Production of lactic acid bacteria starters for manufacturing food, probiotic, and chemical products requires the application of successive steps: fermentation, concentration, stabilization, and ...storage. Despite process optimization, losses of bacterial viability and functional activities are observed after stabilization and storage steps due to cell exposure to environmental stresses (thermal, osmotic, mechanical, and oxidative). Bacterial membrane is the primary target for injury and its damage is highly dependent on its physical properties and lipid organization. Membrane fluidity is a key property for maintaining cell functionality, and depends on lipid composition and cell environment. Extensive evidence has been reported on changes in membrane fatty acyl chains when modifying fermentation conditions. However, a deep characterization of membrane physical properties and their evolution following production processes is scarcely reported. Therefore, the aims of this mini-review are (i) to define the membrane fluidity and the methods used to assess it and (ii) to summarize the effect of environmental conditions on membrane fluidity and the resulting impact on the resistance of lactic acid bacteria to the stabilization processes. This will make it possible to highlight existing gaps of knowledge and opens up novel approaches for future investigations.
Fructo- and galacto-oligosaccharides (FOS and GOS) are non-digestible oligosaccharides with prebiotic properties that can be incorporated into a wide number of products. This review details the ...general outlines for the production of FOS and GOS, both by enzymatic synthesis using disaccharides or other substrates, and by hydrolysis of polysaccharides. Special emphasis is laid on technological aspects, raw materials, properties, and applications.
The role of S-layer proteins (SLP) on the Pb²⁺sequestrant capacity by Lactobacillus kefir CIDCA 8348 and JCM 5818 was investigated. Cultures in the stationary phase were treated with proteinase K. A ...dot blot assay was carried out to assess the removal of SLP. Strains with and without SLP were exposed to 0–0.5 mM Pb(NO₃)₂. The maximum binding capacity (q ₘₐₓ) and the affinity coefficient (b) were calculated using the Langmuir equation. The structural effect of Pb²⁺on microorganisms with and without SLP was determined using Raman spectroscopy. The bacterial interaction with Pb²⁺led to a broadening in the phosphate bands (1,300–1,200 cm⁻¹region) and strong alterations on amide and carboxylate-related bands (νCOO⁻as and νCOO⁻s). Microorganisms without SLP removed higher percentages of Pb²⁺and had higher q ₘₐₓ than those bearing SLP. Isolated SLP had much lower q ₘₐₓ and also removed lower percentages of Pb²⁺than the corresponding whole microorganisms. The hydrofobicity of both strains dramatically dropped when removing SLP. When bearing SLP, strains do not expose a large amount of charged groups on their surfaces, thus making less efficient the Pb²⁺removal. On the contrary, the extremely low hydrofobicity of microorganisms without SLP (and consequently, their higher capacity to remove Pb²⁺) can be explained on the basis of a greater exposure of charged chemical groups for the interaction with Pb²⁺. The viability of bacteria without SLP was not significantly lower than that of bacteria bearing SLP. However, microorganisms without SLP were more prone to the detrimental effect of Pb²⁺, thus suggesting that SLP acts as a protective rather than as a sequestrant layer.
Galacto-oligosaccharides (GOS) and lactulose are well-recognized prebiotics widely used in functional food and pharmaceutical products, but there is still a lack of knowledge regarding their ...physical–chemical properties. In this study, a physical–chemical approach on two GOS of different composition (GOS Cup Oligo H-70® and GOS Biotempo) and lactulose was assessed. Mid infrared and Raman spectra of the freeze-dried sugars allowed their structural characterization in the amorphous state, lactulose, showing the main spectral differences.
Freeze-dried sugars were then equilibrated at 4°C at relative humidity (RH) ranging from 11% to 80%. Near-infrared reflectance spectra were registered in each condition in the 900- to 1700-nm region. A principal component analysis (PCA) was performed on the three sugars equilibrated at different RH. In all the three sugars, the groups observed explained more than 95% of the variance and were related with the RH of the samples. According to the loading plots of PC1, the main differences related with RH were observed in the 1380- to 1500-nm region.
As the amorphous states are very sensitive to changes in temperature and moisture content, and the moisture content is related with the parameter T-Tg (T: storage temperature; Tg: vitreous transition temperature), an effort was made to determine this parameter directly from the NIR spectra. To this aim, a partial least square model (PLS) was defined. Tg values obtained by differential scanning calorimetry (DSC) were used to calculate the T-Tg values of reference. The model was validated with an independent set of data. The mean of predicted values fitted nicely T-Tg obtained from DSC (correlation=0.966; R2=0.934), thus supporting the use of the PLS model to investigate unknown samples.
The stability of amorphous sugars in foods and pharmaceuticals is of practical and economical importance because it affects different quality attributes of foods, including texture, aroma retention and shelf life. Therefore, predicting T-Tg, a parameter that is independent on the sugar investigated, directly from their NIR spectra is of utmost importance to determine the shelf life of food and food-related products and up to our knowledge has never been determined hereto.
•Amorphous GOS and lactulose were structurally characterized using FTIR and Raman spectroscopies.•NIR allowed the discrimination among GOS and lactulose equilibrated at different RH.•A PLS model to determine T-Tg directly from the NIR spectra was defined and validated.•Predicting T-Tg is of utmost importance to determine the shelf-life of food products.
Malolactic fermentation (MLF) is a process in winemaking responsible for the conversion of l-malic acid to l-lactic acid and CO2, which reduces the total acidity, improves the biological stability, ...and modifies the aroma profile of wine. MLF takes place during or after alcoholic fermentation and is carried out by one or more species of lactic acid bacteria (LAB), which are either present in grapes and cellars or inoculated with malolactic starters during the winemaking process. Although the main bacterium among LAB used in commercial starter cultures for MLF has traditionally been Oenococcus oeni, in the last decade, Lactobacillus plantarum has also been reported as a malolactic starter, and many works have shown that this species can survive and even grow under harsh conditions of wine (i.e., high ethanol content and low pH values). Furthermore, it has been proved that some strains of L. plantarum are able to conduct MLF just as efficiently as O. oeni. In addition, L. plantarum exhibits a more diverse enzymatic profile than O. oeni, which could play an important role in the modification of the wine aroma profile. This enzymatic diversity allows obtaining several starter cultures composed of different L. plantarum biotypes, which could result in distinctive wines. In this context, this review focuses on showing the relevance of L. plantarum as a MLF starter culture in winemaking.
How to cite: Brizuela NS, Tymczyszyn E, Semorile LC, et al. Lactobacillus plantarum as a malolactic starter culture in winemaking: a new (old) player? Electron J Biotechnol 2019;38. https://doi.org/10.1016/j.ejbt.2018.12.002.
The ability of galacto-oligosaccharides (GOS) to protect Lactobacillus delbrueckii subsp. bulgaricus upon freeze drying was analyzed on the basis of their capacity to form glassy structures. Glass ...transition temperatures (Tg) of a GOS matrix at various relative humidities (RH) were determined by DSC. Survival of L. bulgaricus in a glassy GOS matrix was investigated after freezing, freeze drying, equilibration at different RHs and storage at different temperatures. At 32°C, a drastic viability loss was observed. At 20°C, the survival was affected by the water content, having the samples stored at lower RHs, the highest survival percentages. At 4°C, no decay in the cells count was observed after 45days of storage. The correlation between molecular mobility as measured by Proton nuclear magnetic resonance (1H NMR) and loss of viability explained the efficiency of GOS as cryoprotectants. The preservation of microorganisms was improved at low molecular mobility and this condition was obtained at low water contents and low storage temperatures. These results are important in the developing of new functional foods containing pre and probiotics.
► Effect of thermophysical properties of GOS on freeze-dried L. bulgaricus. ► GOS remain in the glassy state at room temperature until RH=33%. ► Survival of microorganisms is affected by the water content and temperature of storage. ► The presence of GOS matrix is not sufficient to prevent membrane damage upon freeze drying. ► The increase of molecular mobility is correlated with loss of viability.
The malolactic fermentation (MLF) of Patagonian Malbec wine inoculated with blend cultures of selected native strains of
and
was monitored during 14 days, analyzing the strains ability to modify the ...content of some organic acids and to change the volatile compounds profile. The performance of the LAB strains was tested as single and blends cultures of both species. An implantation control by RAPD PCR was also carried out to differentiate among indigenous and inoculated strains. The
strains UNQLp11 and UNQLp155 and the
strain UNQOe73.2 were able to remain viable during the monitoring time of MLF, whereas the
strain UNQOe31b showed a decrease of five log CFU at day 14. The four strains assayed showed a similar behavior in wine whether they were inoculated individually or as blend cultures. All strains were able to consume L-malic acid, particularly the
strains, which showed the highest consumption values at day 14, both as single or blend cultures. The changes in the volatile compounds profile of Malbec wine samples, before and after MLF, were determined by HS-SPME and GC-MS technique. Wines inoculated with blend cultures containing strain UNQLp155 showed a decrease in the total alcohols content and an increase in the total esters content. On the other hand, wines inoculated with single cultures of strains UNQLp155, UNQOe31b or UNQOe73.2 showed no significant decrease in the total alcohols concentration but a significant increase in the total esters content. When strain UNQLp11 was inoculated as single or as blend culture with strain UNQOe31b, wines exhibited an increase in the total alcohols content, and a decrease in the total esters content. The content of diethyl succinate showed the greatest increase at final of MLF, and a particular synergistic effect in its synthesis was observed with a blend culture of strains UNQLp155 and UNQOe73.2. These results suggest that the use of blend cultures formulated with strains belonging to
and
species could offer an interesting advantage to induce MLF in Malbec wines, contributing to diversify their aromatic profiles.
In this work, the protective capacity of galacto-oligosaccharides in the preservation of
Lactobacillus delbrueckii subsp.
bulgaricus CIDCA 333 was evaluated.
Lactobacillus bulgaricus was freeze-dried ...or dried over silica gel in the presence of three commercial products containing galacto-oligosaccharides. The freeze-dried samples were stored at 5 and 25
°C for different periods of time. After desiccation, freeze-drying or storage, samples were rehydrated and bacterial plate counts were determined.
According to the results obtained, all galacto-oligosaccharides assays demonstrated to be highly efficient in the preservation of
L. bulgaricus. The higher content of galacto-oligosaccharides in the commercial products was correlated with their higher protective capacity.
Galacto-oligosaccharides are widely known by their prebiotic properties. However, their role as protective molecules have not been reported nor properly explored up to now. In this work the protective capacity of galacto-oligosaccharides in the preservation of
L. bulgaricus, a strain particularly sensitive to any preservation process, was demonstrated.
The novel role of galacto-oligosaccharides as protective molecules opens up several perspectives in regard to their applications. The supplementation of probiotics with galacto-oligosaccharides allows the production of self-protected synbiotic products, galacto-oligosaccharides exerting both a prebiotic and protecting effect.
The aim of this work was to study the protective effect of sucrose, trehalose and glutamate during freezing and freeze-drying of three oenological Lactobacillus plantarum strains previously ...acclimated in the presence of ethanol. The efficiency of protective agents was assessed by analyses of membrane integrity and bacterial cultivability in a synthetic wine after the preservation processes. No significant differences in the cultivability, with respect to the controls cells, were observed after freezing at −80 °C and −20 °C, and pre-acclimated cells were more resistant to freeze-drying than non-acclimated ones. The results of multiparametric flow cytometry showed a significant level of membrane damage after freeze-drying in two of the three strains. The cultivability was determined after incubation in wine-like medium containing 13 or 14% v/v ethanol at 21 °C for 24 h and the results were interpreted using principal component analysis (PCA). Acclimation was the most important factor for preservation, increasing the bacterial resistance to ethanol after freezing and freeze-drying. Freeze-drying was the most drastic method of preservation, followed by freezing at −20 °C. The increase of ethanol concentration from 6 to 10% v/v in the acclimation medium improved the recovery of two of the three strains. In turn, the increase of ethanol content in the synthetic wine led to a dramatic decrease of viable cells in the three strains investigated. The results of this study indicate that a successful inoculation of dehydrated L. plantarum in wine depends not only on the use of protective agents, but also on the cell acclimation process prior to preservation, and on the ethanol content of wine.
In this work maltodextrins were added to commercial galacto-oligosaccharides (GOS) in a 1:1 ratio and their thermophysical characteristics were analyzed. GOS:MD solutions were then used as matrices ...during spray-drying of Lactobacillus plantarum CIDCA 83114. The obtained powders were equilibrated at different relative humidities (RH) and stored at 5 and 20°C for 12 weeks, or at 30°C for 6 weeks. The Tgs of GOS:MD matrices were about 20-30°C higher than those of GOS at RH within 11 and 52%. A linear relation between the spin-spin relaxation time (T2) and T-Tg parameter was observed for GOS:MD matrices equilibrated at 11, 22, 33, and 44% RH at 5, 20, and 30°C. Spray-drying of L. plantarum CIDCA 83114 in GOS:MD matrices allowed the recovery of 93% microorganisms. In contrast, only 64% microorganisms were recovered when no GOS were included in the dehydration medium. Survival of L. plantarum CIDCA 83114 during storage showed the best performance for bacteria stored at 5°C. In a further step, the slopes of the linear regressions provided information about the rate of microbial inactivation for each storage condition (k values). This information can be useful to calculate the shelf-life of spray-dried starters stored at different temperatures and RH. Using GOS:MD matrices as a dehydration medium enhanced the recovery of L. plantarum CIDCA 83114 after spray-drying. This strategy allowed for the first time the spray-drying stabilization of a potentially probiotic strain in the presence of GOS.
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