ABSTRACT This study aimed to comprehensively analyze lipid classes after in vitro digestion of high‐fat commercial dairy products. The research focused on the fatty acid (FA) profiles of triacylglycerols, diacylglycerols, monoacylglycerols (MAG), free FA (FFA), and polar lipids of digested full‐fat cream, full‐fat sour cream, cream cheese, and a Gouda‐type cheese. We employed the INFOGEST 2.0 digestion model. The isolated lipids were fractionated by solid‐phase extraction, and the FA profile of each fraction was analyzed using GC–MS. The digested dairy products revealed similar results for the relative amounts of the analyzed lipid classes. However, the cream digest had a significantly lower amount of MAG than the other products. Furthermore, the proportion of individual FAs and FA groups in the lipid classes was found to vary significantly between the digested dairy products. The replicates of cream cheese and sour cream showed a homogenous lipid digestion, whereas the lipid classes after digestion of cream and the Gouda‐type cheese were more diverse. This could be due to differences in product pH and protein and calcium content. Practical Applications : The findings of our study indicate the importance of considering lipid classes other than FFA, as well as the FA composition of these, when studying the lipid digestion of different dairy products. This is especially true for MAG, which is absorbed in the intestine along with FFA. The observed differences in the lipid profile of the digested dairy matrices explored in our study demonstrate that product pH, homogenization, texture, and calcium and protein content could impact lipid bioaccessibility, which could potentially have physiological implications. These factors should, therefore, be assessed in relation to lipid digestion.

This study focused on identifying amylase-trypsin inhibitors (ATIs) in seven Norwegian-cultivated wheat varieties, including common wheat and ancestral species, and identifying potentially harmful opioid peptides within the ex vivo digesta of these wheats. LC-MS/MS analysis of tryptic peptides from ATI fractions revealed that the common wheat variety B & oslash;rsum exhibited the highest diversity of ATIs (n = 24), while they were less represented in tetraploid emmer (n = 11). Hexaploid wheat Bastian showed low diversity and relative abundance of ATIs. Nevertheless, digestion of Mirakel and Bastian by human gastrointestinal juices released the highest number of opioid-containing peptides, representing both gluten exorphins and gliadorphin. In conclusion, emmer had the lowest levels of ATIs, while einkorn and spelt released the fewest opioid-containing peptides after ex vivo digestion. These results point to the potential lower harmful effects of ancestral wheat compared to common hexaploid wheat varieties for wheat-sensitive individuals.

Kombucha, a fermented tea beverage, is gaining popularity due to its rich content of bioactive compounds and associated health benefits. Kombucha fermentation involves a complex microbial consortium, including acetic acid bacteria, lactic acid bacteria, and yeasts, that works synergistically to enhance its nutritional and functional properties. Key compounds produced during fermentation provide antioxidant, anti-inflammatory, and antimicrobial benefits. Despite its well-documented health-promoting properties, limited research exists on how human digestion influences the stability and functionality of kombucha bioactive components. This study investigated how digestion impacts kombucha made from green and black teas, focusing on free amino acid content, antioxidant activity, antimicrobial potential, and microbiota viability. Results showed that digestion significantly increased free amino acids, as fermentation released peptides suitable for gastrointestinal digestion. However, L-theanine, a beneficial tea compound, was no longer detectable after fermentation and digestion, suggesting limited bioaccessibility. Digested kombucha exhibited higher antioxidant activity and stronger antimicrobial effects compared to undigested tea. Moreover, culture-dependent and PMA-based sequencing confirmed the survival of viable microbial strains through simulated gastrointestinal conditions, suggesting the potential of kombucha as a source of live, functional microbes. These findings support the role of kombucha as a natural functional beverage whose health benefits not only persist but may be enhanced after digestion.

Lactic acid bacteria (LAB) that are autochthonous to the raw material are commonly recognised as the most suitable bacteria for starter cultures for fermentation. In this study, 172 LAB isolated from cereals and pulses were genetically characterised, and 37 of these were whole genome sequenced. Twenty-four LAB were selected for technological characterization. Screening assays were set up to determine the ability of these isolates to produce exopolysaccharides and hydrolyse proteins, reduce phytic acid, and ferment various carbohydrate substrates. The ability to grow and acidify was tested in flour-like media and flour slurries. Chromatographic methods were applied to quantify carbohydrates and organic acids before and after fermentation. The 24 isolates belonging to 8 different species of LAB exhibited a diverse combination of important functional and technological properties. Isolates with the ability to rapidly reduce pH, proliferate at a high rate, produce large amounts of organic acids and metabolise citrate were found among the 24 candidates. The isolates displayed different combinations of carbohydrate fermentation ability, proteolytic activity, phytase activity, and formation of mucoid and threading colonies. Several isolates exhibited one or more desirable traits, with one expressing all the desired properties tested here; Lpb. paraplantarum JP51. These results provide insights into the potential of these isolates for developing autochthonous starter cultures.

This study explores the impact of somatic cell count (SCC) and lactation stage on the coagulation properties and milk composition of Norwegian goat milk. Monthly sampling of forty goats throughout lactation assessed rennet coagulation time (RCT), curd firming time (k20), curd firmness (a30), and milk components such as lactose, protein, and minerals. Results demonstrated that SCC levels above 2000 x 103 cells mL-1 significantly delayed RCT and reduced a30, while k 20 remained unaffected. A strong correlation was observed between SCC, milk composition (such as lactose and minerals), and coagulation properties, with elevated SCC linked to impaired curd formation and increased protein degradation. The lactation stage also influenced milk quality, with mid-lactation showing poorer coagulation performance, particularly during mountain grazing. These findings underscore the critical role of monitoring SCC and lactation stage to maintain optimal coagulation properties and ensure stable cheese production.

Somatic cell count is used by the dairy industry as an indicator of milk quality and udder health. However, in goats, its reliability is significantly masked by noninfectious variables such as the milk secretion process and physiological lactation changes. Additionally, notable individual variability between goats might exist. This study aimed to investigate fluctuations in SCC in individual goats during an entire lactation and to examine the relationship between SCC and milk parameters such as bacterial count and chemical composition. Individual milk samples from 40 Norwegian dairy goats from the university herd were collected monthly across an entire lactation including the pasture period. The goats were categorized based on SCC levels to analyze patterns in chemical components within these groups. Notably, goats exhibited increased SCC and decreased bacterial counts when moved to summer grazing pastures. At that stage, milk samples from goats with the highest SCC (>2,000 x 10(3) cells/mL) showed a distinct decrease in individual bacterial counts. The SCC level affected several milk components (especially lactose, protein, and various minerals), as well as pH. These effects were further amplified when considering the interaction with the lactation stage, influencing a broader range of variables. The results presented in this study provide new insights into the effect of SCC on milk composition and its critical role. These insights underscore the necessity for establishing lactation-specific thresholds for interpretation, as well as for making adjustments in quality payments.

Wine protein haze formation is a problem due to grape proteins aggregation during wine storage. The cell wall components of wine yeasts, particularly high molecular weight mannoproteins, have a protective effect against haze formation, although their involvement remains poorly understood. This study aimed at characterizing glycosylated proteins released by Starmerella bacillaris and Saccharomyces cerevisiae during single and sequential fermentations in a synthetic must, and testing their impact on wine protein stability. Mannoproteins-rich extracts from sequential fermentations showed an increase in the low MW polysaccharide fraction and, when added to an unstable wine, had a greater effect on protein stability than S. cerevisiae extracts. Shotgun proteomics approaches revealed that the identified cell wall proteins exclusively found in sequential fermentations were produced by both S. bacillaris (MKC7, ENG1) and S. cerevisiae (Bgl2p). Moreover, sequential fermentations significantly increased the expression of Scw4p and 1,3-beta-glucanosyltransferase (GAS5), produced by S. cerevisiae. Finally, some of the key proteins identified might play a positive role in increasing wine protein stability.

Increasing plant-based food consumption as a sustainable and health-oriented alternative to meat is pivotal. Pulses are rich in proteins, minerals, and vitamins; however, they also contain antinutritional compounds, impairing their nutritional value. This study addresses this challenge through the development and application of four distinct microbial consortia in pulse-based fermentations, featuring lactic acid bacteria or a combination of lactic and propionic acid bacteria. Microbial starters significantly reduced galacto-oligosaccharides in all pulse materials, concurrently degrading vicine and convicine in faba beans, while the impact on tannins in faba beans and lentil was moderate. Fermentation with lactic acid and propionic acid bacteria consortia exhibited notable vitamin B12 production, and the effect on the content of phenolic compounds of the studied pulses was also evidenced. Additionally, genomic analyses discerned distinctive profiles among the samples, elucidating the microbial community dynamics shaping fermentation outcomes. The results of this study proved how fermentation can advance the development of pulse-based products with improved nutritional and sustainability attributes.

Sourdough breadmaking on an industrial scale requires robust, well-performing starters that bring attractive characteristics to the product. The active nature of liquid starters provides a faster fermentation process compared to their dehydrated counterparts. However, liquid sourdough starters require meticulous management in order to maintain stability and functionality during cold storage at 4 °C.This study investigated the stability of three liquid sourdough starters during storage and also the impact of prolonged fermentation, the addition of diastatic malted wheat flour, and a neutralising agent (CaCO3).The sourdough starters were evaluated for their microbial viability and metabolic activity at three individual time points during 16 weeks of cold storage. The microbial composition was analysed using culture-dependent and culture-independent methods, and metabolic changes were investigated using chromatographic methods.Two types of sourdough starter showed satisfying viability of lactic acid bacteria (> 7 log CFU/g) and metabolic stability throughout 16 weeks of cold storage. The introduction of malted wheat flour and CaCO3 caused a decline in viability to <7 log CFU/g within 8 weeks in the third sourdough starter type and additionally revealed an ongoing metabolic activity of this sourdough starter during cold storage.Prolonged fermentation influenced the free amino acid profile, whereas adjusting the sourdough starter formula resulted in a different fungal microbiota and increased levels of fermentable substrates (maltose), organic acids (lactic acid), and aromatic compounds (alcohol and aldehydes).These findings provide stakeholders and researchers in sourdough fermentation technology with new insights concerning the stability of cold-stored liquid sourdough starters.

UHT milk is stable towards spoilage for months, but chemical and enzymatic reactions occur during storage. This study evaluated whether changes over storage (at 4, 20 or 30 degrees C, for 30-90 d) of regular as well as lactose-reduced UHT milk were discernible by untrained and trained sensory panelists, and which physicochemical parameters they were associated with. Most changes were of low magnitude, such as significant increases in non-protein nitrogen (in both milk types) and lactulose (in regular UHT milk). However, the color of lactose-reduced UHT milk noticeably changed due to browning. Trained panelists evaluated lactose-reduced samples stored at 30 degrees C as significantly different in color than those stored at lower temperatures and untrained panelists correctly distinguished between fresh and stored lactose-reduced UHT milk in a tetrad-test already after 30 d. Further research is needed to evaluate if consumers detect such changes and therefore dispose of UHT- milk.

Ribulose-1,5-bisphosphate-carboxylase/oxygenase (RuBisCO) from alfalfa is a potentially climate-friendly alternative protein with a promising amino acid composition. The balance between yield and purity is a challenge for alternative plant proteins, partly due to the naturally occurring antinutrients. Therefore, measuring the in vitro protein digestibility (IVPD) of RuBisCO with various purity levels is of interest. It was hypothesized that the digestibility of RuBisCO from alfalfa might vary with different processing histories and levels of refinement. To test this hypothesis, RuBisCO from alfalfa with 4 different processing histories were subjected to the INFOGEST IVPD protocol and measurement of free N-terminals and peptidomics. The result showed that the digestibility of RuBisCO was high regardless of the processing history and purity, as demonstrated by 77-99% sequence coverage in the gastric phase. In intestinal phase, increase of free N-terminals and lower sequence coverage (< 10%) indicated that the proteins were hydrolyzed to smaller peptides.

Six cattle breeds native to Norway, have for almost half a century been at risk of extinction. Due to their small population sizes, they have hardly been improved by breeding for many decades. Still, the endangered breeds represent a source of genetic diversity with special milk qualities compared to the modern breed, Norwegian red (NRF). This study reports for the first time a detailed overview of their milk composition. Milk from seven native breeds, in total 200 individuals, were included in the study. Rare genetic variants of αs1- and αs2-casein, and β-casein A1 and κ-casein B were more prevalent in milk form the endangered breeds compared to NRF. Moreover, milk from these six breeds showed better renneting properties and lower incidences of non-coagulating milk, compared to the NRF milk, which showed better acid coagulation properties. This study shows the potential for native breeds in small-scale production of high-quality rennet cheeses.

Fermentation has recently been rediscovered as an attractive technique to process legumes, as it can improve the nutritional quality and value of the end product. This study investigated the dynamics and stability of the microbial communities in spontaneously fermented sourdoughs made from flours of two cultivars of faba beans and two cultivars of peas. Sourdoughs were established by the backslopping technique, and the microbial development at 22 degrees C and 30 degrees C was followed by culture dependent and culture independent methods. The utilization of substrates and formation of metabolites were also determined by high-performance liquid chromatography. A stable pH was reached in all the sourdoughs after 11-15 days of daily backslopping. Lactic acid bacteria and yeast from pH stable sourdoughs were isolated, characterized and identified. The fermentation temperature influenced the development of the microbial community and the substrate utilization during spontaneous fermentation. In the 30 degrees C fermentations, one species dominated (Lactiplantibacillus plantarum/pentosus), a lower pH was achieved, and the available substrates were more extensively converted. The 22 degrees C fermentation resulted in a more diverse microbial community (Lactiplantibacillus, Leuconostoc, Pediococcus), a higher pH, and more residual substrates were available after fermentation. Yeasts were only detected in one of the pea sourdoughs fermented at 30 degrees C, with Saccharomyces cerevisiae being the dominant species. Nearly all sourdoughs were depleted of maltose after 24 h fermentation cycles, and higher levels of lactic and acetic acid were detected in 30 degrees C fermen-tations. This research adds to our understanding of the autochthonous microbial community present in faba beans and peas as well as their natural capacity to establish themselves and ferment legume flours. These findings enhance the possibilities of utilizing and improving plant based protein sources.

The biotechnological reuse of winery by-products has great potential to increase the value and sustainability of the wine industry. Recent studies revealed that yeast biomass can be an exciting source of bioactive peptides with possible benefits for human health, and its incorporation in plant-based foods is considered innovative and sustainable. In this study, we aimed to identify, through in silico analyses, potential bioactive peptides from yeast extracts after in vitro digestion. Wine lees from a non-Saccharomyces oenological yeast, Starmerella bacillaris FRI751, Saccharomyces cerevisiae EC1118, and sequential fermentation performed with both strains (SEQ) were recovered in a synthetic must. Cellular pellets were enzymatically treated with zymolyase, and the yeast extracts were submitted to in vitro gastrointestinal digestions. LC-MS/MS sequenced the hydrolyzed peptides, and their potential bioactivity was inferred. S. bacillaris FRI751 fermentation showed 132 peptide sequences, S. cerevisiae EC1118 60, SEQ 89. A total of 243 unique peptide sequences were identified across the groups. Furthermore, based on the peptide sequence, the FRI751 extract showed the highest potential antihypertensive with 275 bioactive fragments. Other bioactivities, such as antimicrobial and immunomodulatory, were also identified in all yeast extracts. A potential antiobesity bioactive peptide VVP was identified only in the yeast extract from S. bacillaris single strain. The wine lees from S. bacillaris single strain and SEQ fermentation are a richer source of potential bioactive peptides than those from S. cerevisiae fermentation. This study opens new possibilities in the valorization of winemaking by-products.

We investigated the digestive peptide profiles of pea and faba bean proteins to evaluate possible bioactive peptides (BAPs), and to test immune-modulating properties of selected peptides using in vitro cell model. More than 400 unique peptides were released upon gastrointestinal digestion of pea and faba bean proteins and identified by high-resolution mass spectrometry (HPLC-ESI-MS/MS). Among these, 24 peptides were potentially bioactive as predicted by in silico analysis. Also, up to 275 recognized BAPs and eight known allergens were identified within the peptide sequences of the digests. Four selected peptides significantly decreased the IL-8 response in Caco-2 cells, with a specific pea peptide, DKPWWPK, reducing the IL-8 response up to 40 % during IL-1β induction. Combined with other nutritional benefits, the immune-modulatory properties of legume proteins point to a regular introduction of legumes in the diet, as they represent a still scarcely exploited food category, especially in the Nordic countries.

Several factors influence the composition of milk. Among these, genetic variation within and between cattle breeds influences milk protein composition, protein heterogeneity, and their posttranslational modifications. Such variations may further influence technological properties, which are of importance for the utilization of milk into dairy products. Furthermore, these potential variations may also facilitate the production of differentiated products (e.g., related to specific breeds or specific genetic variants). The objective of this study was to investigate the genetic variation and relative protein composition of the major proteins in milk from 6 native Norwegian dairy breeds representing heterogeneity in geographical origin, using the modern Norwegian breed, Norwegian Red, as reference. In total, milk samples from 144 individual cows were collected and subjected to liquid chromatography-electrospray ionization/mass spectrometry-based proteomics for identification of genetic and posttranslational modification isoforms of the 4 caseins (αS1-CN, αS2-CN, β-CN, κ-CN) and the 2 most abundant whey proteins (α-lactalbumin and β-lactoglobulin). Relative quantification of these proteins and their major isoforms, including phosphorylations of αS1-CN and glycosylation of κ-CN, were determined based on UV absorbance. The presence and frequency of genetic variants of the breeds were found to be very diverse and it was possible to identify rare variants of the CN, which, to our knowledge, have not been identified in these breeds before. Thus, αS1-CN variant D was identified in low frequency in 3 of the 6 native Norwegian breeds. In general, αS1-CN was found to be quite diverse between the native breeds, and the even less frequent A and C variants were furthermore detected in 1 and 5 of the native breeds, respectively. The αS1-CN variant C was also identified in samples from the Norwegian Red cattle. The variant E of κ-CN was identified in 2 of the native Norwegian breeds. Another interesting finding was the identification of αS2-CN variant D, which was found in relatively high frequencies in the native breeds. Diversity in more common protein genetic variants were furthermore observed in the protein profiles of the native breeds compared with milk from the high-yielding Norwegian Reds, probably reflecting the more diverse genetic background between the native breeds.

The demand for sustainably produced proteins is increasing with the world population and is prompting a dietary shift toward plant sourced proteins. Vegetable proteins have lower digestibility and biological value compared to animal derived counterparts. We explored sprouting of chickpea seeds as a strategy for improving digestibility. Protein evolution associated with by the sprouting process was assessed by proteomics. The sprouting induced breakdown of seed storage proteins and doubled the release of free alpha-amino nitrogen in sprouted chickpea flour. During sprouting, several enzymes involved in plant development were newly expressed. An ex vivo model of gastroduodenal and jejunal digestion was applied to assess the bioaccessibility of the protein digests. Proteins from chickpea sprouts showed a greater susceptibility to digestion with a 10% increase in alpha amino nitrogen. Peptides with potential immunoreactivity or bioactivity were catalogued in both digested chickpea sprouts and seeds using an in-silico approach. Peptides belonging to the non-specific transfer proteins, which are allergens in pulses, and peptides belonging to an IgE-binding hemagglutinin protein could only be identified in the digested chickpea sprouts. The observation collected paved the way to immune-based evaluations to assess the effect of germination on the allergenic potential.