PUMA publications for /user/bastianhttps://puma.ub.uni-stuttgart.de/user/bastianPUMA RSS feed for /user/bastian2024-03-29T14:16:02+01:00L-valine production during growth of pyruvate dehydrogenase complex-deficient Corynebacterium glutamicum in the presence of ethanol or by inactivation of the transcriptional regulator SugRhttps://puma.ub.uni-stuttgart.de/bibtex/205af8a7b28d60ba8272685f0427eaadb/bastianbastian2018-02-09T13:18:17+01:00Complex Corynebacterium Dehydrogenase Ethanol, Factors, Gene Knockout Pyruvate Techniques, Transcription 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/1581b084966b1a19b10f06b5ed069aa58/author/0"><span itemprop="name">B. Blombach</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Annette Arndt" itemprop="url" href="/person/1581b084966b1a19b10f06b5ed069aa58/author/1"><span itemprop="name">A. Arndt</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Marc Auchter" itemprop="url" href="/person/1581b084966b1a19b10f06b5ed069aa58/author/2"><span itemprop="name">M. Auchter</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/1581b084966b1a19b10f06b5ed069aa58/author/3"><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">75 </span></span>(<span itemprop="issueNumber">4</span>):
<span itemprop="pagination">1197--1200</span></em> </span>(<em><span>February 2009<meta content="February 2009" itemprop="datePublished"/></span></em>)</span>Fri Feb 09 13:18:17 CET 2018Appl. Environ. Microbiol.feb41197--1200L-valine production during growth of pyruvate dehydrogenase complex-deficient {Corynebacterium} glutamicum in the presence of ethanol or by inactivation of the transcriptional regulator {SugR}752009Complex Corynebacterium Dehydrogenase Ethanol, Factors, Gene Knockout Pyruvate Techniques, Transcription Valine, glutamicum, myown Pyruvate dehydrogenase complex-deficient strains of Corynebacterium glutamicum produce L-valine from glucose only after depletion of the acetate required for growth. Here we show that inactivation of the DeoR-type transcriptional regulator SugR or replacement of acetate by ethanol already in course of the growth phase results in efficient L-valine production.Carbon flux analysis by 13C nuclear magnetic resonance to determine the effect of CO2 on anaerobic succinate production by Corynebacterium glutamicumhttps://puma.ub.uni-stuttgart.de/bibtex/22e74e76dec66d069069ac7fbe5bedf94/bastianbastian2018-02-09T13:18:17+01:00Acid, Anaerobiosis, Carbon Corynebacterium Dioxide Glucose, Isotope Isotopes, Labeling, Magnetic Resonance Spectroscopy, Succinic glutamicum, 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/138e7d0611f298c2eedc8bd9086a28955/author/0"><span itemprop="name">D. Radoš</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="David L. Turner" itemprop="url" href="/person/138e7d0611f298c2eedc8bd9086a28955/author/1"><span itemprop="name">D. Turner</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Luís L. Fonseca" itemprop="url" href="/person/138e7d0611f298c2eedc8bd9086a28955/author/2"><span itemprop="name">L. Fonseca</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Ana Lúcia Carvalho" itemprop="url" href="/person/138e7d0611f298c2eedc8bd9086a28955/author/3"><span itemprop="name">A. Carvalho</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Bastian Blombach" itemprop="url" href="/person/138e7d0611f298c2eedc8bd9086a28955/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/138e7d0611f298c2eedc8bd9086a28955/author/5"><span itemprop="name">B. Eikmanns</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Ana Rute Neves" itemprop="url" href="/person/138e7d0611f298c2eedc8bd9086a28955/author/6"><span itemprop="name">A. Neves</span></a></span>, </span> and <span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Helena Santos" itemprop="url" href="/person/138e7d0611f298c2eedc8bd9086a28955/author/7"><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">Appl. Environ. Microbiol.</span>, </em> <em><span itemtype="http://schema.org/PublicationVolume" itemscope="itemscope" itemprop="isPartOf"><span itemprop="volumeNumber">80 </span></span>(<span itemprop="issueNumber">10</span>):
<span itemprop="pagination">3015--3024</span></em> </span>(<em><span>May 2014<meta content="May 2014" itemprop="datePublished"/></span></em>)</span>Fri Feb 09 13:18:17 CET 2018Appl. Environ. Microbiol.may103015--3024Carbon flux analysis by 13C nuclear magnetic resonance to determine the effect of {CO}2 on anaerobic succinate production by {Corynebacterium} glutamicum802014Acid, Anaerobiosis, Carbon Corynebacterium Dioxide Glucose, Isotope Isotopes, Labeling, Magnetic Resonance Spectroscopy, Succinic glutamicum, myown Wild-type Corynebacterium glutamicum produces a mixture of lactic, succinic, and acetic acids from glucose under oxygen deprivation. We investigated the effect of CO2 on the production of organic acids in a two-stage process: cells were grown aerobically in glucose, and subsequently, organic acid production by nongrowing cells was studied under anaerobic conditions. The presence of CO2 caused up to a 3-fold increase in the succinate yield (1 mol per mol of glucose) and about 2-fold increase in acetate, both at the expense of l-lactate production; moreover, dihydroxyacetone formation was abolished. The redistribution of carbon fluxes in response to CO2 was estimated by using (13)C-labeled glucose and (13)C nuclear magnetic resonance (NMR) analysis of the labeling patterns in end products. The flux analysis showed that 97\% of succinate was produced via the reductive part of the tricarboxylic acid cycle, with the low activity of the oxidative branch being sufficient to provide the reducing equivalents needed for the redox balance. The flux via the pentose phosphate pathway was low ({\textasciitilde}5\%) regardless of the presence or absence of CO2. Moreover, there was significant channeling of carbon to storage compounds (glycogen and trehalose) and concomitant catabolism of these reserves. The intracellular and extracellular pools of lactate and succinate were measured by in vivo NMR, and the stoichiometry (H(+):organic acid) of the respective exporters was calculated. This study shows that it is feasible to take advantage of natural cellular regulation mechanisms to obtain high yields of succinate with C. glutamicum without genetic manipulation.Current knowledge on isobutanol production with Escherichia coli, Bacillus subtilis and Corynebacterium glutamicumhttps://puma.ub.uni-stuttgart.de/bibtex/20f88c2605071e1c350ed8a296eece367/bastianbastian2018-02-09T13:18:17+01:00Acids, Alcohol Bacillus Bacterial Butanols, Carboxy-Lyases, Corynebacterium Dehydrogenase, Engineering Escherichia Industrial Keto Metabolic Microbiology, Proteins, Recombinant coli, glutamicum, myown subtilis, <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/133960b6afe49761f6398812d65d1660e/author/0"><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/133960b6afe49761f6398812d65d1660e/author/1"><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">Bioeng Bugs</span>, </em> <em><span itemtype="http://schema.org/PublicationVolume" itemscope="itemscope" itemprop="isPartOf"><span itemprop="volumeNumber">2 </span></span>(<span itemprop="issueNumber">6</span>):
<span itemprop="pagination">346--350</span></em> </span>(<em><span>December 2011<meta content="December 2011" itemprop="datePublished"/></span></em>)</span>Fri Feb 09 13:18:17 CET 2018Bioeng Bugsdec6346--350Current knowledge on isobutanol production with {Escherichia} coli, {Bacillus} subtilis and {Corynebacterium} glutamicum22011Acids, Alcohol Bacillus Bacterial Butanols, Carboxy-Lyases, Corynebacterium Dehydrogenase, Engineering Escherichia Industrial Keto Metabolic Microbiology, Proteins, Recombinant coli, glutamicum, myown subtilis, Due to steadily rising crude oil prices great efforts have been made to develop designer bugs for the fermentative production of higher alcohols, such as 2-methyl-1-butanol, 3-methyl-1-butanol and 2-Methyl-1-propanol (isobutanol), which all possess quality characteristics comparable to traditional oil based fuels. The common metabolic engineering approach uses the last two steps of the Ehrlich pathway, catalyzed by 2-ketoacid decarboxylase and an alcohol dehydrogenase converting the branched chain 2-ketoacids of L-isoleucine, L-leucine, and L-valine into the respective alcohols. This strategy was successfully used to engineer well suited and industrially employed bacteria, such as Escherichia coli, Bacillus subtilis and Corynebacterium glutamicum for the production of higher alcohols. Among these alcohols, isobutanol is currently the most promising one regarding final titer and yield. This article summarizes the current knowledge and achievements on isobutanol production with E. coli, B. subtilis and C. glutamicum regarding the metabolic engineering approaches and process conditions.Valorization of pyrolysis water: a biorefinery side stream, for 1,2-propanediol production with engineered Corynebacterium glutamicumhttps://puma.ub.uni-stuttgart.de/bibtex/27ae233f38758c480db193ed5e7bbf7f7/bastianbastian2018-02-09T13:18:17+01:00(propylene 1,2-propanediol Bioeconomy, Biorefinery, Corynebacterium Fast Growth-coupled Lignocellulose, Metabolic Pyrolysis biotransformation, engineering, glutamicum, glycol), myown pyrolysis, water <span data-person-type="author" class="authorEditorList "><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Julian Lange" itemprop="url" href="/person/1dc474d4c00bd416fa2ec00ea366a50da/author/0"><span itemprop="name">J. Lange</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Felix Müller" itemprop="url" href="/person/1dc474d4c00bd416fa2ec00ea366a50da/author/1"><span itemprop="name">F. Müller</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Kerstin Bernecker" itemprop="url" href="/person/1dc474d4c00bd416fa2ec00ea366a50da/author/2"><span itemprop="name">K. Bernecker</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Nicolaus Dahmen" itemprop="url" href="/person/1dc474d4c00bd416fa2ec00ea366a50da/author/3"><span itemprop="name">N. Dahmen</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Ralf Takors" itemprop="url" href="/person/1dc474d4c00bd416fa2ec00ea366a50da/author/4"><span itemprop="name">R. Takors</span></a></span>, </span> and <span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Bastian Blombach" itemprop="url" href="/person/1dc474d4c00bd416fa2ec00ea366a50da/author/5"><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">Biotechnol Biofuels</span>, </em> </span>(<em><span>2017<meta content="2017" itemprop="datePublished"/></span></em>)</span>Fri Feb 09 13:18:17 CET 2018Biotechnol Biofuels277Valorization of pyrolysis water: a biorefinery side stream, for 1,2-propanediol production with engineered {Corynebacterium} glutamicum102017(propylene 1,2-propanediol Bioeconomy, Biorefinery, Corynebacterium Fast Growth-coupled Lignocellulose, Metabolic Pyrolysis biotransformation, engineering, glutamicum, glycol), myown pyrolysis, water Background: A future bioeconomy relies on the efficient use of renewable resources for energy and material product supply. In this context, biorefineries have been developed and play a key role in converting lignocellulosic residues. Although a holistic use of the biomass feed is desired, side streams evoke in current biorefinery approaches. To ensure profitability, efficiency, and sustainability of the overall conversion process, a meaningful valorization of these materials is needed. Here, a so far unexploited side stream derived from fast pyrolysis of wheat straw-pyrolysis water-was used for production of 1,2-propanediol in microbial fermentation with engineered Corynebacterium glutamicum.
Results: A protocol for pretreatment of pyrolysis water was established and enabled growth on its major constituents, acetate and acetol, with rates up to 0.36 ± 0.04 h-1. To convert acetol to 1,2-propanediol, the plasmid pJULgldA expressing the glycerol dehydrogenase from Escherichia coli was introduced into C. glutamicum. 1,2-propanediol was formed in a growth-coupled biotransformation and production was further increased by construction of C. glutamicum Δpqo ΔaceE ΔldhA Δmdh pJULgldA. In a two-phase aerobic/microaerobic fed-batch process with pyrolysis water as substrate, this strain produced 18.3 ± 1.2 mM 1,2-propanediol with a yield of 0.96 ± 0.05 mol 1,2-propanediol per mol acetol and showed an overall volumetric productivity of 1.4 ± 0.1 mmol 1,2-propanediol L-1 h-1.
Conclusions: This study implements microbial fermentation into a biorefinery based on pyrolytic liquefaction of lignocellulosic biomass and accesses a novel value chain by valorizing the side stream pyrolysis water for 1,2-PDO production with engineered C. glutamicum. The established bioprocess operated at maximal product yield and accomplished the so far highest overall volumetric productivity for microbial 1,2-PDO production with an engineered producer strain. Besides, the results highlight the potential of microbial conversion of this biorefinery side stream to other valuable products.Identification of the agr Peptide of Listeria monocytogeneshttps://puma.ub.uni-stuttgart.de/bibtex/2322dd59578e4d23c50f6c970a88667b9/bastianbastian2018-02-09T13:18:17+01:00Listeria accessory autoinducing gene monocytogenes, myown peptide peptide, regulator, sensing <span data-person-type="author" class="authorEditorList "><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Marion Zetzmann" itemprop="url" href="/person/12b076a41023c61c2980c3276780be2c6/author/0"><span itemprop="name">M. Zetzmann</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Andrés Sánchez-Kopper" itemprop="url" href="/person/12b076a41023c61c2980c3276780be2c6/author/1"><span itemprop="name">A. Sánchez-Kopper</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Mark S. Waidmann" itemprop="url" href="/person/12b076a41023c61c2980c3276780be2c6/author/2"><span itemprop="name">M. Waidmann</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Bastian Blombach" itemprop="url" href="/person/12b076a41023c61c2980c3276780be2c6/author/3"><span itemprop="name">B. Blombach</span></a></span>, </span> and <span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Christian U. Riedel" itemprop="url" href="/person/12b076a41023c61c2980c3276780be2c6/author/4"><span itemprop="name">C. Riedel</span></a></span></span>. </span><span class="additional-entrytype-information"><span itemtype="http://schema.org/PublicationIssue" itemscope="itemscope" itemprop="isPartOf"><em><span itemprop="journal">Front Microbiol</span>, </em> </span>(<em><span>2016<meta content="2016" itemprop="datePublished"/></span></em>)</span>Fri Feb 09 13:18:17 CET 2018Front Microbiol989Identification of the agr {Peptide} of {Listeria} monocytogenes72016Listeria accessory autoinducing gene monocytogenes, myown peptide peptide, regulator, sensing Listeria monocytogenes (Lm) is an important food-borne human pathogen that is able to strive under a wide range of environmental conditions. Its accessory gene regulator (agr) system was shown to impact on biofilm formation and virulence and has been proposed as one of the regulatory mechanisms involved in adaptation to these changing environments. The Lm agr operon is homologous to the Staphylococcus aureus system, which includes an agrD-encoded autoinducing peptide that stimulates expression of the agr genes via the AgrCA two-component system and is required for regulation of target genes. The aim of the present study was to identify the native autoinducing peptide (AIP) of Lm using a luciferase reporter system in wildtype and agrD deficient strains, rational design of synthetic peptides and mass spectrometry. Upon deletion of agrD, luciferase reporter activity driven by the PII promoter of the agr operon was completely abolished and this defect was restored by co-cultivation of the agrD-negative reporter strain with a producer strain. Based on the sequence and structures of known AIPs of other organisms, a set of potential Lm AIPs was designed and tested for PII-activation. This led to the identification of a cyclic pentapeptide that was able to induce PII-driven luciferase reporter activity and restore defective invasion of the agrD deletion mutant into Caco-2 cells. Analysis of supernatants of a recombinant Escherichia coli strain expressing AgrBD identified a peptide identical in mass and charge to the cyclic pentapeptide. The Lm agr system is specific for this pentapeptide since the AIP of Lactobacillus plantarum, which also is a pentapeptide yet with different amino acid sequence, did not induce PII activity. In summary, the presented results provide further evidence for the hypothesis that the agrD gene of Lm encodes a secreted AIP responsible for autoregulation of the agr system of Lm. Additionally, the structure of the native Lm AIP was identified.Stereospecificity of Corynebacterium glutamicum 2,3-butanediol dehydrogenase and implications for the stereochemical purity of bioproduced 2,3-butanediolhttps://puma.ub.uni-stuttgart.de/bibtex/2d2fbd13b8243050d76f6784383286149/bastianbastian2018-02-09T13:18:17+01:002,3-Butanediol, Acetoin, Acetolactate Alcohol Butanediol Butylene Carboxy-Lyases, Corynebacterium Engineering Escherichia Glycols, Lactococcus Magnetic Metabolic Oxidoreductases, Proteins, Recombinant Resonance Specificity, Spectroscopy, Stereospecificity, Substrate Synthase, coli, dehydrogenase, 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/1dd4270d58d31233ae25eebd4b8cf903e/author/0"><span itemprop="name">D. Radoš</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="David L. Turner" itemprop="url" href="/person/1dd4270d58d31233ae25eebd4b8cf903e/author/1"><span itemprop="name">D. Turner</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Teresa Catarino" itemprop="url" href="/person/1dd4270d58d31233ae25eebd4b8cf903e/author/2"><span itemprop="name">T. Catarino</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Eugenia Hoffart" itemprop="url" href="/person/1dd4270d58d31233ae25eebd4b8cf903e/author/3"><span itemprop="name">E. Hoffart</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Ana Rute Neves" itemprop="url" href="/person/1dd4270d58d31233ae25eebd4b8cf903e/author/4"><span itemprop="name">A. Neves</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Bernhard J. Eikmanns" itemprop="url" href="/person/1dd4270d58d31233ae25eebd4b8cf903e/author/5"><span itemprop="name">B. Eikmanns</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Bastian Blombach" itemprop="url" href="/person/1dd4270d58d31233ae25eebd4b8cf903e/author/6"><span itemprop="name">B. Blombach</span></a></span>, </span> and <span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Helena Santos" itemprop="url" href="/person/1dd4270d58d31233ae25eebd4b8cf903e/author/7"><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">Appl. Microbiol. Biotechnol.</span>, </em> <em><span itemtype="http://schema.org/PublicationVolume" itemscope="itemscope" itemprop="isPartOf"><span itemprop="volumeNumber">100 </span></span>(<span itemprop="issueNumber">24</span>):
<span itemprop="pagination">10573--10583</span></em> </span>(<em><span>December 2016<meta content="December 2016" itemprop="datePublished"/></span></em>)</span>Fri Feb 09 13:18:17 CET 2018Appl. Microbiol. Biotechnol.dec2410573--10583Stereospecificity of {Corynebacterium} glutamicum 2,3-butanediol dehydrogenase and implications for the stereochemical purity of bioproduced 2,3-butanediol10020162,3-Butanediol, Acetoin, Acetolactate Alcohol Butanediol Butylene Carboxy-Lyases, Corynebacterium Engineering Escherichia Glycols, Lactococcus Magnetic Metabolic Oxidoreductases, Proteins, Recombinant Resonance Specificity, Spectroscopy, Stereospecificity, Substrate Synthase, coli, dehydrogenase, glutamicum, lactis, myown The stereochemistry of 2,3-butanediol (2,3-BD) synthesis in microbial fermentations is important for many applications. In this work, we showed that Corynebacterium glutamicum endowed with the Lactococcus lactis genes encoding α-acetolactate synthase and decarboxylase activities produced meso-2,3-BD as the major end product, meaning that (R)-acetoin is a substrate for endogenous 2,3-butanediol dehydrogenase (BDH) activity. This is curious in view of the reported absolute stereospecificity of C. glutamicum BDH for (S)-acetoin (Takusagawa et al. Biosc Biotechnol Biochem 65:1876-1878, 2001). To resolve this discrepancy, the enzyme encoded by butA Cg was produced in Escherichia coli and purified, and the stereospecific properties of the pure protein were examined. Activity assays monitored online by 1H-NMR using racemic acetoin and an excess of NADH showed an initial, fast production of (2S,3S)-2,3-BD, followed by a slow (∼20-fold lower apparent rate) formation of meso-2,3-BD. Kinetic parameters for (S)-acetoin, (R)-acetoin, meso-2,3-BD and (2S,3S)-BD were determined by spectrophotometric assays. V max values for (S)-acetoin and (R)-acetoin were 119 ± 15 and 5.23 ± 0.06 μmol min-1 mg protein-1, and K m values were 0.23 ± 0.02 and 1.49 ± 0.07 mM, respectively. We conclude that C. glutamicum BDH is not absolutely specific for (S)-acetoin, though this is the preferred substrate. Importantly, the low activity of BDH with (R)-acetoin was sufficient to support high yields of meso-2,3-BD in the engineered strain C. glutamicum ΔaceEΔpqoΔldhA(pEKEx2-als,aldB,butA Cg ). Additionally, we found that the BDH activity was nearly abolished upon inactivation of butA Cg (from 0.30 ± 0.03 to 0.004 ± 0.001 μmol min-1 mg protein-1), indicating that C. glutamicum expresses a single BDH under the experimental conditions examined.CO2 - Intrinsic Product, Essential Substrate, and Regulatory Trigger of Microbial and Mammalian Production Processeshttps://puma.ub.uni-stuttgart.de/bibtex/2ef8131870fb5db07e873c177cfee4609/bastianbastian2018-02-09T13:18:17+01:00bicarbonate, carbon carboxylation, decarboxylation dioxide, myown process, production regulation, <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/1843b15643d0c3ef44a9efb1e4d337222/author/0"><span itemprop="name">B. Blombach</span></a></span>, </span> and <span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Ralf Takors" itemprop="url" href="/person/1843b15643d0c3ef44a9efb1e4d337222/author/1"><span itemprop="name">R. Takors</span></a></span></span>. </span><span class="additional-entrytype-information"><span itemtype="http://schema.org/PublicationIssue" itemscope="itemscope" itemprop="isPartOf"><em><span itemprop="journal">Front Bioeng Biotechnol</span>, </em> </span>(<em><span>2015<meta content="2015" itemprop="datePublished"/></span></em>)</span>Fri Feb 09 13:18:17 CET 2018Front Bioeng Biotechnol108{CO}2 - {Intrinsic} {Product}, {Essential} {Substrate}, and {Regulatory} {Trigger} of {Microbial} and {Mammalian} {Production} {Processes}32015bicarbonate, carbon carboxylation, decarboxylation dioxide, myown process, production regulation, Carbon dioxide formation mirrors the final carbon oxidation steps of aerobic metabolism in microbial and mammalian cells. As a consequence, [Formula: see text] dissociation equilibria arise in fermenters by the growing culture. Anaplerotic reactions make use of the abundant [Formula: see text] levels for refueling citric acid cycle demands and for enabling oxaloacetate-derived products. At the same time, CO2 is released manifold in metabolic reactions via decarboxylation activity. The levels of extracellular [Formula: see text] depend on cellular activities and physical constraints such as hydrostatic pressures, aeration, and the efficiency of mixing in large-scale bioreactors. Besides, local [Formula: see text] levels might also act as metabolic inhibitors or transcriptional effectors triggering regulatory events inside the cells. This review gives an overview about fundamental physicochemical properties of [Formula: see text] in microbial and mammalian cultures effecting cellular physiology, production processes, metabolic activity, and transcriptional regulation.Importance of NADPH supply for improved L-valine formation in Corynebacterium glutamicumhttps://puma.ub.uni-stuttgart.de/bibtex/2dfb0ffb355b1bb961e53e70b66a327fd/bastianbastian2018-02-09T13:18:17+01:00Biological, Cell Corynebacterium Culture Fermentation, Glucose-6-Phosphate Isomerase, Media, Metabolic Metabolome Models, NADP, Networks Pathways, Techniques, Valine, and glutamicum, myown <span data-person-type="author" class="authorEditorList "><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Tobias Bartek" itemprop="url" href="/person/1ea026818f74ee7c252525689e9c0c32c/author/0"><span itemprop="name">T. Bartek</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Bastian Blombach" itemprop="url" href="/person/1ea026818f74ee7c252525689e9c0c32c/author/1"><span itemprop="name">B. Blombach</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Enrico Zönnchen" itemprop="url" href="/person/1ea026818f74ee7c252525689e9c0c32c/author/2"><span itemprop="name">E. Zönnchen</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Pia Makus" itemprop="url" href="/person/1ea026818f74ee7c252525689e9c0c32c/author/3"><span itemprop="name">P. Makus</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Siegmund Lang" itemprop="url" href="/person/1ea026818f74ee7c252525689e9c0c32c/author/4"><span itemprop="name">S. Lang</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Bernhard J. Eikmanns" itemprop="url" href="/person/1ea026818f74ee7c252525689e9c0c32c/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="Marco Oldiges" itemprop="url" href="/person/1ea026818f74ee7c252525689e9c0c32c/author/6"><span itemprop="name">M. Oldiges</span></a></span></span>. </span><span class="additional-entrytype-information"><span itemtype="http://schema.org/PublicationIssue" itemscope="itemscope" itemprop="isPartOf"><em><span itemprop="journal">Biotechnol. Prog.</span>, </em> <em><span itemtype="http://schema.org/PublicationVolume" itemscope="itemscope" itemprop="isPartOf"><span itemprop="volumeNumber">26 </span></span>(<span itemprop="issueNumber">2</span>):
<span itemprop="pagination">361--371</span></em> </span>(<em><span>April 2010<meta content="April 2010" itemprop="datePublished"/></span></em>)</span>Fri Feb 09 13:18:17 CET 2018Biotechnol. Prog.apr2361--371Importance of {NADPH} supply for improved {L}-valine formation in {Corynebacterium} glutamicum262010Biological, Cell Corynebacterium Culture Fermentation, Glucose-6-Phosphate Isomerase, Media, Metabolic Metabolome Models, NADP, Networks Pathways, Techniques, Valine, and glutamicum, myown Cofactor recycling is known to be crucial for amino acid synthesis. Hence, cofactor supply was now analyzed for L-valine to identify new targets for an improvement of production. The central carbon metabolism was analyzed by stoichiometric modeling to estimate the influence of cofactors and to quantify the theoretical yield of L-valine on glucose. Three different optimal routes for L-valine biosynthesis were identified by elementary mode (EM) analysis. The modes differed mainly in the manner of NADPH regeneration, substantiating that the cofactor supply may be crucial for efficient L-valine production. Although the isocitrate dehydrogenase as an NADPH source within the tricarboxylic acid cycle only enables an L-valine yield of Y(Val/Glc) = 0.5 mol L-valine/mol glucose (mol Val/mol Glc), the pentose phosphate pathway seems to be the most promising NADPH source. Based on the theoretical calculation of EMs, the gene encoding phosphoglucoisomerase (PGI) was deleted to achieve this EM with a theoretical yield Y(Val/Glc) = 0.86 mol Val/mol Glc during the production phase. The intracellular NADPH concentration was significantly increased in the PGI-deficient mutant. L-Valine yield increased from 0.49 +/- 0.13 to 0.67 +/- 0.03 mol Val/mol Glc, and, concomitantly, the formation of by-products such as pyruvate was reduced.Application of metabolic engineering for the biotechnological production of L-valinehttps://puma.ub.uni-stuttgart.de/bibtex/2d92973003e51ad7cf70cdb6cc5fad5e6/bastianbastian2018-02-09T13:18:17+01:00Aerobiosis, Anaerobiosis, Biosynthetic Corynebacterium Engineering Escherichia Metabolic Pathways, Valine, coli, glutamicum, myown <span data-person-type="author" class="authorEditorList "><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Marco Oldiges" itemprop="url" href="/person/166e252cc9c947b28a4c2c4f54e174564/author/0"><span itemprop="name">M. Oldiges</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Bernhard J. Eikmanns" itemprop="url" href="/person/166e252cc9c947b28a4c2c4f54e174564/author/1"><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/166e252cc9c947b28a4c2c4f54e174564/author/2"><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. Microbiol. Biotechnol.</span>, </em> <em><span itemtype="http://schema.org/PublicationVolume" itemscope="itemscope" itemprop="isPartOf"><span itemprop="volumeNumber">98 </span></span>(<span itemprop="issueNumber">13</span>):
<span itemprop="pagination">5859--5870</span></em> </span>(<em><span>July 2014<meta content="July 2014" itemprop="datePublished"/></span></em>)</span>Fri Feb 09 13:18:17 CET 2018Appl. Microbiol. Biotechnol.jul135859--5870Application of metabolic engineering for the biotechnological production of {L}-valine982014Aerobiosis, Anaerobiosis, Biosynthetic Corynebacterium Engineering Escherichia Metabolic Pathways, Valine, coli, glutamicum, myown The branched chain amino acid L-valine is an essential nutrient for higher organisms, such as animals and humans. Besides the pharmaceutical application in parenteral nutrition and as synthon for the chemical synthesis of e.g. herbicides or anti-viral drugs, L-valine is now emerging into the feed market, and significant increase of sales and world production is expected. In accordance, well-known microbial production bacteria, such as Escherichia coli and Corynebacterium glutamicum strains, have recently been metabolically engineered for efficient L-valine production under aerobic or anaerobic conditions, and the respective cultivation and production conditions have been optimized. This review summarizes the state of the art in L-valine biosynthesis and its regulation in E. coli and C. glutamicum with respect to optimal metabolic network for microbial L-valine production, genetic strain engineering and bioprocess development for L-valine production, and finally, it will shed light on emerging technologies that have the potential to accelerate strain and bioprocess engineering in the near future.L-valine production with pyruvate dehydrogenase complex-deficient Corynebacterium glutamicumhttps://puma.ub.uni-stuttgart.de/bibtex/25bac540150e10acba07b2f8205f6f070/bastianbastian2018-02-09T13:18:17+01:00Acid, Alanine Complex, Corynebacterium Dehydrogenase Fermentation, Isoleucine, Lysine, Pyruvate Pyruvic 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/1d744d92c5af8f45409137c58d022ef1c/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/1d744d92c5af8f45409137c58d022ef1c/author/1"><span itemprop="name">M. Schreiner</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Jirí Holátko" itemprop="url" href="/person/1d744d92c5af8f45409137c58d022ef1c/author/2"><span itemprop="name">J. Holátko</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Tobias Bartek" itemprop="url" href="/person/1d744d92c5af8f45409137c58d022ef1c/author/3"><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/1d744d92c5af8f45409137c58d022ef1c/author/4"><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/1d744d92c5af8f45409137c58d022ef1c/author/5"><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">73 </span></span>(<span itemprop="issueNumber">7</span>):
<span itemprop="pagination">2079--2084</span></em> </span>(<em><span>April 2007<meta content="April 2007" itemprop="datePublished"/></span></em>)</span>Fri Feb 09 13:18:17 CET 2018Appl. Environ. Microbiol.apr72079--2084L-valine production with pyruvate dehydrogenase complex-deficient {Corynebacterium} glutamicum732007Acid, Alanine Complex, Corynebacterium Dehydrogenase Fermentation, Isoleucine, Lysine, Pyruvate Pyruvic Valine, glutamicum, myown Corynebacterium glutamicum was engineered for the production of L-valine from glucose by deletion of the aceE gene encoding the E1p enzyme of the pyruvate dehydrogenase complex and additional overexpression of the ilvBNCE genes encoding the L-valine biosynthetic enzymes acetohydroxyacid synthase, isomeroreductase, and transaminase B. In the absence of cellular growth, C. glutamicum DeltaaceE showed a relatively high intracellular concentration of pyruvate (25.9 mM) and produced significant amounts of pyruvate, L-alanine, and L-valine from glucose as the sole carbon source. Lactate or acetate was not formed. Plasmid-bound overexpression of ilvBNCE in C. glutamicum DeltaaceE resulted in an approximately 10-fold-lower intracellular pyruvate concentration (2.3 mM) and a shift of the extracellular product pattern from pyruvate and L-alanine towards L-valine. In fed-batch fermentations at high cell densities and an excess of glucose, C. glutamicum DeltaaceE(pJC4ilvBNCE) produced up to 210 mM L-valine with a volumetric productivity of 10.0 mM h(-1) (1.17 g l(-1) h(-1)) and a maximum yield of about 0.6 mol per mol (0.4 g per g) of glucose.Harnessing novel chromosomal integration loci to utilize an organosolv-derived hemicellulose fraction for isobutanol production with engineered Corynebacterium glutamicumhttps://puma.ub.uni-stuttgart.de/bibtex/2bdffd9923d4263adf616891ea145dcde/bastianbastian2018-02-09T13:18:17+01:00imported myown <span data-person-type="author" class="authorEditorList "><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Julian Lange" itemprop="url" href="/person/1e128b514e13083ebf23412da7da2ffe3/author/0"><span itemprop="name">J. Lange</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Felix Müller" itemprop="url" href="/person/1e128b514e13083ebf23412da7da2ffe3/author/1"><span itemprop="name">F. Müller</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Ralf Takors" itemprop="url" href="/person/1e128b514e13083ebf23412da7da2ffe3/author/2"><span itemprop="name">R. Takors</span></a></span>, </span> and <span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Bastian Blombach" itemprop="url" href="/person/1e128b514e13083ebf23412da7da2ffe3/author/3"><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">Microb Biotechnol</span>, </em> <em><span itemtype="http://schema.org/PublicationVolume" itemscope="itemscope" itemprop="isPartOf"><span itemprop="volumeNumber">11 </span></span>(<span itemprop="issueNumber">1</span>):
<span itemprop="pagination">257--263</span></em> </span>(<em><span>January 2018<meta content="January 2018" itemprop="datePublished"/></span></em>)</span>Fri Feb 09 13:18:17 CET 2018Microb Biotechnoljan1257--263Harnessing novel chromosomal integration loci to utilize an organosolv-derived hemicellulose fraction for isobutanol production with engineered {Corynebacterium} glutamicum112018imported myown A successful bioeconomy depends on the manifestation of biorefineries that entirely convert renewable resources to valuable products and energies. Here, the poorly exploited hemicellulose fraction (HF) from beech wood organosolv processing was applied for isobutanol production with Corynebacterium glutamicum. To enable growth of C. glutamicum on HF, we integrated genes required for D-xylose and l-arabinose metabolization into two of 16 systematically identified and novel chromosomal integration loci. Under aerobic conditions, this engineered strain CArXy reached growth rates up to 0.34 ± 0.02 h-1 on HF. Based on CArXy, we developed the isobutanol producer strain CIsArXy, which additionally (over)expresses genes of the native l-valine biosynthetic and the heterologous Ehrlich pathway. CIsArXy produced 7.2 ± 0.2 mM (0.53 ± 0.02 g L-1 ) isobutanol on HF at a carbon molar yield of 0.31 ± 0.02 C-mol isobutanol per C-mol substrate (d-xylose + l-arabinose) in an anaerobic zero-growth production process.Engineering 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.Application of a genetically encoded biosensor for live cell imaging of L-valine production in pyruvate dehydrogenase complex-deficient Corynebacterium glutamicum strainshttps://puma.ub.uni-stuttgart.de/bibtex/29345950b723cb5b2b8e6a41af065cae9/bastianbastian2018-02-09T13:18:17+01:00Biosensing Complex, Corynebacterium Dehydrogenase Fluorescence, Microfluidics Online Phenotype, Pyruvate Systems, Techniques, Valine, glutamicum, myown <span data-person-type="author" class="authorEditorList "><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Nurije Mustafi" itemprop="url" href="/person/15c2b0eaacc8d66ee79a8fdb8e64e9082/author/0"><span itemprop="name">N. Mustafi</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Alexander Grünberger" itemprop="url" href="/person/15c2b0eaacc8d66ee79a8fdb8e64e9082/author/1"><span itemprop="name">A. Grünberger</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Regina Mahr" itemprop="url" href="/person/15c2b0eaacc8d66ee79a8fdb8e64e9082/author/2"><span itemprop="name">R. Mahr</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Stefan Helfrich" itemprop="url" href="/person/15c2b0eaacc8d66ee79a8fdb8e64e9082/author/3"><span itemprop="name">S. Helfrich</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Katharina Nöh" itemprop="url" href="/person/15c2b0eaacc8d66ee79a8fdb8e64e9082/author/4"><span itemprop="name">K. Nöh</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Bastian Blombach" itemprop="url" href="/person/15c2b0eaacc8d66ee79a8fdb8e64e9082/author/5"><span itemprop="name">B. Blombach</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Dietrich Kohlheyer" itemprop="url" href="/person/15c2b0eaacc8d66ee79a8fdb8e64e9082/author/6"><span itemprop="name">D. Kohlheyer</span></a></span>, </span> and <span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Julia Frunzke" itemprop="url" href="/person/15c2b0eaacc8d66ee79a8fdb8e64e9082/author/7"><span itemprop="name">J. Frunzke</span></a></span></span>. </span><span class="additional-entrytype-information"><span itemtype="http://schema.org/PublicationIssue" itemscope="itemscope" itemprop="isPartOf"><em><span itemprop="journal">PLoS ONE</span>, </em> <em><span itemtype="http://schema.org/PublicationVolume" itemscope="itemscope" itemprop="isPartOf"><span itemprop="volumeNumber">9 </span></span>(<span itemprop="issueNumber">1</span>):
<span itemprop="pagination">e85731</span></em> </span>(<em><span>2014<meta content="2014" itemprop="datePublished"/></span></em>)</span>Fri Feb 09 13:18:17 CET 2018PLoS ONE1e85731Application of a genetically encoded biosensor for live cell imaging of {L}-valine production in pyruvate dehydrogenase complex-deficient {Corynebacterium} glutamicum strains92014Biosensing Complex, Corynebacterium Dehydrogenase Fluorescence, Microfluidics Online Phenotype, Pyruvate Systems, Techniques, Valine, glutamicum, myown The majority of biotechnologically relevant metabolites do not impart a conspicuous phenotype to the producing cell. Consequently, the analysis of microbial metabolite production is still dominated by bulk techniques, which may obscure significant variation at the single-cell level. In this study, we have applied the recently developed Lrp-biosensor for monitoring of amino acid production in single cells of gradually engineered L-valine producing Corynebacterium glutamicum strains based on the pyruvate dehydrogenase complex-deficient (PDHC) strain C. glutamicum ΔaceE. Online monitoring of the sensor output (eYFP fluorescence) during batch cultivation proved the sensor's suitability for visualizing different production levels. In the following, we conducted live cell imaging studies on C. glutamicum sensor strains using microfluidic chip devices. As expected, the sensor output was higher in microcolonies of high-yield producers in comparison to the basic strain C. glutamicum ΔaceE. Microfluidic cultivation in minimal medium revealed a typical Gaussian distribution of single cell fluorescence during the production phase. Remarkably, low amounts of complex nutrients completely changed the observed phenotypic pattern of all strains, resulting in a phenotypic split of the population. Whereas some cells stopped growing and initiated L-valine production, others continued to grow or showed a delayed transition to production. Depending on the cultivation conditions, a considerable fraction of non-fluorescent cells was observed, suggesting a loss of metabolic activity. These studies demonstrate that genetically encoded biosensors are a valuable tool for monitoring single cell productivity and to study the phenotypic pattern of microbial production strains.Metabolic 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.Acetohydroxyacid synthase, a novel target for improvement of L-lysine production by Corynebacterium glutamicumhttps://puma.ub.uni-stuttgart.de/bibtex/2ececc5edfb975851e24e89f723cab58c/bastianbastian2018-02-09T13:18:17+01:00Acetolactate Acid, Bacterial Butyrates Corynebacterium Deletion, Enzyme Expression Gene Inhibitors, Isoleucine, Kinetics, Leucine, Lysine, Profiling, Proteins, Pyruvic Sequence Synthase, 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/19cd149b8364b60f0642067f88c464b45/author/0"><span itemprop="name">B. Blombach</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Stephan Hans" itemprop="url" href="/person/19cd149b8364b60f0642067f88c464b45/author/1"><span itemprop="name">S. Hans</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Brigitte Bathe" itemprop="url" href="/person/19cd149b8364b60f0642067f88c464b45/author/2"><span itemprop="name">B. Bathe</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/19cd149b8364b60f0642067f88c464b45/author/3"><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">75 </span></span>(<span itemprop="issueNumber">2</span>):
<span itemprop="pagination">419--427</span></em> </span>(<em><span>January 2009<meta content="January 2009" itemprop="datePublished"/></span></em>)</span>Fri Feb 09 13:18:17 CET 2018Appl. Environ. Microbiol.jan2419--427Acetohydroxyacid synthase, a novel target for improvement of {L}-lysine production by {Corynebacterium} glutamicum752009Acetolactate Acid, Bacterial Butyrates Corynebacterium Deletion, Enzyme Expression Gene Inhibitors, Isoleucine, Kinetics, Leucine, Lysine, Profiling, Proteins, Pyruvic Sequence Synthase, Valine, glutamicum, myown The influence of acetohydroxy acid synthase (AHAS) on L-lysine production by Corynebacterium glutamicum was investigated. An AHAS with a deleted C-terminal domain in the regulatory subunit IlvN was engineered by truncating the ilvN gene. Compared to the wild-type AHAS, the newly constructed enzyme showed altered kinetic properties, i.e., (i) an about twofold-lower K(m) for the substrate pyruvate and an about fourfold-lower V(max); (ii) a slightly increased K(m) for the substrate alpha-ketobutyrate with an about twofold-lower V(max); and (iii) insensitivity against the inhibitors L-valine, L-isoleucine, and L-leucine (10 mM each). Introduction of the modified AHAS into the L-lysine producers C. glutamicum DM1729 and DM1933 increased L-lysine formation by 43\% (30 mM versus 21 mM) and 36\% (51 mM versus 37 mM), respectively, suggesting that decreased AHAS activity is linked to increased L-lysine formation. Complete inactivation of the AHAS in C. glutamicum DM1729 and DM1933 by deletion of the ilvB gene, encoding the catalytic subunit of AHAS, led to L-valine, L-isoleucine, and L-leucine auxotrophy and to further-improved L-lysine production. In batch fermentations, C. glutamicum DM1729 Delta ilvB produced about 85\% more L-lysine (70 mM versus 38 mM) and showed an 85\%-higher substrate-specific product yield (0.180 versus 0.098 mol C/mol C) than C. glutamicum DM1729. Comparative transcriptome analysis of C. glutamicum DM1729 and C. glutamicum DM1729 Delta ilvB indicated transcriptional differences for about 50 genes, although not for those encoding enzymes involved in the L-lysine biosynthetic pathway.Metabolic engineering to guide evolution – creating a novel mode for L-valine production with Corynebacterium glutamicumhttps://puma.ub.uni-stuttgart.de/bibtex/2ed655f55cd2b75b38ea9fea95bcde143/bastianbastian2018-03-20T16:36:43+01:00myown <span data-person-type="author" class="authorEditorList "><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Andreas Schwentner" itemprop="url" href="/person/162ce9a577fb96f3cdd6a5b25e870de44/author/0"><span itemprop="name">A. Schwentner</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="André Feith" itemprop="url" href="/person/162ce9a577fb96f3cdd6a5b25e870de44/author/1"><span itemprop="name">A. Feith</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Eugenia Münch" itemprop="url" href="/person/162ce9a577fb96f3cdd6a5b25e870de44/author/2"><span itemprop="name">E. Münch</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Tobias Busche" itemprop="url" href="/person/162ce9a577fb96f3cdd6a5b25e870de44/author/3"><span itemprop="name">T. Busche</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Christian Rückert" itemprop="url" href="/person/162ce9a577fb96f3cdd6a5b25e870de44/author/4"><span itemprop="name">C. Rückert</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Jörn Kalinowski" itemprop="url" href="/person/162ce9a577fb96f3cdd6a5b25e870de44/author/5"><span itemprop="name">J. Kalinowski</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Ralf Takors" itemprop="url" href="/person/162ce9a577fb96f3cdd6a5b25e870de44/author/6"><span itemprop="name">R. Takors</span></a></span>, </span> and <span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Bastian Blombach" itemprop="url" href="/person/162ce9a577fb96f3cdd6a5b25e870de44/author/7"><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">Metabolic Engineering</span>, </em> </span>(<em><span>2018<meta content="2018" itemprop="datePublished"/></span></em>)</span>Tue Mar 20 16:36:43 CET 2018Metabolic EngineeringMetabolic engineering to guide evolution – creating a novel mode for L-valine production with Corynebacterium glutamicum2018myown Evolutionary approaches are often undirected and mutagen-based yielding numerous mutations, which need elaborate screenings to identify relevant targets. We here apply Metabolic engineering to Guide Evolution (MGE), an evolutionary approach evolving and identifying new targets to improve microbial producer strains. MGE is based on the idea to impair the cell's metabolism by metabolic engineering, thereby generating guided evolutionary pressure. It consists of three distinct phases: (i) metabolic engineering to create the evolutionary pressure on the applied strain followed by (ii) a cultivation phase with growth as straightforward screening indicator for the evolutionary event, and (iii) comparative whole genome sequencing (WGS), to identify mutations in the evolved strains, which are eventually re-engineered for verification. Applying MGE, we evolved the PEP and pyruvate carboxylase-deficient strain C. glutamicum Δppc Δpyc to grow on glucose as substrate with rates up to 0.31 ± 0.02h−1 which corresponds to 80% of the growth rate of the wildtype strain. The intersection of the mutations identified by WGS revealed isocitrate dehydrogenase (ICD) as consistent target in three independently evolved mutants. Upon re-engineering in C. glutamicum Δppc Δpyc, the identified mutations led to diminished ICD activities and activated the glyoxylate shunt replenishing oxaloacetate required for growth. Intracellular relative quantitative metabolome analysis showed that the pools of citrate, isocitrate, cis-aconitate, and L-valine were significantly higher compared to the WT control. As an alternative to existing L-valine producer strains based on inactivated or attenuated pyruvate dehydrogenase complex, we finally engineered the PEP and pyruvate carboxylase-deficient C. glutamicum strains with identified ICD mutations for L-valine production by overexpression of the L-valine biosynthesis genes. Among them, C. glutamicum Δppc Δpyc ICDG407S (pJC4ilvBNCE) produced up to 8.9 ± 0.4gL-valine L−1, with a product yield of 0.22 ± 0.01gL-valine per g glucose.Metabolic engineering to guide evolution – creating a novel mode for L-valine production with Corynebacterium glutamicumUsing gas mixtures of CO, CO2 and H2 as microbial substrates: the do's and don'ts of successful technology transfer from laboratory to production scalehttps://puma.ub.uni-stuttgart.de/bibtex/29f828e956400a30c5ed5cbab1c5b2e32/bastianbastian2018-06-05T10:57:11+02:00myown <span data-person-type="author" class="authorEditorList "><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="R Takors" itemprop="url" href="/person/132d45b33300f6eea2c206fc2ca8fb050/author/0"><span itemprop="name">R. Takors</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="M Kopf" itemprop="url" href="/person/132d45b33300f6eea2c206fc2ca8fb050/author/1"><span itemprop="name">M. Kopf</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="J Mampel" itemprop="url" href="/person/132d45b33300f6eea2c206fc2ca8fb050/author/2"><span itemprop="name">J. Mampel</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="W Bluemke" itemprop="url" href="/person/132d45b33300f6eea2c206fc2ca8fb050/author/3"><span itemprop="name">W. Bluemke</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="B Blombach" itemprop="url" href="/person/132d45b33300f6eea2c206fc2ca8fb050/author/4"><span itemprop="name">B. Blombach</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="B Eikmanns" itemprop="url" href="/person/132d45b33300f6eea2c206fc2ca8fb050/author/5"><span itemprop="name">B. Eikmanns</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="F R Bengelsdorf" itemprop="url" href="/person/132d45b33300f6eea2c206fc2ca8fb050/author/6"><span itemprop="name">F. Bengelsdorf</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="D Weuster-Botz" itemprop="url" href="/person/132d45b33300f6eea2c206fc2ca8fb050/author/7"><span itemprop="name">D. Weuster-Botz</span></a></span>, </span> and <span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="P Dürre" itemprop="url" href="/person/132d45b33300f6eea2c206fc2ca8fb050/author/8"><span itemprop="name">P. Dürre</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 Biotechnol</span>, </em> </span>(<em><span>May 2018<meta content="May 2018" itemprop="datePublished"/></span></em>)</span>Tue Jun 05 10:57:11 CEST 2018Microb BiotechnolmayUsing gas mixtures of CO, CO2 and H2 as microbial substrates: the do's and don'ts of successful technology transfer from laboratory to production scale2018myown The reduction of CO2 emissions is a global effort which is not only supported by the society and politicians but also by the industry. Chemical producers worldwide follow the strategic goal to reduce CO2 emissions by replacing existing fossil-based production routes with sustainable alternatives. The smart use of CO and CO2 /H2 mixtures even allows to produce important chemical building blocks consuming the said gases as substrates in carboxydotrophic fermentations with acetogenic bacteria. However, existing industrial infrastructure and market demands impose constraints on microbes, bioprocesses and products that require careful consideration to ensure technical and economic success. The mini review provides scientific and industrial facets finally to enable the successful implementation of gas fermentation technologies in the industrial scale.Using gas mixtures of CO, CO2 and H2 as microbial substrates: the do's and don'ts of successful technology transfer from laboratory to production s... - PubMed - NCBIThe RamA regulon: complex regulatory interactions in relation to central metabolism in Corynebacterium glutamicumhttps://puma.ub.uni-stuttgart.de/bibtex/276995fb4085e4b25c4a243d7873c547c/bastianbastian2018-06-05T10:58:03+02:00myown <span data-person-type="author" class="authorEditorList "><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="A Shah" itemprop="url" href="/person/1bf5ab14f7c5456840570d9580a126c09/author/0"><span itemprop="name">A. Shah</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="B Blombach" itemprop="url" href="/person/1bf5ab14f7c5456840570d9580a126c09/author/1"><span itemprop="name">B. Blombach</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="R Gauttam" itemprop="url" href="/person/1bf5ab14f7c5456840570d9580a126c09/author/2"><span itemprop="name">R. Gauttam</span></a></span>, </span> and <span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="B J Eikmanns" itemprop="url" href="/person/1bf5ab14f7c5456840570d9580a126c09/author/3"><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> </span>(<em><span>May 2018<meta content="May 2018" itemprop="datePublished"/></span></em>)</span>Tue Jun 05 10:58:03 CEST 2018Appl Microbiol BiotechnolmayThe RamA regulon: complex regulatory interactions in relation to central metabolism in Corynebacterium glutamicum2018myown Corynebacterium glutamicum is an industrial workhorse used for the production of amino acids and a variety of other chemicals and fuels. Within its regulatory repertoire, C. glutamicum possesses RamA which was initially identified as essential transcriptional regulator of acetate metabolism. Further studies revealed its relevance for ethanol and propionate catabolism and also identified RamA to function as global regulator in the metabolism of C. glutamicum. Thereby, RamA acts as transcriptional activator or repressor of genes encoding enzymes which are involved in carbon uptake, central carbon metabolism, and cell wall synthesis. RamA controls the expression of target genes either directly and/or indirectly by constituting feed-forward loop type of transcriptional motifs with other regulators such as GlxR, SugR, RamB, and GntR1. In this review, we summarize the current knowledge on RamA, its regulon, and its regulatory interplay with other transcriptional regulators coordinating the metabolism of C. glutamicum.The RamA regulon: complex regulatory interactions in relation to central metabolism in Corynebacterium glutamicum. - PubMed - NCBICell-Free Protein Synthesis From Fast-Growing Vibrio natriegenshttps://puma.ub.uni-stuttgart.de/bibtex/29b389b784a2e3f61cdcd4cf352c1fb38/bastianbastian2018-06-05T11:00:35+02:00myown <span data-person-type="author" class="authorEditorList "><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Jurek Failmezger" itemprop="url" href="/person/1da61cce3b9357a4f2288dee9db64b6c4/author/0"><span itemprop="name">J. Failmezger</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Steffen Scholz" itemprop="url" href="/person/1da61cce3b9357a4f2288dee9db64b6c4/author/1"><span itemprop="name">S. Scholz</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Bastian Blombach" itemprop="url" href="/person/1da61cce3b9357a4f2288dee9db64b6c4/author/2"><span itemprop="name">B. Blombach</span></a></span>, </span> and <span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Martin Siemann-Herzberg" itemprop="url" href="/person/1da61cce3b9357a4f2288dee9db64b6c4/author/3"><span itemprop="name">M. Siemann-Herzberg</span></a></span></span>. </span><span class="additional-entrytype-information"><span itemtype="http://schema.org/PublicationIssue" itemscope="itemscope" itemprop="isPartOf"><em><span itemprop="journal">Frontiers in Microbiology</span>, </em> </span>(<em><span>June 2018<meta content="June 2018" itemprop="datePublished"/></span></em>)</span>Tue Jun 05 11:00:35 CEST 2018Frontiers in MicrobiologyjunCell-Free Protein Synthesis From Fast-Growing Vibrio natriegens92018myown Frontiers | Cell-Free Protein Synthesis From Fast-Growing Vibrio natriegens | MicrobiologyDeciphering the Adaptation of Corynebacterium glutamicum in Transition from Aerobiosis via Microaerobiosis to Anaerobiosishttps://puma.ub.uni-stuttgart.de/bibtex/2efc842f7261d7d5e9a8a0f28d762da56/bastianbastian2018-06-22T08:19:36+02:00myown <span data-person-type="author" class="authorEditorList "><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Julian Lange" itemprop="url" href="/person/1a1ed304b14cfcc5cc42f06e692d82a7b/author/0"><span itemprop="name">J. Lange</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Eugenia Münch" itemprop="url" href="/person/1a1ed304b14cfcc5cc42f06e692d82a7b/author/1"><span itemprop="name">E. Münch</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Jan Müller" itemprop="url" href="/person/1a1ed304b14cfcc5cc42f06e692d82a7b/author/2"><span itemprop="name">J. Müller</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Tobias Busche" itemprop="url" href="/person/1a1ed304b14cfcc5cc42f06e692d82a7b/author/3"><span itemprop="name">T. Busche</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Jörn Kalinowski" itemprop="url" href="/person/1a1ed304b14cfcc5cc42f06e692d82a7b/author/4"><span itemprop="name">J. Kalinowski</span></a></span>, </span><span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Ralf Takors" itemprop="url" href="/person/1a1ed304b14cfcc5cc42f06e692d82a7b/author/5"><span itemprop="name">R. Takors</span></a></span>, </span> and <span><span itemtype="http://schema.org/Person" itemscope="itemscope" itemprop="author"><a title="Bastian Blombach" itemprop="url" href="/person/1a1ed304b14cfcc5cc42f06e692d82a7b/author/6"><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">Genes</span>, </em> <em><span itemtype="http://schema.org/PublicationVolume" itemscope="itemscope" itemprop="isPartOf"><span itemprop="volumeNumber">9 </span></span>(<span itemprop="issueNumber">6</span>):
<span itemprop="pagination">297</span></em> </span>(<em><span>2018<meta content="2018" itemprop="datePublished"/></span></em>)</span>Fri Jun 22 08:19:36 CEST 2018Genes6297Deciphering the Adaptation of Corynebacterium glutamicum in Transition from Aerobiosis via Microaerobiosis to Anaerobiosis92018myown Zero-growth processes are a promising strategy for the production of reduced molecules and depict a steady transition from aerobic to anaerobic conditions. To investigate the adaptation of Corynebacterium glutamicum to altering oxygen availabilities, we conceived a triple-phase fermentation process that describes a gradual reduction of dissolved oxygen with a shift from aerobiosis via microaerobiosis to anaerobiosis. The distinct process phases were clearly bordered by the bacteria&rsquo;s physiologic response such as reduced growth rate, biomass substrate yield and altered yield of fermentation products. During the process, sequential samples were drawn at six points and analyzed via RNA-sequencing, for metabolite concentrations and for enzyme activities. We found transcriptional alterations of almost 50% (1421 genes) of the entire protein coding genes and observed an upregulation of fermentative pathways, a rearrangement of respiration, and mitigation of the basic cellular mechanisms such as transcription, translation and replication as a transient response related to the installed oxygen dependent process phases. To investigate the regulatory regime, 18 transcriptionally altered (putative) transcriptional regulators were deleted, but none of the deletion strains showed noticeable growth kinetics under an oxygen restricted environment. However, the described transcriptional adaptation of C. glutamicum resolved to varying oxygen availabilities provides a useful basis for future process and strain engineering.