PUMA publications for /tag/complex,%20Proteins,%20bacterialhttps://puma.ub.uni-stuttgart.de/tag/complex,%20Proteins,%20bacterialPUMA RSS feed for /tag/complex,%20Proteins,%20bacterial2024-03-28T17:39:52+01:00Engineering Corynebacterium glutamicum for the production of 2,3-butanediolhttps://puma.ub.uni-stuttgart.de/bibtex/29ce34124eaffe2a799aade38deec413d/bastianbastian2018-02-09T13:18:17+01:00Bacterial Bioreactors, Butylene Complex, Corynebacterium Dehydrogenase Dehydrogenase, Engineering Family, Glucose, Glycols, L-Lactate Lactococcus Metabolic Multigene Oxygen, Proteins, Pyruvate glutamicum, lactis, myown <span data-person-type="author" class="authorEditorList "><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Dušica Radoš" itemprop="url" href="/person/1518ec5750d920964df1a659788edff11/author/0"><span itemprop="name">D. Radoš</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Ana Lúcia Carvalho" itemprop="url" href="/person/1518ec5750d920964df1a659788edff11/author/1"><span itemprop="name">A. Carvalho</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Stefan Wieschalka" itemprop="url" href="/person/1518ec5750d920964df1a659788edff11/author/2"><span itemprop="name">S. Wieschalka</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Ana Rute Neves" itemprop="url" href="/person/1518ec5750d920964df1a659788edff11/author/3"><span itemprop="name">A. Neves</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Bastian Blombach" itemprop="url" href="/person/1518ec5750d920964df1a659788edff11/author/4"><span itemprop="name">B. Blombach</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Bernhard J. Eikmanns" itemprop="url" href="/person/1518ec5750d920964df1a659788edff11/author/5"><span itemprop="name">B. Eikmanns</span></a></span>, </span> and <span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Helena Santos" itemprop="url" href="/person/1518ec5750d920964df1a659788edff11/author/6"><span itemprop="name">H. Santos</span></a></span></span>. </span><span class="additional-entrytype-information"><span itemtype="http://schema.org/PublicationIssue" itemscope="itemscope" itemprop="isPartOf"><em><span itemprop="journal">Microb. Cell Fact.</span>, </em> </span>(<em><span>October 2015<meta content="October 2015" itemprop="datePublished"/></span></em>)</span>Fri Feb 09 13:18:17 CET 2018Microb. Cell Fact.oct171Engineering {Corynebacterium} glutamicum for the production of 2,3-butanediol142015Bacterial Bioreactors, Butylene Complex, Corynebacterium Dehydrogenase Dehydrogenase, Engineering Family, Glucose, Glycols, L-Lactate Lactococcus Metabolic Multigene Oxygen, Proteins, Pyruvate glutamicum, lactis, myown BACKGROUND: 2,3-Butanediol is an important bulk chemical with a wide range of applications. In bacteria, this metabolite is synthesised from pyruvate via a three-step pathway involving α-acetolactate synthase, α-acetolactate decarboxylase and 2,3-butanediol dehydrogenase. Thus far, the best producers of 2,3-butanediol are pathogenic strains, hence, the development of more suitable organisms for industrial scale fermentation is needed. Herein, 2,3-butanediol production was engineered in the Generally Regarded As Safe (GRAS) organism Corynebacterium glutamicum. A two-stage fermentation process was implemented: first, cells were grown aerobically on acetate; in the subsequent production stage cells were used to convert glucose into 2,3-butanediol under non-growing and oxygen-limiting conditions.
RESULTS: A gene cluster, encoding the 2,3-butanediol biosynthetic pathway of Lactococcus lactis, was assembled and expressed in background strains, C. glutamicum ΔldhA, C. glutamicum ΔaceEΔpqoΔldhA and C. glutamicum ΔaceEΔpqoΔldhAΔmdh, tailored to minimize pyruvate-consuming reactions, i.e., to prevent carbon loss in lactic, acetic and succinic acids. Producer strains were characterized in terms of activity of the relevant enzymes in the 2,3-butanediol forming pathway, growth, and production of 2,3-butanediol under oxygen-limited conditions. Productivity was maximized by manipulating the aeration rate in the production phase. The final strain, C. glutamicum ΔaceEΔpqoΔldhAΔmdh(pEKEx2-als,aldB,Ptuf butA), under optimized conditions produced 2,3-butanediol with a 0.66 mol mol(-1) yield on glucose, an overall productivity of 0.2 g L(-1) h(-1) and a titer of 6.3 g L(-1).
CONCLUSIONS: We have successfully developed C. glutamicum into an efficient cell factory for 2,3-butanediol production. The use of the engineered strains as a basis for production of acetoin, a widespread food flavour, is proposed.Corynebacterium glutamicum tailored for high-yield L-valine productionhttps://puma.ub.uni-stuttgart.de/bibtex/29723d6ce30daaf4910269ea54837c2d7/bastianbastian2018-02-09T13:18:17+01:00Bacterial Biomass, Biosynthetic Complex, Corynebacterium Dehydrogenase Engineering, Expression, Fermentation Gene Genetic Ketol-Acid Oxidoreductases, Pathways, Proteins, Pyruvate Reductoisomerase, Transaminases, Valine, glutamicum, 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/1e1d424fe4d9f617e1a0837a7567bea3a/author/0"><span itemprop="name">B. Blombach</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Mark E. Schreiner" itemprop="url" href="/person/1e1d424fe4d9f617e1a0837a7567bea3a/author/1"><span itemprop="name">M. Schreiner</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Tobias Bartek" itemprop="url" href="/person/1e1d424fe4d9f617e1a0837a7567bea3a/author/2"><span itemprop="name">T. Bartek</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Marco Oldiges" itemprop="url" href="/person/1e1d424fe4d9f617e1a0837a7567bea3a/author/3"><span itemprop="name">M. Oldiges</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/1e1d424fe4d9f617e1a0837a7567bea3a/author/4"><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">79 </span></span>(<span itemprop="issueNumber">3</span>):
<span itemprop="pagination">471--479</span></em> </span>(<em><span>June 2008<meta content="June 2008" itemprop="datePublished"/></span></em>)</span>Fri Feb 09 13:18:17 CET 2018Appl. Microbiol. Biotechnol.jun3471--479Corynebacterium glutamicum tailored for high-yield {L}-valine production792008Bacterial Biomass, Biosynthetic Complex, Corynebacterium Dehydrogenase Engineering, Expression, Fermentation Gene Genetic Ketol-Acid Oxidoreductases, Pathways, Proteins, Pyruvate Reductoisomerase, Transaminases, Valine, glutamicum, myown We recently engineered the wild type of Corynebacterium glutamicum for the growth-decoupled production of L: -valine from glucose by inactivation of the pyruvate dehydrogenase complex and additional overexpression of the ilvBNCE genes, encoding the L-valine biosynthetic enzymes acetohydroxyacid synthase, isomeroreductase, and transaminase B. Based on the first generation of pyruvate-dehydrogenase-complex-deficient C. glutamicum strains, a second generation of high-yield L-valine producers was constructed by successive deletion of the genes encoding pyruvate:quinone oxidoreductase, phosphoglucose isomerase, and pyruvate carboxylase and overexpression of ilvBNCE. In fed-batch fermentations at high cell densities, the newly constructed strains produced up to 410 mM (48 g/l) L-valine, showed a maximum yield of 0.75 to 0.86 mol/mol (0.49 to 0.56 g/g) of glucose in the production phase and, in contrast to the first generation strains, excreted neither pyruvate nor any other by-product tested.Increased glucose utilization in Corynebacterium glutamicum by use of maltose, and its application for the improvement of L-valine productivityhttps://puma.ub.uni-stuttgart.de/bibtex/21df12e8a60a9e4096c1d87d23eeb3e5c/bastianbastian2018-02-09T13:18:17+01:00Bacterial Complex, Corynebacterium Dehydrogenase Glucose, Maltose, Phosphoenolpyruvate Phosphotransferase Proteins, Pyruvate Sugar System Valine, 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/1ad8f9fd57bd8d9502e4a4cb86d864384/author/0"><span itemprop="name">F. Krause</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Alexander Henrich" itemprop="url" href="/person/1ad8f9fd57bd8d9502e4a4cb86d864384/author/1"><span itemprop="name">A. Henrich</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Bastian Blombach" itemprop="url" href="/person/1ad8f9fd57bd8d9502e4a4cb86d864384/author/2"><span itemprop="name">B. Blombach</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Reinhard Krämer" itemprop="url" href="/person/1ad8f9fd57bd8d9502e4a4cb86d864384/author/3"><span itemprop="name">R. Krämer</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Bernhard J. Eikmanns" itemprop="url" href="/person/1ad8f9fd57bd8d9502e4a4cb86d864384/author/4"><span itemprop="name">B. Eikmanns</span></a></span>, </span> and <span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Gerd M. Seibold" itemprop="url" href="/person/1ad8f9fd57bd8d9502e4a4cb86d864384/author/5"><span itemprop="name">G. Seibold</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">1</span>):
<span itemprop="pagination">370--374</span></em> </span>(<em><span>January 2010<meta content="January 2010" itemprop="datePublished"/></span></em>)</span>Fri Feb 09 13:18:17 CET 2018Appl. Environ. Microbiol.jan1370--374Increased glucose utilization in {Corynebacterium} glutamicum by use of maltose, and its application for the improvement of {L}-valine productivity762010Bacterial Complex, Corynebacterium Dehydrogenase Glucose, Maltose, Phosphoenolpyruvate Phosphotransferase Proteins, Pyruvate Sugar System Valine, glutamicum, myown Corynebacterium glutamicum efficiently utilizes maltose as a substrate. We show here that the presence of maltose increases glucose utilization by raising the expression of ptsG, which encodes the glucose-specific EII permease of the phosphotransferase system. Consequently, the L-valine productivity of a pyruvate dehydrogenase complex-deficient C. glutamicum strain was improved by the presence of maltose.