LdhA) and introduction of a attenuated variant of the acetohydroxyacid synthase (△C-T IlvN). The latter modification abolished overflow metabolism towards L: -valine and shifted the product spectrum to pyruvate production. In shake flasks, the resulting strain C. glutamicum △aceE △pqo △ldhA △C-T ilvN produced about 190 mM pyruvate with a Y (P/S) of 1.36 mol per mol of glucose; however, it still secreted significant amounts of L: -alanine. Additional deletion of genes encoding the transaminases AlaT and AvtA reduced L: -alanine formation by about 50\%. In fed-batch fermentations at high cell densities with adjusted oxygen supply during growth and production (0-5\% dissolved oxygen), the newly constructed strain C. glutamicum △aceE △pqo △ldhA △C-T ilvN △alaT △avtA produced more than 500 mM pyruvate with a maximum yield of 0.97 mol per mole of glucose and a productivity of 0.92 mmol g ((CDW)) (-1) h(-1) (i.e., 0.08 g g((CDW)) (-1) h(-1)) in the production phase.