PUMA publications for /user/bastian/carboxy-lyases,%20engineering%20coli,https://puma.ub.uni-stuttgart.de/user/bastian/carboxy-lyases,%20engineering%20coli,PUMA RSS feed for /user/bastian/carboxy-lyases,%20engineering%20coli,2024-03-28T11:57:18+01:00Current 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.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.