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Quesada-Chanto, A.; Schmid-Meyer, A. C.; Schroeder, A. G.; Carvalho-Jonas, Maria F.; Blanco, I.; Jonas, Rainer

doi:10.1023/a:1008868907251

Cited by 26 publications

(4 citation statements)

References 16 publications

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“…Further strain engineering resulted in high-level propionate production, with titers comparable to those achieved by anaerobic batch cultivation of Propionibacteria [2,8,15,30]. Though glucose can be rapidly dissimilated by propionogenic E. coli, it was deemed an unsuitable carbon source primarily due to overflow metabolism of acetate [24,38].…”

Section: Discussionmentioning

confidence: 99%

Engineering Escherichia coli for high-level production of propionate

Akawi

Srirangan

Liu

et al. 2015

Journal of Industrial Microbiology and Biotechnology

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Mounting environmental concerns associated with the use of petroleum-based chemical manufacturing practices has generated significant interest in the development of biological alternatives for the production of propionate. However, biological platforms for propionate production have been limited to strict anaerobes, such as Propionibacteria and select Clostridia. In this work, we demonstrated high-level heterologous production of propionate under microaerobic conditions in engineered Escherichia coli. Activation of the native Sleeping beauty mutase (Sbm) operon not only transformed E. coli to be propionogenic (i.e., propionate-producing) but also introduced an intracellular "flux competition" between the traditional C2-fermentative pathway and the novel C3-fermentative pathway. Dissimilation of the major carbon source of glycerol was identified to critically affect such "flux competition" and, therefore, propionate synthesis. As a result, the propionogenic E. coli was further engineered by inactivation or overexpression of various genes involved in the glycerol dissimilation pathways and their individual genetic effects on propionate production were investigated. Generally, knocking out genes involved in glycerol dissimilation (except glpA) can minimize levels of solventogenesis and shift more dissimilated carbon flux toward the C3-fermentative pathway. For optimal propionate production with high C3:C2-fermentative product ratios, glycerol dissimilation should be channeled through the respiratory pathway and, upon suppressed solventogenesis with minimal production of highly reduced alcohols, the alternative NADH-consuming route associated with propionate synthesis can be critical for more flexible redox balancing. With the implementation of various biochemical and genetic strategies, high propionate titers of more than 11 g/L with high yields up to 0.4 g-propionate/g-glycerol (accounting for ~50 % of dissimilated glycerol) were achieved, demonstrating the potential for industrial application. To our knowledge, this represents the most effective engineered microbial system for propionate production with titers and yields comparable to those achieved by anaerobic batch cultivation of various native propionate-producing strains of Propionibacteria.

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Section: Discussionmentioning

confidence: 99%

Engineering Escherichia coli for high-level production of propionate

Akawi

Srirangan

Liu

et al. 2015

Journal of Industrial Microbiology and Biotechnology

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“…Although the B 12 production pathway in P. freudenreichii is anaerobic, yield increments have been reported for P. freudenreichii grown under aerobic conditions [ 5 ]. On the other hand, Quesada-Chanto et al [ 6 ] and Menon et al [ 7 ] found decreased B 12 production when oxygen was present. The presented studies on B 12 production have in common that relatively high amounts of oxygen are used, resulting in decreased cytochrome synthesis [ 5 , 8 ] potentially caused by diminished -aminolevulinic acid dehydratase activity [ 7 ], resulting in lower growth rates and at higher oxygen levels even in diminished growth.…”

Section: Introductionmentioning

confidence: 99%

Microaerobic metabolism of lactate and propionate enhances vitamin B12 production in Propionibacterium freudenreichii

et al. 2022

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Background Propionibacterium freudenreichii is used in biotechnological applications to produce vitamin B12. Although cultured mainly in anaerobic conditions, microaerobic conditions can greatly enhance biomass formation in P. freudenreichii. Since B12 yields may be coupled to biomass formation, microaerobic conditions show great potential for increasing B12 yields in P. freudenreichii. Results Here we show biomass formation increases 2.7 times for P. freudenreichii grown in microaerobic conditions on lactate versus anaerobic conditions (1.87 g/L vs 0.70 g/L). Consumption of lactate in microaerobic conditions resulted first in production of pyruvate, propionate and acetate. When lactate was depleted, pyruvate and propionate were oxidised with a concomitant sixfold increase in the B12 titer compared to anaerobic conditions, showing potential for propionate and pyruvate as carbon sources for B12 production. Consequently, a fed-batch reactor with anaerobically precultured lactate-grown cells was fed propionate in microaerobic conditions resulting in biomass increase and production of B12. Vitamin yields increased from 0.3 $$\mu g$$ μ g B12 per mmol lactate in anaerobic conditions to 2.4 $$\mu g$$ μ g B12 per mmol lactate and 8.4 $$\mu g$$ μ g B12 per mmol propionate in microaerobic conditions. Yield per cell dry weight (CDW) increased from 41 $$\mu g$$ μ g per g CDW in anaerobic conditions on lactate to 92 $$\mu g$$ μ g per g CDW on lactate and 184 $$\mu g$$ μ g per g CDW on propionate in microaerobic conditions. Conclusions Here we have shown both B12 yield per substrate and per CDW were highest on cells oxidising propionate in microaerobic conditions, showing the potential of propionate for biotechnological production of vitamin B12 by P. freudenreichii.

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“…Because of these components, P. shermanii can use low concentrations of oxygen present in the medium to obtain energy and produce biomass. 40,41 In general, the oxygen supply to P. shermanii cultures will have more pronounced effect on biomass than on vitamin B12 formation.…”

Section: Bioreactor Kinetics Of P Shermanii Atcc 13673 Culturesmentioning

confidence: 99%

“…For instance, maintaining the volumetric oxygen transfer coefficient (k l a) of 10/hr, which is a low value, in batch cultures of P. freudenreichii CDB10015 growing in synthetic medium containing 80 g/L of glucose and 100 g/L of corn step liquor as major components, at 35 C for 72 hr, produced similar amounts of biomass (12.5 g/L), propionic acid (7.0 g/L), acetic acid (13.5 g/L), and vitamin B12 specific yields (0.2 mg/g) as obtained in our work. 40 The relatively high production of acetic acid is another characteristic of P. freudenreichii metabolism under aerobic metabolism. 36 The propionic acid produced under the anaerobe phase is completely converted to pyruvate in presence of dissolved oxygen.…”

Section: Bioreactor Kinetics Of P Shermanii Atcc 13673 Culturesmentioning

confidence: 99%

Biosynthesis of vitamin B12 by Propionibacterium freudenreichii subsp. shermanii ATCC 13673 using liquid acid protein residue of soybean as culture medium

Assis

Matte

Aschidamini

et al. 2020

Biotechnology Progress

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Vitamin B12 deficiency still persists, mainly caused by low intake of animal food products affecting vegetarians, vegans, and populations of underdeveloped countries. In this study, we investigate the biosynthesis of vitamin B12 by potential probiotic bacterium using an agroindustry residue, the liquid acid protein residue of soybean (LAPRS), as a low-cost, animal derivate-free alternative culture medium. Cultures of Propionibacterium freudenreichii subsp. shermanii ATCC 13673 growing in LAPRS for vitamin B12 biosynthesis were studied using the Plackett-Burman experimental approach, followed by a central composite design 2 2 to optimize the concentration of significant variables. We also performed a proteolytic treatment of LAPRS and evaluated the optimized-hydrolyzed medium influence on the microbial growth and metabolism in shaker flask and bioreactor experiments. In this all-plant source medium, P. freudenreichii subsp. shermanii produced high concentrations of cells and high amounts of vitamin B12 (0.6 mg/g cells) after process optimization. These results suggest the possibility of producing vitamin B12 by a potential probiotic bacterium in a very cheap, animal derivate-free medium to address the needs of specific population groups, at the same time reducing the production costs of this essential vitamin.

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Untitled

Cited by 26 publications

References 16 publications

Engineering Escherichia coli for high-level production of propionate

Engineering Escherichia coli for high-level production of propionate

Microaerobic metabolism of lactate and propionate enhances vitamin B12 production in Propionibacterium freudenreichii

Biosynthesis of vitamin B12 by Propionibacterium freudenreichii subsp. shermanii ATCC 13673 using liquid acid protein residue of soybean as culture medium

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