Effect of transglutaminase crosslinking combined with lactic fermentation on the potential allergenicity and conformational structure of soy protein.

BACKGROUND: Food allergies are a growing concern worldwide, with soy proteins being important allergens that are widely used in various food products. This study investigated the potential of transglutaminase (TGase) and lactic acid bacteria (LAB) treatments to modify the allergenicity and structural properties of soy protein isolate (SPI), aiming to develop safer soy-based food products. RESULTS: Treatment with TGase, LAB or their combination significantly reduced the antibody reactivity of ß-conglycinin and the immunoglobulin E (IgE) binding capacity of soy protein, indicating a decrease in allergenicity. TGase treatment led to the formation of high-molecular-weight aggregates, suggesting protein crosslinking, while LAB treatment resulted in partial protein hydrolysis. These structural changes were confirmed by Fourier transform infrared spectroscopy, which showed a decrease in ß-sheet content and an increase in random coil and ß-turn contents. In addition, changes in intrinsic fluorescence and ultraviolet spectroscopy were also observed. The alterations in protein interaction and the reduction in free sulfhydryl groups highlighted the extensive structural modifications induced by these treatments. CONCLUSION: The synergistic application of TGase and LAB treatments effectively reduced the allergenicity of SPI through significant structural modifications. This approach not only diminished antibody reactivity of ß-conglycinin and IgE binding capacity of soy protein but also altered the protein's primary, secondary and tertiary structures, suggesting a comprehensive alteration of SPI's allergenic potential. These findings provide a promising strategy for mitigating food allergy concerns and lay the foundation for future research on food-processing techniques aimed at allergen reduction. © 2024 Society of Chemical Industry.

Effects of acidification by traditional Jiaozi starter and neutralization with alkali (Na2CO3) on whole wheat dough properties.

BACKGROUND: Whole wheat steamed bread has been recommended for its potential nutritional benefits to human health. Given the positive role of both organic acid and alkali in improving dough development and product quality, the present study investigated the effects of neutralization by addition of alkali (Na2CO3) after dough acidification with traditional Jiaozi starter on the properties of whole wheat dough. RESULTS: The population of yeast and lactic acid bacteria and the acidification level of the dough increased significantly after fermentation with Jiaozi. Incorporation of alkali greatly improved the leavening capacity of the remixed dough and the quality of steamed bread. Jiaozi fermentation and alkali addition changed the water distribution patterns (T2) and affected the secondary structures of gluten protein, starch crystallinity and pasting properties. The storage modulus (G') of the dough increased significantly with the alkali addition, which could be attributed to the promoted cross-linking of the gluten structure and the altered hydration state of the macromolecules. CONCLUSION: The results of the present study indicate that a combination of Jiaozi fermentation and alkali addition could improve the technological properties of whole wheat dough and the quality of steamed bread. The results will help us to further explore the potential application of moderate acidification and alkali addition in the production of leavened whole wheat products. © 2024 Society of Chemical Industry.

Interaction between lactic acid bacteria and Polygonatum sibiricum saponins and its application to microencapsulated co-delivery.

This study aimed to investigate the interaction between L.casei and L.bulgaricus with Polygonatum sibiricum saponins (PSS) and to explore the co-microencapsulation to reduce their loss rate during storage and consumption. 1% PSS was added to the culture broth, and it was found that the growth and metabolism of the strains were accelerated, especially in the compound probiotic group, indicating that PSS has potential for prebiotics. LC-MS observed significant differences in the composition and content of saponins in PSS. The metabolomics results suggest that the addition of PSS resulted in significant changes in the metabolites of probiotics. In addition, it was found that the combination of probiotics and PSS may have stronger hypoglycemic ability (ɑ-glucosidase, HepG2). Finally, a co-microencapsulated delivery system was constructed using zein and isomaltooligosaccharide. This system can achieve more excellent resistance of probiotics and PSS in gastrointestinal fluids, effectively transporting both to the small intestine.

Optimization of asymmetric bioreduction conditions of 1-indanone by Leuconostoc mesenteroides N6 using a face-centered design-based multi-objective optimization model.

There has been an increasing interest in biocatalysts over the past few decades in order to obtain high efficiency, high yield, and environmentally benign procedures aiming at the manufacture of pharmacologically relevant chemicals. Lactic Acid Bacteria (LAB), a microbial group, can be employed as biocatalysts while performing asymmetric reduction of prochiral ketones. In this study, Leuconostoc mesenteroides N6 was used for the asymmetric bioreduction 1-indanone. And then, a novel and innovative face-centered design-based multi-objective optimization model was used to optimize experimental conditions. Also, the experimental design factors were defined as agitation speed, incubation period, pH, and temperature for optimization to acquire the maximum enantiomeric excess (ee) and conversion rate (cr) values. When using the face-centered design-based multi-objective optimization model, the optimum culture conditions corresponded to 96.34 and 99.42%, ee and cr responses, respectively, were pH = 5.87, incubation temperature = 35 °C, incubation period = 50.88 h, and agitation speed = 152.60 rpm. Notably, the validation experiment under the optimum model conditions confirmed the model results. This study demonstrated the importance of the optimization and the efficiency of the face-centered design-based multi-objective model.

Biological activities and applications of exopolysaccharides produced by lactic acid bacteria: a mini-review.

Exopolysaccharides (EPSs) are naturally occurring high-molecular-weight carbohydrates that have been widely studied for their biological activities, including antioxidant, immunomodulatory, anticancer and gut microbiota regulation activities. Polysaccharides are abundant in nature and can be derived from animals, plants, algae, and microorganisms, but among polysaccharides with potential uses, EPSs from microorganisms have the advantages of a short production cycle, high yield, and independence of production from season and climate and thus have broad prospects. While the safety of the producing microorganism can represent a problem in application of microbial EPSs, lactic acid bacteria (LAB) have been used by humans for thousands of years, and they and their products are generally recognized as safe. This makes LAB excellent sources for exopolysaccharides. EPS-producing LAB are readily found in nature. Through screening of strains, optimization of culture conditions, and improvement of the growth medium, the yield of EPSs from LAB can be increased and the scope of application broadened. This review summarizes EPSs from LAB in terms of structure, function and applications, as well as yield optimization, and introduces recent research on the biological activities and practical applications of LAB EPSs, aiming to provide references for researchers in related areas.

Insights into lactic acid bacteria cryoresistance using FTIR microspectroscopy.

Freezing is widely used for bacterial cell preservation. However, resistance to freezing can greatly vary depending on bacterial species or growth conditions. Our study aims at identifying cellular markers of cryoresistance based on the comparison of three lactic acid bacteria (LAB) exhibiting different tolerance to freezing: Carnobacterium maltaromaticum CNCM I-3298, Lactobacillus delbrueckii subsp. bulgaricus ATCC 11842, and Lactobacillus delbrueckii subsp. bulgaricus CFL1. A thorough characterization of their cytoplasmic membrane properties was carried out by measuring their fatty acid composition, membrane fluidity, and lipid phase transition upon cooling from 50 to -50 °C. Vitrification temperatures of the intra- and extra-cellular compartments were also quantified by differential scanning calorimetry. Additionally, the cell biochemical characterization was carried out using a recently developed Fourier transform infrared (FTIR) micro-spectroscopic approach allowing the analysis of live bacteria in an aqueous environment. The multivariate analysis of the FTIR spectra of fresh and thawed cells enabled the discrimination of the three bacteria according to their lipid, protein, and cell wall peptidoglycan components. It also revealed freezing-induced modifications of these three cellular components and an increase in bacteria heterogeneity for the two strains of L. bulgaricus, the freeze-sensitive bacteria. No cellular damage was observed for C. maltaromaticum, the freeze-resistant bacteria. Comparison of the results obtained from the different analytical methods confirmed previously reported cryoresistance markers and suggested new ones, such as changes in the absorbance of specific infrared spectral bands. FTIR microspectroscopy could be used as a rapid and non-invasive technique to evaluate the freeze-sensitivity of LAB.

Food wastes as natural sources of lactic acid bacterial exopolysaccharides for the functional food industry: A review.

Exopolysaccharides (EPSs) synthesized by lactic acid bacteria (LAB), have recently received much interest because of their various functional features in several industries. Food wastes (FWs) have become a major source of worry, as they can cause serious environmental contamination if improperly disposed. The utilization of these FWs is an excellent choice (approach) for producing value-added products such as EPSs, which will efficiently remediate wastes. The overall EPSs yield for the selected producers is strain-specific, and is heavily influenced by the nutritional and growing conditions used. This review emphasizes what is currently known about LAB's ability to generate economically relevant EPSs from FWs. In addition, a concise overview of the food industry, packaging, pharmaceutical and clinical applications application is discussed.

Dairy associations for the targeted control of opportunistic Candida.

Antifungal and antibacterial activities of twenty-six combinations of lactic acid bacteria, propionibacteria, acetic acid bacteria and dairy yeasts inoculated in whey and milk were investigated. Associations including acetic acid bacteria were shown to suppress growth of the opportunistic yeast Candida albicans in well-diffusion assays. The protective effect of milk fermented with the two most promising consortia was confirmed in Caco-2 cell culture infected with C. albicans. Indeed, these fermented milks, after heat-treatment or not, suppressed lactate dehydrogenase release after 48 h while significant increase in LDH release was observed in the positive control (C. albicans alone) and with fermented milk obtained using commercial yogurt starter cultures. The analysis of volatile compounds in the cell-free supernatant using solid phase microextraction (SPME) coupled to gas chromatography-mass spectrometry (GC-MS) showed accumulation of significant amount of acetic acid by the consortium composed of Lactobacillus delbrueckii 5, Lactobacillus gallinarum 1, Lentilactobacillus parabuchneri 3, Lacticaseibacillus paracasei 33-4, Acetobacter syzygii 2 and Kluyveromyces marxianus 19, which corresponded to the zone of partial inhibition of C. albicans growth during well-diffusion assays. Interestingly, another part of anti-Candida activity, yielding small and transparent inhibition zones, was linked with the consortium cell fraction. This study showed a correlation between anti-Candida activity and the presence of acetic acid bacteria in dairy associations as well as a significant effect of two dairy associations against C. albicans in a Caco-2 cell model. These two associations may be promising consortia for developing functional dairy products with antagonistic action against candidiasis agents.

The efficacy of three double-microencapsulation methods for preservation of probiotic bacteria.

Lactic acid bacteria (LAB) are used as a probiotic alternative to antibiotics in livestock production. Microencapsulation technology is widely used for probiotic preservation. A variety of microencapsulation protocols have been proposed and compared based on chemicals and mechanical procedures. This study aimed to develop a double-encapsulated coating from alginate (1.5%) and chitosan (0.5%) by extrusion, emulsion, and spray drying methods using the LAB strains Lactobacillus plantarum strains 31F, 25F, 22F, Pediococcus pentosaceus 77F, and P. acidilactici 72N, and to monitor the basic probiotic properties of the encapsulated prototypes. The final products from each microencapsulation protocol were analysed for their appearance, probiotic properties and viable cell count. Using the spray drying method, particles smaller than 15 µm in diameter with a regular spherical shape were obtained, whereas the other methods produced larger (1.4-52 mm) and irregularly shaped microcapsules. After storage for 6 months at room temperature, the LAB viability of the spray-dried particles was the highest among the three methods. In all the LAB strains examined, the encapsulated LAB retained their probiotic properties in relation to acid-bile tolerance and antibacterial activity. This study highlights the efficacy of double-coating microencapsulation for preserving LAB properties and survival rate, and demonstrates its potential for probiotic application in livestock farms.

Techno-functional properties of HoPS from lactic acid bacteria of different origins as potential food additives.

Homopolysaccharides (HoPS) produced by lactic acid bacteria (LAB) are highly versatile, biocompatible and safe compounds. In this work, six HoPS from different species of Weisella and Leuconostoc were identified as thermally stable dextrans, with endothermic crystalline deformations between 214 and 239 °C. These dextrans proved to have greater solubility and capacities to retain water and oil than similar polymers in other reports. Furthermore, a surface morphology study presented cubic grumps, stratify mesh with irregular grumps, and highly compact filaments. Assays in vitro revealed moderate antioxidant, browning and foaming activities as well as technological properties, such as anti-syneresis, emulsifying and flocculating activities, even at low concentrations. Taking into account bipolymers' microstructure, functionalities and performance in both, aqueous and hydrophobic matrixes, plus their capacity to maintain themselves at elevated temperatures, we consider these HoPS beneficial and natural food additives.

Profiling of Potential Antibacterial Compounds of Lactic Acid Bacteria against Extremely Drug Resistant (XDR) Acinetobacter baumannii.

A total of 20 of isolates of lactic acid bacteria (LAB) were selected and screened for antagonistic activity against clinical strains of 30 clinical isolates of extremely drug-resistant (XDR) Acinetobacter baumannii using the well diffusion assay method. Results showed that 50% of the highly LAB strains possessed inhibitory activity against (up to 66%) of the XDR A. baumannii strains tested. The supernatant of the twenty LAB strains was subjected to gas chromatography mass spectrometry (GCMS) revealed that the common compound found in the active isolates against XDR A. baumannii was 3-Isobutyl-2,3,6,7,8,8a-hexahydropyrrolo[1,2-a]pyrazine-1,4-dione, a known potential diketopiperazine group. The molecular docking study against potential antibacterial targets with selected ligands was performed to predict the binding mode of interactions, which is responsible for antibacterial activity. The docking analysis of the potent compounds supported the potential antibacterial activity exhibiting high inhibition constant and binding affinity in silico.

Co-Microencapsulated Black Rice Anthocyanins and Lactic Acid Bacteria: Evidence on Powders Profile and In Vitro Digestion.

Two multi-functional powders, in terms of anthocyanins from black rice (Oryza sativa L.) and lactic acid bacteria (Lactobacillus paracasei, L. casei 431®) were obtained through co-microencapsulation into a biopolymer matrix composed of milk proteins and inulin. Two extracts were obtained using black rice flour as a raw material and hot water and ethanol as solvents. Both powders (called P1 for aqueous extract and P2 for ethanolic extract) proved to be rich sources of valuable bioactives, with microencapsulation efficiency up to 80%, both for anthocyanins and lactic acid bacteria. A higher content of anthocyanins was found in P1, of 102.91 ± 1.83 mg cyanindin-3-O-glucoside (C3G)/g dry weight (DW) when compared with only 27.60 ± 17.36 mg C3G/g DW in P2. The morphological analysis revealed the presence of large, thin, and fragile structures, with different sizes. A different pattern of gastric digestion was observed, with a highly protective effect of the matrix in P1 and a maximum decrease in anthocyanins of approximatively 44% in P2. In intestinal juice, the anthocyanins decreased significantly in P2, reaching a maximum of 97% at the end of digestion; whereas in P1, more than 45% from the initial anthocyanins content remained in the microparticles. Overall, the short-term storage stability test revealed a release of bioactive from P2 and a decrease in P1. The viable cells of lactic acid bacteria after 21 days of storage reached 7 log colony forming units (CFU)/g DW.

Evaluation of the probiotic and postbiotic potential of lactic acid bacteria from artisanal dairy products against pathogens.

INTRODUCTION: Probiotic and postbiotic potential of thirty-two strains of lactic acid bacteria (LAB), obtained earlier from artisanal dairy sources in Pakistan, have been investigated against major multi-drug resistant (MDR) and food borne pathogenic bacteria. METHODOLOGY: LAB strains were identified by 16S rRNA gene sequencing and their antibacterial activity was assessed by the microdilution method. Four LAB isolates, Weissella confusa PL6, Enterococcus faecium PL7, and Lactobacillus delbrueckii PL11 and PL13 were shortlisted. Their ability to degrade lactose and safety for human consumption in terms of hemolysis and antibiotic susceptibility were assessed in vitro. The antibacterial components in the cell-free supernatants (CFSs) of isolate cultures were characterized biochemically by HPLC. RESULTS: Acid neutralization but not protease treatment abolished the antibacterial activity of CFSs. Lactic, acetic and propionic acids were the main acids in the CFSs, and acid production peaked in the stationary phase of growth. The antibacterial activity of the LAB cultures resulted from secretion of organic acids that lowered the pH. The strains exhibited variable ability to degrade lactose and were non-hemolytic and susceptible to the most common antibiotics. CONCLUSIONS: These LAB strains are probiotic candidates for further investigation of their postbiotic role in naturally preserving processed foods and for attenuation of lactose intolerance.

A dynamic resazurin microassay allowing accurate quantification of cells and suitable for acid-forming bacteria.

A resazurin micro-assay was developed to quantify acidifying bacteria. The resorufin fluorescent signal was measured over time and the determined time to reach the max slope (TMS) was plotted against CFU (colony forming unit) counts. This dynamic assay enabled to quantify nine lactic acid bacteria and a Bacillus licheniformis strain despite the increasing acidity of the medium.

Suitability of lactic acid bacteria and deriving antibacterial preparations to enhance shelf-life and consumer safety of emulsion type sausages.

Ready-to-eat (RTE) sliced emulsion type sausages are sensitive to recontamination with Listeria (L.) monocytogenes during processing and packaging steps. Since Listeria spp. are able to grow on those products under cold storage conditions, taking steps to reduce the recontamination risk and implementing antibacterial hurdles contribute to consumer safety and increase the product quality. With this study data about the suitability of culture broth, cell-free supernatant (CFS) or concentrated bacteriocin preparations (CFSconc) of bacteriocin-producing lactic acid bacteria (LAB) obtained from fermented sausages from Germany as protective culture or antibacterial additive were provided. In different challenge tests, the potential of selected LAB or their preparations were investigated for their potential to reduce growth of L. monocytogenes and/or Brochothrix (B.) thermosphacta on sliced RTE emulsion type sausages under modified atmosphere or vacuum during refrigerated storage for a 21-day period. Applied LAB culture broth and CFS could not reduce the growth of L. monocytogenes or B. thermosphacta. On the other hand, samples treated with CFSconc obtained from Pediococcus spp. strains showed a significant inhibition (p < 0.05) of more than 1.5 log10 of the applied L. monocytogenes strains during the storage period. The growth of B. thermosphacta could not be influenced. Thereby, the need for concentrating preparations was shown to be important to obtain a suitable antibacterial preparation that would contribute to consumer safety and food quality when applied as a protective additive.

Lactic acid bacterial exopolysaccharides strongly bind histamine and can potentially be used to remove histamine contamination in food.

The symptoms of foodborne histamine poisoning are similar to those of IgE-mediated food allergies. In this study, we investigated the histamine-binding capacity of lactic acid bacteria (LAB) strains as potential preventive agents against histamine poisoning. Histamine biosorption capacity was determined for 16 LAB strains. Leuconostoc mesenteroides TOKAI 51 m, Lactobacillus paracasei TOKAI 65 m, Lactobacillus plantarum TOKAI 111 m and Pediococcus pentosaceus TOKAI 759 m showed especially high biosorption rates and reached saturation within 30 min. Adsorption isotherms showed better conformance to the Freundlich model than to the Langmuir model. Analyses after heat, periodic acid and guanidine hydrochloride treatments suggested that histamine was bound to the bacterial cell surface. HPLC analysis revealed that exopolysaccharides produced by Lact. paracasei TOKAI 65 m strongly bound to histamine. In the detachment test with 1 mol l-1 HCl solution, the dissociation rate of histamine for Lact. paracasei TOKAI 65 m was <10 %. This strain is presumably a suitable candidate for use against histamine poisoning.

Postbiotics produced by lactic acid bacteria: The next frontier in food safety.

There are many critical challenges in the use of primary and secondary cultures and their biological compounds in food commodities. An alternative is the application of postbiotics from the starter and protective lactic acid bacteria (LAB). The concept of postbiotics is relatively new and there is still not a recognized definition for this term. The word "postbiotics" is currently used to refer to bioactive compounds, which did not fit to the traditional definitions of probiotics, prebiotics, and paraprobiotics. Therefore, the postbiotics may be presently defined as bioactive soluble factors (products or metabolic byproducts), produced by some food-grade microorganisms during the growth and fermentation in complex microbiological culture (in this case named cell-free supernatant), food, or gut, which exert some benefits to the food or the consumer. Many LAB are considered probiotic and their postbiotic compounds present similar or additional health benefits to the consumer; however, this review aimed to address the most recent applications of the postbiotics with food safety purposes. The potential applications of postbiotics in food biopreservation, food packaging, and biofilm control were reviewed. The current uses of postbiotics in the reduction and biodegradation of some food safety-related chemical contaminants (e.g., biogenic amines) were considered. We also discussed the safety aspects, the obstacles, and future perspectives of using postbiotics in the food industry. This work will open up new insights for food applications of postbiotics prepared from LAB.

Aflatoxin B1 and Sterigmatocystin Binding Potential of Lactobacilli.

Due to global climate change, mould strains causing problems with their mycotoxin production in the tropical-subtropical climate zone have also appeared in countries belonging to the temperate zone. Biodetoxification of crops and raw materials for food and feed industries including the aflatoxin B1 (AFB1) binding abilities of lactobacilli is of growing interest. Despite the massive quantities of papers dealing with AFB1-binding of lactobacilli, there are no data for microbial binding of the structurally similar mycotoxin sterigmatocystin (ST). In addition, previous works focused on the detection of AFB1 in extracts, while in this case, analytical determination was necessary for the microbial biomass as well. To test binding capacities, a rapid instrumental analytical method using high-performance liquid chromatography was developed and applied for measurement of AFB1 and ST in the biomass of the cultured bacteria and its supernatant, containing the mycotoxin fraction bound by the bacteria and the fraction that remained unbound, respectively. For our AFB1 and ST adsorption studies, 80 strains of the genus Lactobacillus were selected. Broths containing 0.2 µg/mL AFB1and ST were inoculated with the Lactobacillus test strains. Before screening the strains for binding capacities, optimisation of the experiment parameters was carried out. Mycotoxin binding was detectable from a germ count of 107 cells/mL. By studying the incubation time of the cells with the mycotoxins needed for mycotoxin-binding, co-incubation for 10 min was found sufficient. The presence of mycotoxins did not affect the growth of bacterial strains. Three strains of L. plantarum had the best AFB1 adsorption capacities, binding nearly 10% of the mycotoxin present, and in the case of ST, the degree of binding was over 20%.

Lipidomic analysis of lactic acid bacteria strains by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

Analysis by MALDI-TOF mass spectrometry and gas chromatography-mass spectrometry was used to characterize the lipid profile of 3 lactic acid bacteria strains. By gas chromatography coupled with mass spectrometry, 23 fatty acids were identified. Dominant acids were palmitic (C16:0), oleic (C18:1), and α-linoleic acid (C18:3n-3) for Lactobacillus paracasei; for Lactococcus lactis they were palmitic (C16:0), gondoic (C20:1), myristoleic (C14:1), and eicosadienoic acid (C20:2), respectively; and in the case of Lactobacillus curvatus were C18:1, C18:2n-6, and C16:0, respectively. The effect of the medium on fatty acid composition was also determined. In addition, the fatty acid profile was also compared using MALDI MS analysis. The MALDI-TOF MS was used for qualitative analysis and identification of bacterial lipids. Phosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, phosphatidylcholine, triacylglycerols, and ceramides were the most abundant species in lactic acid bacteria. One hundred different combinations of fatty acids in polar and nonpolar lipids have been identified, including 11 phospholipids (18 phosphatidylglycerol, 16 phosphatidylethanolamine, 10 phosphatidylinositol, 8 phosphatidylcholine, 4 lyso-phosphatidylethanolamine, 3 lyso-phosphatidylcholine, 3 phosphatidylserine, 1 lyso-phosphatidic acid, 1 lyso-phosphatidylglycerol, 1 lyso-phoshatidylinositol, and 1 phosphatidic acid), 23 triacylglycerols, 9 ceramides, and 2 sphingomyelin. The most abundant fatty acids identified were C16:0, C16:1, C18:0, and C18:3. Obtained lipid profiles allowed to distinguish the tested bacterial strains.

Physicochemical properties of soy protein hydrolysate and its formulation and stability with encapsulated probiotic under in vitro gastrointestinal environment.

The objective of this study was to prepare protein isolate from defatted soybean and identify an optimal hydrolysis protocol to create improved hydrolysates and ascertain the optimum encapsulation technique for probiotics. Soy protein isolate (SPI) was prepared using an alkaline extraction procedure for solubility within a neutral, beverage-specific pH range. The soy protein hydrolysate (SPH) was prepared from aqueous extracted SPI using pepsin. The physicochemical properties of the SPH were investigated by solubility, degree of hydrolysis (DH), surface hydrophobicity, and electrophoresis. Hydrolysates from 2, 2.5, and 3 hr of hydrolysis time achieved the suitable DH between 2.5% to 5.0%. The 2.5 to 3 hr hydrolysates were also significantly more soluble than SPI at all pH levels from 85% to 95% solubility. Surface hydrophobicity of the hydrolysates ranged from 15 to 20 S0 values. Alginate (1%), resistant starch (2%), and probiotic culture (0.1%) were used as an encapsulation agent to protect probiotics. Alginate microcapsules were observed to be 1 mm in size using environmental scanning electron microscopy. The dried SPH and encapsulated probiotics with alginate in a dry powder formulation were tested for its gastrointestinal resistance and probiotic viability under in vitro simulated digestion. Approximately 1-log decrease was observed for all experimental groups after simulated digestion (final log colony forming units [CFU]/mL range: 6.55 to 6.19) with free probiotics having the lowest log CFU/mL (6.10 ± 0.10) value. No significant difference was observed among experimental groups for probiotic viability (P = 0.445). The findings of this research will provide an understanding of formulation for easily digestible protein and encapsulated probiotics. PRACTICAL APPLICATION: The findings of this research provide an understanding of improved formulation for more suitable soy protein hydrolysate and viability of encapsulated probiotics in gastrointestinal environment. Probiotics with the prebiotics in an encapsulated environment provide a technology for the enhancement of probiotics viability and for applications in suitable products for health and wellness.