Advances in molecular enzymology of ß-1,3-glucanases: A comprehensive review.

ß-1,3-Glucanases are essential enzymes involved in the hydrolysis of ß-1,3-glucans, with significant biological and industrial relevance. These enzymes are derived from diverse sources, including bacteria, fungi, plants, and animals, each exhibiting unique substrate specificities and biochemical properties. This review provides an in-depth analysis of the natural sources and ecological roles of ß-1,3-glucanases, exploring their enzymatic properties such as optimal pH, temperature, molecular weight, isoelectric points, and kinetic parameters, which are crucial for understanding their functionality and stability. Advances in molecular enzymology are discussed, focusing on gene cloning, expression in systems like Escherichia coli and Pichia pastoris, and structural-functional relationships. The reaction mechanisms and the role of non-catalytic carbohydrate-binding modules in enhancing substrate hydrolysis are examined. Industrial applications of ß-1,3-glucanases are highlighted, including the production of ß-1,3-glucooligosaccharides, uses in the food industry, biological control of plant pathogens, and nutritional roles. This review aims to provide a foundation for future research, improving the efficiency and robustness of ß-1,3-glucanases for various industrial applications.

A novel coupled fermentation system for low-molecular-weight xanthan gum with diverse biological activities.

Xanthan gum (XG) is a bacterial exopolysaccharide widely used in various industries due to its stability and rheological properties. Low-molecular-weight xanthan gum (LXG) exhibits enhanced properties and broader applications, but current degradation methods are limited. This study introduces an innovative coupled fermentation system for the efficient production of LXG. Endo-xanthanase from Microbacterium sp. XT11 was expressed in Pichia pastoris GS115, exhibiting optimal activity at pH 6.0 and 40 °C, with broad pH tolerance. The optimized coupled fermentation system used bean sprouts juice as nitrogen source, the inoculation quantity of X. campestris: P. pastoris was 1: 3, and the pH was controlled at 6.0. In the bioreactor, the total sugar concentration reached 12.12 g/L, the reducing sugar concentration reached 5.32 g/L, and the endo-xanthanase activity increased to 1150.26 U/L, which were 2.13, 2.3, and 3.71 times higher than those at the shake flask level, respectively. The prepared LXG had a molecular weight of 1093 Da and a monosaccharide ratio of 2.0:1.57:0.89 (glucose, mannose, and glucuronic acid). Bioactivity analysis revealed its antioxidant and prebiotic properties, promoting the growth of beneficial intestinal microbiota and metabolite production. This suggests the potential of LXG as a functional ingredient in intestinal health-focused foods and supplements.

Glycerol-driven adaptive evolution for the production of low-molecular-weight Welan gum: Characterization and activity evaluation.

Through adaptive laboratory evolution (ALE) of Sphingomonas sp. ATCC 31555, fermentation for production of low-molecular-weight welan gum (LMW-WG) was performed using glycerol as sole carbon source. During ALE, GPC-MALS analysis revealed a gradual decrease in WG molecular weight with the increase of adaptation cycles, accompanied by changes in solution conformation. LMW-WG was purified and structurally analyzed using GPC-MALS, monosaccharide composition analysis, infrared spectroscopy, NMR analysis, atomic force microscopy, and scanning electron microscopy. Subsequently, LMW-WG obtains hydration, transparency, antioxidant activity, and rheological properties. Finally, an in vitro simulation colon reactor was used to evaluate potential prebiotic properties of LMW-WG as dietary fiber. Compared with WG produced using sucrose as substrate, LMW-WG exhibited a fourfold reduction in molecular weight while maintaining moderate viscosity. Structurally, L-Rha nearly completely replaced L-Man. Furthermore, LMW-WG demonstrated excellent hydration, antioxidant activity, and high transparency. It also exhibited resistance to saliva and gastrointestinal digestion, showcasing a favorable colonization effect on Bifidobacterium, making it a promising symbiotic agent.

Improvement of antibacterial activity of polysaccharides via chemical modification: A review.

Antibiotic residue and bacterial resistance induced by antibiotic abuse have seriously threatened food safety and human healthiness. Thus, the development and application of safe, high-efficiency, and environmentally friendly antibiotic alternatives are urgently necessary. Apart from antitumor, antivirus, anti-inflammatory, gut microbiota regulation, immunity improvement, and growth promotion activities, polysaccharides also have antibacterial activity, but such activity is relatively low, which cannot satisfy the requirements of food preservation, clinical sterilization, livestock feeding, and agricultural cultivation. Chemical modification not only provides polysaccharides with better antibacterial activity, but also promotes easy operation and large-scale production. Herein, the enhancement of the antibacterial activity of polysaccharides via acetylation, sulfation, phosphorylation, carboxymethylation, selenation, amination, acid graft, and other chemical modifications is reviewed. Meanwhile, a new trend on the application of loading chemically modified polysaccharides into nanostructures is discussed. Furthermore, possible limitations and future recommendations for the development and application of chemically modified polysaccharides with better antibacterial activity are suggested.

New Horizons in Probiotics: Unraveling the Potential of Edible Microbial Polysaccharides through In Vitro Digestion Models.

In vitro digestion models, as innovative assessment tools, possess advantages such as speed, high throughput, low cost, and high repeatability. They have been widely applied to the investigation of food digestion behavior and its potential impact on health. In recent years, research on edible polysaccharides in the field of intestinal health has been increasing. However, there is still a lack of systematic reviews on the application of microbial-derived edible polysaccharides in in vitro intestinal models. This review thoroughly discusses the limitations and challenges of static and dynamic in vitro digestion experiments, while providing an in-depth introduction to several typical in vitro digestion models. In light of this, we focus on the degradability of microbial polysaccharides and oligosaccharides, with a particular emphasis on edible microbial polysaccharides typically utilized in the food industry, such as xanthan gum and gellan gum, and their potential impacts on intestinal health. Through this review, a more comprehensive understanding of the latest developments in microbial polysaccharides, regarding probiotic delivery, immobilization, and probiotic potential, is expected, thus providing an expanded and deepened perspective for their application in functional foods.

Effects of In Vitro Fermentation of Polysialic Acid and Sialic Acid on Gut Microbial Community Composition and Metabolites in Healthy Humans.

The influence of polysialic acid (PSA) and sialic acid (SA) on the gut microbial community composition and metabolites in healthy humans was investigated using a bionic gastrointestinal reactor. The results indicated that PSA and SA significantly changed the gut microbiota and metabolites to different degrees. PSA can increase the relative abundances of Faecalibacterium and Allisonella, whereas SA can increase those of Bifidobacterium and Megamonas. Both can significantly increase the content of short-chain fatty acids. The results of metabolome analysis showed that PSA can upregulate ergosterol peroxide and gallic acid and downregulate the harmful metabolite N-acetylputrescine. SA can upregulate 4-pyridoxic acid and lipoic acid. PSA and SA affect gut microbiota and metabolites in different ways and have positive effects on human health. These results will provide a reference for the further development of PSA- and SA-related functional foods and health products.

Optimization of welan gum extraction and purification using lysozyme and alkaline protease.

Welan gum, a natural polysaccharide produced by Sphingomonas sp. ATCC 31555, has attracted considerable attention in the scientific community due to its desirable properties. However, challenges, such as high viscosity, residual bacterial cells, carotenoids, and protein complexation, hinder the widespread application of welan gum. In this study, we established a method for the extraction and purification of welan gum using a synergistic approach with lysozyme and alkaline protease. Lysozyme hydrolysis conditions were optimized by applying response surface methodology, and the best results for bacterial cell removal were achieved at 11 000 U/g, 44 °C, and pH 9 after 3 h of treatment. Subsequently, we evaluated protein hydrolysis through computer simulation and identified alkaline protease as the most suitable enzyme. Through experimental investigations, we found that the optimal conditions for alkaline protease hydrolysis were 7500 U/g, 50 °C, pH 10, and 600 rpm. These conditions resulted in a sugar recovery rate of 76.1%, carotenoid removal rate of 89.5%, bacterial removal rate of 95.2%, and protein removal rate of 87.3% after 3 h of hydrolysis. The purified welan gum exhibited high transparency and purity. Structural characterization and antioxidant activity evaluation revealed that enzymatically purified welan gum has potential application prospects. Our study provides valuable insights into the optimal method for the enzymatic extraction and purification of welan gum. Such a method is conducive to the development of the multiple potential applications of welan gum. KEY POINTS: • A novel process for the synergistic purification of welan gum using lysozyme and alkaline protease was established. • In silico virtual digestion was employed to select the purification enzyme. • Welan gum with high transparency and purity was obtained.

Lipid Metabolism-Related Gene Markers Used for Prediction Prognosis, Immune Microenvironment, and Tumor Stage of Pancreatic Cancer.

Recently, more and more evidence shows that lipid metabolism disorder has been observed in tumor, which impacts tumor cell proliferation, survival, invasion, metastasis, and response to the tumor microenvironment (TME) and tumor treatment. However, hitherto there has not been sufficient research to demonstrate the role of lipid metabolism in pancreatic cancer. This study contrives to get an insight into the relationship between the characteristics of lipid metabolism and pancreatic cancer. We collected samples of patients with pancreatic cancer from the Gene Expression Omnibus (GEO), the Therapeutically Applicable Research to Generate Effective Treatments (TARGET), and the International Cancer Genome Consortium (ICGC) databases. Firstly, we implemented univariate regression analysis to get prognosis-related lipid metabolism genes screened and a construction of protein-protein interaction (PPI) network ensued. Then, contingent on our screening results, we explored the molecular subtypes mediated by lipid metabolism-related genes and the correlated TME cell infiltration. Additionally, we studied the disparately expressed genes among disparate lipid metabolism subtypes and established a scoring model of lipid metabolism-related characteristics using the least absolute shrinkage and selection operator (LASSO) regression analysis. At last, we explored the relationship between the scoring model and disease prognosis, tumor stage, tumor microenvironment, and immunotherapy. Two subtypes, C1 and C2, were identified, and lipid metabolism-related genes were studied. The result indicated that the patients with subtype C2 have a significantly lower survival rate than that of the patients with subtype C1, and we found difference in abundance of different immune-infiltrating cells. The gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses revealed the association of these differentially expressed genes with functions and pathways related to lipid metabolism. Finally, we established a scoring model of lipid metabolism-related characteristics based on the disparately expressed genes. The results show that our scoring model have a substantial effect on forecasting the prognosis of patients with pancreatic cancer. The lipid metabolism model is an important biomarker of pancreatic cancer. Using the model, the relationship between disease prognosis, molecular subtypes, TME cell infiltration characteristics, and immunotherapy in pancreatic cancer patients could be explored.

Effect of monosaccharide composition and proportion on the bioactivity of polysaccharides: A review.

Polysaccharides have been widely used in pharmaceutical and food industries due to their diverse bioactivity, high safety, and few or no side effects. However, inability to quickly produce, screen, and synthesize bioactive polysaccharides is the limiting factor for their development and application. Structural features determine and influence the bioactivity of polysaccharides. Among them, monosaccharide is the basic unit of polysaccharide, which not only affects electrification, functional group, and bioactivity of polysaccharide but also is one of the simplest polysaccharide indexes to be detected. At present, effects of monosaccharide composition and proportion on anti-inflammatory, antioxidant, antitumor, immunomodulatory, antibacterial, and prebiotic activities of polysaccharides are reviewed. Further problems need to be considered during regulation and analysis of monosaccharide composition and proportion of polysaccharides. Overall, present work will provide help and reference for production, development, and structure-function investigation of polysaccharides based on their monosaccharide.

Antibacterial activity of a polysaccharide isolated from Artemisia argyi leaf against Staphylococcus aureus and mechanism investigation.

Abuse of antibiotics has led to excessive amounts of antibiotic residues in food and environment, thus enhancing pathogenic bacterium resistance and threatening human health. Therefore, searching and developing safe and green antibiotic alternatives are necessary. In this study, an Artemisia argyi leaf polysaccharide (AALP) fraction was extracted and analyzed. Chemical composition analysis showed that the carbohydrate, uronic acid, protein, and polyphenol content in AALP were 68.3 % ± 4.13 %, 9.4 % ± 0.86 %, 1.79 % ± 0.27 %, and 0.16 % ± 0.035 %, respectively. Chromatographic results suggested that AALP contained rhamnose, arabinose, glucosamine, galactose, glucose, xylose, mannose, galacturonic acid, and glucuronic acid in a molar ratio of 9.26, 1.35, 1.18, 3.04, 48.51, 2.33, 31.26, 3.93, and 9.08; the weight average molecular weight, number average molecular weight, and polydispersity of AALP were 5.41 kDa, 4.63 kDa, and 1.168, respectively. Fourier transform infrared spectroscopy indicated that AALP constituted the polysaccharide-specific groups of CH, CO, and OH. Meanwhile, AALP showed a dose-dependent inhibitory effect on Staphylococcus aureus in the inhibition zone assay, and the minimal inhibitory concentration was 1.25 mg/mL. Furthermore, AALP disrupted the cell wall, depolarized the inner membrane potential, and inhibited the activities of succinate dehydrogenase and malate dehydrogenase in S. aureus.

A Comparative Study of the Effects of Whole Cereals and Refined Cereals on Intestinal Microbiota.

Cereals are one of the most important foods on which human beings rely to sustain basic life activities and are closely related to human health. This study investigated the effects of different steamed buns on intestinal microbiota. Three steamed buns were prepared using refined flour (RF), 1:1 mixed flour (MF), and whole wheat flour (WF). In vitro digestion simulations were conducted using a bionic gastrointestinal reactor (BGR) to examine their influence on intestinal microbiota. The results showed that at 0.5% addition, butyric acid and short-chain fatty acids in WF were significantly different from those in RF and MF (p < 0.05). WF also promoted the proliferation of beneficial microbiota, such as Megamonas and Subdoligranulum. At 0.5%, 1.0%, and 1.5% additions of WF, acetic acid and short-chain fatty acids at 1.5% WF increased by 1167.5% and 11.4% from 0.5% WF, respectively, and by 20.2% and 7.6% from 1.0% WF, respectively. WF also promoted the proliferation of Bifidobacterium, Lactobacillus, and Bacteroides and inhibited the growth of pathogenic microbiota, such as Streptococcus, Enterococcus, and Klebsiella. These findings support the consumption of whole cereals and offer insights into the development of new functional foods derived from wheat.

Promoting substrates uptake and curdlan synthesis of Agrobacterium sp. by attenuating the exopolysaccharide encapsulation.

During curdlan production by Agrobacterium sp., the secreted exopolysaccharide (EPS) gradually encapsulated Agrobacterium sp., accompanied by cell aggregation, resulted in inhibited substrate uptake and curdlan synthesis. To relieve the EPS encapsulation effect, the shake-flask culture medium was quantitatively supplemented with 2 % to 10 % endo-ß-1,3-glucanase (BGN), while obtaining curdlan with a decreased weight-average molecular weight ranging from 18.99 × 104 Da to 3.20 × 104 Da. In a 7-L bioreactor, the 4 % BGN supplement substantially attenuated the EPS encapsulation, resulting in increased glucose consumption and curdlan yield to 66.41 g/L and 34.53 g/L after fermentation of 108 h, which improved 43 % and 67 %, respectively compared with the control. The disruption of EPS encapsulation with BGN treatment accelerated the regeneration of ATP and UTP, resulting in sufficient uridine diphosphate glucose for curdlan synthesis. The upregulation of related genes at the transcription level reveals that the respiratory metabolic intensity, the energy regeneration efficiency, and the curdlan synthetase activity were enhanced. This study presents a simple and novel strategy of relieving the effects of EPS encapsulation on the metabolism of Agrobacterium sp. for the high-yield and value-added production of curdlan, which could be potentially applied in producing other EPSs.

New two-stage pH combined with dissolved oxygen control strategy for cyclic ß-1,2 glucans synthesis.

On the basis of a novel two-stage pH combined with dissolved oxygen (DO) control strategy in fed-batch fermentation, this research addresses the influence of pH on cyclic ß-1,2-glucans (CßGs) biosynthesis and melanin accumulation during the production of CßGs by Rhizobium radiobacter ATCC 13,333. Under these optimal fermentation conditions, the maximum cell concentration and CßGs concentration in a 7-L stirred-tank fermenter were 7.94 g L-1 and 3.12 g L-1, which were the maximum production reported for R. radiobacter. The melanin concentration of the fermentation broth was maintained at a low level, which was beneficial to the subsequent separation and purification of the CßGs. In addition, a neutral extracellular oligosaccharide (COGs-1) purified by the two-stage pH combined with DO control strategy fermentation medium was structurally characterized. Structural analyses indicated that COGs-1 was a family of unbranched cyclic oligosaccharides composed of only ß-1,2-linked D-glucopyranose residues with degree of polymerization between 17 and 23, namely CßGs. This research provides a reliable source of CßGs and structural basis for further studies of biological activity and function. KEY POINTS: • A two-stage pH combined with DO control strategy was proposed for CßGs production and melanin biosynthesis by Rhizobium radiobacter. • The final extracellular CßGs production reached 3.12 g L-1, which was the highest achieved by Rhizobium radiobacter. • The existence of CßGs could be detected by TLC quickly and accurately.

In Vitro Digestion and Fecal Fermentation of Low-Gluten Rice and Its Effect on the Gut Microbiota.

Low-gluten rice is part of a special diet for chronic kidney disease patients, but its digestive mechanism in the gastrointestinal tract is unclear. In this study, low-gluten rice (LGR), common rice (CR), and rice starch (RS) were used as experimental samples, and their digestion and bacterial fermentation were simulated using an in vitro gastrointestinal reactor to investigate the mechanism of the effect of LGR on human health. The starch digestibility of CR was higher than that of LGR, with statistically significant differences. LGR has growth-promoting and metabolic effects on Akkermansia muciniphila. Among the beneficial metabolites, the concentration of short-chain fatty acids (SCFAs) from LGR reached 104.85 mmol/L, an increase of 44.94% (versus RS) and 25.33% (versus CR). Moreover, the concentration of lactic acid reached 18.19 mmol/L, an increase of 60.55% (versus RS) and 25.28% (versus CR). Among the harmful metabolites, the concentration of branched-chain fatty acids (BCFAs) in LGR was 0.29 mmol/L and the concentration of ammonia was 2.60 mmol/L, which was 79.31% and 16.15% lower than CR, respectively. A significant increase in the concentration of the beneficial intestinal bacteria Bacteroides and Bifidobacterium occurred from LGR. The 16s rDNA sequencing showed that the abundance of the Bacteroidetes and Firmicutes increased and the abundance of the Proteobacteria and Fusobacteria decreased. Thus, LGR has positive effects on digestion and gut microbiota structure and metabolism in humans.

The effects of high-fat foods on gut microbiota and small molecule intestinal gases: release kinetics and distribution in vitro colon model.

Profiling intestinal gases and their responses to dietary changes can reveal the products and functions of the gut microbiota and their influence on human health. High-fat foods (HFF) can alter the gut microbiota and its metabolites, posing a potential health risk. However, little is known about the effects of HFF on intestinal gas distribution. Therefore, in this study, we used human fecal microorganisms as strains, an in vitro three-chamber colon model and an intestinal gas array sensor as tools. We performed in vitro fermentation using HFF as the fermentation substrate to reveal the effects of HFF on the kinetics of intestinal gas production and changes in the gut microbiota and its metabolites. We found that dietary fatty acids stimulated the production of H2S and volatile organic compounds in the colon, promoted Firmicutes abundance, and decreased Bacteroidetes abundance. These results highlight the potential role of HFF in altering the gut microbiota and intestinal gas, which can lead to health hazards.

Analysis of sleep problem in children aged 1-3 years with autism spectrum disorder in Zhejiang province, China.

Background: High prevalence of sleep problems have been founded in children with Autism Spectrum Disorder (ASD), with rates ranging from 50 to 80%. We aimed to study the sleep status and the occurrence of sleep problems in children with autism spectrum disorder (ASD) aged 1-3 years, and to provide reference for guiding early comprehensive intervention for ASD children from the perspective of sleep. Methods: From January 1 to December 31, 2021, 74 ASD children who met the diagnostic criteria of "Diagnostic and Statistical Manual of Mental Disorders-5 (DSM-V)" served as case group while 84 typically-developing children of the same sex and age served as control group. An original Children's Sleep Habit Questionnaire was adopted to compare the sleep status of children in the two groups and to conduct statistical analysis on related factors. Results: The incidence of sleep problems in the case group (78.4%) was significantly higher than that in the control group (34.5%) (P < 0.001). Compared with the children in the control group, children in the case group had later bedtime (P < 0.05) and less sleep duration (P < 0.05), and required longer time to fall asleep (P < 0.001) The incidence of sleep problems in children who could fall asleep autonomously in the case group was significantly lower than that in children who needed parental help (P < 0.05). In the case group, the longer the screen exposure time, the higher the incidence of sleep problems (P < 0.05). Conclusions: The incidence of sleep problems in ASD children aged 1-3 years is also high, mainly manifested in late bedtime, difficulty falling asleep, frequent night awakenings and less sleep duration. Both sleep patterns and screen exposure can impact their sleep. In the early comprehensive intervention of ASD children, it is necessary to pay full heed to their sleep status and take timely intervention measures in order to improve the quality of life for the ASD children and their families.

Fabrication of a smart drug delivery system based on hollow Ag2S@mSiO2 nanoparticles for fluorescence-guided synergistic photothermal chemotherapy.

A novel near-infrared (NIR) light-triggered smart nanoplatform has been developed for cancer targeting and imaging-guided combined photothermal-chemo treatment. Notably, Ag2S has a dual function of photothermal therapy and fluorescence imaging, which greatly simplifies the structure of the system. It can emit fluorescence at 820 nm under an excitation wavelength of 560 nm. The phase-change molecule of 1-tetradecanol (TD) is introduced as a temperature-sensitive gatekeeper to provide the nanocarrier with controlled release capability of doxorubicin (DOX). The nanocarrier (HAg2S@mSiO2-TD/DOX) shows a high drug loading capacity of 26.3% and exhibits an apparent NIR-responsive DOX release property. Under NIR irradiation, the photothermal effect of HAg2S nanocores facilitated the release of DOX through the melting of TD. The cytotoxicity test shows that the nanocarriers have good biocompatibility. As the same time, the synergistic combination leads to a better cancer inhibition effect than individual therapy alone in vitro. Cell uptake tests indicate that the carriers have excellent fluorescence imaging ability and high cellular uptake for HepG2 cells. This work provides a new strategy for the fabrication of smart nanocarriers with simple structures for fluorescence-mediated combination cancer therapy. Fabrication of a smart drug delivery system based on hollow Ag2S@mSiO2 nanoparticles for fluorescence-guided synergistic photothermal chemotherapy.

Encapsulation of polyphenols in pH-responsive micelles self-assembled from octenyl-succinylated curdlan oligosaccharide and its effect on the gut microbiota.

An amphiphilic polymer based on octenyl succinic anhydride-modified curdlan oligosaccharide (MCOS) was synthesized. The critical micelle concentration of MCOS was 3.91 µg·mL-1. MCOS could self-assemble into spherical micelles with a particle size of 230.1 nm and a zeta potential of - 37.9 mV. When used for polyphenol encapsulation, the loading capacity of curcumin and quercetin-co-encapsulated micelles was higher than that of single-polyphenol encapsulated micelles. In vitro gastrointestinal release test showed that the MCOS micelle presented a pH-dependent release, released a little polyphenol in simulated gastric fluid, but presented sustained release in the simulated intestinal fluid. The gastrointestinal-digested polyphenol-loaded micelles exhibited excellent antioxidant ability. In vitro human fecal fermentation indicated that the MCOS carrier could promote the production of short-chain fatty acids by gut microbiota and exhibited the highest relative abundance of Megamonas. In addition, the supplementation of curcumin and quercetin-co-loaded MCOS micelles increased the relative abundance of Bifidobacterium and inhibited the growth of Escherichia_Shigella. These findings indicated that the MCOS carrier can be potentially used for the colon-targeted delivery of hydrophobic polyphenols due to its pH-responsive property.

Antioxidant and anti-inflammatory properties of an aminoglycan-rich exopolysaccharide from the submerged fermentation of Bacillus thuringiensis.

Proteins from Bacillus thuringiensis are widely used as biopesticides but little is known about its exopolysaccharides. The exopolysaccharide BPS-2 was extracted from B. thuringiensis IX-01 after high-cell-density fermentation. BPS-2 is a heteropolysaccharide (molecular weight 29.36 kDa) composed of D-galactosamine, arabinose, glucosamine, glucose, and mannose in molar ratios 5.53: 1.77:4.74:3.24:1. In vitro upper gastrointestinal simulations showed that BPS-2 has strong anti-digestive capacity, with scavenging of DPPH, hydroxyl, ABTS, and superoxide anions radicals of 31.34 ± 1.67 %, 32.43 ± 3.01 %, 34.31 ± 2.12 %, and 48.53 ± 3.55 %, respectively, after BPS-2 entered the colon. It significantly inhibited production of lipopolysaccharide-induced nitric oxide and multiple pro-inflammatory cytokines and had proliferative effects on RAW 264.7 cells. BPS-2 inhibited malondialdehyde secretion and elevated activities of glutathione peroxidase, superoxide dismutase, and total antioxidants, significantly improving the antioxidant status of inflammation model cells. This first report of the in vitro anti-inflammation and antioxidant properties of BPS-2 from B. thuringiensis provides a basis for biopharmaceutical applications.

Emulsifying properties and bioavailability of clove essential oil Pickering emulsions stabilized by octadecylaminated carboxymethyl curdlan.

In the present study, clove essential oil (CEO) Pickering emulsions were stabilized by octadecylamine-modified carboxymethyl curdlan (CMCD-ODA) at different pH values. The droplet size and negatively charged zeta potential of the CMCD-ODA emulsions decreased as the pH increased from 3.0 to 11.0. Rheology results indicated that the CMCD-ODA polymer/emulsion prepared at pH 5.0 showed higher apparent viscosity and viscoelasticity than other pH conditions, which might prevent droplets from flocculating. The Pickering emulsions obtained at pH 5.0 were spherical droplets with a uniform size distribution and a mean diameter of 9.54 µm, and they exhibited excellent stability during 28 days of storage. The morphological structures of the emulsions investigated by confocal laser scanning microscopy and scanning electron microscopy indicated that the CMCD-ODA Pickering emulsion obtained at pH 5.0 was stabilized by loading amphiphilic CMCD-ODA polymer around the spherical oil droplets and forming a weak gel network structure. The CEO-loaded CMCD-ODA emulsions had higher antioxidant capacity than free CEO after 28 days of storage at pH 5.0. Given the good emulsion stability, antioxidant activity, and great antibacterial effect, the CEO-loaded carboxymethyl curdlan Pickering emulsion has promising applications in food, cosmetic, and biomedicine industries.