Application of Strain Selection Technology in Alcoholic Beverages: A Review.

The diversity of alcohol beverage microorganisms is of great significance for improving the brewing process and the quality of alcohol beverage products. During the process of making alcoholic beverages, a group of microorganisms, represented by yeast and lactic acid bacteria, conducts fermentation. These microorganisms have complex synergistic or competitive relationships, and the participation of different microorganisms has a major impact on the fermentation process and the flavor and aroma of the product. Strain selection is one of the key steps. Utilizing scientific breeding technology, the relationship between strains can be managed, the composition of the alcoholic beverage microbial community can be improved, and the quality and flavor of the alcoholic beverage products can be increased. Currently, research on the microbial diversity of alcohol beverages has received extensive attention. However, the selection technology for dominant bacteria in alcohol beverages has not yet been systematically summarized. To breed better-quality alcohol beverage strains and improve the quality and characteristics of wine, this paper introduces the microbial diversity characteristics of the world's three major brewing alcohols: beer, wine, and yellow wine, as well as the breeding technologies of related strains. The application of culture selection technology in the study of microbial diversity of brewed wine was reviewed and analyzed. The strain selection technology and alcohol beverage process should be combined to explore the potential application of a diverse array of alcohol beverage strains, thereby boosting the quality and flavor of the alcohol beverage and driving the sustainable development of the alcoholic beverage industry.

Endophytic fungi in buckwheat seeds: exploring links with flavonoid accumulation.

Buckwheat is a famous edible and medicinal coarse cereal which contain abundant of bioactive flavonoids, such as rutin. In this study, the composition and diversity of endophytic fungi in eight different buckwheat seeds were analyzed by high-throughput sequencing of ITS rDNA. Results showed that, the fungal sequences reads were allocated to 272 OTUs, of them, 49 OTUs were shared in eight buckwheat seeds. These endophytic fungi could be classified into 6 phyla, 19 classes, 41 orders, 79 families, 119 genera, and 191 species. At genus level, Alternaria sp. was the domain fungal endophyte. Besides, fungal endophytes belonged to the genera of Epicocum, Cladosporium, Botrytis, Filbobasidium, Stemphylium, and Vishniacozyma were highly abundant in buckwheat seeds. The total flavonoids and rutin contents in tartary buckwheat cultivars (CQ, XQ, CH, K2) were much higher than those in common buckwheat cultivars (HT, T2, T4, T8). For tartary buckwheat cultivars, the total flavonoids and rutin contents were ranging from 2.6% to 3.3% and 0.9% to 1.3%, respectively. Accordingly, the tartary buckwheat samples displayed stronger antioxidant activity than the common buckwheat. Spearman correlation heat map analysis was successfully found that certain fungal species from the genera of Alternaria, Botryosphaeria, Colletorichum and Diymella exhibited significant positive correlation with flavonoids contents. Results of this study preliminary revealed the fungi-plant interaction relationship at secondary metabolite level, and could provide novel strategy for increasing the flavonoids accumulation of buckwheat seeds, as well as improving their quality.

Exploring the Potential of Plant-Derived Exosome-like Nanovesicle as Functional Food Components for Human Health: A Review.

Plant-derived exosome-like nanovesicles (PELNs) are bilayer membrane-enclosed nanovesicles secreted by plant cells, serving as carriers of various substances such as proteins, RNA, and metabolites. The mounting evidence suggests that PELN plays a crucial role in transmembrane signaling, nutrient transportation, apoptosis, and regulation of gut microbiota composition. This makes it a promising "dark nutrient" for plants to modulate human physiology and pathogenesis. A comprehensive understanding of PELN formation, uptake, and functional mechanisms can offer novel insights into plant nutrition and functional properties, thereby facilitating the precise development of plant-based foods and drugs. This article provides a summary of PELN extraction and characterization, as well as absorption and delivery processes. Furthermore, it focuses on the latest discoveries and underlying physiological mechanisms of PELN's functions while exploring future research directions.

The Quality Evaluation of Highland Barley and Its Suitability for Chinese Traditional Tsampa Processing.

The physicochemical traits of highland barley prominently affect the quality of Tsampa. To find out the relevance between the physicochemical properties of raw material and the texture parameters of processed products, twenty-five physicochemical traits and ten quality parameters for seventy-six varieties of highland barley were measured and analyzed. The results showed that there was a significant difference between the physicochemical indexes for highland barleys of various colors. The dark highland barley generally has more fat, protein, total dietary fiber, phenolic, Mg, K, Ca, and Zn and less amylose, Fe, Cu, and Mo than light colored barley. Then, these highland barleys were made into Tsampa. A comprehensive quality evaluation model based on the color and texture parameters of Tsampa was established through principal component analysis. Then, cluster analysis was used to classify the tested samples into three edible quality grades predicated on the above evaluation model. At last, the regression analysis was applied to establish a Tsampa quality predictive model according to the physicochemical traits of the raw material. The results showed that amylose, protein, ß-Glucan, and a* and b* could be used to predict the comprehensive quality of Tsampa. The predicted results indicated that 11 of 14 validated samples were consistent with the actual quality, and the accuracy was above 78.57%. Our study built the approach of the appropriate processing varieties evaluation. It may provide reference for processing specific highland barley.

Analogy of silicon and boron in plant nutrition.

Silicon (Si) and boron (B) are a class of elements called metalloids, which have properties like metals and non-metals. Si is classified as a quasi-essential element, while B is a micronutrient element for plants. Nowadays, numerous discoveries have shown the analogy of silicon and boron in plant nutrition. In this minireview, the molecular mechanisms for the transport of these two metalloids are compared. We also discussed the chemical forms of Si and B and their functional similarity in response to environmental stresses in plants. In conclusion, it can be proposed that cell wall-bound silicon rather than silica might partially replace boron for plant growth, development, and stress responses, and the underlying mechanism is the Si contribution to B in its structural function.

Role of MicroRNAs in Dietary Interventions for Obesity and Obesity-Related Diseases.

Obesity and related metabolic syndromes pose a serious threat to human health and quality of life. A proper diet is a safe and effective strategy to prevent and control obesity, thus maintaining overall health. However, no consensus exists on the connotations of proper diet, and it is attributed to various factors, including "nutritional dark matter" and the "matrix effect" of food. Accumulating evidence confirms that obesity is associated with the in vivo levels of miRNAs, which serve as potential markers and regulatory targets for obesity onset and progression; food-derived miRNAs can regulate host obesity by targeting the related genes or gut microbiota across the animal kingdom. Host miRNAs mediate food nutrient-gut microbiota-obesity interactions. Thus, miRNAs are important correlates of diet and obesity onset. This review outlines the recent findings on miRNA-mediated food interventions for obesity, thereby elucidating their potential applications. Overall, we provide new perspectives and views on the evaluation of dietary nutrition, which may bear important implications for dietary control and obesity prevention.

Genome-Wide Identification of Histone Deacetylases and Their Roles Related with Light Response in Tartary Buckwheat (Fagopyrum tataricum).

Histone deacetylases (HDACs), known as histone acetylation erasers, function crucially in plant growth and development. Although there are abundant reports focusing on HDACs of Arabidopsis and illustrating their important roles, the knowledge of HDAC genes in Tartary buckwheat (Polygonales Polygonaceae Fagopyrum tataricum (L.) Gaertn) is still scarce. In the study, a total of 14 HDAC genes were identified and divided into three main groups: Reduced Potassium Dependency-3/His-52 tone Deacetylase 1 (RPD3/HDA1), Silent Information Regulator 2 (SIR2), and the plant-53 specific HD2. Domain and motif composition analysis showed there were conserved domains and motifs in members from the same subfamilies. The 14 FtHDACs were distributed asymmetrically on 7 chromosomes, with three segmental events and one tandem duplication event identified. The prediction of the cis-element in promoters suggested that FtHDACs probably acted in numerous biological processes including plant growth, development, and response to environmental signals. Furthermore, expression analysis based on RNA-seq data displayed that all FtHDAC genes were universally and distinctly expressed in diverse tissues and fruit development stages. In addition, we found divergent alterations in FtHDACs transcript abundance in response to different light conditions according to RNA-seq and RT-qPCR data, indicating that five FtHDACs might be involved in light response. Our findings could provide fundamental information for the HDAC gene family and supply several targets for future function analysis of FtHDACs related with light response of Tartary buckwheat.

Analysis of synonymous codon usage patterns in mitochondrial genomes of nine Amanita species.

Introduction: Codon basis is a common and complex natural phenomenon observed in many kinds of organisms. Methods: In the present study, we analyzed the base bias of 12 mitochondrial core protein-coding genes (PCGs) shared by nine Amanita species. Results: The results showed that the codons of all Amanita species tended to end in A/T, demonstrating the preference of mitochondrial codons of Amanita species for a preference for this codon. In addition, we detected the correlation between codon base composition and the codon adaptation index (CAI), codon bias index (CBI), and frequency of optimal codons (FOP) indices, indicating the influence of base composition on codon bias. The average effective number of codons (ENC) of mitochondrial core PCGs of Amanita is 30.81, which is <35, demonstrating the strong codon preference of mitochondrial core PCGs of Amanita. The neutrality plot analysis and PR2-Bias plot analysis further demonstrated that natural selection plays an important role in Amanita codon bias. In addition, we obtained 5-10 optimal codons (ΔRSCU > 0.08 and RSCU > 1) in nine Amanita species, and GCA and AUU were the most widely used optimal codons. Based on the combined mitochondrial sequence and RSCU value, we deduced the genetic relationship between different Amanita species and found large variations between them. Discussion: This study promoted the understanding of synonymous codon usage characteristics and evolution of this important fungal group.

Study on morphological traits, nutrient compositions and comparative metabolomics of diploid and tetraploid Tartary buckwheat sprouts during sprouting.

Tartary buckwheat (TB) sprout is a kind of novel nutritional vegetable, but its consumption was limited by low biomass and thin hypocotyl. The tetraploid TB sprouts was considered to be able to solve this issue. However, the nutritional quality of tetraploid TB sprouts and differences between conventional (diploid) and tetraploid TB sprouts remain unclear. In this study, the morphological traits, nutrient compositions and metabolome changes of diploid and tetraploid TB sprouts were analyzed. The water, pigments and minerals contents of TB sprouts increased during sprouting, while the contents of total soluble protein, reducing sugar, cellulose, and total phenol decreased. Compared with diploid sprouts, tetraploid sprouts had higher biomass and thicker hypocotyl. Tetraploid sprouts had higher ash and carotenoid contents, but had lower phenol and flavonoid accumulation. 677 metabolites were identified in TB sprouts by UPLC-MS analysis, including 62 diseases-resistance metabolites and 43 key active ingredients. Some key bioactive metabolites, such as rimonabant, quinapril, 1-deoxynojirimycin and miglitol, were identified. 562 differential expressed metabolites (DEMs) were identified during sprouting with seven accumulation patterns, and five hormones were found to be involved in sprout development. Additionally, 209 DEMs between diploid and tetraploid sprouts were found, and some key bioactive metabolites were induced by chromosome doubling such as mesoridazine, amaralin, atractyloside A, rhamnetin and Qing Hau Sau. This work lays a basis for the development and utilization of TB sprouts and provides evidence for the selection of tetraploid varieties to produce sprouts with high biomass and quality.

Insect Cell-Based Models: Cell Line Establishment and Application in Insecticide Screening and Toxicology Research.

During the past decades, research on insect cell culture has grown tremendously. Thousands of lines have been established from different species of insect orders, originating from several tissue sources. These cell lines have often been employed in insect science research. In particular, they have played important roles in pest management, where they have been used as tools to evaluate the activity and explore the toxic mechanisms of insecticide candidate compounds. This review intends to first briefly summarize the progression of insect cell line establishment. Then, several recent studies based on insect cell lines coupled with advanced technologies are introduced. These investigations revealed that insect cell lines can be exploited as novel models with unique advantages such as increased efficiency and reduced cost compared with traditional insecticide research. Most notably, the insect cell line-based models provide a global and in-depth perspective to study the toxicology mechanisms of insecticides. However, challenges and limitations still exist, especially in the connection between in vitro activity and in vivo effectiveness. Despite all this, recent advances have suggested that insect cell line-based models promote the progress and sensible application of insecticides, which benefits pest management.

Isolation and purification of Tartary buckwheat polysaccharides and their effect on gut microbiota.

Tartary buckwheat (Fagopyrum tataricum) is rich in polysaccharides that can be utilized by the gut microbiota (GM) and provide several health benefits. However, the mechanisms underlying the action of these polysaccharides remain unclear to date. In this study, Tartary buckwheat polysaccharides (TBP) were purified, and five fractions were obtained. The composition of these fractions was determined using ion chromatography. Different TBP components were investigated regarding their probiotic effect on three species of Bifidobacteria and Lactobacillus rhamnosus. In addition, the effect of TBP on GM and short-chain fatty acids (SCFAs) was evaluated. Results showed that the probiotic effect of TBP fraction was dependent on their composition. The polysaccharides present in different fractions had specific probiotic effects. TBP-1.0, mainly composed of fucose, glucose, and d-galactose, exhibited the strongest proliferation effect on L. rhamnosus, while TBP-W, rich in glucose, d-galactose, and fructose, had the best promoting effect on Bifidobacterium longum and Bifidobacterium adolescentis growth. Furthermore, TBP-0.2, composed of d-galacturonic acid, d-galactose, xylose, and arabinose, exhibited its highest impact on Bifidobacterium breve growth. The composition of GM was significantly altered by adding TBP during fecal fermentation, with an increased relative abundance of Lactococcus, Phascolarctobacterium, Bacteroidetes, and Shigella. Simultaneously, the level of SCFA was also significantly increased by TBP. Our findings indicate that Tartary buckwheat can provide specific dietary polysaccharide sources to modulate and maintain GM diversity. They provide a basis for Tartary buckwheat commercial utilization for GM modulation.

Fast and sensitive UHPLC-QqQ-MS/MS method for simultaneous determination of typical α,ß-unsaturated aldehydes and malondialdehyde in various vegetable oils and oil-based foods.

This study aimed to develop and evaluate a method for simultaneously analyzing malondialdehyde (MDA) and two typical toxic α,ß-unsaturated aldehydes, 4-hydroxy-2-hexenal (HHE), and 4-hydroxy-2-nonenal (HNE), using ultra-high-performance liquid chromatography with triple quadrupole mass spectrometry (UHPLC-QqQ-MS/MS). The method possessed widely linear range (10-1000 ng/mL). The limit of detection (LOD) and limit of quantitation (LOQ) of MDA, HHE, and HNE were 2.0 and 5.0 ng/g, 2.0 and 5.0 ng/g, and 0.1 and 0.3 ng/g, respectively; the recovery rates all fall into 95.56-104.22 %. The method was sufficiently precise (<5%), and not affected by the analysis matrix. Application to 17 food products revealed total MDA, HHE, and HNE contents were 0.11-3.56, 0.05-3.32, and 0.09-3.70 µg/g, respectively. It will be useful in future research on the influence of food composition and main substrate structure on the generation and distribution of these three aldehydes and the implementation of corresponding control methods.

Bioactive compounds, health benefits, and industrial applications of Tartary buckwheat (Fagopyrum tataricum).

Tartary buckwheat belongs to the family Polygonaceae, which is a traditionally edible and medicinal plant. Due to its various bioactive compounds, the consumption of Tartary buckwheat is correlated to a wide range of health benefits, and increasing attention has been paid to its potential as a functional food. This review summarizes the main bioactive compounds and important bioactivities and health benefits of Tartary buckwheat, emphasizing its protective effects on metabolic diseases and relevant molecular mechanisms. Tartary buckwheat contains a wide range of bioactive compounds, such as flavonoids, phenolic acids, triterpenoids, phenylpropanoid glycosides, bioactive polysaccharides, and bioactive proteins and peptides, as well as D-chiro-inositol and its derivatives. Consumption of Tartary buckwheat and Tartary buckwheat-enriched products is linked to multiple health benefits, e.g., antioxidant, anti-inflammatory, antihyperlipidemic, anticancer, antidiabetic, antiobesity, antihypertensive, and hepatoprotective activities. Especially, clinical studies indicate that Tartary buckwheat exhibits remarkable antidiabetic activities. Various tartary buckwheat -based foods presenting major health benefits as fat and blood glucose-lowering agents have been commercialized. Additionally, to address the safety concerns, i.e., allergic reactions, heavy metal and mycotoxin contaminations, the quality control standards for Tartary buckwheat and its products should be drafted and completed in the future.

Study on dietary intake, risk assessment, and molecular toxicity mechanism of benzo[α]pyrene in college students in China Bashu area.

As an extremely strong polycyclic aromatic hydrocarbon carcinogen, benzo[α]pyrene (BaP) is often produced during food processing at high temperatures. Recently, food safety, as well as toxicity mechanism and risk assessment of BaP, has received extensive attention. We first constructed the database of BaP pollution concentration in Chinese daily food with over 104 data items; collected dietary intake data using online survey; then assessed dietary exposure risk; and finally revealed the possible toxicity mechanism through four comparative molecular dynamics (MD) simulations. The statistical results showed that the concentration of BaP in olive oil was the highest, followed by that in fried meat products. The margins of exposure and incremental lifetime cancer risk both indicated that the dietary exposure to BaP of the participants was generally safe, but there were still some people with certain carcinogenic risks. Specifically, the health risk of the core district population was higher than that of the noncore district in Bashu area, and the female postgraduate group was higher than the male group with bachelor degree or below. From MD trajectories, BaP binding does not affect the global motion of individual nucleic acid sequences, but local weak noncovalent interactions changed greatly; it also weakens molecular interactions of nucleic acid with Bacillus stearothermophilus DNA polymerase I large fragment (BF), and significantly changes the cavity structure of recognition interface. This work not only reveals the possible toxicity mechanism of BaP, but also provides theoretical guidance for the subsequent optimization of food safety standards and reference of rational diet.

Effects of Storage Temperature, Packaging Material and Wash Treatment on Quality and Shelf Life of Tartary Buckwheat Microgreens.

Tartary buckwheat microgreens (TBM) are popular worldwide products but display an extremely short shelf life. Thus, the effects of storage temperature, packaging material, and wash treatment on the quality and shelf life were analyzed. Headspace composition, weight loss, electrolyte leakage, microbial population and sensory quality were investigated during storage. Results showed that shelf life and quality of TBM decreased with the increment of storage temperature when stored at 5-25 °C. During 5 °C storage, LDPE bags were the best packaging materials for preserving the quality of LDPE, PE and HDPE bags. On the basis of 5 °C and LDPE packages, ClO2 + citric acid wash treatment could further inhibit quality deterioration and extend the shelf life. The results demonstrated bioactive constituents and antioxidant capacity were significantly affected by storage time. The study provides insights into developing optimal packaging and storage conditions for TBM.

Mitochondria-Mediated Apoptosis and Autophagy Participate in Buprofezin-Induced Toxic Effects in Non-Target A549 Cells.

Buprofezin (BUP) is an insecticide used for control of sucking pests. Its widespread use has raised concerns about possible adverse effects on the environment, and especially human health. The mechanism of toxicity of BUP, with respect to human health, is still unclear. Consequently, human A549 cells were employed to clarify the cytotoxicity and toxic mechanism of BUP at the molecular and cellular levels. The outcomes revealed BUP latent toxicity to A549 in a time- and dose-related way. Moreover, BUP induced mitochondrial dysfunction associated with mitochondrial membrane potential collapse, mitochondrial calcium overload, and ROS aggregation, ultimately resulting in the apoptosis and autophagy of A549 cells. Symbolic apoptotic and autophagic modifications were detected, including leakage of cyt-c, elevation of Bax/Bcl-2, activation of cas-9/-3, constitution of autophagic vacuoles, promotion of Beclin-1, conversion of LC3-II, and reduction of p62. Additionally, in total, 1216 differentially expressed genes (DEGs) were defined after BUP treatment. Several apoptosis- and autophagy-related genes, such as BCL2, ATG5, and ATG16, down- or upregulated at the RNA transcription level, and functional DEGs enrichment analysis showed their involvement in the metabolism of xenobiotics by cytochrome P450, mTOR signalling pathway, and AMPK signalling pathway. Results confirmed that BUP could induce cytotoxicity associated with mitochondria-mediated programmed cell death in A549 cells.

Unravelling the Polytoxicology of Chlorfenapyr on Non-Target HepG2 Cells: The Involvement of Mitochondria-Mediated Programmed Cell Death and DNA Damage.

Chlorfenapyr (CHL) is a type of insecticide with a wide range of insecticidal activities and unique targets. The extensive use of pesticides has caused an increase in potential risks to the environment and human health. However, the potential toxicity of CHL and its mechanisms of action on humans remain unclear. Therefore, human liver cells (HepG2) were used to investigate the cytotoxic effect and mechanism of toxicity of CHL at the cellular level. The results showed that CHL induced cellular toxicity in HepG2 cells and induced mitochondrial damage associated with reactive oxygen species (ROS) accumulation and mitochondrial calcium overload, ultimately leading to apoptosis and autophagy in HepG2 cells. Typical apoptotic changes occurred, including a decline in the mitochondrial membrane potential, the promotion of Bax/Bcl-2 expression causing the release of cyt-c into the cytosol, the activation of cas-9/-3, and the cleavage of PARP. The autophagic effects included the formation of autophagic vacuoles, accumulation of Beclin-1, transformation of LC3-II, and downregulation of p62. Additionally, DNA damage and cell cycle arrest were detected in CHL-treated cells. These results show that CHL induced cytotoxicity associated with mitochondria-mediated programmed cell death (PCD) and DNA damage in HepG2 cells.

Research Progress in Human AP Endonuclease 1: Structure, Catalytic Mechanism, and Inhibitors.

The high stability of phosphodiester bonds is considered to be one of the important reasons for the genetic role of nucleic acids, and their cleavage is also the core of many key biochemical processes, including DNA replication/ repair, and RNA processing/ degradation. As an important part of the base excision repair (BER) pathway, human apurinic/ apyrimidinic endonuclease 1 (APE1) is indispensable for the repair of abasic sites and other DNA damage, including ionizing radiation, DNA covalently bonding induced by cytotoxic antitumor drugs, etc. For tumor cells, the DNA repair activity of APE1 may lead to the occurrence of radiotherapy and chemotherapy resistance. The overexpression of APE1 often poses a serious threat to the effectiveness of tumor treatment, indicating a longer time, a much larger dose, less effective chemotherapy, and poor prognosis. It is of great urgency to design novel APE1 inhibitors. Rational design and modification of inhibitor molecules are closely related to the research progress of both structural biology and catalytic mechanism. In this review, the structure, catalytic mechanism, inhibitors, and other important biochemical information regarding APE1 are summarized, which will help in the design and modification of drug molecules targeting APE1.

In Vitro Effects of Tartary Buckwheat-Derived Nanovesicles on Gut Microbiota.

Evidence suggests that plant-derived nanovesicles may play a significant role in human health. Tartary buckwheat has several physiological activities; however, its underlying health-promoting mechanism remains unclear. In this study, first, Tartary buckwheat-derived nanovesicles (TBDNs) were collected, their structures were analyzed, and microRNA sequencing was performed. Next, target prediction and functional verification were conducted. Finally, the effects of TBDNs on gut microbiota and short-chain fatty acid levels were evaluated. The average size of TBDNs was 141.8 nm diameter. Through the sequencing analyses, 129 microRNAs, including 11 novel microRNAs were identified. Target gene prediction showed that some microRNAs could target functional genes in Escherichia coli and Lactobacillus rhamnosus-related physiological processes. TBDNs significantly promoted the growth of E. coli and L. rhamnosus, enhanced the diversity of fecal microorganisms and increased the short-chain fatty acid levels. These findings provided a new nutritional perspective for Tartary buckwheat and were conducive to promote the development and utilization of Tartary buckwheat.

Quantitative N-glycoproteome analysis of bovine milk and yogurt.

Post-translational modification structure of food's proteins might be changed during processing, thereby affecting the nutritional characteristics of the food product. In this study, differences in protein N-glycosylation patterns between milk and yogurt were quantitatively compared based on glycopeptide enrichment, liquid chromatography separation, and tandem mass spectrometry analysis. A total of 181 N-glycosites were identified, among which 142 were quantified in milk and yogurt. Significant alterations in the abundance of 13 of these N-glycosites were evident after the fermentation of milk into yogurt. Overall, the N-glycosylation status of the majority of milk proteins remained relatively unchanged in yogurt, suggesting that their conformations, activities, and functions were maintained despite the fermentation process. Among the main milk proteins, N241 of cathepsin D and N358 of lactoperoxidase were markedly reduced after undergoing lactic acid fermentation to produce yogurt. Furthermore, a comparative analysis of current and previously reported N-glycoproteomic data revealed heterogeneity in the N-glycosylation of milk proteins. To sum up, a quantitative comparison of the N-glycoproteomes of milk and yogurt was presented here for the first time, providing evidence that the fermentation process of yogurt could cause changes in the N-glycosylation of certain milk proteins.