PUMA publications for /tag/bacterial,%20andhttps://puma.ub.uni-stuttgart.de/tag/bacterial,%20andPUMA RSS feed for /tag/bacterial,%20and2024-03-29T07:07:06+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.Corynebacterium glutamicum tailored for efficient isobutanol productionhttps://puma.ub.uni-stuttgart.de/bibtex/2a0a5fcfba6017eb6a61292baef557af7/bastianbastian2018-02-09T13:18:17+01:00Anaerobiosis, Bacterial Bacterial, Butanols, Chromosomes, Corynebacterium Escherichia Fungal Glucose, Lactococcus Metabolic Networks Pathways, Plasmids, Proteins Proteins, Recombinant Saccharomyces and cerevisiae, coli, glutamicum, lactis, myown <span data-person-type="author" class="authorEditorList "><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Bastian Blombach" itemprop="url" href="/person/10ff226fcd2ff0614810a1890f8f46f8e/author/0"><span itemprop="name">B. Blombach</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Tanja Riester" itemprop="url" href="/person/10ff226fcd2ff0614810a1890f8f46f8e/author/1"><span itemprop="name">T. Riester</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Stefan Wieschalka" itemprop="url" href="/person/10ff226fcd2ff0614810a1890f8f46f8e/author/2"><span itemprop="name">S. Wieschalka</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Christian Ziert" itemprop="url" href="/person/10ff226fcd2ff0614810a1890f8f46f8e/author/3"><span itemprop="name">C. Ziert</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Jung-Won Youn" itemprop="url" href="/person/10ff226fcd2ff0614810a1890f8f46f8e/author/4"><span itemprop="name">J. Youn</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Volker F. Wendisch" itemprop="url" href="/person/10ff226fcd2ff0614810a1890f8f46f8e/author/5"><span itemprop="name">V. Wendisch</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/10ff226fcd2ff0614810a1890f8f46f8e/author/6"><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">77 </span></span>(<span itemprop="issueNumber">10</span>):
<span itemprop="pagination">3300--3310</span></em> </span>(<em><span>May 2011<meta content="May 2011" itemprop="datePublished"/></span></em>)</span>Fri Feb 09 13:18:17 CET 2018Appl. Environ. Microbiol.may103300--3310Corynebacterium glutamicum tailored for efficient isobutanol production772011Anaerobiosis, Bacterial Bacterial, Butanols, Chromosomes, Corynebacterium Escherichia Fungal Glucose, Lactococcus Metabolic Networks Pathways, Plasmids, Proteins Proteins, Recombinant Saccharomyces and cerevisiae, coli, glutamicum, lactis, myown We recently engineered Corynebacterium glutamicum for aerobic production of 2-ketoisovalerate by inactivation of the pyruvate dehydrogenase complex, pyruvate:quinone oxidoreductase, transaminase B, and additional overexpression of the ilvBNCD genes, encoding acetohydroxyacid synthase, acetohydroxyacid isomeroreductase, and dihydroxyacid dehydratase. Based on this strain, we engineered C. glutamicum for the production of isobutanol from glucose under oxygen deprivation conditions by inactivation of l-lactate and malate dehydrogenases, implementation of ketoacid decarboxylase from Lactococcus lactis, alcohol dehydrogenase 2 (ADH2) from Saccharomyces cerevisiae, and expression of the pntAB transhydrogenase genes from Escherichia coli. The resulting strain produced isobutanol with a substrate-specific yield (Y(P/S)) of 0.60 ± 0.02 mol per mol of glucose. Interestingly, a chromosomally encoded alcohol dehydrogenase rather than the plasmid-encoded ADH2 from S. cerevisiae was involved in isobutanol formation with C. glutamicum, and overexpression of the corresponding adhA gene increased the Y(P/S) to 0.77 ± 0.01 mol of isobutanol per mol of glucose. Inactivation of the malic enzyme significantly reduced the Y(P/S), indicating that the metabolic cycle consisting of pyruvate and/or phosphoenolpyruvate carboxylase, malate dehydrogenase, and malic enzyme is responsible for the conversion of NADH + H+ to NADPH + H+. In fed-batch fermentations with an aerobic growth phase and an oxygen-depleted production phase, the most promising strain, C. glutamicum ΔaceE Δpqo ΔilvE ΔldhA Δmdh(pJC4ilvBNCD-pntAB)(pBB1kivd-adhA), produced about 175 mM isobutanol, with a volumetric productivity of 4.4 mM h⁻¹, and showed an overall Y(P/S) of about 0.48 mol per mol of glucose in the production phase.