The production of malolactic starter cultures requires the obtention of suitably large biomass at low‐cost. In this work it was possible to obtain a good amount of biomass, at laboratory scale, of ...two enological strains of Lb. plantarum, by formulating a culture medium based on whey permeate (WP), a by‐product of the cheese industry usually disposed as waste, when this was supplemented with yeast extract (Y), salts (S) and Tween 80 (T) (WPYST). Bacteria grown in WPYST medium exhibited good tolerance to stress conditions of synthetic wine (pH 3.5, ethanol 13% vol/vol). However, when WPYST was added with 8% vol/vol ethanol, cultures inoculated in synthetic wine, showed a lower viability and capacity to consume L‐malic acid than when they were cultured in WPYST without ethanol. Subsequently, strains grown in WPYST were inoculated in sterile wine samples (final stage of alcoholic fermentation) of the red varietals Merlot and Pinot noir, and incubated at laboratory scale. Cultures from WPYST, inoculated in Pinot noir wine, showed a better performance than bacteria grown in MRS broth, and exhibited a consumption of L‐malic acid higher than 90%. However, cultures from WPYST or from MRS broth, inoculated in sterile Merlot wine, showed a lower survival. This study allowed the formulation of a low‐cost culture medium, based on a by‐product of the food industry, which showed to be adequate for the growth of two enological strains of Lb. plantarum, suggesting their potentiality for application in the elaboration of malolactic starter cultures.
The ability of Patagonian L. plantarum and O. oeni strains to change the volatile profile of a sterile Pinot noir wine was studied through fermentation assays, at laboratory scale. Two strains of ...each LAB species were selected based on data regarding to their ability to survive in wine and to consume l-malic acid. Both O. oeni strains but only one L. plantarum (UNQLp 11) strain were able to remain viable, consuming l-malic acid through the fermentation assay with a concomitant increase of l-lactic acid. The volatile profile of Pinot noir wine, before and after LAB inoculation, was measured by using HS-SPME gas chromatography technique. This analysis showed that alcohols were the main volatile compounds after alcoholic fermentation and that after fermentation with the selected LAB strains, a decrease in the volatile alcohols concentration and an increase in the volatile esters concentration could be observed. The O. oeni UNQOe 73.2 strain produced the most notable change in the volatile profile, with the production of some important odorant esters at higher concentration, compared to O. oeni UNQOe 31b strain. Although, L. plantarum UNQLp 11 strain showed a better performance in the consumption of l-malic acid, this strain had a low capacity to modify the volatile compounds profile after incubation in red wine. The results found in the present work showed that different strains selected as potential malolactic starters could have different behavior when are incubated in real wine. Although L. plantarum UNQLp 11 strain showed a good consumption of l-malic acid, the O. oeni UNQOe 73.2 strain exhibited superior capacity to improve the flavor of wine due to its esterase activity that produce an increase of fruity and creamy odorants.
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•Changes in volatile profile of inoculated Pinot noir wine was studied.•Selected L. plantarum and O. oeni strains were used in the vinification assays.•Inoculation diminished alcohols concentration and increased esters concentration.•All strains assayed were able to modify the wine volatile profile.•O. oeni strain UNQOe 73.2 produced the most notable change in wine volatile profile.
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.
Several lactic acid bacteria (LAB) species have been recognized as probiotics and are of considerable interest due to their potential ability to confer health benefits upon consumption. In the animal ...feed sector, probiotics offer an alternative to the use of antibiotic growth promoters. The preservation and incorporation of probiotics into dry products requires carefully meeting several criteria and overcoming technological challenges to maintain their functionality. Drying is a crucial step in the process, but the probiotic properties of the resulting powder and the final cell viability in the food product are significantly influenced by the type of protective compounds and drying techniques employed. In light of the growing demand for functional animal products, this review focuses on the damages incurred during microorganism dehydration processes for food incorporation, and explores strategies to minimize such damages. It provides an overview of the effects of probiotic products in the animal feed industry, including their incorporation in low-moisture food matrices and key considerations for success. Additionally, it highlights postbiotics as an attractive alternative for live probiotic cells with many technological advantages.
The performance of Patagonian Lactobacillus plantarum and Oenococcus oeni strains as malolactic starter cultures was compared. Two autochthonous strains of each species were selected, based on the ...presence of aroma-related genes, and inoculated in sterile wine of high ethanol content. The effects of initial inoculum size and acclimation treatment on the efficiency of malolactic fermentation (MLF) were analyzed for each strain. O. oeni strains were able to successfully conduct the MLF only when the inoculum concentration was higher than 1.108 CFU/mL and cells were acclimated in sublethal ethanol concentrations. The increase of ethanol concentration in the acclimation medium also improved the kinetics of malic acid consumption. Successful MLF with L. plantarum strains required lower inocula and no acclimation treatment. In addition, these strains showed a better profile of aroma-related genes than O. oeni. L. plantarum strains appeared to be more efficient than O. oeni strains as candidates for malolactic starter cultures to be used in Patagonian red wines.
•L. plantarum and O. oeni strains were isolated and fermentation examined.•L. plantarum was more efficient at MLF than O. oeni in Patagonian wines.•Standardization of acclimation and inoculum was necessary for successful MLF by O. oeni.•Acclimation in high ethanol content improved the malic acid consumption of O. oeni.•MLF with L. plantarum strains required lower inocula and no pre-acclimation treatment.
The aim of this work was to obtain freeze-dried biomass of the native Patagonian
Lactiplantibacillus plantarum
strain UNQLp 11 from a whey permeate (WP)-based medium and compare it with the growth in ...commercial MRS broth medium. Survival and activity of the freeze-dried
Lb. plantarum
strain were investigated after inoculation in wine as a starter culture for malolactic fermentation (MLF). The effect of storage and rehydration condition of the dried bacteria and the nutrient supplementation of wine were also studied. The freeze-dried cultures from WP and those grown in MRS showed similar survival results. Rehydration in MRS broth for 24 h and the addition of a rehydration medium to wine as nutrient supplementation improved the survival under wine harsh conditions and guaranteed the success of MLF. Storage at 4 °C under vacuum was the best option, maintaining high cell viability for at least 56 days, with malic acid consumption higher than 90% after 7 days of inoculation in a wine-like medium. These results represent a significant advance for sustainable production of dried malolactic starter cultures in an environmentally friendly process, which is low cost and easy to apply in winemaking under harsh physicochemical conditions.
Graphic Abstract
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.
The aim of this study was to evaluate fifty-three
Lactobacillus plantarum
isolates obtained from a Patagonian red wine, molecularly identified and typified using RAPD analysis, in order to select ...starter cultures for malolactic fermentation (MLF). The results obtained suggest a considerable genetic diversity, taking into account that all
L. plantarum
isolates were obtained from one cellar and one vintage. Based on the capacity to tolerate a concentration of 14 % ethanol in MRS broth for 2 days, eight isolates were selected for the subsequent analysis. The incidence of various wine stress factors (ethanol, acid pH, lysozyme and sulfur dioxide) on isolates growth was studied. Besides, glucosidase and tannase activities were evaluated, and the presence of genes involved in the synthesis of biogenic amines was examined by PCR. A previously characterized indigenous
Oenococcus oeni
strain was included with comparative purposes. Differences in technologically relevant characteristics were observed among the eight
L. plantarum
selected isolates, revealing an isolate-dependent behavior. Detectable glucosidase and tannase activities were found in all isolates. The presence of genes encoding histidine and tyrosine descarboxylases and putrescine carbamoyltransferase was not detected. The ability of
L. plantarum
isolates to grow and consume
l
-malic acid in simulated laboratory-scale vinifications revealed that two of them could be considered as possible MLF starter cultures for Patagonian red wines. These isolates will be subjected to further analysis, for a final winery technological characterization.
Spontaneous malolactic fermentation (MLF) in the North Patagonian region of Argentina may have to bear low environmental temperatures, an important stress factor for the lactic acid bacteria (LAB) ...that conduct this process. Thus, inducing MLF by inoculation of a native starter culture able to lead this biological process at low temperatures is encouraged because it would allow preserving the regional character of the wine. The goal of this work was to obtain autochthonous Oenococcus oeni strains able to consume l-malic acid when MLF takes place at temperatures lower than 15 °C. Among the psychrotrophic isolates obtained from a Pinot noir wine, sixteen O. oeni strains were identified by sequencing the 16S rRNA gene and typing by RAPD PCR, allowing the differentiation of five genotypic groups. After the evaluation of the viability and metabolic performance of the strains in sterile wine at low temperatures, strains UNQOe4 and UNQOe19 were selected to evaluate their behavior in a non-sterile wine at 4 and 10 °C. The strains selected showed good implantation capacity and were able to successfully conduct MLF, suggesting that their use as starter cultures is suitable to improve this biological process in a wine region where environmental temperatures can be low.
•O. oeni strains were isolated at low temperature from a Patagonian Pinot noir wine.•Psychrotrophic O. oeni were able to consume l-malic acid in sterile wine.•O. oeni strains were able to implant and induce MLF in wine at low temperatures.•Selected psychrotrophic O. oeni strains could be used as starter cultures of MLF.
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.
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