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We extracted, purified, and characterized three neutral and three acidic polysaccharides from the roots, stems, and leaves of Aralia continentalis Kitigawa. The results of the analysis of monosaccharide composition indicated that the polysaccharides from the roots and stems were more similar to each other than they were to the polysaccharides from the leaves. The in vitro antioxidant results demonstrated that the acidic polysaccharides had stronger antioxidant activity than the neutral fractions. Therefore, we investigated the primary purified acidic polysaccharide fractions (WACP(R)-A-c, WACP(S)-A-c, and WACP(L)-A-d) by NMR and enzymatic analysis. The structural analytical results indicated that WACP(R)-A-c contained homogalacturonan (HG); WACP(S)-A-c contained HG and rhamnogalacturonan II (RG-II), and WACP(L)-A-d contained HG, RG-II, and rhamnogalacturonan I (RG-I) domains. Our findings offer insights into the screening of natural polysaccharide-based antioxidants and provide a theoretical basis for the application of A. continentalis.

Three β-glucosidases from Bifidobacterium adolescentis ATCC15703, namely, BaBgl1A, BaBgl3A, and BaBgl3B, were overexpressed in Escherichia coli. The recombinant β-glucosidases were sufficiently purified using Ni2+ affinity chromatography, and BaBgl1A exhibited the best purification efficiency with a purification factor of 2.3-fold and specific activity of 71.2 U/mg. Three recombinant β-glucosidases acted on p-nitrophenyl-β-glucopyranoside (pNPβGlc) at around pH 7.0 and 30–50°C. The results of the substrate specificity assay suggested that BaBgl1A acted exclusively as β-1,2-glucosidase, while BaBgl3A and BaBgl3B acted mostly as β-1,3-glucosidase and β-1,4-glucosidase, respectively. The substrate specificity of the three recombinant enzymes was further studied using the ginsenosides Rb1 and Rd as substrates. The results of thin-layer chromatography and high-performance liquid chromatography analyses showed that BaBgl1A exhibited the highest bioconversion ability on Rb1 and Rd, where it hydrolyzed the outer C-3 glucose moieties of Rb1 and Rd into the rare ginsenosides Gypenoside XVII and F2; BaBgl3A exhibited medium bioconversion ability on Rb1, where it hydrolyzed both the outer C-3 and C-20 glucose moieties of Rb1 into Gyp XVII and Rd; and BaBgl3B was not active on Rb1 and Rd. These β-glucosidases will act as new biocatalytic tools for transforming ginsenosides and preparing active glycosides and aglycone.

A neutral polysaccharide (SSPS-N) and an acidic polysaccharide (SSPS-A) were isolated from Okara by anion-exchange chromatography. The physical and chemical properties of polysaccharides were studied by HPLC, FT-IR, and SEM. The results showed that SSPS-N might be a galactomannans with a lamella-like structure, and SSPS-A might be an arabinogalactan with a complete porous structure. Furthermore, a new neutral soybean soluble polysaccharide (SSPS-N-b) with a molecular weight of 8.6 kDa was isolated from SSPS-N, and its fine structure was analyzed by GC-MS, enzymatic analysis, and NMR. The results showed that SSPS-N-b was a mixture of β-1,4galactan and glucomannan, which (1→4)βd-linked mannose and (1→4)βd-linked glucose were connected alternately, and it was in parallel with β-1,4-galactan. The antioxidant activity results showed that β-1,4-Galactan in SSPS-N-b was identified as the main activity domain. This result will be utilized for the development of SSPS as novel functional foods and medicine.

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