The antiobesity molecular mechanisms of mulberry leave components were analyzed based on intestinal micro-ecology and metabolomics. An obesity model was established by feeding rats with a high-calorie diet. Rats were divided into seven groups: the obesity model control (MC), positive control (PC), mulberry leaf powder (MLP), mulberry leaf fiber (MLF), mulberry leaf polyphenols (MLPS), mulberry leaf fiber and polyphenols mixture (MLM), and normal control (NC), and fed daily for 6 consecutive weeks. The results demonstrated that the MLM group had the best efficiency on weight loss, indicating synergistic interactions between MLPS and MLF. The reduction of Firmicutes abundance, and the downstream Clostridiales, Lachnespiraceae, was a key pathway for the antiobesity effects. The increased abundances of Lactobacillus vaginalis and Lactobacillus gasseri might result in lipid metabolism disorder. The test groups regulated the amino acid and oligopeptides metabolic disorder tents to normal levels compared with the MC and NC groups.

Applying the intermolecular co-pigmentation to improve the stability of mulberry anthocyanins is an important co-pigment method. Seven co-pigments, ferulic acid (FA), caffeic acid (CA), p-hydroxybenzoic acid (HBA), protocatechuic acid (PA), gallic acid (GA), vanillic acid (VA) and vanillin (VN) were selected to investigate mulberry anthocyanin co-pigmentation thermal reaction kinetics. The strongest co-pigment reactions were observed for FA at a molar ratio of 1:20, pH 3.5 and 20 °C, with the highest hyperchromic effects (52.94%), equilibrium constant (K) values (3.51) and negative values of Gibbs free energy (ΔG°) (−3.06 KJ/mol). Co-pigments that contained more free hydroxyl groups facilitated the co-pigmentation, and methyl contributed more to color enhancement, with respect to the hydrogen group. Ultra Performance Liquid Chromatography-Quadrupole-Time Of Flight-Mass/Mass Spectrometry (UPLC-Q-TOF-MS/MS) results indicated that FA and CA formed different anthocyanin derivatives with mulberry anthocyanin. The Fourier Transform Infrared Spectroscopy (FTIR) and molecular docking confirmed that hydrogen bonding, π–π stacking and hydrophobic interaction were formed between anthocyanins and three prevalent co-pigments (FA, CA and VA). CA and C3G could form four hydrogen bonds and two π–π stackings; this was the most stable system among three phenolic acid–C3G complexes. Due to the functional effect of phenolic acids, the addition of FA and CA not only enhanced the stability and color intensity of mulberry anthocyanins but also the functionality of the processing product.

A water-soluble oligosaccharide termed EMOS-1a was prepared by enzymatic hydrolysis of polysaccharides purified from mulberries by column chromatography. The chemical structure of the purified fraction was investigated by ultraviolet spectroscopy, Fourier-transform infrared spectroscopy, and gas chromatography–mass spectrometry, which indicated that galactose was the main constituent of EMOS-1a. Chemical analyses showed that the uronic acid and sulfate content of EMOS-1a were 5.6% and 8.35%, respectively, while gel permeation chromatography showed that EMOS-1a had an average molecular weight of 987 Da. The antioxidant activities of EMOS-1a were next investigated, and EMOS-1a exhibited concentration-dependent 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity, Trolox equivalent antioxidant capacity, and ferric reducing antioxidant power. The level of proliferation of Lactobacillus rhamnosus reached 1420 ± 16% when 4% (w/v) EMOS-1a was added, where the number of colonies in MRS (de Man, Rogosa, and Sharpe) medium with no added oligosaccharide was defined as 100% proliferation. These results indicate that the oligosaccharide EMOS-1a could be used as a natural antioxidant in prebiotic preparations.