Engineering biotin prototrophic Corynebacterium glutamicum strains for amino acid, diamine and carotenoid production

Peters‐Wendisch, Petra; Götker, Susanne; Heider, Sabine A. E.; Reddy, Gajendar Komati; Nguyen, Anh Q.; Stansen, K Corinna; Wendisch, Volker F.

doi:10.1016/j.jbiotec.2014.01.023

Cited by 30 publications

(28 citation statements)

References 85 publications

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“…For the detection of carbohydrates and organic acids, the analyte was separated with a column for organic acids (length, 300 mm; inside diameter, 8 mm; particle size, 10 m; pore diameter, 25 Å; CS-Chromatographie), and a G1362A refractive index detector (RID) (1200 series; Agilent Technologies) was used. Derivatization and quantification were carried out as described previously (47). Measurement of enzyme activities.…”

Section: Methodsmentioning

confidence: 99%

Metabolic Engineering of an ATP-Neutral Embden-Meyerhof-Parnas Pathway in Corynebacterium glutamicum: Growth Restoration by an Adaptive Point Mutation in NADH Dehydrogenase

Reddy

Lindner

Wendisch

2015

Appl Environ Microbiol

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Corynebacterium glutamicum uses the Embden-Meyerhof-Parnas pathway of glycolysis and gains 2 mol of ATP per mol of glucose by substrate-level phosphorylation (SLP). To engineer glycolysis without net ATP formation by SLP, endogenous phosphorylating NAD-dependent glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was replaced by nonphosphorylating NADPdependent glyceraldehyde-3-phosphate dehydrogenase (GapN) from Clostridium acetobutylicum, which irreversibly converts glyceraldehyde-3-phosphate (GAP) to 3-phosphoglycerate (3-PG) without generating ATP. As shown recently (S. Takeno, R. Murata, R. Kobayashi, S. Mitsuhashi, and M. Ikeda, Appl Environ Microbiol 76:7154 -7160, 2010, http://dx.doi.org/10.1128/AEM.01464-10), this ATP-neutral, NADPH-generating glycolytic pathway did not allow for the growth of Corynebacterium glutamicum with glucose as the sole carbon source unless hitherto unknown suppressor mutations occurred; however, these mutations were not disclosed. In the present study, a suppressor mutation was identified, and it was shown that heterologous expression of udhA encoding soluble transhydrogenase from Escherichia coli partly restored growth, suggesting that growth was inhibited by NADPH accumulation. Moreover, genome sequence analysis of second-site suppressor mutants that were able to grow faster with glucose revealed a single point mutation in the gene of non-proton-pumping NADH:ubiquinone oxidoreductase (NDH-II) leading to the amino acid change D213G, which was shared by these suppressor mutants. Since related NDH-II enzymes accepting NADPH as the substrate possess asparagine or glutamine residues at this position, D213G, D213N, and D213Q variants of C. glutamicum NDH-II were constructed and were shown to oxidize NADPH in addition to NADH. Taking these findings together, ATP-neutral glycolysis by the replacement of endogenous NAD-dependent GAPDH with NADP-dependent GapN became possible via oxidation of NADPH formed in this pathway by mutant NADPH-accepting NDH-II D213G and thus by coupling to electron transport phosphorylation (ETP).A TP generation occurs naturally by substrate-level phosphorylation (SLP), electron transport phosphorylation (ETP), photophosphorylation, and decarboxylation phosphorylation. The Embden-Meyerhof-Parnas (EMP) pathway of glycolysis supplies ATP by SLP in the absence of terminal electron acceptors or anaerobic conditions, as well as supplying direct precursors for biomass formation (1). Bacteria and archaea possess one or more of multiple biologically feasible routes for glucose catabolism, such as the EMP pathway, the Entner-Doudoroff (ED) pathway, and the phosphoketolase pathway, which yield zero to 3 ATP molecules per glucose molecule and exist in different variants. In natural habitats, a trade-off between the rate and the yield of ATP production is observed for heterotrophic organisms; faster but less efficient ATP production may be advantageous under conditions characterized by a low abundance of growth substrates (2, 3). For example, Zymomonas mobilis ferments suga...

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Section: Methodsmentioning

confidence: 99%

Metabolic Engineering of an ATP-Neutral Embden-Meyerhof-Parnas Pathway in Corynebacterium glutamicum: Growth Restoration by an Adaptive Point Mutation in NADH Dehydrogenase

Reddy

Lindner

Wendisch

2015

Appl Environ Microbiol

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“…For elution a buffer gradient consisted of 0.25% sodium acetate, pH 6.0 as the polar phase and methanol as the nonpolar phase was used. Diaminohexane was used as an internal standard (Peters-Wendisch et al, 2014).…”

Section: Quantification Of Acetylputrescine Putrescinementioning

confidence: 99%

“…Based on recombinant C. glutamicum strains, http://dx.doi.org/10.1016/j.jbiotec.2014.10.035 0168-1656/© 2014 Elsevier B.V. All rights reserved. putrescine production from the alternative carbon sources crude glycerol (Meiswinkel et al, 2013b), hemicellulosic hydrolysates (Meiswinkel et al, 2013a) and glucosamine (Uhde et al, 2013) as well as putrescine production by a C. glutamicum rendered biotin prototrophic (Peters-Wendisch et al, 2014) were described.…”

Section: Introductionmentioning

confidence: 99%

Elimination of polyamine N-acetylation and regulatory engineering improved putrescine production by Corynebacterium glutamicum

Nguyen

Schneider

Wendisch

2015

Journal of Biotechnology

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“…The Gram-positive Corynebacterium glutamicum is known for its use in the million-ton-scale production of the amino acids L-glutamate and L-lysine (Eggeling and Bott 2015;Peters-Wendisch et al 2014). C. glutamicum has been engineered for access to alternative carbon sources (Leßmeier et al 2015;Wendisch 2014) and for the production of a variety of different products (Wendisch 2014), like the diamine 1,5-diaminopentane (Mimitsuka et al 2007), a monomeric polyamide precursor commonly known as cadaverine (Kind et al 2010;Zahoor et al 2012).…”

Section: Introductionmentioning

confidence: 99%

Production of carbon-13-labeled cadaverine by engineered Corynebacterium glutamicum using carbon-13-labeled methanol as co-substrate

Leßmeier

Pfeifenschneider

Carnicer

et al. 2015

Appl Microbiol Biotechnol

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Methanol, a one-carbon compound, can be utilized by a variety of bacteria and other organisms as carbon and energy source and is regarded as a promising substrate for biotechnological production. In this study, a strain of non-methylotrophic Corynebacterium glutamicum, which was able to produce the polyamide building block cadaverine as non-native product, was engineered for co-utilization of methanol. Expression of the gene encoding NAD+-dependent methanol dehydrogenase (Mdh) from the natural methylotroph Bacillus methanolicus increased methanol oxidation. Deletion of the endogenous aldehyde dehydrogenase genes ald and fadH prevented methanol oxidation to carbon dioxide and formaldehyde detoxification via the linear formaldehyde dissimilation pathway. Heterologous expression of genes for the key enzymes hexulose-6-phosphate synthase and 6-phospho-3-hexuloisomerase of the ribulose monophosphate (RuMP) pathway in this strain restored growth in the presence of methanol or formaldehyde, which suggested efficient formaldehyde detoxification involving RuMP key enzymes. While growth with methanol as sole carbon source was not observed, the fate of 13C-methanol added as co-substrate to sugars was followed and the isotopologue distribution indicated incorporation into central metabolites and in vivo activity of the RuMP pathway. In addition, 13C-label from methanol was traced to the secreted product cadaverine. Thus, this synthetic biology approach led to a C. glutamicum strain that converted the non-natural carbon substrate methanol at least partially to the non-native product cadaverine.

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Engineering biotin prototrophic Corynebacterium glutamicum strains for amino acid, diamine and carotenoid production

Cited by 30 publications

References 85 publications

Metabolic Engineering of an ATP-Neutral Embden-Meyerhof-Parnas Pathway in Corynebacterium glutamicum: Growth Restoration by an Adaptive Point Mutation in NADH Dehydrogenase

Metabolic Engineering of an ATP-Neutral Embden-Meyerhof-Parnas Pathway in Corynebacterium glutamicum: Growth Restoration by an Adaptive Point Mutation in NADH Dehydrogenase

Elimination of polyamine N-acetylation and regulatory engineering improved putrescine production by Corynebacterium glutamicum

Production of carbon-13-labeled cadaverine by engineered Corynebacterium glutamicum using carbon-13-labeled methanol as co-substrate

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