Lyophilization of Probiotics Preserving Viability and Extending Shelf Life

Electrospray drying of anaerobic bacteria into a stable powder for animal feed applications, using low heat and controlled atmosphere conditions to preserve viability and stability of the microorganisms. The method involves preparing a bacterial culture, forming a slurry with a carrier, applying an electrostatic charge, atomizing the slurry, and drying the droplets in a controlled atmosphere chamber. The resulting powder contains encapsulated bacteria with less than 15% moisture content, suitable for use in animal feed to prevent lactic acidosis and promote digestive health.

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A spray-dried composition for delivering probiotics in food products, comprising a prebiotic, a probiotic, and a coating material. The composition is prepared by spray drying a solution containing the prebiotic, probiotic, and coating material, and can be tailored to various food matrices. The composition exhibits improved probiotic viability and stability compared to conventional drying methods, enabling the delivery of live microbes in adequate amounts to exert a functional effect within the body.

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A water-soluble, lyophilized microbial soil amendment that enhances plant growth and soil health through a diverse population of beneficial microorganisms. The amendment is produced by combining brewer's yeast and processed honey as a nutrient-rich food source, which supports the growth of a broad spectrum of beneficial microbes. The yeast and honey mixture is formulated to maintain a stable pH environment that promotes balanced microbial growth, and is then lyophilized to create a powder that is 100% soluble in water. The resulting product can be easily rehydrated and applied to soil using conventional agricultural sprayers.

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A system for deploying microorganisms to a target environment comprises a plurality of coated micromagmatics formed by adhering a microorganism to a glass microparticle. The system can be deployed in various ways, including as an aerosol, liquid suspension, or solid matrix. The micromagmatics can be lyophilized and stored for later use, and can be engineered for specific applications such as subsurface deployment or wastewater treatment. The system enables efficient and easy deployment of beneficial microorganisms for bioremediation, agriculture, and other applications.

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Methods for preparing bacterial cultures that improve their robustness against downstream processing steps such as freezing and drying. The methods involve adjusting the pH of the bacterial culture to a range of 6.0-8.0 after fermentation termination, rather than maintaining the acidic pH typical of fermentation. This pH adjustment enables the bacteria to withstand freezing and drying processes more effectively, resulting in improved viability and activity retention.

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Methods for cryoprotecting and lyoprotecting cells, such as mesenchymal stem cells, for long-term storage. The methods involve suspending cells in a composition containing urea, a monosaccharide, and an aqueous component, and then freezing or lyophilizing the composition. The composition can also include a bulking agent, such as sucrose and mannitol, and a hydrogel, such as hyaluronan. The frozen or lyophilized cells can be reconstituted in a reconstitution agent to form a viable cell population.

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Probiotics, such as

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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

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Shelf-stable, heat-treated beverage containing encapsulated probiotics that can be stored at ambient temperatures for extended periods without spoilage. The beverage contains microparticles with live probiotics encapsulated within. The microparticles are made by coating a core of sub-microparticles containing the probiotics with denatured protein using a fluidized bed process. This prevents leakage and degradation of the probiotics during heat treatment and storage. The encapsulated probiotics survive UHT processing and maintain viability for 24 months at room temperature.

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A method for producing a freeze-dried probiotic composition comprising Streptococcus salivarius, particularly S. salivarius K12, ENT-K12, M18, and M18DF, through large-scale fermentation in a controlled environment. The method involves cultivating the bacteria in a growth medium containing dextrose, yeast extract, and ascorbate, with controlled pH and oxygen levels, followed by concentration, lyoprotectant addition, and freeze-drying to produce a stable powder. The composition is suitable for use in various probiotic products, including oral care and dietary supplements.

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Compositions for preserving anaerobic microorganisms, including cryopreservatives, comprising a viscosity modifier, an energy source, and a redox potential in the range of -50 mV to -500 mV. The compositions maintain viability and facilitate rapid recovery of anaerobic microorganisms during storage and administration, enabling effective modulation of the gastrointestinal microbiome.

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Probiotics have become increasingly recognized for their potential health-promoting properties; however, the viability of probiotics can be affected by storage and transportation processes as well as the stressful environment of the human digestive tract, preventing them from achieving effective concentration (10

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Microencapsulated microbial cultures with enhanced storage stability at elevated temperatures, comprising a microbial culture entrapped in a coacervate comprising a non-homogeneous encapsulation matrix with a high ratio of matrix material to core material, wherein the matrix material includes carbohydrates, proteins, and antioxidants. The microencapsulated cultures exhibit preserved viability over extended periods of storage at temperatures up to 37°C, enabling applications in products where refrigerated storage is not feasible.

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A probiotic granule comprising a core of probiotic bacteria coated with a single continuous layer of a hydrophobic solid dispersion containing a water-soluble polymeric stress absorber. The stress absorber is dispersed within a hydrophobic solid component such as fat, wax, or fatty acid, and provides mechanical protection and controlled dissolution of the granule. The granule enables prolonged survival of the probiotics during storage and passage through the gastrointestinal tract, and can be used in a variety of food products.

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A probiotic microcapsule preparation method that produces uniform microcapsules with controlled particle size, high encapsulation efficiency, and resistance to gastric acids and high temperatures. The method involves coating probiotic bacteria with a hydroxypropyl methylcellulose solution containing a coating material, followed by lyophilization. The resulting microcapsules exhibit improved survival ratios and stability compared to conventional methods.

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Probiotics, comprising living microorganisms like Lactobacillus fermentum MT308789 and Lactobacillus oris MT308790, offer significant health benefits when consumed regularly. However, the efficacy of probiotics heavily relies on their viability and stability during storage. Various storage methods, including agar slant, glycerol stocks, lyophilization, and dry form, are utilized to preserve probiotic viability. In this study, we evaluated the efficacy of these storage methods in maintaining the viability of probiotic strains. Our findings demonstrate that lyophilization emerged as the most effective method, yielding the highest viabilities for both strains [77.51 %]. Glycerol stocks also showed promise for short-to-medium-term storage, while agar slants and dry form storage exhibited suboptimal viability. These results underscore the importance of selecting appropriate storage methods to ensure the delivery of viable and effective probiotic formulations to consumers.

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The selection of appropriate probiotic strains is vital for their successful inclusion in foods. These strains must withstand processing to reach consumers with 10

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A dry composition of lactic acid bacteria (LAB) with enhanced storage stability, comprising LAB cells and a synergistic stabilizer blend of oligofructans, maltodextrin, inulin, and/or pea fiber. The stabilizer blend provides cryoprotection, lyoprotection, and storage stability to the LAB cells, enabling long-term viability and shelf life. The composition is suitable for use in infant formulas, dietary supplements, and other food products.

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Encapsulation of microbial cultures, such as lactic acid bacteria, in a fat matrix to improve their stability and viability during storage and processing. The encapsulated cultures retain viability through pasteurization and subsequent storage at ambient temperature, enabling their direct addition to dairy products without refrigeration. The encapsulation matrix comprises one or more fat components with a melting point of at least 30°C, which protects the cells from heat and prevents post-acidification during storage.

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A solid, porous glass-based substrate for storing and delivering microorganisms, particularly beneficial bacteria, in a lyophilized biofilm format. The substrate, which can be a natural or synthetic foamed glass or glass ceramic, supports the growth of microorganisms into a dense biofilm that can be lyophilized and stored for later deployment. The lyophilized biofilm can be easily transported and applied to a site of use, where it can regrow and provide beneficial effects such as bioremediation.

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Lyophilization is a crucial procedure for maintaining probiotic viability. This study aimed to screen the optimal protective agent for lyophilization and explore its potential protective mechanism on Lactiplantibacillus plantarum L5, which has a sleep-promoting function. The results showed that sodium L-glutamate is the optimal protective agent, with a survival rate of over 90%. After storage at 4C for 6 months, the viable bacteria count remained stable at 4.801011 CFU/g. Under the protection of sodium L-glutamate, the lyophilized powder maintained a gastric juice tolerance rate of over 90%, and the auto-agglutination rate and acid-producing activity were similar to those before lyophilization. Scanning electron microscope and fourier infrared spectroscopy observations showed that the cellular structure of the lyophilized powder remained relatively stable. Finally, we found that the pyruvate content eventually accumulated through transcriptomic analysis, which may account for the high survival rate of L. plantarum L5 during the lyophilization process.

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Two lactic acid bacteria strains with probiotic potential (Lactiplantibacillus plantarum MIUG BL21 and Lactiplantibacillus paraplantarum MIUG BL74) were used to produce paraprobiotics and postbiotics, respectively. The paraprobiotics were obtained by inactivation of Lactiplantibacillus plantarum MIUG BL21 strain by using a combined ohmic-thermal treatments. The postbiotics were produced by Lactiplantibacillus paraplantarum MIUG BL74 by fermentation of a bovine colostrum-based medium. Paraprobiotics were added before fermentation, whereas the fermented medium was used to develop four customized freeze-dried biotics formulas. The freeze-dried biotics showed a probiotics survival rate varying between 75% and 84%, whereas different polyphenolic profile was highlighted, with the highest total content of 8843 g/g dry weight (DW) in the variant based on whey protein isolate and -cyclodextrin. The biotics highlighted a wide range of free and bounded phenolic profile, with theaflavin as the main representative in samples containing whey protein isolate and carboxymethylcellulose (3841.11 ... Read More

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Viability loss of probiotics often occur during processing, storage and gastrointestinal transit. In this study, the viability of freeze-dried

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Bacteria get stressed and damaged during freeze-drying process for commercialization and this result in loss of its effect. Viability is important for its efficacy, but this drying process can deteriorate viability by damaging the integrity of the cell membrane as well. In this study, we propose 0.03 M histidine for rehydration of freeze-dried probiotics to improve their viability. The freeze-dried bacteria mixture with 0.03 M histidine showed augmented survivability during in vitro simulated gastric and duodenum stress conditions and increased viability during 60 min rehydration. It exhibited a significantly increased adherence ability of lyophilized bacteria to the HT-29 cell-line. Therefore, this shows possibility of probiotics commercialization with damage of lyophilization restored and survivability ameliorated.

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Abstract In recent years, more and more scientific community, food producers, and food industry show increased interest in functional foods containing probiotics, which is a big challenge. The consumption of probiotics in the context of a balanced diet through the consumption of functional foods or through the intake of pharmaceutical preparations has proven to contribute to the improvement of human health, even contributing to the prevention of diseases. In order for probiotics to be considered suitable for consumption, they must contain a minimum concentration of viable cells, namely, at least 10 7 colony forming units of beneficial microbes per gram. Ensuring the viability of bacterial cells until the moment of consumption is the overriding priority of functional probiotic food manufacturers. Probiotic bacteria are subject to stress conditions not only during food manufacturing but also during gastrointestinal passage, which limit or even compromise their functionality. This paper first examines all the stressful conditions faced by probiotic cells in their production stages and r... Read More

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The intake of probiotics offers various health benefits; however, their efficacy depends on the maintenance of viability during industrial processing and digestion. Probiotic viability can be compromised during encapsulation, freeze-drying, storage, and digestion, necessitating multiple coatings. This complicates production and raises costs. In this study, CaCO

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A method for lyophilizing pharmaceutical substances that maintains product integrity and stability through a controlled lyophilization process. The method involves optimizing primary drying conditions by dynamically adjusting shelf temperature and chamber pressure based on real-time product temperature measurements, rather than maintaining constant process parameters. This approach enables efficient removal of water vapor while preventing material collapse and degradation. The method also includes a secondary drying step to achieve the desired moisture level.

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These experimental data are concerning the optimization of freeze-drying cycles of a model-type Casei probiotic bacteria with water based formulations using lactose and polymer polyvinylpyrrolidone (PVP) as cryo-/lyo-protectants. After a previous study devoted to the influence of the main freeze-drying parameters (freezing rates; shelf temperature; absolute pressure) on the bacteria survival ratios, this paper reports some morphological data obtained by scanning electron microscopy (SEM) with the final lyophilisates. The numerous and original SEM images realized with different formulations of water binary or ternary mixtures with fixed operating conditions proved to be very useful to explain and to understand the different bacteria survival ratios observed. Thus, with bacteria or micro-organisms systems, this powerful investigation method of characterization of lyophilisates morphology should be more largely used in the future for setting up the optimal formulations parameters (composition) and the connected operating physical parameters of freeze-drying steps (freezing rate, shelf t... Read More

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Freeze drying is a commonly used method for preserving probiotic bacteria and live biotherapeutic products. Before drying, the bacterial cells are formulated with a lyoprotectant, and the design of these two process steps are crucial to achieve a high-quality product. There are several factors that may affect the biological and physicochemical properties of the freeze-dried cells and we have used a Design of Experiment approach to investigate the effects of formulation and freeze-drying parameters on properties and performance of Limosilactobacillus reuteri R2LC. The biological characteristics of the dried bacteria were evaluated by measuring cell survival, metabolic activity and stability, and physicochemical characteristics were studied using visual inspection, differential scanning calorimetry (DSC), scanning electron microscopy (SEM), and analysis of residual moisture content and bacterial aggregation. A comparison between the lyoprotectants trehalose and sucrose showed that the latter gave better freeze-drying survival, metabolic activity, and storage stability. We also want to ... Read More

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Lactic acid bacteria are very important for biotechnology, pharmaceutical and food industry, and are extensively used as probiotic products and starter cultures. Freeze-drying is a widely used and effective process for preservation of lactic acid bacteria in a dried form. To ensure stability and maintain biological activity of lactic acid bacteria, suitable protective media formulations should be used during the freeze-drying process. Twelve oral strains with functional and probiotic properties were subjected to freeze-drying and assessed for viability after the process and after long-term storage. Two lyoprotective media with different composition were used and the freeze-drying process was performed, where retained stability of the studied strains in both lyoprotectors was reported. Viability after four and eight months of storage was assessed, showing that the strains maintained high viability. The two lyoprotective media ensured the obtaining of freeze-dried samples with preserved viability for all studied strains of lactic acid bacteria. Four of the tested strains, L. fermentum ... Read More

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Probiotics have demonstrated oral health benefits by influencing the microbiome and the host. Although promising, their current use is potentially constrained by several restrictions. One such limiting factor lies in the prevailing preparation of a probiotic product. To commercialize the probiotic, a shelf stable product is achieved by temporarily inactivating the live probiotic through drying or freeze drying. Even though a lyophilized probiotic can be kept dormant for an extended period of time, their viability can be severely compromised, making their designation as probiotics questionable. Additionally, does the application of an inactive probiotic directly into the oral cavity make sense? While the dormancy may allow for survival on its way towards the gut, does it affect their capacity for oral colonisation? To evaluate this, 21 probiotic product for oral health were analysed for the number of viable (probiotic), culturable (CFU) and dead (postbiotic) cells, to verify whether the commercial products indeed contain what they proclaim. After isolating and uniformly lyophilizing t... Read More

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A method for preparing high-performance powders through a combination of spray drying, liquid nitrogen quick-freezing, and vacuum freeze-drying. The process enables the precise control of particle size and density, while maintaining the activity and viability of microorganisms like probiotics. The method employs a unique sequence of freeze-drying steps, where the microorganisms are first rapidly cooled to form a slurry, followed by liquid nitrogen quick-freezing to rapidly freeze the slurry, and finally vacuum freeze-drying to remove the remaining water content while preserving the microorganism's structural integrity. This approach enables the production of uniform, freeze-dried powders with controlled particle size and density, making them ideal for applications requiring precise powder characteristics.

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A probiotic preparation method using a double emulsion structure to improve stability and viability of probiotics in foods, beverages, supplements, etc. The method involves dispersing probiotics in a water phase, forming a primary water-in-oil emulsion, and then encapsulating it in a second water phase to create a double emulsion. This protects the probiotics from environmental factors and enables better storage and digestion stability compared to free probiotic solutions.

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Lyophilized formulations of live attenuated Bordetella bacteria, such as BPZE1, that exhibit stability for at least two years when stored at temperatures between -20° and 22.5° C. The formulations are made by harvesting bacteria at an optimal optical density, mixing with a lyophilization buffer containing a cryoprotectant sugar, and lyophilizing the mixture within a 48-hour hold time. The lyophilized formulations exhibit sufficient homogeneity, viability, and potency to be used as a live vaccine.

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A process for lyophilizing bacterial products, particularly anaerobic bacteria, that minimizes cell loss during the process. The process involves freezing the bacterial product under anaerobic conditions, followed by sublimation and secondary drying steps. The anaerobic freezing step is critical in preserving cell viability, as it prevents oxygen-induced inactivation of the bacteria. The process enables efficient and economic lyophilization of bacterial products, including live organisms, without significant loss of viable cells.

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Bifidobacterium breve is a common habitant of the human gut and is used as probiotic in functional foods. B. breve has to cope with multiple stress conditions encountered during processing and passage through the human gut, including high temperature, low pH and exposure to oxygen. Additionally, during industrial processing and in the gut, B. breve could encounter nutrient limitation resulting in reduced growth rates that can trigger adaptive stress responses. For this reason, it is important to develop culture methods that elicit resistance to multiple stresses (robustness) encountered by the bacteria. To investigate the impact of caloric restriction on robustness of the probiotic B. breve NRBB57, this strain was grown in lactose-limited chemostat cultures and in retentostat for 21 days, at growth rates ranging from 0.4 h-1 to 0.00081 h-1. Proteomes of cells harvested at different growth rates were correlated to acid, hydrogen peroxide and heat stress survival capacity. Comparative proteome analysis showed that retentostat-grown cells had significantly increased abundance of a varie... Read More

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In order to expand the range of autoprobiotic release forms, increase consumer attractiveness, prolongate products shelf life and transportation possibilities, a protocol for obtaining an encapsulated form of autoprobiotics was developed. Tests were carried out on encapsulated forms of indigenous bacteria in order to assess the viability of bacterial cells after lyophilization and storage under various conditions. The effect of various cryoprotectants on the lyophilized Lactobacillus spp. and Enterococcus spp. cells viability was studied. The results obtained showed that the use of 10 % sucrose and 1 % gelatin as part of a protective medium retains the original properties of bacterial strains, contributing to their survival during cryopreservation and lyophilization. The most favorable storage conditions and expiration dates for encapsulated autoprobiotics based on Lactobacillus spp. and Enterococcus spp . were determined.

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A method for preparing lyophilized cell preparations, particularly probiotic bacteria, by incorporating a cryogenic freezing step under pressure. The method involves dissolving gases in the cell composition before freezing, which is achieved through a dense gas zone generated by evaporation of a cryogenic fluid, increased pressure, or a combination of both. The resulting frozen composition is then lyophilized to produce a stable dry powder. This method enables improved cell viability and membrane integrity in lyophilized preparations, particularly for fragile strains, and enables long-term storage at ambient temperature.

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Designing stable dried functional food ingredients and foods containing live probiotic cultures maintaining high viable cell loads at the time of consumption is a challenge for the industry. The aim of the present study was the development of stable freeze-dried functional food ingredients with enhanced shelf-life during long storage. Zea flakes, pistachios, and raisins were used as immobilization supports for the wild-type presumptive probiotic strains Pediococcus acidilactici SK and Lactiplantibacillus plantarum F4, while L. plantarum B282 was used as a reference strain. Cell survival was monitored during storage at room and refrigerated temperatures for up to 6 months. Levels of freeze-dried cultures were maintained up to 7.2 logcfu/g after 6 months storage at room temperature and up to 8.5 logcfu/g at refrigerator temperature, in contrast to free cell levels that ranged <7 logcfu/mL, suggesting the positive effects of immobilization and freeze-drying on cell viability. Of note, levels of freeze-dried immobilized P. acidilactici SK cells on zea flakes and pistachios remained st... Read More

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In recent years, probiotics have expanded from their traditional classification as health promoting food to the development of live biotherapeutic products (LBP). Traditional probiotics are marketed as food/dietary supplements while LBPs are drug products intended for treatment or prevention of diseases. This type of products offers several advantages over traditional drugs, but also entail potential challenges with development, manufacturing, and demonstration of clinical safety. To obtain a sufficient quality, LBPs are typically produced by cultivation in a bioreactor, followed by formulation and lyophilization. In the first part of the project, the impact of lyophilization parameters on physicochemical and biological properties of Limosilactobacillus reuteri R2LC was evaluated. Using sucrose as a lyoprotectant gave a better freeze-drying survival, vitality and storage stability than using trehalose. A high concentration (20%) of sucrose sometimes resulted in a collapsed structure and 15% gave the overall best properties of the lyophilized bacteria. Interestingly, vitality was po... Read More

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In the probiotics manufacturing process, lyophilization damages the cell membrane of microorganisms, reducing their viability and delaying reactivation of their cellular metabolism. The growth phase of microorganisms at the time of lyophilization is thought to significantly affect their viability during storage and reactivation in the intestine. However, no study has systematically investigated the differences in overcoming mechanisms in microorganisms harvested at different growth phases against lyophilization. Therefore, in this study, we comparatively assessed the resistance mechanism of Bifidobacterium animalis ssp. lactis and Lacticaseibacillus rhamnosus harvested at exponential and stationary growth phases. We found that B. animalis ssp. lactis and L. rhamnosus harvested at the stationary phase had shorter lag phase in growth and higher survival rates during storage than those harvested at the exponential phase. As a result of metabolome analysis, the cells harvested at the stationary phase accumulated trehalose and arabinose, which function as cryoprotectants and protect cells... Read More

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Probiotics are of great economic importance mainly because of the various health benefits it offers. It is important for the probiotics to maintain its viability and health-promoting properties during the manufacturing process and storage. Freeze- drying is a common method to preserve the probiotics. The probiotic bacteria were freeze-dried using different cryoprotectants and its effect on the stability and viability of the probiotics were studied. The freeze dried culture was stored at a temperature of -20C and 4C to check for the survival rate. The plate count method was used to determine the survival rate of bacterial cells before and after freeze-drying. The inactivation rate constant k, was determined. Production of vitamins - Riboflavin and Cobalamin in probiotic bacteria was analysed by HPLC and the presence was confirmed by FTIR. Yoghurt was prepared using monoculture and co-culture of Lactobacillus sp. and Lactococcus sp. The proximate analysis indicated that the yoghurt was good source of protein, fat and energy.

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Research background. Increasing awareness of the importance of nutrition in health promotion and disease prevention has driven to the development of foods for special medical purposes (FSMPs). In this study, the probiotic strain Lacticaseibacillus paracasei ssp. paracasei (Lacticaseibacillus casei 431) was incorporated into FSMPs to develop an innovative product. The aim was to investigate the influence of the FSMP matrix on the specific probiotic properties of L. casei 431 in vitro. Experimental approach. A series of in vitro experiments were performed as part of the probiotic approach. After evaluation of antibiotic susceptibility profiles, functional properties such as survival under simulated gastrointestinal tract (GIT) conditions, bile salt deconjugation activities, cholesterol assimilation, antagonistic activity against spoilage bacteria and adhesion to Caco-2 cell line monolayers and extracellular matrix proteins were investigated. Results andconclusions. The L. casei 431 strain, both the lyophilised strain and the strain isolated from the FSMP matrix, effectively survived... Read More

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The present study investigated Spirulina platensis protein isolate (SPI) for its ability to preserve the biological activity of L. rhamnosus GG (LGG) cells under diverse physicochemical stressors associated with the lyophilisation process, storage, and in vitro digestion. The lyoprotective and stabilising effects of SPI were compared to whey protein isolate (WPI) and pea protein isolate (PPI). The microstructural, physicochemical, and thermal properties of the probiotic lyophilisates during freeze-drying, storage, and in vitro digestion were also assessed. Overall, SPI demonstrated superior lyoprotective and storage stabilising effects compared to WPI, although it was less efficient than PPI. Elevated storage temperature and relative humidity (RH) conditions accelerated LGG inactivation rates, especially in the case of WPI, which was primarily attributed to increased LGGs metabolic activity. Fermentation of the probiotic freeze-drying media impaired LGG's ability to withstand the physicochemical stressors during lyophilization and storage. However, the pre-fermentation step improved ... Read More

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The unregulated use of antibiotics has emerged as a major driving force of antibiotic resistance and public health hazard, particularly in developing countries. This necessitates to find out a solution that benefits humans and animals. Probiotics are one such alternative; they are simply known as beneficial bacteria that exert health benefits to living beings. Probiotic microorganisms are crucial for the biotechnology and food industries. They are frequently employed as starters for the production of probiotic goods, food (such as yogurt, cheese and fermented meats), and food as well as for green chemistry uses. Bacteria can be conveniently preserved using lyophilization. It permits long-term storage and inexpensive distribution at suprazero temperatures while limiting viability and functionality losses by lowering water activity to values below 0.2. This study aimed to determine the efficacy of lyophilized probiotic after lyophilization. All the lyophilized isolates were found to be Gram positive and showed no catalase activity. The isolates fermented the sugars viz. D-glucose, Lact... Read More

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This review aims to bring a more general view of the technological and biological challenges regarding production and use of probiotic bacteria in promoting human health. After a brief description of the current concepts, the challenges for the production at an industrial level are presented from the physiology of the central metabolism to the ability to face the main forms of stress in the industrial process. Once produced, these cells are processed to be commercialized in suspension or dried forms or added to food matrices. At this stage, the maintenance of cell viability and vitality is of paramount for the quality of the product. Powder products requires the development of strategies that ensure the integrity of components and cellular functions that allow complete recovery of cells at the time of consumption. Finally, once consumed, probiotic cells must face a very powerful set of physicochemical mechanisms within the body, which include enzymes, antibacterial molecules and sudden changes in pH. Understanding the action of these agents and the induction of cellular tolerance mec... Read More

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Methods for preserving bacteria, particularly anaerobic bacteria, by flash freezing and lyophilizing the bacteria in a specialized vial, resulting in high viability preservation of at least 10% of colony forming units over extended periods of time. The flash freezing step involves rapid cooling to temperatures below -70°C, followed by lyophilization in a controlled environment. The method enables long-term preservation of bacteria with minimal loss of viability, particularly for anaerobic species that are challenging to preserve.

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Amino acids, which are important compatible solutes, play a significant role in probiotic lyophilization. However, studies on the functions of Bifidobacterium during freeze-drying are limited. Therefore, in this study, we compared the freeze-drying survival rate of Bifidobacterium longum CCFM 1029 cultivated in different media containing different kinds of compatible solutes. We found that the addition of 21 g/L proline to the culture media substantially improved the freeze-drying survival rate of B. longum CCFM 1029 from 18.61 0.42% to 38.74 1.58%. Interestingly, this change has only been observed when the osmotic pressure of the external culture environment is increased. Under these conditions, we found that proline accumulation in this strain increased significantly. This change also helped the strain to maintain its membrane integrity and the activity of some key enzymes during freeze-drying. Overall, these results show that the addition of proline can help the strain resist a tough environment during lyophilization. The findings of this study provide preliminary data for pro... Read More

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A method for lyophilizing cells that remain viable after reconstitution, comprising freezing a composition comprising cells, an aqueous component, a polyol, a sugar, and a polysaccharide, and removing at least 90% of the aqueous component from the frozen composition to produce the lyophilized cells. The lyophilized cells can be reconstituted with a reconstitution agent to form a viable cell population, with at least 1% of the cells remaining viable.

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Lyophilization is one of the most used methods for bacterial preservation. In this process, the cryoprotectant not only largely decreases cellular damage but also plays an important part in the conservation of viability during freeze-drying. This study investigated using cryoprotectant and a mixture of the cryoprotectant to maintain probiotic activity. Seven probiotic strains were considered: (Limosilactobacillus reuteri KUKPS6103; Lacticaseibacillus rhamnosus KUKPS6007; Lacticaseibacillus paracasei KUKPS6201; Lactobacillus acidophilus KUKPS6107; Ligilactobacillus salivarius KUKPS6202; Bacillus coagulans KPSTF02; Saccharomyces cerevisiae subsp. boulardii KUKPS6005) for the production of a multi-strain probiotic and the complex medium for the lyophilized synbiotic production. Cholesterol removal, antioxidant activity, biofilm formation and gamma aminobutyric acid (GABA) production of the probiotic strains were analyzed. The most biofilm formation occurred in L. reuteri KUKPS6103 and the least in B. coagulans KPSTF02. The multi-strain probiotic had the highest cholesterol removal. All ... Read More

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