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Limosilactobacillus reuteri DSM 17938 Produce Bioactive Components during Formulation in Sucrose

Ermann Lundberg, Ludwig ; Mata Forsberg, Manuel ; Lemanczyk, James ; Sverremark-Ekström, Eva ; Sandström, Corine ; Roos, Stefan and Håkansson, Sebastian LU (2024) In Microorganisms 12(10).
Abstract

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... (More)

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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author
; ; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
exopolysaccharides, formulation, immunomodulation, lyoconversion, metabolites, probiotics
in
Microorganisms
volume
12
issue
10
article number
2058
publisher
MDPI AG
external identifiers
  • pmid:39458367
  • scopus:85207677654
ISSN
2076-2607
DOI
10.3390/microorganisms12102058
language
English
LU publication?
yes
id
61da3d92-aba6-428d-a20e-64e07dfd6f35
date added to LUP
2024-12-10 11:04:55
date last changed
2025-07-09 04:05:59
@article{61da3d92-aba6-428d-a20e-64e07dfd6f35,
  abstract     = {{<p>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.</p>}},
  author       = {{Ermann Lundberg, Ludwig and Mata Forsberg, Manuel and Lemanczyk, James and Sverremark-Ekström, Eva and Sandström, Corine and Roos, Stefan and Håkansson, Sebastian}},
  issn         = {{2076-2607}},
  keywords     = {{exopolysaccharides; formulation; immunomodulation; lyoconversion; metabolites; probiotics}},
  language     = {{eng}},
  number       = {{10}},
  publisher    = {{MDPI AG}},
  series       = {{Microorganisms}},
  title        = {{Limosilactobacillus reuteri DSM 17938 Produce Bioactive Components during Formulation in Sucrose}},
  url          = {{http://dx.doi.org/10.3390/microorganisms12102058}},
  doi          = {{10.3390/microorganisms12102058}},
  volume       = {{12}},
  year         = {{2024}},
}