Effects of heat, cold, acid and bile salt adaptations on the stress tolerance and protein expression of kefir-isolated probiotic Lactobacillus kefiranofaciens M1

Chen, Ming‐Ju; Tang, Hung-Jie; Chiang, Meng‐Tsan

doi:10.1016/j.fm.2017.03.020

Cited by 100 publications

(63 citation statements)

References 77 publications

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“…This strain, which was shown to adapt hyperosmotic constraint and to transport GB, overexpressed proteins involved in carbohydrates transport and utilization, electron transfer chain and ATP synthesis (Table 2). Enolase, as an example, was previously shown to participate in multiple stress adaptation in P. freudenreichii [27] and in other food-grade bacteria [46]. This glycolytic enzyme is also a component of RNA degradosome, which is involved in RNA processing and gene regulation, in accordance with its role in stress tolerance acquisition in many bacteria [47].…”

Section: Proteomic Patterns May Explain Differential Adaptation and Smentioning

confidence: 94%

Benefits and drawbacks of osmotic adjustment in Propionibacterium freudenreichii

Bonnassié

Rabah

Leverrier

et al. 2019

Journal of Proteomics

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Propionibacterium freudenreichii is a beneficial bacterium used as a cheese starter and as a probiotic. Indeed, selected strains of P. freudenreichii combine both technological and health-promoting abilities. Moreover, during large-scale industrial production of dried bacteria and during consumption, P. freudenreichii may undergo different stressful processes. Osmotic adaptation was shown to enhance P. freudenreichii tolerance towards stresses, which are encountered during freeze-drying and during digestion. In this report, we compared the osmoadaptation molecular mechanisms of two P. freudenreichii strains. Both osmotolerance and osmoadaptation were strain-dependent and had different effects on multiple stress tolerance, depending on the presence of osmoprotectants. Availability of glycine betaine (GB) restored the growth of one of the two strains. In this strain, osmotic preadaptation enhanced heat, oxidative and acid stresses tolerance, as well as survival upon freezedrying. However, addition of GB in the medium had deleterious effects on stress tolerance, while restoring optimal growth under hyperosmotic constraint. In the other strain, neither salt nor GB enhanced stress tolerance, which was constitutively low. Accordingly, whole cell proteomics revealed that mechanisms triggered by salt in the presence and in the absence of GB are different between strains. Osmotic adjustment may thus have deleterious effects on industrial abilities of P. freudenreichii. Biological significance: Propionibacteria are found in various niches including fodder, silage, rumen, milk and cheeses. This means adaptation towards different ecological environments with different physicochemical parameters. Propionibacterium freudenreichii, in particular, is furthermore used both as dairy starter and as probiotic and is thus submitted to high scale industrial production. Production and subsequent stabilization still need optimization. Drying processes like freeze-drying are stressful. Osmotic adjustments may modulated tolerance towards drying. However, they are strain-dependent, medium-dependent and may either reduce or increase stress tolerance. A case-by-case study, for each strain-medium thus seems necessary. In this work, we identify key proteins involved in osmoadaptation and give new insights into adaptation mechanisms in P. freudenreichii. This opens new perspectives for the selections of strains and for the choice of the growth medium composition. Union) status [1,2]. P. freudenreichii is consumed in high amounts, mainly in Swiss-type cheeses, but also in functional probiotic food supplements. Probiotics are "live microorganisms that, when administered in adequate amounts, confer a health benefit on the host" [3]. Modification of the life style, including Western diet (which is high in

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Section: Proteomic Patterns May Explain Differential Adaptation and Smentioning

confidence: 94%

Benefits and drawbacks of osmotic adjustment in Propionibacterium freudenreichii

Bonnassié

Rabah

Leverrier

et al. 2019

Journal of Proteomics

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“…Additionally, the stable pH value and TA in the fermented products made with CISCP during the fermentation cycles suggested that the immobilized LAB gradually adapted to the acid environment during each successive batch fermentation, contributing to increasing LAB survivability and maintaining their metabolic activity over long-term successive fermentation. Other studies have confirmed that acid adaptation can enhance the pH resistance characteristics of microorganisms (Kourkoutas et al, 2005;Chen et al, 2017). It is also worth noting that fermented milk made from CISCP had higher pH and lower TA than traditional yogurt products.…”

Section: Chemical and Physical Properties Of Fermented Milkmentioning

confidence: 82%

A novel immobilized cell system involving Taiwanese kefir microorganisms and sugar cane pieces for fermented milk production

Wang²,

Chen³

2020

Journal of Dairy Science

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The objective of this study was to develop a novel immobilized system using kefir lactic acid bacteria and sugar cane pieces for the production of fermented milk. Lactobacillus kefiranofaciens HL1, Lactobacillus kefiri HL2, Leuconostoc mesenteroides HL3, and Lactococcus lactis HL4 were isolated from Taiwanese kefir grains and immobilized on pieces of sugar cane using adsorption. Scanning electron micrographs of the cellimmobilized sugar cane pieces (CISCP) showed that the microorganisms were embedded within the porous structures of the sugar cane pieces. During 28 cycles of repeated batch fermentation, viable cells on both sugar cane pieces and fermented products were maintained at 10 log cfu/g (cfu/mL). Random amplified polymorphic DNA PCR analysis revealed that Leu. mesenteroides HL3 (29-43%) and Lc. lactis HL4 (31-49%) were predominant on the CISCP, and the fermented samples had 79% Lc. lactis HL4. When tracking fermentation parameters, the data on the microbial, chemical, and physical properties of the fermented milk suggested that the CISCP had stable fermentative ability over the course of successive fermentations. We found an enhancement of the acid-producing ability of CISCP as the number of fermentations increased, with a significant growth in titratable acidity from 0.65 to 0.81% by the end.

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“…This variation in experiment‐to‐experiment counts was also observed – to a lesser extent – in the development phase and can be attributed to slight variations in the culturing conditions, batch‐to‐batch variation at manufacturing and/or standard error in the plate count method. Culture conditions have been shown to influence probiotic survival (Mills et al ; Chen et al ), and it has been shown to be able to precondition the cells to GI stress by incubating or growing them under, for example, low acid conditions (Mills et al ; Chen et al ). Lower stress during processing or preconditioning during cell culture leads to a hypothesized lower stress for the cell, and a higher ability to survive during gastrointestinal transit.…”

Section: Resultsmentioning

confidence: 99%

Probiotic survival during a multi‐layered tablet development as tested in a dynamic, computer‐controlledin vitromodel of the stomach and small intestine (TIM‐1)

Venema

Verhoeven

Verbruggen

et al. 2019

Lett Appl Microbiol

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The aim of the research was to develop a galenical formulation for the combination of the three probiotic strains Lactobacillus gasseri PA 16/8, Bifidobacterium longum SP 07/3 and Bifidobacterium bifidum MF 20/5 that would lead to the presence of a high amount of viable cells in the small intestine, the presumed site of action of these strains. This was tested in a validated, dynamic in vitro model of the stomach and small intestine (TIM‐1), simulating human adults after intake of a meal. Experiments were performed both in the gastric compartment of the model, as well as in the complete system (stomach + small intestine). Survival of the strains in an unformulated probiotic powder after transit through the gastric compartment was 5·3% for the bifidobacteria and 1% for L. gasseri. After transit through the complete gastrointestinal tract, this dropped to 2% for bifidobacteria and 0·1% for Lactobacillus. After several rounds of optimization, an enteric‐coated tablet was developed that increased the delivery of viable cells reaching the small intestine to 72% (gastric survival) for bifidobacteria, and 53% (gastric) for L. gasseri. Also survival in the small intestine increased by about an order of magnitude. The final galenical formulation was tested in two applications: adults and elderly, both of which have their own physiological parameters. These experiments corroborated the results obtained in the development phase of the project. In conclusion, the developed enteric coating led to a 20‐ to 40‐fold increase in the delivery of viable cells to the small intestine.Significance and Impact of the StudyPredictive GI in vitro models are very helpful and reliable tools for the development of new galenical formula containing probiotics, and in the current example helped to deliver >10‐fold higher numbers of viable cells to the small intestine, presumably leading to improved functionality of the strains.

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Effects of heat, cold, acid and bile salt adaptations on the stress tolerance and protein expression of kefir-isolated probiotic Lactobacillus kefiranofaciens M1

Cited by 100 publications

References 77 publications

Benefits and drawbacks of osmotic adjustment in Propionibacterium freudenreichii

Benefits and drawbacks of osmotic adjustment in Propionibacterium freudenreichii

A novel immobilized cell system involving Taiwanese kefir microorganisms and sugar cane pieces for fermented milk production

Probiotic survival during a multi‐layered tablet development as tested in a dynamic, computer‐controlledin vitromodel of the stomach and small intestine (TIM‐1)

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