Comparative genomics and metabolomics analyses of the adaptation mechanism in Ketogulonicigenium vulgare-Bacillus thuringiensis consortium

Nan, Jun; Ding, Ming-Zhu; Zou, Yang; Gao, Feng; Yang, Yuan

doi:10.1038/srep46759

Cited by 12 publications

(5 citation statements)

References 52 publications

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“…Native microbial communities have usually evolved over thousands of years toward an extremely efficient use of the available resources, thereby heavily relying on cooperation and cross-feeding among the community members [ 1 , 3 , 5 ]. Recent advances in evolving synthetic co-cultures of strains of the same species [ 6 , 40 ] or mixed species communities [ 41 , 42 ] underlined the potential of increasing product cross-feeding, which could be of high biotechnological value. In this work, we successfully evolved a synthetic community composed of amino acid auxotrophic strains [ 9 ], identified the relevant mutations and used these to increase l -arginine production also in monoculture.…”

Section: Discussionmentioning

confidence: 99%

Discovery of novel amino acid production traits by evolution of synthetic co-cultures

et al. 2023

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Background Amino acid production features of Corynebacterium glutamicum were extensively studied in the last two decades. Many metabolic pathways, regulatory and transport principles are known, but purely rational approaches often provide only limited progress in production optimization. We recently generated stable synthetic co-cultures, termed Communities of Niche-optimized Strains (CoNoS), that rely on cross-feeding of amino acids for growth. This setup has the potential to evolve strains with improved production by selection of faster growing communities. Results Here we performed adaptive laboratory evolution (ALE) with a CoNoS to identify mutations that are relevant for amino acid production both in mono- and co-cultures. During ALE with the CoNoS composed of strains auxotrophic for either l-leucine or l-arginine, we obtained a 23% growth rate increase. Via whole-genome sequencing and reverse engineering, we identified several mutations involved in amino acid transport that are beneficial for CoNoS growth. The l-leucine auxotrophic strain carried an expression-promoting mutation in the promoter region of brnQ (cg2537), encoding a branched-chain amino acid transporter in combination with mutations in the genes for the Na+/H+-antiporter Mrp1 (cg0326-cg0321). This suggested an unexpected link of Mrp1 to l-leucine transport. The l-arginine auxotrophic partner evolved expression-promoting mutations near the transcriptional start site of the yet uncharacterized operon argTUV (cg1504-02). By mutation studies and ITC, we characterized ArgTUV as the only l-arginine uptake system of C. glutamicum with an affinity of KD = 30 nM. Finally, deletion of argTUV in an l-arginine producer strain resulted in a faster and 24% higher l-arginine production in comparison to the parental strain. Conclusion Our work demonstrates the power of the CoNoS-approach for evolution-guided identification of non-obvious production traits, which can also advance amino acid production in monocultures. Further rounds of evolution with import-optimized strains can potentially reveal beneficial mutations also in metabolic pathway enzymes. The approach can easily be extended to all kinds of metabolite cross-feeding pairings of different organisms or different strains of the same organism, thereby enabling the identification of relevant transport systems and other favorable mutations.

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Section: Discussionmentioning

confidence: 99%

Discovery of novel amino acid production traits by evolution of synthetic co-cultures

et al. 2023

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“…Heterologous expression of Vitreoscilla hemoglobin-VHb 6NSL (11) 87.5 (Liu et al 2021) the strains. Furthermore, the adaptation mechanism of B. thuringiensis weakened its sporulation and increased its growth rate, which positively affected K. vulgare growth and productivity towards 2-KLG (4) (Ma et al 2014;Jia et al 2017). In addition, when G. oxydans are in co-culture with B. endophyticus and K. vulgare, a negative impact on its carbon metabolism (PPP-pentose phosphate pathway) occurred, which affected cells growth and protein synthesis.…”

Section: G Oxydans Ab-pqq-vhbmentioning

confidence: 99%

The industrial versatility of Gluconobacter oxydans: current applications and future perspectives

Alves-Ribeiro

Oliveira

Lima

et al. 2022

World J Microbiol Biotechnol

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Gluconobacter oxydans is a well-known acetic acid bacterium that has long been applied in the biotechnological industry. Its extraordinary capacity to oxidize a variety of sugars, polyols, and alcohols into acids, aldehydes, and ketones is advantageous for the production of valuable compounds. Relevant G. oxydans industrial applications are in the manufacture of L-ascorbic acid (vitamin C), miglitol, gluconic acid and its derivatives, and dihydroxyacetone. Increasing efforts on improving these processes have been made in the last few years, especially by applying metabolic engineering. Thereby, a series of genes have been targeted to construct powerful recombinant strains to be used in optimized fermentation. Furthermore, low-cost feedstocks, mostly agro-industrial wastes or byproducts, have been investigated, to reduce processing costs and improve the sustainability of G. oxydans bioprocess. Nonetheless, further research is required mainly to make these raw materials feasible at the industrial scale. The current shortage of suitable genetic tools for metabolic engineering modifications in G. oxydans is another challenge to be overcome. This paper aims to give an overview of the most relevant industrial G. oxydans processes and the current strategies developed for their improvement.

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“…In the case of the industrial production of 2‐keto‐ L ‐gulonic acid (KGA), the precursor molecule of vitamin C, the fermentation process is often conducted with a coculture , . Ketogulonicigenium vulgare is the microorganism converting sorbose into KGA and variations are widely encountered in this biotechnological process .…”

Section: Implementation Of a Microbial Consortiummentioning

confidence: 99%

Fermentation Titer Optimization and Impact on Energy and Water Consumption during Downstream Processing

et al. 2018

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A common focus of fermentation process optimization is the product titer. Different strategies to boost fermentation titer target whole‐cell biocatalyst selection, process control, and medium composition. Working at higher product concentrations reduces the water that needs to be removed in the case of aqueous systems and, therefore, lowers the cost of downstream separation and purification. Different approaches to achieve higher titer in fermentation are examined. Energy and water consumption data collected from different Cargill fermentation plants, i.e., ethanol, lactic acid, and 2‐keto‐L‐gulonic acid, confirm that improvements in fermentation titer play a decisive role in downstream economics and environmental footprint.

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Comparative genomics and metabolomics analyses of the adaptation mechanism in Ketogulonicigenium vulgare-Bacillus thuringiensis consortium

Cited by 12 publications

References 52 publications

Discovery of novel amino acid production traits by evolution of synthetic co-cultures

Discovery of novel amino acid production traits by evolution of synthetic co-cultures

The industrial versatility of Gluconobacter oxydans: current applications and future perspectives

Fermentation Titer Optimization and Impact on Energy and Water Consumption during Downstream Processing

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