Fermentation characteristics and transit tolerance of probiotic Lactobacillus casei Zhang in soymilk and bovine milk during storage

Lactobacillus casei Zhang is a novel strain that was screened out of koumiss collected in Inner Mongolia, and our previous research showed that L. casei Zhang has health benefits such as cholesterol-reducing and immunomodulating effects. The fermentation characteristics of L. casei Zhang in soymilk and bovine milk and the transit tolerance of L. casei Zhang in fermented milk products during refrigerated storage for 28 d were assessed. A faster decrease in pH and faster growth of L. casei Zhang during fermentation were observed in soymilk compared with bovine milk at various inoculation rates, probably because of the low pH buffering capacity of soymilk. The fermented bovine milk samples had much higher final titratable acidity (TA) values (between 0.80 and 0.93%) than the soymilk samples (between 0.40 and 0.46%). Dramatic increases in TA values in the fermented soymilk samples during storage were observed, and the TA values of the fermented soymilk samples changed from <0.56% to values between 0.86 and 0.98%. On the other hand, only slight increases in TA were observed in the bovine milk samples during the 28 d of storage. The survival rates of freshly prepared cultures of L. casei Zhang in simulated gastric juice at pH 2.0 and 2.5 were 31 and 69%, respectively, and the delivery of L. casei Zhang through fermented soymilk and bovine milk significantly improved the viability of L. casei Zhang in simulated gastric transit. Lactobacillus casei Zhang showed good tolerance to simulated gastric juice and intestinal juice in the fermented soymilk and bovine milk samples, and maintained high viability (>10⁸ cfu/g) during storage at 4°C for 28 d. Our results indicated that both soymilk and bovine milk could serve as vehicles for delivery of probiotic L. casei Zhang, and further research is needed to elucidate the mechanism of the change in pH and TA of L. casei Zhang in fermented milk samples during fermentation and storage and to understand the difference between soy- and milk-based systems.     

The nutrient requirements of Lactobacillus acidophilus LA-5 and their application to fermented milk

Lactobacillus acidophilus LA-5 is a suitable probiotic for food application, but because of its slow growth in milk, an increase in its efficiency is desired. To shorten the time required for fermentation, the nutrient requirements of L. acidophilus LA-5 were analyzed, including the patterns of consumption of amino acids, purines, pyrimidines, vitamins, and metal ions. The nutrients required by L. acidophilus LA-5 were Asn, Asp, Cys, Leu, Met, riboflavin, guanine, uracil, and Mn²⁺, and when they were added to milk, the fermentation time of fermented milk prepared by L. acidophilus LA-5 alone was shortened by 9 h, with high viable cell counts that were maintained during storage of nutrient-supplemented fermented milk compared with the control. For fermented milk prepared by fermentation with Streptococcus thermophilus, Lactobacillus delbrueckii ssp. bulgaricus, and L. acidophilus LA-5, viable cell counts of L. acidophilus LA-5 increased 1.3-fold and were maintained during storage of nutrient-supplemented fermented milk compared with the control. Adding nutrients had no negative effect on the quality of the fermented milk. The results indicated that suitable nutrients enhanced the growth of L. acidophilus LA-5 and increased its viable cell counts in fermented milk prepared by L. acidophilus LA-5 alone and mixed starter culture, respectively.     

Competition mechanisms of lactic acid bacteria and bifidobacteria: Fermentative metabolism and colonization

Enzyme activities (α- and β-glucosidases, α- and β-galactosidases and β-fructofuranosidase) and organic acid production of four strains of lactic acid bacteria (LAB; Streptococcus thermophilus STY-31, Lactobacillus delbrueckii subsp. bulgaricus LBY-27, Lactobacillus casei LC-01 and Lactobacillus acidophilus LA-5) and Bifidobacterium lactis BB-12 were tested on milk and MRS fermentation broth with glucose, lactose or fructooligosaccharides (FOS) as carbon source. The highest β-galactosidase activity was found in L. acidophilus growing on milk. As compared to milk, α-glucosidase activity was increased with FOS by B. lactis, L. acidophilus and L. casei. The analysis of organic acids and short-chain fatty acids in the medium growth showed that lactate and acetate were the major fermentation metabolites produced by LAB and bifidobacteria, respectively. However, a metabolic shift towards more acetate and formate production, at the expense of lactate production, was observed during growth of L. casei on FOS. When grown on FOS as sole carbon source, L. acidophilus showed the highest production of lactate among the species tested. In addition, L. acidophilus demonstrated resistance to colonization against the intestinal pathogens Escherichia coli and Salmonella enterica in competition assays.     

Study on the Influence of Tea Extract on Probiotics in Skim Milk: From Probiotics Propagation to Metabolite

In this study, the influence of tea extract (TE) on the growth of probiotics in skim milk was examined. Lactobacillus plantarum ST‐III, Bifidobacterium bifidum Bb02, Lactobacillus acidophilus NCFM, and Lactobacillus rhamnosus GG were used in this study. The introduction of TE in milk significantly stimulated the propagation and acidification of L. rhamnosus GG and L. acidophilus NCFM. The antioxidant capacities and the total free amino acid contents of all fermented milk products were enhanced by the addition of TE; however, there were different antioxidant properties and free amino acid contents of fermented milk samples fermented by different bacteria. With a 9% (w/w) level, the fermentation with L. rhamnosus GG and L. acidophilus NCFM showed larger numbers of viable cells and faster acidifying rates, as well as excellent antioxidant capacity and abundant free amino acids.  The stimulative effects of TE on probiotics can be considered for industrial purposes and has practical implications for commercial applications.     

Amino acid catabolism by Lactococcus lactis during milk fermentation

Two wild-type Lactococcus lactis strains isolated from naturally Tunisian fermented milk (Leben), and one laboratory strain, were used to investigate the ability of L. lactis to transform amino acids into aroma compounds during milk fermentation. The α-ketoacid acceptor used for leucine transamination, the first step of catabolism, was identified by gas chromatography/mass spectrometry analysis of the ¹⁵N-labelled amino acids that formed from ¹⁵N-labelled leucine in fermented milk. Furthermore, the amino acids produced or catabolized by the laboratory strain via transamination were identified by comparing the free amino acids in milk fermented with the wild-type strain and the double mutant for aromatic and branched-chain aminotransferases, which cannot transaminate amino acids. The three L. lactis strains strongly catabolized leucine and valine during milk fermentation. The principal amino acid formed via leucine and valine transamination was glutamate indicating that α-ketoglutarate was the principal α-ketoacid acceptor and was generated during milk fermentation.     

Survival and activity of 5 probiotic lactobacilli strains in 2 types of flavored fermented milk

The viability of 5 probiotic lactobacilli strains (Lactobacillus acidophilus LA-5, Lactobacillus casei L01, Lactobacillus casei LAFTI L26, Lactobacillus paracasei Lcp37, and Lactobacillus rhamnosus HN001) was assessed in 2 types of probiotic flavored drink based on fermented milk during 21 days of refrigerated storage (5°C). Also, changes in biochemical parameters (pH, titrable acidity, and redox potential) during fermentation as well as the sensory attributes of final product were determined. Among the probiotic strains, L. casei LAFTI L26 exhibited the highest retention of viability during refrigerated storage period, while L. acidophilus LA-5 showed the highest loss of viability during this period. The decline in cell count of probiotic bacteria in strawberry fermented milk was significantly greater compared to peach fermented milk. In an overall approach, peach fermented milk containing L. casei LAFTI L26 was selected as the optimal treatment in this study in both aspects of viability and sensory accpeptibility.     

Lactobacillus casei starter culture improves vitamin content,increases acidity and decreases nitrite concentration during sauerkraut fermentation

To investigate the effects of Lactobacillus casei on Chinese sauerkraut fermentation, L. casei 11MZ‐5‐1 was inoculated into Chinese sauerkraut. Physicochemical indexes were measured, and the microbial dynamics during the fermentation were analysed by quantitative real‐time PCR (qRT‐PCR) and polymerase chain reaction‐denaturing gradient gel electrophoresis (PCR‐DGGE). The result showed that inoculation with L. casei 11MZ‐5‐1 lowered the pH of the fermentation system more rapidly than in the control model (CK). The content of vitamin C (V_C) (44.64 ± 2.12 mg kg^?1) was higher and nitrite (under 0.76 mg kg^?1) was lower than CK (P < 0.05). The numbers of 16S rRNA gene copies in the experimental model (BA) were lower than in CK at the end fermentation time. According to the PCR‐DGGE analyses, 22 and 17 specific bands were detected in CK and BA, respectively. L. paracasei and L. casei were predominant during the fermentation in BA. The relative abundance and diversity indices of bacteria in BA were 8.23 ± 0.25 and 2.01 ± 0.06, respectively, lower than in the CK (P < 0.05). So, the L. casei 11MZ‐5‐1 inoculations could effectively inhibit the microbial diversity in the fermentation system. The fermented cabbage with L. casei 11MZ‐5‐1 was more favourably estimated by consumers in terms of colour, crispness, sourness, aroma, bitterness, stink, stale flavour and overall acceptability.     

The biological activity of fermented milk produced by Lactobacillus casei ATCC 393 during cold storage

Lactobacillus casei ATCC 393 is an important probiotic strain widely known in dairy technology. However, its capability to produce bioactive peptides from milk proteins has not been studied. The viability of the Lb. casei ATCC 393 strain and some physicochemical properties in fermented milk throughout storage for 21 days at 4 °C was evaluated; biological activity, i.e., antioxidant, angiotensin converting enzyme inhibitory and anticancer activities of water soluble extract and its filtrate (< 2 kDa; F1) were determined. Lb. casei counts remained over 9 log cfu g⁻¹ during the storage period in fermented milk. These bioactivities were increased significantly (P < 0.01) during storage. F1 of fermented milk after three weeks of storage showed the highest bioactivity impact. De novo sequencing assay for peptide identification was applied to the mass spectrum of F1. The promising capability of Lb. casei ATCC 393 to release bioactive peptides from milk proteins was demonstrated.     

Characteristics of carbonated fermented milk and survival of probiotic bacteria

The carbonation of pasteurised milk was evaluated as a method for improving bacterial viability in fermented milk added with probiotic bacteria (Lactobacillus acidophilus and/or Bifidobacterium bifidum). The behaviour of microorganisms during fermentation and cold storage, and the biochemical and sensory properties of the products were assessed. In AT (Streptococcus thermophilus/L. acidophilus) and ABT (S. thermophilus/L. acidophilus/B. bifidum) products, the fermentation times to decrease the pH to 5 were significantly lowered when CO₂ or lactic acid was added to milk. The higher acidity levels of carbonated (as a result of production of carbonic acid) and lactic acidified samples enhanced growth and metabolic activity of the starter during fermentation and was the reason for this reduction in incubation time. Cell counts of S. thermophilus, L. acidophilus and B. bifidum gradually decreased through the cold storage of carbonated and non-acidified fermented milk, although the counts were always higher than 10⁶ viable cells g⁻¹. The CO₂ did not exert any influence on the viability of S. thermophilus and L. acidophilus in AT fermented milk stored at 4°C but the presence of B. bifidum and CO₂ in ABT-type products was associated with lower viability of L. acidophilus during the refrigerated storage. The higher acetate concentrations of ABT products made with non-acidified milk as compared with the carbonated products could have contributed to major survival of L. acidophilus in the former. The use of milk acidified with CO₂ had no detrimental effects on the sensory properties of ABT fermented milk. Therefore, we concluded that the carbonation of pasteurised milk prior to the starter addition could be satisfactorily used to reduce the manufacture time of fermented milk.     

Characterization of the angiotensin-converting enzyme inhibitory activity of fermented milks produced with Lactobacillus casei

Our study assayed angiotensin-converting enzyme (ACE) inhibitory activity and fermentation characteristics of 41 food-originated Lactobacillus casei strains in fermented milk production. Twenty-two of the tested strains produced fermented milks with a high ACE inhibitory activity of over 60%. Two strains (IMAU10408 and IMAU20411) expressing the highest ACE inhibitory activity were selected for further characterization. The heat stability (pasteurization at 63°C for 30 min, 75°C for 25 s, and 85°C for 20 s) and resistance to gastrointestinal proteases (pepsin, trypsinase, and sequential pepsin/trypsinase treatments) of the ACE inhibitory activity in the fermented milks produced with IMAU10408 and IMAU20411 were determined. Interestingly, such activity increased significantly after the heat or protease treatment. Because of the shorter milk coagulation time of L. casei IMAU20411 (vs. IMAU10408), it was selected for optimization experiments for ACE inhibitory activity production. Our results show that fermentation temperature of 37°C, inoculum density of 1 × 10⁶ cfu/g, and fermentation time of 12 h were optimal for maximizing ACE inhibitory activity. Finally, the metabolite profiles of L. casei IMAU20411 after 2 and 42 h of milk fermentation were analyzed by ultra-HPLC electron spray ionization coupled with time-of-flight mass spectrometry. Nine differential abundant metabolites were identified, and 2 of them showed a strong and positive correlation with fermented milk ACE inhibitory activity. To conclude, we have identified a novel ACE inhibitory L. casei strain, which has potential for use as a probiotic in fermented milk production.     

Probiotic viability and storage stability of yogurts and fermented milks prepared with several mixtures of lactic acid bacteria

Currently, the food industry wants to expand the range of probiotic yogurts but each probiotic bacteria offers different and specific health benefits. Little information exists on the influence of probiotic strains on physicochemical properties and sensory characteristics of yogurts and fermented milks. Six probiotic yogurts or fermented milks and 1 control yogurt were prepared, and we evaluated several physicochemical properties (pH, titratable acidity, texture, color, and syneresis), microbial viability of starter cultures (Lactobacillus delbrueckii ssp. bulgaricus and Streptococcus thermophilus) and probiotics (Lactobacillus acidophilus, Lactobacillus casei, and Lactobacillus reuteri) during fermentation and storage (35 d at 5°C), as well as sensory preference among them. Decreases in pH (0.17 to 0.50 units) and increases in titratable acidity (0.09 to 0.29%) were observed during storage. Only the yogurt with S. thermophilus, L. delbrueckii ssp. bulgaricus, and L. reuteri differed in firmness. No differences in adhesiveness were determined among the tested yogurts, fermented milks, and the control. Syneresis was in the range of 45 to 58%. No changes in color during storage were observed and no color differences were detected among the evaluated fermented milk products. Counts of S. thermophilus decreased from 1.8 to 3.5 log during storage. Counts of L. delbrueckii ssp. bulgaricus also decreased in probiotic yogurts and varied from 30 to 50% of initial population. Probiotic bacteria also lost viability throughout storage, although the 3 probiotic fermented milks maintained counts ≥10⁷ cfu/mL for 3 wk. Probiotic bacteria had variable viability in yogurts, maintaining counts of L. acidophilus ≥10⁷ cfu/mL for 35 d, of L. casei for 7 d, and of L. reuteri for 14 d. We found no significant sensory preference among the 6 probiotic yogurts and fermented milks or the control. However, the yogurt and fermented milk made with L. casei were better accepted. This study presents relevant information on physicochemical, sensory, and microbial properties of probiotic yogurts and fermented milks, which could guide the dairy industry in developing new probiotic products.     

Reduced biogenic amine contents in sauerkraut via addition of selected lactic acid bacteria

The contents of free amino acids and biogenic amines in spontaneously fermented sauerkraut, inoculated or not with specific lactic acid bacterium strains, were monitored throughout 45 d of storage. The strains tested were Lactobacillus plantarum 2142, Lactobacillus casei subsp. casei 2763 and Lactobacillus curvatus 2771. In both the control and the experiments, the total amino acid contents increased with time – and the predominant ones were aspartic acid, arginine and glutamic acid. However, upon inoculation with either strain, the total biogenic amine contents remained considerably lower than those of the control (especially in the cases of L. casei subsp. casei and L. curvatus); every single biogenic amine was actually below the 100 ppm-threshold. The dominant biogenic amines in the control were putrescine – and tyramine and histamine, to a lesser extent; the putrescine content was 10-fold lower if inoculation had taken place with either lactobacillus tested; and histamine and tyramine were essentially absent during storage, whereas they ranked above 200 ppm in the control by 45 d. Hence, an efficient food-grade biological tool was made available that constrains buildup of dangerous biogenic amines in fermented vegetables during storage.     

Survival of Lactobacillus casei and functional characteristics of reduced sugar red beetroot yoghurt with natural sugar substitutes

In this study, probiotic gel fermentation and functional characteristics of reduced sugar yoghurt with red beetroot bioactive components and steviol glycosides from Stevia rebaudiana extracts as natural source of noncaloric sweeteners were investigated. A significant increase in Lactobacillus casei viability and fermented gel network and a decrease in syneresis were determined in yoghurt sweetened with steviol glycosides and high content of red beetroot bioactive components such as total phenolics, anthocyanins and total antioxidant capacity (DPPH and FRAP). The survival of L. casei was maintained within probiotic and therapeutic levels (>9 log₁₀ CFU/g), due to the possible prebiotic effect of steviol glycosides and fermentable fibres and phenolic compounds of red beetroot during 28 days of cold storage of the probiotic yoghurts. Colour properties (L*, a*, b*, ΔE*, C*, h*) and such sensory properties as smoothness, sweetness astringency were significantly affected. The highest organic acid content was found in red beetroot enriched yoghurt with stevia, while the most dominant organic acids were butyric, lactic, malic and citric. As a result, it was determined that fermented milk products, which are supported by phytochemical and steviol glycoside interactions, have adequate nutritional effects, high probiotic viability and acceptable sensory properties     

Optimization of the Fermentation of Cantaloupe Juice by Lactobacillus casei NRRL B-442

This study was carried out to optimize the fermentation pH and temperature and also the fermentation time for development of a new probiotic fermented drink with cantaloupe juice. The fermented juice was subjected to a storage study during 42?days at 4?°C. The initial pH and temperature influenced the growth of Lactobacillus casei in cantaloupe juice. However, on the microbial viability only the effect of temperature was significant. Optimum conditions for a satisfactory growth and viability of L. casei in cantaloupe juice were as follows: pH?=?6.1 and temperature?=?31?°C. The fermentation time of 8 hours was selected as the optimal fermentation time to prepare the probiotic cantaloupe juice. The cell viability was 8.3 log CFU mL^?1 at the end of the fermentation. This level was kept over the 42?days of refrigerated storage. The consistent growth and viability of the probiotic microorganism in cantaloupe juice during fermentation and storage suggested that melon is a suitable vehicle for L. casei delivering. The fermented juice arises as a good alternative to the traditional dairy based products.     

Mesopic fermentation contributes more to the formation of important flavor compounds and increased growth of Lactobacillus casei Zhang than does high temperature during milk fermentation and storage

Probiotic fermented milk is more and more popular due to their positive health associated properties. However, fermentation temperature and other process conditions may affect the growth and metabolism of probiotic strains, thereby affecting quality of the final products. In this study, the growth behaviors and metabolomic profiles of yogurts induced by Lactobacillus casei Zhang at fermentation termination (FT) and d 10 of storage (S10d) under different fermentation temperatures at 37°C (low) and 42°C (high) were analyzed and compared using liquid chromatography-mass spectrometry (MS)- and gas chromatography-MS-based metabolomics approaches. At 37°C, the growth of L. casei Zhang at FT and S10d was significantly increased, and the potential relationship between riboflavin, starch, and sucrose metabolism and growth of L. casei Zhang may be mutually promoting. Fermentation temperature (37°C and 42°C) affected volatile and nonvolatile metabolomic profiles and pathways. The levels of acetaldehyde, 2,3-butanedione, acetoin, butyric acid, decanoic acid, hexanoic acid, and octanoic acid were significantly higher at 37°C than at 42°C at FT and S10d. This indicates that the low temperature (37°C) most likely contributes more to the formation of important flavor compounds during the fermentation process and production of short-chain fatty acids during storage.     

Effect of alsD deletion and overexpression of nox and alsS on diacetyl and acetoin production by Lacticaseibacillus casei during milk fermentation

Diacetyl and acetoin are key aroma components of fermented milk but are produced in low concentrations by starter cultures. In this study, we expressed NADH oxidase, acetolactate synthase, and inactivated acetolactate decarboxylase in Lacticaseibacillus casei TCS to generate recombinant L. casei strains, and investigated the effects of the genes encoding these enzymes on diacetyl and acetoin production during milk fermentation. In the single-gene recombinant strains tested, diacetyl concentrations were highest in milk fermented by L. casei TCSI-nox (nox gene overexpressed, 3.68 mg/kg), whereas acetoin concentrations were highest in milk fermented by L. casei TCS-ΔalsD (alsD gene deleted, 32.94 mg/kg). Moreover, diacetyl and acetoin concentrations were higher in the inducible strains than in the corresponding constitutive strains (e.g., TCSI-nox vs. TCSC-nox, and TCSI-ΔalsD-nox vs. TCSC-ΔalsD-nox). This phenomenon was also reflected in the protein expression levels and enzyme activities. In the double-gene recombinant strains tested, the highest concentrations of diacetyl and acetoin were produced by L. casei TCSI-ΔalsD-nox (nox overexpressed and alsD deleted, 4.66 mg/kg, 69.62 mg/kg, respectively). The triple-gene recombinant L. casei TCS-ΔalsD-nox-alsS produced the highest concentrations of diacetyl and acetoin, which were 2.38 and 11.19 times, respectively, the concentrations produced by the original strain. These results show that the nox, alsS, and alsD genes make key contributions to the biosynthesis of diacetyl and acetoin by L. casei. The modification of multiple genes had a synergistic effect, leading to greatly increased synthesis of diacetyl and acetoin by L. casei during its fermentation of milk.     

Potato juice fermented with Lactobacillus casei as a probiotic functional beverage

This study was conducted to evaluate probiotic potato juice as a potential substrate for the production of Lactobacillus casei, and the change in the functionality of potato juice was monitored during fermentation. L. casei grew well in potato juice without nutrient supplementation, and lactic acid bacteria of fermented ‘Haryoung’ juice reached 1.7×10⁹ CFU/mL after a 48 h fermentation. DPPH radical scavenging activities of the potato juices decreased after a 72 h fermentation, but fermented colored potato juice still maintained >50% radical scavenging activity. The survival rate of L. casei fermented in’ Haryoung’ juice was 89.0% after exposure to an acidic condition, and L. casei in all fermented potato juice samples showed the ability (50–85%) to survive in the presence of bile. These results suggest that fermented potato juice might serve as a probiotic functional beverage for vegetarians or consumers who are allergic to dairy products.     

Probiotic beverage from cashew apple juice fermented with Lactobacillus casei

This study optimized the conditions of Lactobacillus casei NRRL B-442 cultivation in cashew apple juice, as well as, determined the proper inoculum amount and fermentation time. Moreover, it was investigated the survivability ability of L. casei in cashew apple juice during refrigerated storage (4 °C) for 42 days. The optimum conditions for probiotic cashew apple juice production were initial pH 6.4, fermentation temperature of 30 °C, inoculation level of 7.48 Log CFU/mL (L. casei) and 16 h of fermentation process. It was observed that the L. casei grew during the refrigerated storage. Viable cell counts were higher than 8.00 Log CFU/mL throughout the storage period (42 days). The values of lightness, yellowness and total color change increased and the values of redness reduced along the fermentation and refrigerated storage periods. The fermented juice with L. casei is a good and healthy alternative functional food containing probiotics. Cashew apple juice showed to be as efficient as dairy products for L. casei growth.     

Growth and viability of Lactobacillus acidophilus NRRL B-4495, Lactobacillus casei NRRL B-1922 and Lactobacillus plantarum NRRL B-4496 in milk supplemented with cysteine,ascorbic acid and tocopherols

The effect of three reducing agents (RAs), l-cysteine, ascorbic acid, and tocopherols, on the growth of Lactobacillus acidophilus, Lactobacillus casei, or Lactobacillus plantarum during milk fermentation was evaluated. pH, redox potential, and Lactobacillus counts were determined until pH ≈ 4.6. Further, the study aimed to optimise the concentration of the RAs by formulating the fermented milks with the same RA at different concentrations (0–250 mg L⁻¹ for l-cysteine or ascorbic acid and 0–15 mg L⁻¹ for tocopherols) using a Box–Behnken experimental design. After 45 days of refrigerated storage, the viability of each Lactobacillus species was maximised. We observed that the effect of RA on Lactobacillus is species dependent; ascorbic acid and tocopherols reduced the fermentation time (29%–43%), whereas l-cysteine enhanced the Lactobacillus counts (≥1 log₁₀ cfu mL⁻¹). Lactobacillus species differ in terms of oxygen sensitivity.     

A new synbiotic dairy food containing lactobionic acid and Lactobacillus casei

As well as its beneficial health properties, the incorporation of the prebiotic lactobionic acid (LBA) in fermented dairy products can provide a technological advantage due to its gelling capacities. This study aimed to develop a new functional dairy product containing LBA synthesised within the process itself by the bacterium Pseudomonas taetrolens. A probiotic Lactobacillus casei strain was introduced through a sequential fermentation system. After incubation, in the case of the most effective experimental procedure, a synbiotic fermented milk with 30 g/L of prebiotic LBA was obtained, together with an active L. casei population of 10⁹ cfu/mL and <1% lactose content.