Comparative proteomic analysis of donor human milk treated by high-pressure processing or Holder pasteurization on undigested proteins across dynamic simulated preterm infant digestion.

High-pressure processing (HPP) of donor human milk (DM) minimally impacts the concentration and bioactivity of some important bioactive proteins including lactoferrin, and bile salt-stimulated lipase (BSSL) compared to Holder pasteurization (HoP), yet the impact of HPP and subsequent digestion on the full array of proteins detectable by proteomics remains unclear. We investigated how HPP impacts undigested proteins in DM post-processing and across digestion by proteomic analysis. Each pool of milk (n = 3) remained raw, or was treated by HPP (500 MPa, 10 min) or HoP (62.5 °C, 30 min), and underwent dynamic in vitro digestion simulating the preterm infant. In the meal, major proteins were minimally changed post-processing. HPP-treated milk proteins better resisted proximal digestion (except for immunoglobulins, jejunum 180 min) and the extent of undigested proteins after gastric digestion of major proteins in HPP-treated milk was more similar to raw (e.g., BSSL, lactoferrin, macrophage-receptor-1, CD14, complement-c3/c4, xanthine dehydrogenase) than HoP.

Action of microbial transglutaminase (MTGase) on the processing properties of glutinous rice flour and the quality attributes of sweet dumplings and in vitro digestion.

The development and manufacture of high-quality starch are a new research focus in food science. Here, transglutaminase was used in the wet processing of glutinous rice flour to prepare customized sweet dumplings. Transglutaminase (0.2 %) lowered protein loss in wet processing and reduced the crystallinity and viscosity of glutinous rice flour. Moreover, it lowered the cracking and cooking loss of sweet dumplings after freeze-thaw cycles, and produced sweet dumplings with reduced hardness and viscosity, making them more suitable for people with swallowing difficulties. Additionally, in sweet dumplings with 0.2 % transglutaminase, the encapsulation of starch granules by the protein slowed down the digestion and reduced the final hydrolysis rate, which are beneficial for people with weight and glycemic control issues. In conclusion, this study contributes to the production of tasty, customized sweet dumplings.

Bioenergy recovery and carbon emissions benefits of short-term bio-thermophilic pretreatment on low organic sewage sludge anaerobic digestion: A pilot-scale study.

Sewage sludge in cities of Yangzi River Belt, China, generally exhibits a lower organic content and higher silt contentdue to leakage of drainage system, which caused low bioenergy recovery and carbon emission benefits in conventional anaerobic digestion (CAD). Therefore, this paper is on a pilot scale, a bio-thermophilic pretreatment anaerobic digestion (BTPAD) for low organic sludge (volatile solids (VS) of 4%) was operated with a long-term continuous flow of 200 days. The VS degradation rate and CH4 yield of BTPAD increased by 19.93% and 53.33%, respectively, compared to those of CAD. The analysis of organic compositions in sludge revealed that BTPAD mainly improved the hydrolysis of proteins in sludge. Further analysis of microbial community proportions by high-throughput sequencing revealed that the short-term bio-thermophilic pretreatment was enriched in Clostridiales, Coprothermobacter and Gelria, was capable of hydrolyzing acidified proteins, and provided more volatile fatty acid (VFA) for the subsequent reaction. Biome combined with fluorescence quantitative polymerase chain reaction (PCR) analysis showed that the number of bacteria with high methanogenic capacity in BTPAD was much higher than that in CAD during the medium temperature digestion stage, indicating that short-term bio-thermophilic pretreatment could provide better methanogenic conditions for BTPAD. Furthermore, the greenhouse gas emission footprint analysis showed that short-term bio-thermophilic pretreatment could reduce the carbon emission of sludge anaerobic digestion system by 19.18%.

Effect of lysozyme combined with hydrothermal pretreatment on excess sludge and anaerobic digestion.

Anaerobic digestion (AD) is widely employed for sludge stabilization and waste reduction. However, the slow hydrolysis process hinders methane production and leads to prolonged sludge issues. In this study, an efficient and eco-friendly lysozyme pre-treatment method was utilized to address these challenges. By optimizing lysozyme dosage, hydrolysis and cell lysis were maximized. Furthermore, lysozyme combined with hydrothermal pretreatment enhanced overall efficiency. Results indicate that: (1) When lysozyme dosage reached 90 mg/g TS after 240 min of pretreatment, SCOD, soluble polysaccharides, and protein content reached their maxima at 855.00, 44.09, and 204.86 mg/L, respectively. This represented an increase of 85.87%, 365.58%, and 259.21% compared to the untreated sludge. Three-dimensional fluorescence spectroscopy revealed the highest fluorescence intensity in the IV region (soluble microbial product), promoting microbial metabolic activity. (2) Lysozyme combined with hydrothermal pretreatment significantly increased SCOD, soluble proteins, and polysaccharide release from sludge, reducing SCOD release time. Orthogonal experiments identified Group 3 as the most effective for SCOD and soluble polysaccharide release, while Group 9 released the most soluble proteins. The significance order of factors influencing SCOD, soluble proteins, and polysaccharide release is hydrothermal temperature > hydrothermal time > enzymatic digestion time.(3) The lysozyme-assisted hydrothermal pretreatment group exhibited the fastest release and the highest SCOD concentration of 8,135.00 mg/L during anaerobic digestion. Maximum SCOD consumption and cumulative gas production increased by 95.89% and 130.58%, respectively, compared to the control group, allowing gas production to conclude 3 days earlier.

Human milk fat globule delivers entrapped probiotics to the infant's gut and acts synergistically to ameliorate oxidative and pathogenic stress.

The human milk fat globule membrane (hMFGM) and Lactobacillus modulate the infant's gut and benefit health. Hence, the current study assesses the probiotic potential of Lactiplantibacillus plantarum (MRK3), Limosilactobacillus ferementum (MK1) isolated from infant feces, and its interaction with hMFGM during conditions mimicking infant digestive tract. Both strains showed high tolerance to gastrointestinal conditions, cell surface hydrophobicity, and strong anti-pathogen activity against Staphylococcus aureus. During digestion, hMFGM significantly exhibited xanthine oxidase activity, membrane roughness, and surface topography. In the presence of hMFGM, survival of MRK3 was higher than MK1, and electron microscopic observation revealed successful entrapment of MRK3 in the membrane matrix throughout digestion. Interestingly, probiotic-membrane matrix interaction showed significant synergy to alleviate oxidative stress and damage induced by cell-free supernatant of Escherichia coli in Caco-2 cells. Our results show that a probiotic-encapsulated membrane matrix potentially opens the functional infant formula development pathway.

Enhancing the High-Solid Anaerobic Digestion of Horticultural Waste by Adding Surfactants.

The influence of adding surfactants on the performance of high-solid anaerobic digestion of horticultural waste was extensively investigated in batch systems. Adding Tween series and polyethylene glycol series non-ionic surfactants had positive effects on biogas production, resulting in 370.1 mL/g VS and 256.6 mL/g VS with Tween 60 and polyethylene glycol 300 at a surfactant-to-grass mass ratio of 0.20, while the biogas production of anaerobic digestion without surfactants was 107.54 mL/g VS. The optimal and economically feasible choice was adding Tween 20 at a ratio of 0.08 g/g grass in high-solid anaerobic digestion. A kinetics model reliably represented the relationship between surfactant concentration and biogas production. The mechanism of surfactants working on lignocellulose was investigated. The improvement in high-solid anaerobic digestion by adding surfactants was attributed to the interaction between lignocelluloses and surfactants and the extraction of biodegradable fractions from the porous structure. An economic analysis showed that adding Tween 20 was likely to make a profit and be more feasible than adding Tween 60 and polyethylene glycol 300. This study confirms the enhancement in biogas production from horticultural waste by adding non-ionic surfactants.

Quantitation of Host Cell Proteins by Capillary LC/IMS/MS/MS in Combination with Rapid Digestion on Immobilized Trypsin Column Under Native Conditions.

Host cell protein (HCP) impurities are considered a critical quality attribute of biopharmaceuticals because of their potential to compromise safety and efficacy, and LC/MS-based analytical methods have been developed to identify and quantify individual proteins instead of employing enzyme-linked immunosorbent assay to assess total HCP levels. Native digestion enables highly sensitive detection of HCPs but requires overnight incubation to generate peptides, limiting the throughput of sample preparation. In this study, we developed an approach employing native digestion on a trypsin-immobilized column to improve the sensitivity and throughput. We examined suitable databases for the identification of HCPs derived from Chinese hamster ovary (CHO) cells and selected RefSeq's Chinese Hamster as the optimal database. Then, we investigated methods to identify HCPs with greater efficiency than that of denatured in-solution digestion. Native in-column digestion not only reduced the digestion time from overnight to 10 min but also increased the number of quantified HCPs from 154 to 226. In addition to this rapid digestion methodology, we developed high-throughput LC/MS/MS with a monolithic silica column and parallel reaction monitoring-parallel accumulation-serial fragmentation. The optimized system was validated with synthetic peptides derived from high-risk HCPs, confirming excellent linearity, precision, accuracy, and low limit of detection (LOD) and limit of quantification (LOQ) (1-3 ppm). The optimized digestion and analysis method enabled high-throughput quantification of HCPs, and is expected to be useful for quality control and characterization of HCPs in antibody drugs.

Influences of released humic acids during thermal hydrolysis on sludge anaerobic digestion: New insights from the molecular weight of humic acids.

Humic acids (HAs) would be excessively released during thermal hydrolysis pretreatment (THP) and deeply disturb anaerobic digestion (AD) of waste activated sludge (WAS). The molecular weights of HAs could affect HAs entering microbial cells, binding with digestive enzymes and participating in electron transfer, thereby determining its influences on sludge AD. Results in this study confirmed the different influences of HAs from diverse sources on sludge AD indeed had significant correlations with their molecular weights. The presence of commercial HAs (SAHA) inhibited methane production by 53.3% at 0.5 g/L while HAs extracted from raw sludge (WNHA) increased methane production by 20.5% at the same concentration, which attribute to the comprehensive impacts from their differences in functional group compositions and molecular weights. Moreover, comparing to WNHA, the HAs extracted from thermally hydrolyzed sludge (THHA) showed unchanged functional group compositions but reduced methane generation facilitation to 5.1%, which only be due to its decreased molecular weights. In-depth research indicated that HAs influences on enzymes were closely relative to its molecular weight. HAs with greater molecular weights presented more significant inhibition to extracellular enzymes while micromolecular HAs affected intracellular enzymes more. Furthermore, macromolecular HAs promoted sludge solubilization and acidification but hindered hydrolysis and methanogenesis, whereas micromolecular HAs promoted acidification but inhibited methanogenesis. This study underscored the importance of changes in molecular weight of HAs during sludge THP, offering insights into previous discrepancies in reports on HAs effects on sludge AD.

Anaerobic digestates in agricultural soils: A systematic review of their effects on antibiotic resistance genes.

Tackling the dissemination of antibiotic resistance is one of the main global challenges. Manures from animal production are a recognized source of antibiotic resistance genes (ARGs) and mobile genetic elements (MGEs) requiring appropriate treatment methods. One of the main approaches for manure treatment is anaerobic digestion (AD). Meta-analyses have demonstrated that AD can significantly reduce the load of ARGs. However, antibiotics, ARGs and MGEs still remain in the final product (digestate). A sustainable agricultural use of digestates under the One Health framework requires wide assessments of their effects in the soil resistome. The objective of this review was to present the state of the art of digestate effects on ARGs of agricultural soils, focusing exclusively on digestates from animal manures. A systematic review was conducted. The examination of the resulting literature indicated that although temporal decays are observed for a variety of ARGs in single-application and repeated-applications experiments, for certain ARGs the pre-treatment or control levels are not restored. However, the low number of studies and the heterogeneous experimental conditions preclude a clear understanding of the fate of ARGs in soil and their risk for agroecosystems. The inclusion of multiple MGEs and the assessment of the long-term influence of digestates on soil properties and microbial communities could be keystones for a better understanding of the risks associated with digestate-induced changes in the soil resistome.

Green tech soil analysis: a comparison of potentially toxic element contents among urban, agricultural, and gold mining areas in the Eastern Brazilian Amazon.

Artisanal gold mining can lead to soil contamination with potentially toxic elements (PTEs), necessitating soil quality monitoring due to environmental and human health risks. However, determining PTE levels through acid digestion is time-consuming, generates chemical waste, and requires significant resources. As an alternative, portable X-ray fluorescence (pXRF) offers a faster, more cost-effective, and sustainable analysis. This study compared total As, Ba, Cr, Cu, Fe, Mn, Ni, Pb, Sr, Ti, V, and Zn obtained from pXRF with their pseudo-total contents obtained through acid digestion (USEPA method 3051A) in areas influenced by artisanal gold mining in the Eastern Amazon, Brazil. pXRF data and machine learning algorithms were used to predict extractable Cu, Fe, Mn, and Zn. Linear regression models were fitted to compare the two methods, and random forest and support vector machine techniques were used to predict extractable contents. The best regression model fits for the pseudo-total PTE contents were those for Cu, Fe, Mn and Pb in agricultural areas (R2 > 0.80); Fe and Mn in gold mining residue (R2 > 0.70); and Ba, Cu and Mn in urban areas (R2 > 0.80). The best models for predicting the extractable PTE contents were those for Cu (R2 = 0.72; RMSE = 2.58 mg dm-3) and Zn (R2 = 0.71; RMSE = 1.44 mg dm-3) in agricultural areas and for Zn (R2 = 0.72; RMSE = 0.43 mg dm-3) in gold mining residue. The results demonstrated that pXRF can characterize and predict PTE contents in mining-impacted areas, offering a sustainable approach to soil quality analysis.

A new paradigm for whole-chain low-carbon utilization of food waste secondary waste based on multivariate evaluations.

The rapid growth of China's food and beverage industry has led to a significant increase in food waste, presenting major challenges for its disposal. Anaerobic digestion is the primary treatment method, but its by-products-biogas slurry (BS) and biogas residue (BR)-pose additional treatment challenges. This study proposes and evaluates three management scenarios for these by-products: (1) BS sewage treatment with BR incineration (S1), (2) BS land application with BR composting (S2), and (3) BS sewage treatment with BR composting (S3). The scenarios were comprehensively assessed using material flow analysis, life cycle assessment, and economic benefit analysis. The findings show that S2 achieves the highest carbon utilization efficiency (80.16%) and delivers superior environmental and economic benefits. Scaling up S2 by 2030 could reduce national carbon emissions by 1817.8 kilotons and generate $11.14 billion in economic profit. This study offers a novel model for the sustainable, low-carbon utilization of food waste by-products, providing valuable insights for future organic waste management.

Metabolic pathway analysis of methane from methanol as substrate in microbial consortium.

Simplified anaerobic digestion (SAD) of substrates facilitates microbial methanogenic pathways. In this study, a methane-producing microbial consortium from cow dung was enriched to determine the metabolism and metabolic pathway in the SAD of methanol. The results showed that methanol as a sole substrate produced 167 mL of methane at 10 days significantly higher than 58 mL, 17.6 mL, and 4 mL generated when methanol was combined with sodium formate, sodium formate alone, or sodium acetate. The relative abundance of Methanobacterium, Candidatus_Methanomethylophilus, Methanomassiliicoccus, and Methanosarcina was increased by 5.96 %, 3.77 %, 2.85 %, and 0.14 % in the methanol substrate of AD, respectively. Macrogenome sequencing indicates that methanol is converted into Methyl-CoM in the presence of Methanosarcina, which combines with Coenzyme B to produce methane. This study revealed that methanol is converted into methane by a simple pathway.

Response of food waste anaerobic digestion to the dimensions of micron-biochar under 30 g VS/L organic loading rate: Focus on gas production and microbial community structure.

Biochar modification is an effective approach to enhance its ability to promote anaerobic digestion (AD). Focusing on the physical properties of biochar, the impact of different particle sizes of biochar on AD of food waste (FW) at high organic loading rate (OLR) was investigated. Four biochar with different sizes (40-200 mesh) were prepared and used in AD systems at OLR 30 g VS/L. The research results found that biochar with a volume particle size of 102 µm (RBC-P140) had top-performance in promoting cumulative methane production, increasing by 13.20% compared to the control group. The analysis results of the variety in volatile acids and alkalinity in the system did not show a correlation with the size of biochar, but small size has the potential to improve the environmental tolerance of the system to high acidity. Microbial community analysis showed that the abundance of aceticlastic methanogen and the composition of zoogloea were optimized through relatively small-sized biochar. Through revealing the effect of biochar particle size on AD system at high OLR, this work provided theoretical guidance for regulating fermentation systems using biochar.

Mechanistic insights into Fe3O4-mediated inhibition of H2S gas production in sludge anaerobic digestion.

The addition of iron-based conductive materials has been extensively validated as a highly effective approach to augment methane generation from anaerobic digestion (AD) process. In this work, it was additionally discovered that Fe3O4 notably suppressed the production of hazardous H2S gas during sludge AD. As the addition of Fe3O4 increased from 0 to 20 g/L, the accumulative H2S yields decreased by 89.2 % while the content of element sulfur and acid volatile sulfide (AVS) respectively increased by 55.0 % and 30.4 %. Mechanism analyses showed that the added Fe3O4 facilitated sludge conductive capacity, and boosted the efficiency of extracellular electron transfer, which accelerated the bioprocess of sulfide oxidation. Although Fe3O4 can chemically oxidize sulfide to elemental sulfur, microbial oxidation plays a major role in reducing H2S accumulation. Moreover, the released iron ions reacted with soluble sulfide, which promoted the chemical equilibrium of sulfide species from H2S to metal sulfide. Microbial analysis showed that some SRBs (i.e., Desulfomicrobium and Defluviicoccus) and SOB (i.e., Sulfuritalea) changed into keystone taxa (i.e., connectors and module hubs) in the reactor with Fe3O4 addition, showing that the functions of sulfate reduction and sulfur oxidation may play important roles in Fe3O4-present system. Fe3O4 presence also increased the content of functional genes encoding sulfide quinone reductase and flavocytochrome c sulfidedehydrogenase (e.g., Sqr and Fcc) that could oxidize sulfide to sulfur. The impact of other iron-based conductive material (i.e., zero-valent iron) was also verified, and the results showed that it could also significantly reduce H2S production. These findings provide new insights into the effect of iron-based conductive materials on anaerobic process, especially sulfur conversion.

Detailed Analysis of Prebiotic Fructo- and Galacto-Oligosaccharides in the Human Small Intestine.

Galacto-oligosaccharides (GOS) and fructo-oligosaccharides (FOS) are food ingredients that improve human health, but their degradation throughout the human small intestine is not well understood. We studied the breakdown kinetics of FOS and GOS in the intestines of seven healthy Dutch adults. Subjects were equipped with a catheter in the distal ileum or proximal colon and consumed 5 g of chicory-derived FOS (degree of polymerization (DP) DP2-10), and 5 g of GOS (DP2-6). Postprandially, intestinal content was frequently collected until 350 min and analyzed for mono-, di-, and oligosaccharides. FOS and GOS had recoveries of 96 ± 25% and 76 ± 28%, respectively. FOS DP ≥ 2 and GOS DP ≥ 3 abundances in the distal small intestine or proximal colon matched the consumed doses, while GOS dimers (DP2) had lower recoveries, namely 22.8 ± 11.1% for ß-D-gal-(1↔1)-α-D-glc+ß-D-gal-(1↔1)-ß-D-glc, 19.3 ± 19.1% for ß-D-gal-(1 → 2)-D-glc+ß-D-gal-(1 → 3)-D-glc, 43.7 ± 24.6% for ß-D-gal-(1 → 6)-D-gal, and 68.0 ± 38.5% for ß-D-gal-(1 → 4)-D-gal. Lactose was still present in the distal small intestine of all of the participants. To conclude, FOS DP ≥ 2 and GOS DP ≥ 3 were not degraded in the small intestine of healthy adults, while most prebiotic GOS DP2 was hydrolyzed in a structure-dependent manner. We provide evidence on the resistances of GOS with specific ß-linkages in the human intestine, supporting the development of GOS prebiotics that resist small intestine digestion.

In vitro digestion study comparing a predigested glycerolysis product versus long-chain polyunsaturated fatty acid-rich oils (LCPUFA) as a strategy for administering LCPUFA to preterm neonates.

BACKGROUND & AIMS: Maintaining an adequate supply of arachidonic acid (ARA) and docosahexaenoic acid (DHA) is essential for optimal growth of preterm infants. This study aims to evaluate and compare the digestibility and bioaccessibility of ARA and DHA oils compared to their predigested product through an in vitro digestion model. METHODS: An in vitro gastrointestinal digestion model was used in two stages: gastric digestion and intestinal digestion. Samples of two polyunsaturated rich oils (ARA and DHA oils) and their predigested product (2:1, ARA: DHA) produced by enzymatic glycerolysis have been digested for 120 min. The final digestion product obtained was composed of three phases: an upper oily phase (OP) containing the undigested species, an intermediate micellar phase (MP) containing digested and bioaccessible lipids, and a precipitate phase (PP) with insoluble compounds. The reaction was monitored by taking aliquots and their subsequent lipid extraction and analysis. RESULTS: Poorer digestibility for ARA and DHA oils was observed based on the percentage of the oily phase (26.7% and 20%, respectively) found compared to the glycerolysis product (GP) oily phase (13.9%). The highest micellar phase was found in the GP (approx. 83%). On the other hand, the monoglyceride (MAG) content was lower in the digestion product (DP) from ARA and DHA oils, 4.3% and 9.2%, respectively, compared to the MAG observed in the DP of GP (15%). CONCLUSION: Considering the percentage of oily phase, micellar phase, and the MAG content, it can be concluded that the GP is more digestible and ARA and DHA are more bioaccessible than in its precursor oils.

Anti-solvent precipitation for encapsulation of oyster protein hydrolysate nanoparticles: Effect on off-flavor elimination and bioaccessibility.

In this study, to investigate the potential for delivering bioactive substances, various cereal proteoglycan nanoparticles were prepared using anti-solvent precipitation for the delivery of oyster protein hydrolysate (OPH). The hydrogen bonding and hydrophobic interaction were the driving forces for OPH embedding. OPH-Loaded Zein/NaCas/Gum Arabic composite nanoparticles could self-assemble into spherical (191 nm) particles with strong stability, uniform distribution (about PDI = 0.2), and high negative charge (about -28 mV). The secondary structure of OPH-Loaded zein was converted from α-helix to random coil with heat treatment. The off-flavor, including (E, E)-2,4-heptadienal, octadecane, and 1-octanol in OPH-Loaded Zein nanoparticles significantly decreased. Furthermore, these nanoparticles containing OPH had excellent oxidation resistance, in vitro digestibility, and non-toxicity in Caco-2 and HT-29 cells. Overall, this work provides an efficient strategy for OPH delivery, suggesting that OPH could be ingested as a nutraceutical in powder form while also promoting the development of high-flavor-quality oyster products.

Simultaneous Production of Biogas and Electricity from Anaerobic Digestion of Pine Needles: Sustainable Energy and Waste Management.

Power scarcity and pollution can be overcome with the use of green energy forms like ethanol, biogas, electricity, hydrogen, etc., especially energy produced from renewable and industrial feedstocks. In hilly areas, pine needles are the most abundant biomass that has a low possibility of valorization due to high lignin content. On the other hand, anaerobic digestion (AD) of lignin and animal waste has low biogas yield due to poor conductivity. This study focuses on the simultaneous production of biogas and electricity through the co-digestion of cow dung and pine needles. The digester was initially established and stabilized in the lab to ensure a continuous supply of inoculum throughout the experiment. The optimization process involved the determination of an ideal cow dung-to-water ratio and selecting the appropriate conductive material that can enhance the energy generation from the feedstock. Afterward, both batch and continuous anaerobic digestion experiments were conducted. The results revealed that the addition of powdered graphite (5 mM), activated charcoal (15 mM), and biochar (25 mM) exhibited maximum voltage of 0.71 ± 0.013 V, 0.56 ± 0.013 V, and 0.49 ± 0.011 V on the 30th, 25th and 20th day of AD, respectively. The batch experiment showed that 5 mM graphite powder enhanced electron transfer in the AD process and generated a voltage of 0.77 ± 0.014 V on the 30th day, indicating an increase of ~1.5-fold as compared to the control (0.56 ± 0.019 V). The results from the continuous AD process showed that the digester with cow dung, pine needle, and a conductive material in combination exhibited the maximum voltage of 0.76 ± 0.012 V on the 21st day of AD, while the digester with cow dung only exhibited a maximum voltage of 0.62 ± 0.015 V on the 22nd day of AD, representing a 1.3-fold increase over the control. Furthermore, the current work used discarded plastic items and electrodes from spent batteries to emphasize waste management and aid in attaining sustainable energy and development goals.

Comparison of in vitro digestive characteristics of proteins from different sources in simulated elderly gastrointestinal conditions.

This study aimed to evaluate the impact of in vitro digestive characteristics of five different protein sources (fish, crab, beef, chicken, and soy) in adults and elderly individuals. Simulated gastrointestinal tract degradation in the elderly significantly reduced the digestibility of all proteins. The Elderly model had difficulties digesting any protein that had a firm structure or was prone to aggregation. Of the five protein types, soy protein was the least easily digested. Ultraviolet spectroscopy and SDS-PAGE analysis revealed that beef and crab proteins contain myoglobin and hemocyanin, respectively, making their structure firm. Simultaneously, alkali-soluble proteins in meat were more likely to be digested by the elderly than salt-soluble proteins. The alkali-soluble protein percentage in fish protein was the highest, including the best in vitro digestion characteristics in the elderly population. This study provides novel and significant recommendations for the elderly's daily intake of various protein sources.

Protein-added kefir: biochemical changes in in vitro digestion stages.

BACKGROUND: While yogurt is the leading fermented milk product, kefir is at the top of the beverage scale. Milk proteins, on the other hand, show specific functions that positively affect healthy nutrition due to the bioactive components, that they provide the necessary amino acids for growth and development. RESULTS: In our study, kefir, a functional product enriched with whey proteins, casein and skimmed milk powder, which are the natural components of milk, was produced. Added-protein kefir samples were applied the in vitro digestion protocol, static method. In order to observe different protein behaviors, samples were taken pre-digestion, at 120th minute and at 240th minute of digestion protocol. ACE and Antioxidant capacity determination analyzes were carried out. While ACE inhibition values were in the range of 78.63-90.30% pre-digestion, they changed in the range of 86.97-96.38% after gastrointestinal digestion. It was determined that the ACE inhibition values of the control sample remained at the lowest level at all stages of digestion and that the difference between all of samples was significant (P < 0.05). Antioxidant activity value ranging from 0.3615-0.5512 meq Ascorbic acid/µg before digestion was determined as 1.3796-1.9313 meq Ascorbic acid/µg after gastrointestinal digestion (P < 0.05). CONCLUSION: Kefir samples containing whey protein stand out with their high potential in terms of both antioxidant activity capacity and ACE inhibition activity at all stages of digestion. Considering their therapeutic effects in fermented products, it is thought that whey proteins among milk proteins will be important alternative sources to enrich the protein content in kefir production. © 2024 Society of Chemical Industry.