Influence of hydrothermal treatment on the structural and digestive changes of actomyosin.

BACKGROUND: Heat treatment induces both structural and digestive change of meat protein. However, little has been revealed regarding the associations between structural changes and digested peptides of myofibrillar proteins. This work investigated the effects of heat treatment on the structures and in vitro digestibility of actomyosin, and the peptidomics of the digests were analyzed using liquid chromatography tandem mass spectrometry (LC-MS/MS). RESULTS: Heat treatment resulted in unfolding and aggregation behavior of actomyosin according to the results of surface hydrophobicity and particle size. Formation of disulfide bonds and increase in carbonyl groups that occurred during heat treatment of actomyosin indicated the oxidation of specific residues. Unfolding behavior could elevate digestibility of actomyosin by exposing residues, based on the identification of peptides in digests of actomyosin using LC-MS/MS. However, the disulfide bond proved to reduce the action of digestive proteases, since the peptides number (increased from 56 to 86 in sample heated at 70 °C for 30 min) and peptides intensity in digests largely increased after the addition of dithiothreitol (DTT). Heating at higher temperature (100 °C) induced severer aggregation and oxidation, which resulted in lower digestibility of actomyosin than that heated at 70 °C by burying or damaging partial cleavage sites for digestive proteases. CONCLUSIONS: This work highlights the huge influence of heat treatment on the multi-scale structures of myofibrillar proteins, which largely changed the peptides composition in protein digests. © 2019 Society of Chemical Industry.

Comparison of nutritional profile between plant-based meat analogues and real meat: A review focusing on ingredients, nutrient contents, bioavailability, and health impacts.

In order to fully understand the nutritional heterogeneity of plant-based meat analogues and real meat, this review summarized their similarities and differences in terms of ingredients, nutrient contents, bioavailability and health impacts. Plant-based meat analogues have some similarities to real meat. However, plant-based meat analogues are lower in protein, cholesterol and VB12 but higher in dietary fiber, carbohydrates, sugar, salt and various food additives than real meat. Moreover, some nutrients in plant-based meat analogues, such as protein and iron, are less bioavailable. There is insufficient evidence that plant-based meat analogues are healthier, which may be related to the specific attributes of these products such as formulation and degree of processing. As things stand, it is necessary to provide comprehensive nutrition information on plant-based meat products so that consumers can make informed choices based on their nutritional needs.

Kappa-carrageenan in a pork-based high-fat diet inhibited lipid bioavailability through interactions with pork protein.

κ-Carrageenan is a soluble dietary fiber widely used in meat products. Although its regulatory effect on glycolipid metabolism has been reported, the underlying mechanism remains unclear. The present study established a pork diet model for in vitro digestion to study how κ-carrageenan affected its digestive behavior and lipid bioavailability. The results revealed that κ-carrageenan addition to a pork-based high-fat diet reduced the rate of lipolysis and increased the number and size of lipid droplets in an in vitro digestion condition. However, κ-carrageenan did not inhibit lipolysis when lipids and κ-carrageenan were mixed directly or with the addition of pork protein. Furthermore, the pork protein in the diet significantly enhanced the inhibitory effect of κ-carrageenan on lipolysis with decreased proteolysis and raised hydrophobicity of protein hydrolysate. Our findings suggest that κ-carrageenan can inhibit dietary lipid bioavailability by interacting with pork protein in meat products or meat-based diets during digestion and indicate the positive role of carrageenan in the food industry to alleviate the excessive accumulation of lipids in the body.

Carrageenan in meat: improvement in lipid metabolism due to Sirtuin1-mediated fatty acid oxidation and inhibited lipid bioavailability.

Kappa-carrageenan (κ-CGN) is widely used in the meat industry. However, its impact on the host metabolism is less revealed. The current study investigated the effect of κ-CGN in pork-based diets on the lipid metabolism of male C57BL/6J mice. The κ-CGN supplement significantly suppressed the increase in body weight by 6.79 g on an average. Supplement of κ-CGN in high-fat diets significantly upregulated the genes and protein expression of Sirtuin1, which was accompanied by the increased gene expression of downstream fatty acids oxidation (Cpt1a and Acadl). The sirtuin1-mediated improvement of lipid metabolism was negatively associated with the levels of bile acids, especially for deoxycholic acid, 3ß-cholic acid, glycodeoxycholic acid and glycolithocholic acid. Moreover, κ-CGN in high-fat diets inhibited lipid digestion and absorption, being associated with the decrease in lipid accumulation and improved serum lipid profile. These results highlighted the role of κ-CGN in alleviating diet-induced adiposity by promoting energy expenditure and suppressing the bioavailability of ingested lipids.

Plant-based meat analogues enhance the gastrointestinal motility function and appetite of mice by specific volatile compounds and peptides.

Eating behavior is critical for maintaining energy homeostasis. Previous studies have found that plant-based meat analogues increased diet intake in mice compared with animal meat under a free feeding mode, however the reasons were unclear. To explore the underlying mechanisms of plant-based meat analogues increasing diet intake, mice were fed animal or plant-based pork and beef analogue diets, respectively. Biochemical and histological analyses were performed to evaluate appetite-regulating hormones and gastrointestinal motility function. Peptiomics and GC-IMS were applied to identify key substances. We found that the intake of plant-based meat analogues significantly enhanced the gastrointestinal motility function of mice. The long-term intake (68 days) of plant-based meat analogues significantly increased the muscle layer thickness of the duodenum and jejunum of mice; the activity of gastrointestinal cells of Cajal were also promoted by upregulating the expression of c-kit related signals as compared to animal meat; plant-based meat analogues intake markedly enhanced the signal intensity of the intestinal neurotransmitter 5-hydroxytryptamine (5-HT) by upregulating the expression of 5-HT synthase and receptors but downregulating its transporter and catabolic enzyme in the intestine. Moreover, plant-based meat analogues intake significantly increased levels of appetite-stimulating factors in the peripheral or hypothalamus but reduced levels of appetite-suppressing factors compared with animal meat. Specific volatile compounds were significantly associated with appetite regulating factors. Among them, 7 substances such as linalool have a potential promoting effect on food intake. Besides, different digestive peptides in gastrointestinal tract may affect eating behavior mainly through the neuroactive ligand-receptor interaction pathway, exerting hormone-like effects or influencing endocrine cell secretion. These findings preliminarily clarified the mechanism of plant-based meat analogues promoting diet intake and provided a theoretical basis for a reasonable diet.

SaaS sRNA promotes Salmonella intestinal invasion via modulating MAPK inflammatory pathway.

Salmonella Enteritidis is a foodborne enteric pathogen that infects humans and animals, utilizing complex survival strategies. Bacterial small RNA (sRNA) plays an important role in these strategies. However, the virulence regulatory network of S. Enteritidis remains largely incomplete and knowledge of gut virulence mechanisms of sRNAs is limited. Here, we characterized the function of a previously identified Salmonella adhesive-associated sRNA (SaaS) in the intestinal pathogenesis of S. Enteritidis. We found that SaaS promoted bacterial colonization in both cecum and colon of a BALB/c mouse model; it was preferentially expressed in colon. Moreover, our results showed that SaaS enhanced damage to mucosal barrier by affecting expressions of antimicrobial products, decreasing the number of goblet cells, suppressing mucin gene expression, and eventually reducing thickness of mucus layer; it further breached below physical barrier by strengthening invasion into epithelial cells in Caco-2 cell model as well as decreasing tight junction expressions. High throughput 16S rRNA gene sequencing revealed that SaaS also altered gut homeostasis by depleting beneficial gut microbiota while increasing harmful ones. Furthermore, by employing ELISA and western blot analysis, we demonstrated that SaaS regulated intestinal inflammation through sequential activation P38-JNK-ERK MAPK signaling pathway, which enabled immune escape at primary infection stage but strengthened pathogenesis at later stage, respectively. These findings suggest that SaaS plays an essential role in the virulence of S. Enteritidis and reveals its biological role in intestinal pathogenesis.

A Small RNA, SaaS, Promotes Salmonella Pathogenicity by Regulating Invasion, Intracellular Growth, and Virulence Factors.

Salmonella enterica serovar Enteritidis is a common foodborne pathogen that infects both humans and animals. The S. Enteritidis virulence regulation network remains largely incomplete, and knowledge regarding the specific virulence phenotype of small RNAs (sRNAs) is limited. Here, we investigated the role of a previously identified sRNA, Salmonella adhesive-associated sRNA (SaaS), in the virulence phenotype of S. Enteritidis by constructing mutant (ΔsaaS) and complemented (ΔsaaS/psaaS) strains. SaaS did not affect S. Enteritidis; it was activated in the simulated intestinal environment (SIE), regulating the expression of virulence target genes. We discovered that it directly binds ssaV mRNA. Caco-2 and RAW 264.7 cell assays revealed that SaaS promoted S. Enteritidis invasion and damage to epithelial cells while suppressing macrophage overgrowth and destruction. Furthermore, a BALB/c mouse model demonstrated that the deletion of SaaS significantly reduced mortality and attenuated the deterioration of pathophysiology, bacterial dissemination into systemic circulation, and systemic inflammation. Our findings indicate that SaaS is required for S. Enteritidis virulence and further highlight its biological role in bacterial pathogenesis. IMPORTANCE Salmonella is a zoonotic pathogen with high virulence worldwide, and sRNAs have recently been discovered to play important roles. We explored the biological characteristics of the sRNA SaaS and developed two cell infection models and a mouse infection model. SaaS is an SIE-responsive sRNA that regulates the expression of virulence-targeted genes. Additionally, it differentially mediates invasion and intracellular growth for survival and infection of the epithelium and macrophages. We further found that SaaS enhanced bacterial virulence by promoting lethality, colonization, and inflammatory response. These findings provide a better understanding of the critical role of sRNA in bacterial virulence.

Reconsidering Meat Intake and Human Health: A Review of Current Research.

Meat consumption is gradually increasing and its impact on health has attracted widespread attention, resulting in epidemiological studies proposing a reduction in meat and processed meat intake. This review briefly summarizes recent advances in understanding the effects of meat or processed meat on human health, as well as the underlying mechanisms. Meat consumption varies widely among individuals, populations, and regions, with higher consumption in developed countries than in developing countries. However, increasing meat consumption may not be the main cause of increasing incidence of chronic disease, since the development of chronic disease is a complex physiological process that involves many factors, including excessive total energy intake and changes in food digestion processes, gut microbiota composition, and liver metabolism. In comparison, unhealthy dietary habits and a sedentary lifestyle with decreasing energy expenditure are factors more worthy of reflection. Meat and meat products provide high-value protein and many key essential micronutrients. In short, as long as excessive intake and overprocessing of meats are avoided, meat remains an indispensable source of nutrition for human health.

Plant-Based Meat Analogues Weaken Gastrointestinal Digestive Function and Show Less Digestibility Than Real Meat in Mice.

Real meat and plant-based meat analogues have different in vitro protein digestibility properties. This study aims to further explore their in vivo digestion and absorption and their effects on the gastrointestinal digestive function of mice. Compared with the real pork and beef, plant-based meat analogues significantly reduced the number of gastric parietal cells, the levels of gastrin/CCKBR, acetylcholine/AchR, Ca2+, CAMK II, PKC, and PKA, the activity of H+, K+-ATPase, and pepsin, the duodenal villus height, and the ratio of villus height to crypt depth and downregulated the expression of most nitrogen nutrient sensors. Peptidomics revealed that plant-based meat analogues released fewer peptides during in vivo digestion and increased the host- and microbial-derived peptides. Moreover, the real beef showed better absorption properties. These results suggested that plant-based meat analogues weaken gastrointestinal digestive function of mice, and their digestion and absorption performance in vivo is not as good as the real meat.

Body weight index indicates the responses of the fecal microbiota, metabolome and proteome to beef/chicken-based diet alterations in Chinese volunteers.

Relationships between meat consumption and gut diseases have been debated for decades, and the gut microbiota plays an important role in this interplay. It was speculated that the gut microbiota and relevant indicators of hosts with different body weight indexes (BMIs) might respond differentially to meat-based diet alterations, since lean and obese hosts have different gut microbiota composition. Forty-five young Chinese volunteers were recruited and assigned to high-, middle- and low-BMI groups. All of the volunteers were given a beef-based diet for 2 weeks and subsequently with a chicken-based diet for another 2 weeks. Body weight and blood indexes were measured, and fecal samples were obtained for 16S rRNA sequencing, metabolome and proteome analyses. The fecal metabolites of the low-BMI volunteers showed greater sensitivity to meat-based diet alterations. In contrast, the fecal proteome profiles and blood indexes of the high- and middle-BMI volunteers indicated greater sensitivity to meat-based diet alterations. Replacing the beef-based diet with the chicken-based diet largely changed operational taxonomic units of Bacteroides genus, and thus probably induced downregulation of immunoglobulins in feces. Compared with the beef-based diet, the chicken-based diet decreased inflammation-related blood indexes, especially in high- and middle-BMI volunteers. This work highlighted the role of BMI as an important factor predicting changes in gut homeostasis in response to meat consumption. Compared with the chicken-based diet, the beef-based diet may induce more allergic and inflammation-related responses in high- and middle- BMI Chinese at the current level.

Real meat and plant-based meat analogues have different in vitro protein digestibility properties.

To explore the nutritional values of meat and meat analogues, the in vitro protein digestion of pork, beef, plant-based pork and beef were evaluated. In the gastric phase, the digestibility of pork was significantly higher than that of the plant-based pork, while the value of beef was lower than that of the plant-based beef. In the intestinal phase, both pork and beef showed higher digestibility than plant-based meat analogues. A greater number of small molecular peptides were identified from pork and beef than from plant-based meat after gastrointestinal digestion. Larger quantities of potential bioactive peptides were released from the meat than from the plant-based meat analogues during digestion. These differences were closely related to protein secondary structure, the formation of disulfide bonds and apparent viscosity of digestion solution. The findings give a new insight into the underlying mechanisms of the different phenotype responses of consumers to meat and plant-based meat.

Transcriptomics and metabolomics reveal the adaption of Akkermansia muciniphila to high mucin by regulating energy homeostasis.

In gut, Akkermansia muciniphila (A. muciniphila) probably exerts its probiotic activities by the positive modulation of mucus thickness and gut barrier integrity. However, the potential mechanisms between A. muciniphila and mucin balance have not been fully elucidated. In this study, we cultured the bacterium in a BHI medium containing 0% to 0.5% mucin, and transcriptome and gas chromatography mass spectrometry (GC-MS) analyses were performed. We found that 0.5% (m/v) mucin in a BHI medium induced 1191 microbial genes to be differentially expressed, and 49 metabolites to be changed. The metabolites of sorbose, mannose, 2,7-anhydro-ß-sedoheptulose, fructose, phenylalanine, threonine, lysine, ornithine, asparagine, alanine and glutamic acid were decreased by 0.5% mucin, while the metabolites of leucine, valine and N-acetylneuraminic acid were increased. The association analysis between transcriptome and metabolome revealed that A. muciniphila gave strong responses to energy metabolism, amino sugar and nucleotide sugar metabolism, and galactose metabolism pathways to adapt to high mucin in the medium. This finding showed that only when mucin reached a certain concentration in a BHI medium, A. muciniphila could respond to the culture environment significantly at the level of genes and metabolites, and changed its metabolic characteristics by altering the effect on carbohydrates and amino acids.

Long-Term Intake of Pork Meat Proteins Altered the Composition of Gut Microbiota and Host-Derived Proteins in the Gut Contents of Mice.

SCOPE: This study is to investigate the effects of long-term intake of pork protein on the composition of gut microbiota and proteins in mice. METHODS AND RESULTS: C57BL/6J mice are fed pork meat protein diets for 240 days, and the composition of gut microbiota and proteins in luminal contents from the duodenum to the colon are analyzed by 16S rRNA gene sequencing and LC-MS/MS. The stewed pork protein diet group has a highly similar microbiota composition to that of the cooked pork protein diet group, but different from the pork emulsion sausage or dry-cured pork protein diet groups. Lachnospiraceae NK4A136, Odoribacter, Defluviitaleaceae UCG-011, Ruminiclostridium 9, Blautia, Lachnoclostridium, and Ruminococcaceae UCG-010 play an important role in response to changes in gut luminal proteins. Specific microbes are significantly correlated with the Cela3b and Cpa1 that are derived from the host and involve protein digestion and absorption. CONCLUSIONS: Pork meat protein diets alter the gut microbiota composition and specific gut microbes may have a great impact on protein digestion and absorption by regulating the secretion of digestive proteins from the host. These findings provide a new insight into the associations of long-term intake of meat protein diet with gut microbiota and host.

Processing Method Altered Mouse Intestinal Morphology and Microbial Composition by Affecting Digestion of Meat Proteins.

Our previous study showed that the intake of meat proteins dynamically affected fecal microbial composition. However, the digestion of processed meat proteins in vivo and its relationship with gut microbiota and host remain unclear. In this study, we collected cecal contents and intestinal tissue from the mice fed with casein, soybean protein (SP), and four processed pork proteins for 8 months, and analyzed the amino acid (AA) files, cecum microbial composition and metabolites, and intestinal morphology. Dry-cured pork protein and stewed pork protein (SPP) groups had significantly higher total AA content in gut content than the other groups, but the content of the SPP group was relatively lower in the serum. The microbial composition of the processed meat protein groups differed from the casein or SP group, which is consistent with changes in AA composition. Emulsion sausage protein and SP diets upregulated the microbial AA metabolism, energy metabolism, signaling molecules and interaction, translation, and digestive system function but downregulated the microbial membrane transport, signal transduction and cell motility function compared to the casein diet. The SPP diets increased concentrations of acetate, propionate, butyrate, and isovalerate by specific gut microbes, but it decreased the relative abundance of Akkermansia. Moreover, the mice fed SP diet had relatively lower crypt depth, higher villus height and V/C ratio in duodenum, with the longer small intestines and the heavier cecum than other diets. These results suggested that processing methods altered bioavailability of meat proteins, which affected the intestinal morphology and the cecum microbial composition and function.

Dietary Proteins Regulate Serotonin Biosynthesis and Catabolism by Specific Gut Microbes.

More than 90% of serotonin is produced in the intestine. Previous studies have shown that different protein diets significantly affect serum serotonin levels. Here, the colonic microbiota and intestinal serotonin were measured to elaborate how protein diets affect serotonin production in a mouse model. The emulsion-type sausage protein and cooked pork protein diets increased the mRNA levels of tryptophan hydroxylase 1 (Tph1) and monoamine oxidase A (Maoa) and serotonin level as well but reduced the number of enterochromaffin cells. However, the soy protein diet increased the number of enterochromaffin cells and Tph1 mRNA level but decreased the Maoa mRNA level and the serotonin content. Specific gut microbes that responded to dietary changes and affected the content of short-chain fatty acids were significantly related to serotonin-associated biomarkers. These results suggest that dietary proteins may regulate serotonin biosynthesis and catabolism by altering specific gut microbes.

Structural Changes and Evolution of Peptides During Chill Storage of Pork.

In this work, we investigated changes in protein structures in vacuum-packed pork during chill storage and its impact on the in vitro protein digestion. Longissimus dorsi muscles were vacuum packed and stored at 4°C for 3 days. Samples were subjected to Raman spectroscopy, in vitro digestion and nano LC-MS/MS. The 3 d samples had lower α-helix content, but higher ß-sheet, ß-turn, and random coil contents than the 0 d samples (P < 0.05). SDS-PAGE revealed significant protein degradation in the 3 d samples and the differences in digested products across the storage time. Proteome analysis indicated that the 3 d samples had the higher susceptibility to digestion. Increasing protein digestibility was mainly attributed to the degradation of myofibrillar proteins. Thus, exposure of more enzymatic sites in loose protein structure during chill storage could increase protein degradation in meat.

Specific Microbiota Dynamically Regulate the Bidirectional Gut-Brain Axis Communications in Mice Fed Meat Protein Diets.

The purpose of this study was to characterize the dynamic changes of different protein diets to gut microbiota and explore the influence on communications between the gut and the brain. C57BL/6J mice were fed casein, soy protein, and four kinds of processed meat proteins at a normal dose of 20% for 8 months. Bacteroidales S24-7 abundance increased from 4 to 8 months, whereas the abundances of six genera including Akkermansia decreased remarkably. Lachnospiraceae Unclassified abundance in the emulsion-type sausage protein and stewed pork protein groups showed an opposite change from 4 to 8 months. Twenty-eight and 48 specific operational taxonomy units in cecum and colon respectively were involved in regulating serotonin, peptide YY, leptin, and insulin levels. Specific microbiota was involved, directly or indirectly through signaling molecules, in the regulation of body metabolism, which may affect the communications between the gut and brain and cause different growth performances.

Phenolic compounds in beer inhibit formation of polycyclic aromatic hydrocarbons from charcoal-grilled chicken wings.

The effect of various beer marinades on formation of polycyclic aromatic hydrocarbons (PAHs) in charcoal-grilled chicken wings (CWs) and the active ingredients in beer contributing to inhibition of PAH formation were studied. The 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging activity and total phenolic content (TPC) of six beers were evaluated. LC-MS analysis indicated a total of 32 phenolic compounds, among which we screened 11 to verify the inhibition of select PAH production. The total stable free radicals and selected PAH content of charcoal-grilled CWs were assayed, revealing a positive correlation. Heineken exhibited the highest phenol content and excellent performance in TPC (393.86 mg gallic acid equivalents (GAE)/L), ability to scavenge free radicals (27.0%), and the most effective inhibition of PAH8 formation (67%). Our study supplies a theoretical foundation for using edible materials rich in phenolic compounds as potential natural inhibitors of PAHs formed during the cooking process.