Global food systems are facing increasing challenges associated with population growth, climate change, environmental pressures, resource limitations, and the demand for safe and nutritious foods. Food biotechnology has emerged as an interdisciplinary field that contributes to addressing these challenges through innovations in microbial processes, bioprocess engineering, food safety technologies, and the development of functional food ingredients. This editorial provides an overview of the research topics presented in this issue of Applied Food Biotechnology. The published articles cover a broad range of subjects, including microbial biotechnology, fermentation technologies, probiotics and postbiotics, enzyme engineering, bioactive compounds, food safety interventions, pathogen control, nanobiotechnology, and sustainable bioprocesses. Several studies examine approaches for improving microbial performance, enhancing food quality, optimizing biotechnological production systems, and increasing the stability and functionality of food ingredients. Other contributions focus on food preservation, antimicrobial strategies, waste valorization, alternative bioresources, and applications relevant to sustainable food production. Collectively, these studies reflect current research directions in food biotechnology and highlight the diversity of biotechnological approaches being explored to support food quality, safety, functionality, and sustainability. The findings presented in this issue may contribute to future developments in research, industry applications, and evidence-based decision-making within the food sector.
Background and Objective: Rosa damascena Mill. possesses bioactive compounds, including flavonoids and anthocyanins, which are addressed for their antioxidant and anti-inflammatory characteristics. This study aimed to develop and optimize a microencapsulation system for rose extract and probiotics. It focused on particle size and morphological characteristics analyzed via particle size analysis, scanning electron microscopy-energy dispersive X-ray spectroscopy and , gas chromatography-mass spectrometry and further assessed the bioavaliability and bioaccessibility of the encapsulated probiotics. Material and Methods: Lacticaseibacillus casei and Bifidobacterium longum were cultivated in De Man, Rogosa, and Sharpe broth. The petals of Rosa damascena Mill. were extracted with ethanol 70% 1:1 aqueous solution. The microencapsulation involved dissolving the extract and probiotics, followed by the addition of chitosan and sodium tripolyphosphate to form stable colloids. The particle size was analyzed using dynamic light scattering and the morphology of microcapsules was investigated using scanning electron microscopy coupled with energy-dispersive X-ray spectroscopy. Detection of ethanol was carried out using gas chromatography-mass spectrometry. Probiotic viability was assessed after storage at 4 degrees C for 0, 14 and 28 d and bioaccessibility was assessed using in vitro gastrointestinal simulation method. Results and Conclusion: The microencapsulation process resulted in spherical microcapsules with a mean particle size of 3107 nm +/- 273.2. Scanning electron microscopy analysis verified uniform morphology, indicating effective encapsulation. The probiotic count for microencapsulated samples was 5.16 +/- 0.37 log cfu ml(-1). Gas chromatography-mass spectrometry data showed that the ethanol content was 2.53% +/- 0.21 (v/v). Microencapsulation of R. damascena Mill. and the probiotics increased the recovery of anthocyanin by 9%. These findings have suggested that combining microencapsulation with probiotic strains provides viable strategy to improve the functional delivery of anthocyanin-rich botanicals in nutraceutical uses.
Background and Objective: Nanobioremediation using various biological entities has emerged as a rapidly evolving field of research. These biologically synthesized nanoparticles are increasingly used in various uses, primarily in the remediation of environmental contaminants and biomedicine. Additionally, the increasing influx of harmful pollutants into the environment, driven by swift technological progress and population expansion, has become a significant issue. A significant number of chemicals have been recognized as endocrine disruptor chemicals, including various pesticides. Therefore, this subject was chosen due to the significant failure rate associated with traditional remediation methods in addressing persistent pesticides. While nanobioremediation has verified effective as a hybrid solution, there is a significant absence of comprehensive reviews focusing on the food chain. Results and Conclusion: Although there are various physical, chemical and biological remediation technologies available, their effectiveness frequently diminishes because of complex processes. Therefore, linking nanoscience and bioremediation strategies can play a promising role in decreasing the endocrine-linked disorders associated with pesticide contamination.
Background and Objective: Fermented foods of camel milk are important components of the diet for people living in arid lands. Shubat is a widely consumed fermented camel milk product in Kazakhstan. However, a comprehensive assessment of its protein quality, specifically amino acid composition, is lacking. This study aimed to assess biological value of the protein component of traditionally prepared shubat from Western Kazakhstan. Material and Methods: Totally, 12 shubat samples were collected from the regions of Atyrau, Aktobe, Mangystau and West Kazakhstan. All samples were set to the national standard ST RK 117-2015. Amino acid composition was assessed using highperformance liquid chromatography with pre-column derivatization. The biological value was assessed by calculating the essential amino acid content; then, amino acid score was compared to the Food and Agriculture Organization/World Health Organization (2011) reference score and the amino acid composition index (UA) based on Harrington's desirability scale. Data were present as mean +/- standard deviation (n = 3). Results and Conclusion: Significant regional variance was observed. Fat and protein contents varied from 5.47 to 6.69% and from 2.14 to 3.21%, respectively. The essential amino acid content constituted 43-46% of total amino acids. The presence of limiting amino acids was a key factor reducing the protein quality. The first limiting amino acid was histidine in samples from Atyrau and West Kazakhstan and leucine in samples from Aktobe and Mangystau. The identification of these region-specific limiting amino acids highlights potential nutritional considerations for populations relying on shubat as a primary protein source. The amino acid composition index (UA) was highest for the sample from Mangystau (0.66) rated as "good," while samples from other regions were rated "satisfactory" (0.56-0.58).
Background and Objective: The global demand for plant-based dairy alternatives is steadily increasing. However, relatively little is known about the fermentation processes of plant-based beverages. The main objective of this study was to investigate the changes in chemical composition, as well as structural, rheological, and organoleptic properties of rice-based beverages, depending on the initial rice-to-water ratio and fermentation time. Material and Methods: Hydromodules with rice-to-water ratios of 1:2 and 1:3 were prepared. Each hydromodule was subjected to enzymatic hydrolysis, followed by fermentation for 8 and 24 hours. The samples were evaluated for physicochemical, rheological, and sensory characteristics. Data analysis and correlation studies were performed using GraphPad Prism and Origin8 software at a significance level of p < 0.05. Results and Conclusion: Fermented samples showed a decrease in pH (by up to 3.5 units) and an increase in titratable acidity (from baseline to 34.2 degrees T and 90.1 degrees T after 8 and 24 hours of fermentation, respectively), which indicates successful acidification due to microbial activity. Fermentation for 8 hours at a 1:2 rice-to-water ratio improved water-holding capacity (98%), as well as textural properties such as cohesiveness (89.2%) and gumminess (17.4%). These results suggest that combining enzymatic prehydrolysis with fermentation using L. bulgaricus can significantly enhance the technological and functional properties of rice-based beverages, offering a promising approach for developing high-quality fermented plant-based products.
Background and Objective: Probiotics, defined as specific strains of live microorganisms, usually bacteria or yeast, play a crucial role in host health by enhancing digestive functions. Traditionally, these organisms were sourced from dairy-based fermented foods such as yogurt, cheese, and kefir; however, the rise in plant-based diets has spurred the popularity of plant-based probiotics, driven by factors such as veganism, lactose intolerance, and dairy allergies. Plant-based probiotics offer a valuable alternative that caters to diverse dietary requirements while also contributing to environmental sustainability through lower greenhouse gas emissions, reduced water consumption, and less land use. This review delves into the development, potential health benefits, future applications, and technological challenges related to the production and marketing of plant-based probiotics. Results and Conclusion: Innovative approaches have led to the creation of functional foods that combine plant sources with robust probiotic strains capable of surviving in plant-based matrices. The integration of probiotics with prebiotic fibers such as chicory and acacia, has proven to enhance viability and performance. Furthermore, plant-based probiotic products are evolving beyond beverages to encompass snacks, ready-to-eat fruits, and nuts. Nonetheless, challenges persist regarding the viability of probiotic strains during storage, the costs associated with large-scale production, and the necessity of consumer education. Regulatory frameworks are being adapted to ensure health claims attributed to probiotic products that are backed by scientific evidence. As a promising frontier in sustainable functional foods, plant-based probiotics cater to a wide range of consumer needs. Future advancements are anticipated to focus on personalized formulations and broader applications within health and wellness, alongside improved techniques to enhance probiotic stability and efficacy.
Background and Objective: Obesity is an increasing public health issue that needs practical and scientifically supported nutritional interventions. This study aimed to formulate functional coffees enriched with fermented lotus leaves (Nelumbo nucifera) using Bacillus subtilis to enhance polyphenol concentration and lipase enzyme activity. Additional components included breadfruit leaves (Artocarpus altilis), lotus seeds, notoginseng flowers (Panax notoginseng), caterpillar fungi (Cordyceps militaris), and collagen, selected for their complementary effects on metabolic functions, immune supports, sensory attributes and market feasibility. Material and Methods: The multiple component formulation was optimized using mixture design integrated with response surface methodology. Efficacy was assessed through a 6-m randomized controlled trial involving 127 overweight adults. The trial used a double-blind placebo-controlled design to ensure reliability and minimize bias. Additionally, a consumer acceptance survey involving 800 participants was carried out to assess repurchase intention and product perception. Results and Conclusion: Fermentation (10(7) CFU.g(-1), 35 degrees C, 65.00-70.00% relative humidity, 72 h) led to a 2.5-fold increase in polyphenol content and doubled lipase enzyme activity. In the clinical trial, participants consuming the nutritional coffee showed average weight increases of 1.40 kg, decreases in low-density lipoprotein cholesterol C by approximately 10.00 mg.dl(-1) and increases in high-density lipoprotein cholesterol by nearly 3.00 mg.dl(-1). These improvements were statistically significant (p < 0.05) and were not associated with serious adverse effects. The consumer survey indicated a 65.00% repurchase intention, suggesting promising market potential. Although the study included limitations such as those of sample size, dropout rate and intervention time, the findings demonstrated metabolic benefits and industrial feasibility. This study provides a solid foundation for the development and commercialization of functional coffee targeting weight management and cardiovascular support. These findings provide valuable insights for researchers and industries worldwide interested in developing innovative functional beverages aimed at managing obesity and improving cardiovascular health.
Background and Objective: Chemical treatments in chitin extraction from shrimp shell wastes have affected the environment. Shrimp shell primarily bonds chitin with inorganic salts, lipids, proteins and pigments. Extraction of chitin from shrimp shells involves protein separation processes. Deproteinization process of chitin from dried shrimp (Litopenaeus vannamei) shells with papain enzyme was optimized and nanochitin as derivative product of chitin was characterized. Material and Methods: Effect of hydrolysis time, temperature and enzyme concentration were optimized using RSM Box-Behnken method to maximize chitin yields. Nanochitin was prepared using dialysis and ultrasonic methods and characterized for physical characteristics using scanning electron microscope, particle size analysis and Fourier transforms infrared spectroscopy. Results and Conclusion: Optimum conditions using enzymatic hydrolysis at 6 h, 50 o and 1.25% papain decreased the protein content from 33.66 to 2.31% and produced high chitin yield (46.03%). Deproteinization using enzymatic hydrolysis method was more efficient than that using fermentation. Data of scanning electron microscope, particle size analysis and Fourier transforms infrared spectroscopy showed that the characteristics of chitin and nanochitin products were similar to those of chemical treatments for chitin products. Conflict of interest: The authors declare no conflict of interest.
Background and Objective: Lactic acid bacteria play a fundamental role in the human diet, particularly in fermented foods such as dairy products. This study aimed to investigate microbial diversity of a traditional milk product (Dhan) prepared from raw milk of lactating dairy cows in Northwestern Algeria, highlighting the significance of this traditional dairy product and providing a detailed characterization of its lactic acid bacterial population. Material and Methods: Nine samples were collected from the available farms in Tiaret and Tissemsilt regions and subsequently cultivated on selective media of MRS agar for Lactobacillus and M17 agar for Lactococcus. Identification was carried out using API Systems (API 50 CHL) and other biochemical assays. Results and Conclusion: The bacterial load lactic acid bacteria counts (log10 CFU g- 1) averaged 4.92 +/- 0.47 on MRS and 4.52 +/- 0.77 on M17 media. Eighteen bacterial isolates were categorized into three genera of Lactobacillus (73%), Lactococcus (16%), and Leuconostoc (11%), with Lactiplantibacillus plantarum as the most prevalent species (seven isolates). Lactobacillus spp. are the dominant lactic acid bacteria in Dhan and further studies are needed to isolate and characterize biotechnological potentials of these isolated species and their possible roles in innovation.
Background and Objective: Cheese is one of the major dairy products with high nutritional value, but its susceptibility to microbial growth and early spoilage remains a challenge for the dairy industry. While chemical additives are widely applied to control microbial contamination, increasing awareness of the potential hazards of synthetic preservatives has led to a growing demand for natural alternatives. This study was designed to evaluate the antimicrobial potential of postbiotics derived from lactiplantibacillus plantarum and Lacticaseibacillus casei against common spoilage and pathogenic microorganisms in cheese, as well as to investigate their impact on the microbiological and chemical properties of cheese during storage. Material and Methods: Postbiotics were extracted from cultures of L. plantarum and L. casei and their antimicrobial activities were tested using standard microbiological assays against Gram-positive and Gram-negative bacteria. To assess practical application, the postbiotics were applied as coatings on cheese samples either alone or in combination with whey protein concentrate (WPC). Microbiological counts, chemical parameters, and sensory evaluation were performed throughout storage. Data were analyzed to determine the comparative effectiveness of treatments. Results and Conclusion: The findings showed that the postbiotic derived from L. plantarum demonstrated stronger antimicrobial effects, particularly against Grampositive bacteria, compared to that from L. casei. However, when combined with WPC, the antimicrobial activity of both postbiotics declined. Despite this limitation, postbiotics applied alone significantly reduced microbial counts during storage without altering the main chemical properties of the cheese. Sensory evaluation confirmed the overall acceptability of postbiotic and postbiotic-WPC treated samples. In conclusion, postbiotics can serve as promising natural antimicrobial agents in cheese preservation, though further optimization is required to enhance their activity when combined with protein-based carriers such as WPC.
Background and Objective: Improving probiotics viability in digestive and storage conditions is challenging for the food and pharmaceutical industries. The present study aimed to increase viability of the microencapsulated probiotic strain of Lactobacillus reuteri ATCC 23272 in Ocarboxymethyl chitosan-coated bionanocomposite. The O-carboxymethyl chitosan was used to coat bionanocomposite containing prebiotics of pectin and inulin in presence of magnesium oxide nanoparticles. Material and Methods: Pectin and inulin were used as prebiotics with magnesium oxide nanoparticles to improve the microgel structure and O-carboxymethyl chitosan for coating the microcapsules for increasing viability and stability of the probiotics. The extrusion efficiency, viability after microwave oven drying, survival in the simulated digestive fluids, viability after heat treatment and survival rate in long-term storage at 4 and 25 degrees C after 42 d were analyzed. Optimization of inulin, pectin and O-carboxymethyl chitosan in O-carboxymethyl chitosancoated alginate-based bionanocomposite was achieved using Design-Expert software and simplex lattice mixture design. Results and Conclusion: Optimal formulation was achieved using O-carboxymethyl chitosan coating polymer (68% w/v), inulin (29.4% w/v) and pectin (2.6% w/v) with magnesium oxide nanoparticles at a constant concentration. Results showed microencapsulation efficiency (96.43%), survival after microwave oven drying (99.45%) and survival in simulated gastrointestinal conditions (88.95%). Probiotic viability entrapped in O-carboxymethyl chitosancoated microcapsules decreased by 1.46 log CFU.g-1 at 80 degrees C for 5 min. Moreover, Ocarboxymethyl chitosan-coated bionanocomposite improved the stability of probiotics by 2.93 and 3.25 log CFU.g-1 at 4 and 25 degrees C after 42 d, compared to alginate beads. Additionally, it was observed that O-carboxymethyl chitosan coating enhanced the stability of probiotics entrapped in bionanocomposite beads. Results demonstrated that O-carboxymethyl chitosan-coated bionanocomposite, as a novel microencapsulation, could significantly increase the shelf life and viability of Lactobacillus reuteri in various harsh conditions, compared to alginate beads.
Background and Objective: Phytase is a phosphatase enzyme. It is essential for hydrolyzing phytic acid, an antinutrient present in plant-based foods that chelates essential minerals and limits their bioavailability. Enzyme techniques used to improve phytase characteristics are discussed including thermostability, catalytic efficiency, and substrate specificity. Results and Conclusion: Phytase efficiency has been significantly improved for industrial applications by different engineering methods, including rational design, computational modeling, directed evolution, glycosylation engineering and semi-rational design. The study further highlights the application of phytase in food processing, including breadmaking, fermented foods, and functional food formulations, to address mineral deficiencies. Future advances in enzyme engineering, such as computational de novo design and enzyme immobilization, could expand commercial and nutritional uses of phytase. The focus of ongoing research on thermostable and highly efficient phytase variants provides significant possibilities for enhancing global food security and sustainability.
Background and Objective: Foodborne pathogens represent a substantial threat to living organisms. Therefore, techniques for prolonging food shelf life while ensuring food its quality are imperative practices that must be adopted. Bacteriocins are broadly addressed as preservatives. This study generally characterized LCI peptide as a beta structure antimicrobial peptide and a novel alternative for extending food shelf life. Material and Methods: The antimicrobial activity of recombinant LCI was assessed against selected Gram-positive and Gram-negative bacterial strains. Temperature, pH and bile salt concentration stability of the antimicrobial peptide were studied. Furthermore, the effect of the peptide on the bacterial membrane was assessed. Results and Conclusion: The study demonstrated that this novel LCI recombinant bacteriocin included antimicrobial characteristics with wide-spectrum activity against Gram-positive and Gram-negative bacteria. The minimum inhibitory concentrations (MICs) were 50 mu g.ml-1 for Micrococcus (M.) luteus ATCC 6633, Staphylococcus (S.) aureus ATCC 6538 and Bacillus (B.) subtilis ATCC 6633 and 100 mu g.ml-1 for Gramnegative bacteria when assessed against Escherichia (E.) coli ATCC 8739, Salmonella (S.) typhimurium ATCC 13311 and Vibrio (V.) parahaemolyticus. Time-kill kinetics demonstrated a bactericidal mechanism of action, showing increased antimicrobial efficacy when reported with acetic acid. Membrane permeabilization assessments indicated that LCI created pores in bacterial membranes in a dose-dependent fashion. The peptide stability assessments revealed its heat resistance up to 100 degrees C for 15 min, while preserving activity in aqueous solutions within pH range of 3-11 and bile salt concentration of 0-2%. These characteristics indicate that LCI may be a viable candidate for antimicrobial uses, especially when used in combination with organic acids.
Background and Objective: The fermentation of Algerian goat milk, a process for the production of valuable dairy products, relies on the synergistic activity of Streptococcus (S.) thermophilus and Lactobacillus (L.) bulgaricus. However, a significant knowledge gap is seen regarding the precise dynamics of these starter cultures within the unique matrix of Algerian goat milks. Specifically, the intricate relationships between their growth patterns and the resulting physicochemical changes, which regulate the distinct biochemical characteristics of fermented products, are poorly understood. So, this study addressed this problem by studying specific contributions of S. thermophilus and L. bulgaricus to goat milk fermentation. Material and Methods: Goat milk was fermented by starter cultures of S. thermophilus and L. bulgaricus (8 h). Bacterial growth and physicochemical parameters, including pH, titratable acidity, viscosity and syneresis, were assessed. Mixed-effects models were used for statistical analysis to assess the relationship between physicochemical changes and bacterial growth. Results and Conclusion: The results showed a strong relationship between L. bulgaricus and the control of acidification, viscosity and syneresis (r = 0.979 for titratable acidity, p < 0.0001). S. thermophilus contributed significantly, particularly to the increases in viscosity (r = 0.773, p < 0.01). The two species significantly decreased the pH, with L. bulgaricus having twice the acidifying effects. By the end of the fermentation process, pH reached 4.12 +/- 0.20, titratable acidity increased to 84.75 +/- 2.19 degrees D and viscosity increased to 6425.00 mPa.s +/- 638.64. The final bacterial counts of S. thermophilus and L. bulgaricus were 519.00 +/- 115.29x10(7) and 65.54 +/- 6.89x10(7) CFU.ml(-1), respect-ively. In addition to providing a robust statistical framework for process control and quality assurance in fermented milk manufacture, this study highlighted the critical role of L. bulgaricus in regulating structural and sensory qualities of fermented goat milks. Results can be used to optimize fermentation processes for goat milk by strategically manipulating the ratio of L. bulgaricus to S. thermophilus. The strong correlation between L. bulgaricus and acidification, viscosity and syneresis (r = 0.979 for titratable acidity, p<0.0001) provides a clear target for controlling key product attributes.
Background and Objective: Diabetes mellitus is a long-term disorder characterized by the body's inability to regulate excessive blood glucose levels. The incidence of diabetes mellitus worldwide has significantly increased in recent decades. In addition to careful regulation of food quantities, improving food quality through the consumption of functional foods that do not trigger glucose spikes is also recommended. Accordingly, the aim of this research is to determine the potency of antidiabetic functional food derived from the fermentation of A2 cow's milk with higher beta-casein content using Lacticaseibacillus rhamnosus RAL43. Material and Methods: This study involved in vitro assays to test the inhibitory activity against diabetes-related enzymes, namely alpha-glucosidase, alpha-amylase, and dipeptidyl peptidase-4 (DPP4), followed by molecular docking simulations. Results and Conclusion: The results of the study showed inhibitory activity against enzymes that trigger blood glucose spikes, namely 66.35% against alpha-glucosidase, 68.87% against alpha-amylase, and 10.69% against DPP4. The results of the analysis showed an increase in the quantity of peptides after fermentation, along with the results of the analysis of L. rhamnosus RAL43 which showed high proteolytic activity during fermentation. After ultrafiltration, it was found that the greatest inhibitory activity came from protein with molecular weight (MW) larger than 10 kDa. The amino acid sequencing process with high-resolution liquid chromatography-mass spectrometry then showed bioactive peptides, including VLVLDTDYK which was previously reported to show DPP4 inhibitory activity, along with many other peptides that display various specific bioactivities. The VLVLDTDYK peptide fragment was successfully docked and positioned in the DPP4 molecule through molecular docking simulations. This study concluded that A2 milk can be a functional substrate to produce specific bioactive peptides that inhibit enzymes that trigger blood glucose spikes that have a negative impact on diabetes. L. rhamnosus RAL43 can be developed as a proteolytic isolate and starter culture to produce foods with functional properties that can aid in blood glucose regulation.
Background and Objective: This study aimed to increase the viable cells and biomass production of a potential probiotic strain of Lactiplantibacillus plantarum DLBSK207 by optimizing concentrations of key nutrients and fermentation conditions using response surface methodology with Box-Behnken design. Material and Methods: The experiments investigated two key variables for medium composition and fermentation conditions. Based on the OFAT result, six factors were selected for the Plackett-Burman Design to evaluate whether the variables had significant effects to the response. The medium contains carbon (glucose) and nitrogen sources (yeast extract and peptone), while the fermentation conditions include initial pH and temperature. The basal medium, consisting of sodium acetate, MgSO4 7H(2)O, K2HPO4, MnSO4 H2O, and Tween 80, was kept constant. Using RSM, the concentrations of glucose, yeast extract, and peptone, as well as the initial pH and temperature, were optimized to maximize viable cell counts and biomass. Results and Conclusion: The optimum medium concentrations determined by RSM were 33.76 g l(-1 )glucose, 32.59 g l(-1) yeast extract, and 28.38 g l(-1)peptone at an initial pH of 6.0 and a temperature of 35 degrees C. Under these optimized conditions, this study achieved a viable cell counts of 9.30 log CFU.ml(-1) and a dry cell weight of 4.319 g l(-1), representing a 1.82-fold increase compared to standard MRS broth. The experimental results were in closely matched the predicted values of 9.30 log CFU.ml(-1) and 4.280 g l(-1). Scaling up the process in a 10-l bioreactor controlled at pH 6.0 resulted in even higher biomass production, reaching a maximum viable cell counts of 9.88 log CFU.ml(-1) and a dry cell weight of 5.819 g l(-1)after 20 h of incubation.
Background and Objective: The demand for cost-effective and thermostable alpha amylases for industrial applications has driven the research to discover new microbial sources. This research aimed to isolate and characterize alpha-amylase-producing bacteria from rice milling wastes and employ Response Surface Methodology (RSM) to improve enzyme production. Material and Methods: Bacterial samples were collected from different agroindustrial wastes and primarily screened using Lugol's iodine method. Secondary isolation was performed by alpha-amylase activity assessment using the DNS assay. Enzyme production was optimized by RSM, with the temperature, pH, and starch concentration as key variables. In addition, the effect of different pH and temperatures was assessed on the alpha-amylase activity. 16S rRNA sequencing and phylogenetic analysis were used for bacterial identification. Results and Conclusion: The isolate was identified as Bacillus subtilis NllST B 627. Optimum conditions for maximum enzyme production (0.21 Umg-1) were starch 5.5 gL-1, temperature 40 degrees C, and pH 7. Temperature was the most significant factor influencing enzyme production, whereas pH and starch concentration showed weaker effects but potentially relevant interactions. The overall model based on response surface curves was statistically significant, indicating that the combination of independent variables significantly influences enzyme production. The enzyme exhibited maximum activity at pH 7, while the lowest activity was observed at pH 5. Also, the enzyme's optimal activity occurred at 40 degrees C, while the lowest catalysis was detected at 60 degrees C. The identified strain exhibits promising properties for application in starch hydrolysis and other industrial purposes. This highlights the potential of rice mill wastes as a sustainable and low-cost resource for microbial enzyme production, and this study is the first to explore Guilan rice mill wastes for alpha-amylase production.
Background and Objective: Identifying a milk-clotting enzyme (MCE) with high xcasein specificity and heat sensitivity remains a challenge in cheese production. Current microbial, plant, and recombinant MCEs often exhibit low clotting activity, poor x-casein specificity, and high thermostability, compromising cheese quality and increasing production costs. In this study, to address this, we developed a computational pipeline combining structural analysis, machine learning, and molecular dynamics simulation to design approximately 160,000 peptides from the Rhizomucor miehei protease-Pepstatin A complex (PDB ID: 2RMP). Material and Methods: Single-site mutagenesis, ML-driven affinity re-prediction, and physicochemical filtering yielded 84 peptides. Their specificity as aspartic proteases were validated via predicted Pepstatin A binding and further screened for cross-reactivity with alpha s1-, alpha s2-, and beta-caseins. Results and Conclusion: Two candidates, Pep1 and Pep2, demonstrated superior xcasein binding affinities (Delta G = -50.20 and -39.07 kcal/mol at 40 degrees C, respectively), lower melting indices (-4.05 and -3.09), and significantly enhanced specificity scores (-10.85) compared to Rhizomucor miehei protease (Delta G = -33.9 kcal/mol at 45 degrees C; melting index = 0.17; specificity score = 0.85). These peptides represent promising vegan- and halal-friendly alternatives to chymosins, pending experimental validation.
Background and Objective: The delivery of probiotics using functional foods or supplements (e.g., capsules, tablets, or sachets) as carriers is an important strategy to provide health benefits, as it is necessary to maintain 10(6)-10(9) colony-forming units (CFU) per gram of the probiotic strain alive and active at the time of consumption for effectiveness. This study investigated the effects of batch and fed-batch fermentation methods on increasing the cell and spore populations of Heyndrickxia coagulans MTCC 5856 (formerly Bacillus coagulans MTCC 5856). Material and Methods: Batch and fed-batch fermentation strategies were applied to evaluate their effects on vegetative growth and sporulation. The protective effects of various cryoprotectants (sorbitol, sucrose, inulin, calcium lactate, manganese chloride, and skim milk) were assessed during freeze-drying. Spore resistance under simulated gastrointestinal conditions and stability in two functional food matrices (pastille and coffee mix) were also evaluated. Results and Conclusion: Skim milk increased sporulation from 63% to 88%, with >80% spore viability in GI conditions and 85-98% stability in food matrices after 6 months. This study provides the first comparative analysis of batch versus fed-batch fermentation on H. coagulans MTCC 5856 sporulation in industrial food matrices, revealing glucose's inhibitory effect and skim milk's superior cryoprotective efficacy. These findings highlight the importance of optimizing fermentation conditions and applying suitable cryoprotectants to enhance the long-term viability and application of H. coagulans in functional food formulations.
This study aimed to enhance probiotics thermal stability and viability in the digestive tract through encapsulation using hybrid fibers of cellulose acetate and polyvinyl alcohol with single-jet electrospinning. This study used Lactiplantibacillus plantarum NIMBB003 as an encapsulated probiotic strain in engineered sandwich nanofibers (cellulose acetate/polyvinyl alcohol and Lactiplantibacillus plantarum/cellulose acetate). Regarding nanostructure, polyvinyl alcohol and cellulose acetate nanofibers were spun independently; when these layers were set on top of each other, they could act as an integrated system. Results of scanning electron microscope images and Fourier transform infrared spectrometry have verified the micro/nanoencapsulation structure of probiotics. The layered structure demonstrated increased protection against environmental factors, particularly heat and acidity. Thermogravimetric analysis verified that cellulose acetate-polyvinyl alcohol and probiotic-cellulose acetate nanofibers maintained the structural stability up to 530 degrees C, while encapsulated probiotics showed 89.8% encapsulation efficiency or 9% improvement, compared to single-layer polyvinyl alcohol and probiotic fibers. Moreover, probiotic survival under simulated gastrointestinal conditions (75 degrees C and stomach acid exposure) was extended to 8 min, whereas unencapsulated probiotics were entirely destroyed within 5 min. Scanning electron microscopy and Fourier transform infrared spectroscopy validated the formation of nanofiber encapsulation and probiotic integration. This engineered nanofiber sandwich structure offers enhanced probiotic protection, making it a promising candidate for food and pharmaceutical uses.
