Culture conditions affect Lactobacillus reuteri DSM 17938 ability to perform glycerol bioconversion into 3-hydroxypropionic acid

Nguyen, Thi-Lan-Phuong; Saulou‐Bérion, Claire; Delettre, Jérôme; Béal, Catherine

doi:10.1016/j.jbiosc.2020.12.011

Cited by 4 publications

(32 citation statements)

References 48 publications

(82 reference statements)

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“…The strain Limosilactobacillus reuteri DSM 17938 (BioGaia AB, Stockholm, Sweden) was stored at −80°C in cryotubes with glycerol 20% (v/v). Bacterial cells were reactivated according to Nguyen et al (2021) 6 by two successive cultures in MRS broth (Biokar Diagnostic, Beauvais, France) at 37°C, 100 rpm for 16 h and then 8 h, respectively.…”

Section: Methodsmentioning

confidence: 99%

“…Cells were inoculated at an initial concentration of 5.1 × 10 −6 g of cell dry weight (CDW) per liter (g CDW L −1 ), corresponding to 5.8 × 10 3 cells ml −1 . Growth conditions were defined from the results obtained by Nguyen et al (2021) 6 . The growth medium was composed of MRS broth supplemented with 20 g L −1 glucose (VWR, Fontenay‐sous‐Bois, France), 25 g L −1 phytone peptone (Merck KGaA, Darmstadt, Germany), 3 g L −1 1,2‐propanediol (1,2‐PDO) (Sigma‐Aldrich, Saint‐Quentin‐Fallavier, France), 0.234 g L −1 betaine (Sigma‐Aldrich) plus 7.455 g L −1 KCl (VWR), and 4 g L −1 Tween 80 (VWR).…”

Section: Methodsmentioning

confidence: 99%

“…The volume variation in the bioreactor was calculated on‐line by considering both glycerol and NH 4 OH addition. Bioconversion was interrupted when no more 3‐HP was produced, which was evaluated by the cessation of base consumption and by the concomitant increase in pO 2 6 …”

Section: Methodsmentioning

confidence: 99%

“…Limosilactobacillus reuteri ( L. reuteri ), that was previously known as Lactobacillus reuteri , 4 has the natural ability to convert glycerol into 3‐HP via the propanediol‐utilization Pdu pathway 5 . With this bacterial species, the growth phase on glucose must be separated from the 3‐HP production step, in order to allow resting cells to convert glycerol into 3‐HP instead of only 1,3‐propanediol (1,3‐PDO) 6 . Two main approaches are applied to improve this biotransformation.…”

Section: Introductionmentioning

confidence: 99%

“…By considering the medium composition, a non‐growing medium is reported to be used for 3‐HP bioproduction in order to prevent the synthesis of undesirable fermentation products (i.e., lactic acid, ethanol, acetic acid) and to balance the redox cofactors during bioconversion 5,9 . Consequently, only glycerol shall be supplied as a carbon source during the fed‐batch step 6,10 . The glycerol concentration was shown to influence the bioproduction of 3‐HPA by L. reuteri ATCC 53608 11 .…”

Section: Introductionmentioning

confidence: 99%

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Environmental conditions during glycerol bioconversion affect 3‐hydroxypropionic acid bioproduction byLimosilactobacillus reuteriDSM17938

Nguyen

Béal

Ghorbal

et al. 2022

Biotechnology Progress

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3-Hydroxypropionic acid (3-HP) is a platform molecule whose biological production was carried out by the bacterium Limosilactobacillus reuteri according to a two-step process: first, a growth phase in batch mode on glucose, then a glycerol bioconversion into 3-HP in fed-batch mode. With the objective of improving 3-HP bioproduction, this study aimed at defining the operating conditions during the bioconversion phase that increases the bioproduction performance. A central composite rotatable design allowed testing various pH levels and specific glycerol feeding rates. By establishing response surfaces, optimal conditions have been identified that were different depending on the considered output variable (final 3-HP quantity, 3-HP production yield and production rate). Of them, 3-HP final quantity and 3-HP production yield were maximized at pH 6.0 and at specific glycerol feeding rates of 60 and 55 mg gly g CDW À1 h À1 , respectively. The specific 3-HP production rate was the highest at the upper limit of the specific substrate feeding rate (80 mg gly g CDW À1 h À1 ) but was not affected by the pH. An additional experiment was carried out at pH 6.0 and a specific glycerol feeding rate of 80 mg gly g CDW À1 h À1 to validate the previous observations. In conclusion, the results showed a significant improvement of 3-HP concentration by 13%, of specific production rate by 34% and of 3-HP volumetric productivity by 39%, as compared to the initial values.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Environmental conditions during glycerol bioconversion affect 3‐hydroxypropionic acid bioproduction byLimosilactobacillus reuteriDSM17938

Nguyen

Béal

Ghorbal

et al. 2022

Biotechnology Progress

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Glycerol and glucose co‐fermentation by 1,3‐propanediol producer Limosilactobacillus reuteri: A kinetic study of batch and continuous cultivations

Avilez,

Vieira,

Bresciani

et al. 2023

Biofuels Bioprod Bioref

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The conversion of glycerol into 1,3‐propanediol (1,3‐PDO) has gained significant attention as a sustainable pathway within the biodiesel production chain. Limosilactobacillus reuteri, a bacterium capable of efficiently converting glycerol to 1,3‐PDO, holds promise for this application. In industrial biotechnology, the development of kinetic models is important for bioreactor design, process analysis and scalability, supporting the advancement of biorefineries and the bioeconomy. This study focuses on investigating the kinetics of co‐fermentation of glucose and glycerol by L. reuteri, utilizing data from previous batch culture experiments. Three different models were evaluated for these batch runs, and the parameters were adjusted using the Levenberg–Marquardt algorithm in conjunction with an integration method. The results from these previous batch experiments indicate that pH did not significantly influence the specific growth rate of the bacterium. However, as expected, reducing the temperature from 37 to 30°C resulted in a 60% reduction in the growth rate. Moreover, continuous co‐fermentation experiments were conducted, and a simple kinetic model was proposed. The advantage of the continuous process is that the productivity of 1,3‐PDO is 3.5 times higher compared with co‐fermentation in batch mode. Notably, this study represents the first attempt to model continuous co‐fermentation under a range of dilution rates (0.25–0.9 h−1), identifying a dilution rate of 0.5 h−1 as the optimal condition to produce 1,3‐PDO, considering productivity and substrate conversion. The findings of this research contribute valuable insights for future investigations on the viability of 1,3‐PDO production from glycerol, particularly within the context of waste valorization.

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Limosilactobacillus reuteri DSM 17938 Produce Bioactive Components during Formulation in Sucrose

Ermann Lundberg,

Mata Forsberg,

Lemanczyk

et al. 2024

Microorganisms

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Improved efficacy of probiotics can be achieved by using different strategies, including the optimization of production parameters. The impact of fermentation parameters on bacterial physiology is a frequently investigated topic, but what happens during the formulation, i.e., the step where the lyoprotectants are added prior to freeze-drying, is less studied. In addition to this, the focus of process optimization has often been yield and stability, while effects on bioactivity have received less attention. In this work, we investigated different metabolic activities of the probiotic strain Limosilactobacillus reuteri DSM 17938 during formulation with the freeze-drying protectant sucrose. We discovered that the strain consumed large quantities of the added sucrose and produced an exopolysaccharide (EPS). Using NMR, we discovered that the produced EPS was a glucan with α-1,4 and α-1,6 glycosidic bonds, but also that other metabolites were produced. The conversion of the lyoprotectant is hereafter designated lyoconversion. By also analyzing the samples with GCMS, additional potential bioactive compounds could be detected. Among these were tryptamine, a ligand for the aryl hydrocarbon receptor, and glycerol, a precursor for the antimicrobial compound reuterin (3-hydroxypropionaldehyde). To exemplify the bioactivity potential of lyoconversion, lyoconverted samples as well as purified EPS were tested in a model for immunomodulation. Both lyoconverted samples and purified EPS induced higher expression levels of IL-10 (2 times) and IL-6 (4–6 times) in peripheral blood mononuclear cells than non-converted control samples. We further found that the initial cultivation of DSM 17938 with sucrose as a sugar substrate, instead of glucose, improved the ability to convert sucrose in the lyoprotectant into EPS and other metabolites. Lyoconversion did not affect the viability of the bacteria but was detrimental to freeze-drying survival, an issue that needs to be addressed in the future. In conclusion, we show that the metabolic activities of the bacteria during the formulation step can be used as a tool to alter the activity of the bacteria and thereby potentially improve probiotic efficacy.

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Culture conditions affect Lactobacillus reuteri DSM 17938 ability to perform glycerol bioconversion into 3-hydroxypropionic acid

Cited by 4 publications

References 48 publications

Environmental conditions during glycerol bioconversion affect 3‐hydroxypropionic acid bioproduction byLimosilactobacillus reuteriDSM17938

Environmental conditions during glycerol bioconversion affect 3‐hydroxypropionic acid bioproduction byLimosilactobacillus reuteriDSM17938

Glycerol and glucose co‐fermentation by 1,3‐propanediol producer Limosilactobacillus reuteri: A kinetic study of batch and continuous cultivations

Limosilactobacillus reuteri DSM 17938 Produce Bioactive Components during Formulation in Sucrose

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