Cascaded valorization of brown seaweed to produce l-lysine and value-added products using Corynebacterium glutamicum streamlined by systems metabolic engineering

Hoffmann, Sarah Lisa; Kohlstedt, Michael; Jungmann, Lukas; Hutter, Michael C.; Poblete‐Castro, Ignacio; Becker, Jörg; Wittmann, Christoph

doi:10.1016/j.ymben.2021.07.010

Cited by 39 publications

(23 citation statements)

References 93 publications

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“…This research group further obtained strain SEA-7 by introducing transhydrogenase PntAB and fructokinase Mak from E. coli , and gapN from S. mutans . The SEA-7 strain was expanded to produce l -lysine from Laminaria digitata extracts and Durvillaea antarctica waste stream with a yield of 0.27 and 0.4 mol/mol, respectively [ 44 ]. Recently, C. glutamicum CgRibo4, a riboflavin-producing strain, was generated via heterologous expression of mtlDBs and mtlAFBs and overexpression of the riboflavin operon ribGCAH .…”

Section: Resultsmentioning

confidence: 99%

“…Therefore, attempts have been made to expand the application of third-generation biomass feedstock for microbial fermentation production of l -ornithine. For instance, we previously developed an engineered strain MTL13 that could produce 54.56 g/L of l -ornithine from mannitol, a sugar alcohol that can be easily extracted from ocean-farmed macroalgae globally [ 20 , 21 ]. This l -ornithine technical parameters is significantly higher than that obtained using second-generation biomass, but there is still room for improvement compared with the performance using glucose as a carbon source.…”

Section: Introductionmentioning

confidence: 99%

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Improvement in l-ornithine production from mannitol via transcriptome-guided genetic engineering in Corynebacterium glutamicum

Nie

et al. 2022

Biotechnol Biofuels

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Background l-Ornithine is an important medicinal intermediate that is mainly produced by microbial fermentation using glucose as the substrate. To avoid competition with human food resources, there is an urgent need to explore alternative carbon sources for l-ornithine production. In a previous study, we constructed an engineered strain, Corynebacterium glutamicum MTL13, which produces 54.56 g/L of l-ornithine from mannitol. However, compared with the titers produced using glucose as a substrate, the results are insufficient, and further improvement is required. Results In this study, comparative transcriptome profiling of MTL01 cultivated with glucose or mannitol was performed to identify novel targets for engineering l-ornithine-producing strains. Guided by the transcriptome profiling results, we modulated the expression of qsuR (encoding a LysR-type regulator QsuR), prpC (encoding 2-methylcitrate synthase PrpC), pdxR (encoding a MocR-type regulator PdxR), acnR (encoding a TetR-type transcriptional regulator AcnR), CGS9114_RS08985 (encoding a hypothetical protein), and CGS9114_RS09730 (encoding a TetR/AcrR family transcriptional regulator), thereby generating the engineered strain MTL25 that can produce l-ornithine at a titer of 93.6 g/L, representing a 71.6% increase as compared with the parent strain MTL13 and the highest l-ornithine titer reported so far for C. glutamicum. Conclusions This study provides novel indirect genetic targets for enhancing l-ornithine accumulation on mannitol and lays a solid foundation for the biosynthesis of l-ornithine from marine macroalgae, which is farmed globally as a promising alternative feedstock.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Improvement in l-ornithine production from mannitol via transcriptome-guided genetic engineering in Corynebacterium glutamicum

Nie

et al. 2022

Biotechnol Biofuels

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“…In C. glutamicum, there are four enzymes that are reported to involve in NADPH regeneration (Fig. 1) (Hoffmann et al 2021;Xu et al 2019). In order to construct an NADPH-auxotrophic C. glutamicum strain, we aimed to inactivate the Zwf and the MalE as well as to replace the native NADP-Icd Cg by NAD-Icd Ca from S. mutans.…”

Section: Construction and Characterization Of An Nadph-auxotrophic C ...mentioning

confidence: 99%

“…In C. glutamicum, four enzymes were reported to involve in NADPH biosynthesis, i.e., glucose-6-phosphate dehydrogenase (Zwf), 6-phosphogluconate dehydrogenase (Gnd), NADP-dependent isocitrate dehydrogenase (NADP-Icd) and malic enzyme (MalE)(Fig. 1) (Hoffmann et al 2021;Xu et al 2019). However, the Zwf and Gnd in pentose phosphate (PP) pathway are the main enzymes for NADPH regeneration (Lindner et al 2018).…”

Section: Introductionmentioning

confidence: 99%

An NADPH-auxotrophic Corynebacterium glutamicum recombinant strain and used it to construct L-leucine high-yielding strain

Chen

Liu²,

Zhang³

et al. 2022

Int Microbiol

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The NADPH-regeneration enzymes in Corynebacterium glutamicum were inactivated to construct an NADPH-auxotrophic C. glutamicum strain by gene knockout and gene replacement. The resultant NADPH-auxotrophic C. glutamicum XL-1 ΔZMI Cg ::I Sm (i.e., strain Leu-1) grew well in the basic medium only with gluconate as carbon source. Replacement of the native glyceraldehyde 3-phosphate dehydrogenase (NAD-GapDH Cg ) by NADP-GapDH Ca from Clostridium acetobutylicum is an effective strategy for producing L-leucine in NADPH-prototrophic strain XL-1 and NADPH-auxotrophic strain Leu-1, whereas the L-leucine yield did not differ signi cantly between these strains (14.1 ± 1.8 g/L v.s. 16.2 ± 1.1 g/L). Enhancing the carbon ux in biosynthetic pathway by recombinant expression plasmid pEC-ABNCE promoted L-leucine production, but the shortage NADPH supply limited the L-leucine yield. The mutated promoters of zwf and icd Cg were introduced into C. glutamicum with NADP-GapDH Ca and pEC-ABNCE increased L-leucine yield (32.8 ± 1.7 g/L) and improved cell growth (OD 562 = 41.3 ± 4.2) because the resultant strain C. glutamicum XL-1 ΔMI Cg ::I Sm G Cg ::G Ca P zwf -D1 P icd -D2/pEC-ABNCE (i.e., strain Leu-9) exhibited the proper intracellular NADPH and NADH level. This is the rst report of constructing an L-leucine high-yielding strain that reasonably supplies NADPH by optimizing the biosynthetic pathway of NADPH from an NADPH-auxotrophic strain.

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“…C. glutamicum has no significant extracellular proteolytic activity avoiding degradation of secreted protein or peptide products ( Becker and Wittmann, 2017 ; Lee and Kim, 2018 ). Moreover, the organism has been engineered for efficient growth and production on different renewable feedstocks and industrial waste streams including raw starches, pentoses from hemicellulosic wastes, mannitol from seaweed hydrolysates and amino sugars derived from chitin ( Hoffmann et al, 2021 ; Matano et al, 2014 ; Meiswinkel et al, 2013 ; Seibold et al, 2006 ).…”

Section: Introductionmentioning

confidence: 99%

Establishing recombinant production of pediocin PA-1 in Corynebacterium glutamicum

Goldbeck

Desef

Ovchinnikov

et al. 2021

Metabolic Engineering

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Bacteriocins are antimicrobial peptides produced by bacteria to inhibit competitors in their natural environments. Some of these peptides have emerged as commercial food preservatives and, due to the rapid increase in antibiotic resistant bacteria, are also discussed as interesting alternatives to antibiotics for therapeutic purposes. Currently, commercial bacteriocins are produced exclusively with natural producer organisms on complex substrates and are sold as semi-purified preparations or crude fermentates. To allow clinical application, efficacy of production and purity of the product need to be improved. This can be achieved by shifting production to recombinant microorganisms. Here, we identify Corynebacterium glutamicum as a suitable production host for the bacteriocin pediocin PA-1. C. glutamicum CR099 shows resistance to high concentrations of pediocin PA-1 and the bacteriocin was not inactivated when spiked into growing cultures of this bacterium. Recombinant C. glutamicum expressing a synthetic pedACD Cgl operon releases a compound that has potent antimicrobial activity against Listeria monocytogenes and Listeria innocua and matches size and mass:charge ratio of commercial pediocin PA-1. Fermentations in shake flasks and bioreactors suggest that low levels of dissolved oxygen are favorable for production of pediocin. Under these conditions, however, reduced activity of the TCA cycle resulted in decreased availability of the important pediocin precursor l -asparagine suggesting options for further improvement. Overall, we demonstrate that C. glutamicum is a suitable host for recombinant production of bacteriocins of the pediocin family.

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Cascaded valorization of brown seaweed to produce l-lysine and value-added products using Corynebacterium glutamicum streamlined by systems metabolic engineering

Cited by 39 publications

References 93 publications

Improvement in l-ornithine production from mannitol via transcriptome-guided genetic engineering in Corynebacterium glutamicum

Improvement in l-ornithine production from mannitol via transcriptome-guided genetic engineering in Corynebacterium glutamicum

An NADPH-auxotrophic Corynebacterium glutamicum recombinant strain and used it to construct L-leucine high-yielding strain

Establishing recombinant production of pediocin PA-1 in Corynebacterium glutamicum

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