Preparation, structural characterization and prebiotic potential of mulberry leaf oligosaccharides.

The present study shows the purification of a main oligosaccharide fraction (MLO 1-2) from the enzymatic hydrolysate of mulberry leaf polysaccharides by DEAE-52 cellulose and gel column chromatography. The physicochemical properties of MLO 1-2 were characterized. The structure of MLO 1-2 was obtained as follows: α-(2-OAc)-Manp-1 → 2-ß-Glcp-1 → 4-ß-Glcp-1 → 4-α-Glcp-1 → 2-α-Glcp-1 → 2-α-Galp-1 → 2-ß-Galp-1 → 2-ß-Galp-1, which was elucidated by methylation and NMR analysis. The molecular weight of MLO 1-2 showed no significant change after simulated saliva, gastric and intestinal digestion. This indicated that MLO 1-2 could pass through the digestive system without being degraded to safely reach the colon to regulate the gut microbiota. Additionally, MLO 1-2, more than glucose or galactooligosaccharides, promoted the proliferation of Bifidobacterium bifidum, B. adolescentis, Lacticaseibacillus rhamnosus and Lactobacillus acidophilus. Furthermore, the acetic and lactic acid concentrations of bacterial cultures inoculated with MLO 1-2 were higher than those inoculated with glucose and galactooligosaccharide (GOS). These results suggest that MLO 1-2 could be an excellent prebiotic for intestinal flora regulation and the promotion of gut health.

The hypoglycemic effect of freeze-dried fermented mulberry mixed with soybean on type 2 diabetes mellitus.

Mulberry has significant hypoglycemic effect and can be used as an auxiliary food for people with type 2 diabetes. However, it is rich in carbohydrate and cannot be consumed directly by diabetic patients. In the study, we fermented the mulberry to reduce the content of glucose and fructose, and added the soybean to reduce the loss of probiotics during fermentation and then determined its hypoglycemic effect. We induced type 2 diabetes mellitus (T2DM) mice by streptozotocin and measured its blood glucose, serum biochemistry, hepatic and pancreatic histopathology, and the diversity of the gut microbiota. After 5 weeks of oral DFMS administration, the glucose tolerance was improved significantly in T2DM mice. Furthermore, there were also significant increases in superoxide dismutase activity and glutathione concentration, and marked reductions in the concentrations of malondialdehyde and free fatty acids. Moreover, DFMS also prevented histopathological changes and the increases in the activities of alanine transaminase and aspartate transaminase. DFMS treatment also markedly increased the richness of the gut microbial community. The abundance of Bacteroidetes was increased, and those of Proteobacteria, Escherichia-Shigella, and Lactobacillus were reduced. In summary, DFMS has a clear hypoglycemic effect in mice with T2DM.

Effects of enzymatic hydrolysis on the structural, rheological, and functional properties of mulberry leaf polysaccharide.

Effects of enzymatic hydrolysis on the structural, rheological, and functional properties of mulberry leaf polysaccharide (MLP) were characterized in this study. The enzymatic hydrolysis of MLP raised the carbonyl, carboxyl, and hydroxyl groups from 7.21 ± 0.86 to 10.08 ± 0.28 CO/100 Glu, 9.40 ± 0.13 to 17.55 ± 0.34 COOH/100 Glu, and 5.71 ± 0.33 to 8.14 ± 0.24 OH/100 Glu, respectively. Meanwhile, an increase in thixotropic performance and structure-recovery capacities were observed in hydrolyzed MLP, while the molecular weight, surface tension, apparent viscosity, and thermal stability were decreased. An improved antioxidant activity of MLP was also achieved after the enzymatic degradation. Moreover, the hydrolyzed MLP showed greater ability to promote the growths of Bifidobacterium bifidum, Bifidobacterium adolescentis, Lactobacillus rhamnosus, and Lactobacillus acidophilus and the production of acetic acid, butyric acid, and lactic acid. The results demonstrate that enzymatic modification is a useful approach for polysaccharide processing.

Effect of 1-Deoxynojirimycin Isolated from Mulberry Leaves on Glucose Metabolism and Gut Microbiota in a Streptozotocin-Induced Diabetic Mouse Model.

1-Deoxynojirimycin (DNJ) exerts hypoglycemic effects. However, the traditional method for DNJ extraction is inefficient, and the hypoglycemic mechanism of DNJ remains unclear. In this study, the mixed fermentation by Lactobacillus fermentum and Saccharomyces cerevisiae was used to enhance DNJ extraction efficiency. It was found that this strategy was more efficient than the traditional method as the yield improved from the original 3.24 mg/g to 5.97 mg/g. The purified DNJ significantly decreased serum glucose (P < 0.01) and insulin levels (P < 0.05), improved serum lipid levels (P < 0.05), and reversed insulin resistance (P < 0.05) in diabetic mice. These changes were caused by up-regulating the protein expression of insulin receptor and glycolysis enzymes (GK, PK, and PFK) (P < 0.05) and down-regulating the protein expression of insulin receptor substrate-1 and gluconeogenesis enzymes (PCB, PEPCK, FBPase, and G-6-Pase) (P < 0.05), thus alleviating glucose tolerance. Additionally, DNJ treatment relieved gut dysbiosis in diabetic mice by promoting the growth of Lactobacillus, Lachnospiraceae NK4A136 group, Oscillibacter, norank Lachnospiraceae, Alistipes, and Bifidobacterium (P < 0.05) and suppressing the growth of Ruminococcaceae UCG-014, Weissella, Ruminococcus, Prevotellaceae Ga6A1 group, Anaerostipes, Klebsiella, Prevotellaceae UCG-001, and Bacteroidales S24-7 group (P < 0.05).

Qualitative and quantitative determination of seven triterpene acids in Eriobotrya japonica Lindl. by high-performance liquid chromatography with photodiode array detection and mass spectrometry.

INTRODUCTION: The leaves of Eriobotrya japonica are used for the treatment of diabetes mellitus, chronic bronchitis, coughs and skin diseases. No method is currently available, however, by which to assess the quality of the crude herb on the basis of the quantitative profile of the main bioactive triterpene acids present. OBJECTIVE: To develop a simple and accurate HPLC-UV (photodiode array detection) method for the simultaneous quantification of seven triterpene acids in the leaves of E. japonica. METHOD: Separations were performed on an Ultimate XB-C18 column by gradient elution using methanol:formic acid. Analytes were identified by HPLC coupled with electrospray ionisation mass spectrometry experiments. The quantitative HPLC-UV method was validated for linearity, precision, accuracy, and limits of detection and quantification. RESULTS: Calibration curves presented good linear regression (r > 0.9992) within test ranges. The precision and accuracy of the method were acceptable with overall intra-day and inter-day variations of 1.35-3.30 and 1.98-4.43%, respectively, and overall recoveries of 95.60-102.67% for the seven compounds analysed. The method was successfully applied to the quantification of seven triterpene acids in eleven samples of E. japonica collected from different provinces of China. CONCLUSION: The developed assay could be considered as a suitable quality control method for E. japonica.