Kinetics of pepsin-induced hydrolysis and the coagulation of milk proteins.

Hydrolysis-induced coagulation of casein micelles by pepsin occurs during the digestion of milk. In this study, the effect of pH (6.7-5.3) and pepsin concentration (0.110-2.75 U/mL) on the hydrolysis of κ-casein and the coagulation of the casein micelles in bovine skim milk was investigated at 37°C using reverse-phase HPLC, oscillatory rheology, and confocal laser scanning microscopy. The hydrolysis of κ-casein followed a combined kinetic model of first-order hydrolysis and putative pepsin denaturation. The hydrolysis rate increased with increasing pepsin concentration at a given pH, was pH dependent, and reached a maximum at pH ∼6.0. Both the increase in pepsin concentration and decrease in pH resulted in a shorter coagulation time. The extent of κ-casein hydrolysis required for coagulation was independent of the pepsin concentration at a given pH and, because of the lower electrostatic repulsion between para-casein micelles at lower pH, decreased markedly from ∼73% to ∼33% when pH decreased from 6.3 to 5.3. In addition, the rheological properties and the microstructures of the coagulum were markedly affected by the pH and the pepsin concentration. The knowledge obtained from this study provides further understanding on the mechanism of milk coagulation, occurring at the initial stage of transiting into gastric conditions with high pH and low pepsin concentration.

Copper-induced oxidative stress, transcriptome changes, intestinal microbiota, and histopathology of common carp (Cyprinus carpio).

Copper (Cu) is a common contaminant in aquatic environments, which could cause physiological dysfunction in aquatic organisms. However, few studies have comprehensively examined the impact of copper toxicity in freshwater fish over the past decade. In this research, the oxidative stress, liver transcriptome, intestinal microbiota, and histopathology of common carp (C. carpio) in response to Cu exposure were studied, by exposing juvenile carp to 0.2 mg/ml Cu2+ for 30 days. The results revealed that Cu2+ could induce significant changes in malondialdehyde (MDA) content and antioxidant enzyme (superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx)) activity. The changes in antioxidant enzyme activities indicate that Cu can induce oxidative stress by generating reactive oxygen species (ROS) content. RNA-seq analysis of the liver identified 1069 differentially expressed genes (DEGs) after treatment with 2.0 mg/L Cu2+. Among the DEGs, 490 genes were upregulated and 579 genes were downregulated. GO functional enrichment analysis revealed that Cu could affect the fatty acid biosynthetic process, carnitine biosynthetic process, and activity of carboxylic acid transmembrane transporter. Meanwhile, the most significantly enriched KEGG pathway also included the lipid metabolism pathway. In addition, Cu2+ exposure increased bacterial richness and changed bacterial composition. At the phylum level, we found that the ratio of Bacteroidetes to Firmicutes was increased in the treatment carps, which can regulate intestinal epithelium function and reduce inflammation and immune responses. At the genus level, the abundances of 11 genera were significantly altered after exposure to Cu2+. The altered composition of the microbial community caused by Cu exposure may play a useful role in compensation of the intestinal lesions by Cu exposure. Furthermore, we found that Cu2+ exposure could cause histological alterations such as structural damage to the liver and intestines. The results of this research contribute to a better understanding of mechanisms related to Cu toxicity in fish.

Dynamic In Vitro Gastric Digestion Behaviour of Commercial Infant Formulae Made with Cow, Goat and Sheep Milk.

There are a wide range of commercial infant formulae available on the market. These are made using milk from different species, such as goat, sheep, and cow. The different protein compositions of these milks and the process used during infant-formulae manufacture, such as heat treatment, may impact the digestion of nutrients. This study compared the effect of protein composition and heat treatment on the in vitro gastric digestion behaviour of commercial infant formulae made with cow, goat, and sheep milk using a dynamic infant human gastric simulator (IHGS). During the simulated dynamic gastric digestion, the goat milk infant formula (GIF) showed earlier signs of aggregate formation compared to cow milk infant formula (CIF) and sheep milk infant formula (SIF). In addition, the microstructures of GIF chyme showed fragmented and porous structures. On the contrary, CIF formed dense protein networks that trapped oil droplets, whereas SIF exhibited a microstructure of smooth oil droplets surrounded by fewer protein networks. The different aggregation behaviours and aggregate structures of the three infant-formulae chyme were related to their different protein compositions, especially the different casein compositions. Furthermore, the open fragile structure of GIF aggregates provided easier access to pepsin, allowing it to hydrolyse protein. The results from the present study provided some information to assist in understanding the coagulation and digestion behaviours of commercial infant formulae made from different species of milk.

Achieving High Ionic Conductivity and Mechanical Strength by a Leather Gel Electrolyte for Flexible Zinc-Ion Batteries.

The continuous advancement in the field of flexible and wearable electronics has led to increased research interest in safe, low-cost, and flexible zinc-ion batteries, particularly with a focus on flexible electrolytes. In this study, we present a leather gel electrolyte (LGE) that offers robust mechanical properties and an excellent electrochemical performance. LGE exhibits an ionic conductivity of 1.36 × 10-2 S cm-1 and achieves a capacity of 303.7 mAh g-1 in flexible zinc-manganese dioxide batteries. Even after 1000 cycles, the capacity retention remains above 90%, demonstrating outstanding performance in protecting the zinc anode. Furthermore, such a flexible battery shows good resistance to damage due to the strong mechanical strength originating from leather. Notably, LGE utilizes green and sustainable leather as a raw material, making it a promising option for sustainable flexible devices.

Pepsin-induced coagulation of casein micelles: Effect of whey proteins and heat treatment.

The effect of whey proteins and heat treatment (90 °C, 5 min) on pepsin-induced hydrolysis of κ-casein, and subsequent coagulation of casein micelles, was investigated at pH 6.3 and 6.0 using reverse-phase HPLC, oscillatory rheology, and confocal laser scanning microscopy. Whey proteins did not affect the hydrolysis of κ-casein but retarded the coagulation process. Heat treatment did not affect the hydrolysis kinetics in whey protein (WP)-free samples, but slightly impaired the hydrolysis rate in WP-containing samples. The coagulation process of WP-free samples was little affected by heat-treatment. However, compared with unheated WP-contained sample at the same pH, the coagulation process of the heated sample was retarded at pH 6.3 but enhanced at pH 6.0. The curd in heated samples with smaller pores had higher water holding capacity. This knowledge provides further understanding on the role of whey proteins and heat treatment on the coagulation mechanisms of milk under gastric conditions.

Role of Ca2+ in the pepsin-induced coagulation and in the dynamic in vitro gastric digestion behavior of casein micelles.

The effect of Ca2+ on pepsin-induced hydrolysis of κ-casein and subsequent coagulation of casein micelles was studied in a micellar casein (MC) solution at pH ≈ 6.0 at 37 °C without stirring. An NaCl-supplemented MC solution was used as a positive control to assess the effect of increased ionic strength after CaCl2 addition. Quantitative determination of the released para-κ-casein during the reaction using reverse-phase high-performance liquid chromatography showed that specific hydrolysis of κ-casein by pepsin was little affected by the addition of either CaCl2 or NaCl. However, rheological properties and microstructures of curds induced by pepsin hydrolysis depended markedly on the addition of salts. Addition of CaCl2 up to 17.5 mM facilitated coagulation, with decreases in coagulation time and critical hydrolysis degree, and increases in firming rate and maximum storage modulus (G'max); further addition of CaCl2 (22.5 mM) resulted in a lower G'max. Increased ionic strength to 52.5 mM by adding NaCl retarded the coagulation and resulted in a looser curd structure. In a human gastric simulator, MC, without the addition of CaCl2, did not coagulate until the pH decreased to ≈5.0 after ≈50 min of digestion. Addition of CaCl2 facilitated coagulation of casein micelles and resulted in more cohesive curds with dense structures during digestion, which slowed the emptying rate of caseins. At the same CaCl2 concentration, a sample with higher ionic strength coagulated more slowly. This study provides further understanding on the effect of divalent (Ca2+) ions and ionic strength on the coagulation of casein micelles and the digestion behavior of milk.

Gut microbiota of two invasive fishes respond differently to temperature.

Temperature variation structures the composition and diversity of gut microbiomes in ectothermic animals, key regulators of host physiology, with potential benefit to host or lead to converse results (i.e., negative). So, the significance of either effect may largely depend on the length of time exposed to extreme temperatures and how rapidly the gut microbiota can be altered by change in temperature. However, the temporal effects of temperature on gut microbiota have rarely been clarified. To understand this issue, we exposed two juvenile fishes (Cyprinus carpio and Micropterus salmoides), which both ranked among the 100 worst invasive alien species in the world, to increased environmental temperature and sampled of the gut microbiota at multiple time points after exposure so as to determine when differences in these communities become detectable. Further, how temperature affects the composition and function of microbiota was examined by comparing predicted metagenomic profiles of gut microbiota between treatment groups at the final time point of the experiment. The gut microbiota of C. carpio was more plastic than those of M. salmoides. Specifically, communities of C. carpio were greatly altered by increased temperature within 1 week, while communities of M. salmoides exhibit no significant changes. Further, we identified 10 predicted bacterial functional pathways in C. carpio that were temperature-dependent, while none functional pathways in M. salmoides was found to be temperature-dependent. Thus, the gut microbiota of C. carpio was more sensitive to temperature changes and their functional pathways were significantly changed after temperature treatment. These results showed the gut microbiota of the two invasive fishes differ in response to temperature change, which may indicate that they differ in colonization modes. Broadly, we have confirmed that the increased short-term fluctuations in temperatures are always expected to alter the gut microbiota of ectothermic vertebrates when facing global climate change.

High-pressure processing of bovine milk: Effects on the coagulation of protein and fat globules during dynamic in vitro gastric digestion.

The effect of high-pressure processing (HPP) on the digestion behavior of skim and whole bovine milks was investigated using a human gastric simulator. Both milks formed clots during gastric digestion. HPP treatment led to the formation of a coagulum with a fragmented and crumbled structure, compared with the coagulum formed from untreated milk. At pressures over 400 MPa, more intense pressure resulted in looser and more fragmented gastric clot structures. The weight of the dried clots and the moisture content in the clots of the skim milk treated at 600 MPa were significantly lower and higher than that of untreated skim milk, respectively. The looser and more fragmented gastric clot structures consequently led to faster hydrolysis of the proteins by pepsin during gastric digestion. The denaturation of the whey proteins induced by HPP may have also altered the resistance of α-lactalbumin and ß-lactoglobulin in the HPP-treated milk samples to pepsin hydrolysis. This study provides insights into the differences among untreated skim milk, untreated whole milk and HPP-treated milk under in vitro gastric digestion conditions. The structure of the clots formed in the gastric environment affects their breakdown and consequently their emptying rate into the intestine.