Structural characterization of a polysaccharide from Abelmoschus esculentus L. Moench (okra) and its hypoglycemic effect and mechanism on type 2 diabetes mellitus.

A novel acidic polysaccharide named AeP-P-1 was prepared from Abelmoschus esculentus L. Moench (okra). AeP-P-1 is a heteropolysaccharide with a molecular weight of 3.02 × 103 kDa and is composed of L-rhamnose, D-galactose, and D-galacturonic acid in the ratio 1.87 : 3.58 : 1.00. Structural characterization based on methylation and 1D/2D NMR analyses indicated that AeP-P-1 is composed of T-linked-Rhap, T-linked-Galp, 1,2,4-linked-Rhap, 1,4-linked-Galp, 1,6-linked-Galp, and 1,3,4-linked-Galp in a molar ratio of 2.42 : 3.36 : 6.46 : 13.31 : 3.12 : 1, respectively. The hypoglycemic effect and mechanism of AeP-P-1 on type 2 diabetes mellitus were also explored. Firstly, AeP-P-1 can reduce blood lipids and liver and kidney damage caused by T2DM. Finally, AeP-P-1 induces the phosphorylation of GSK3ß, maintains the activity of glycogen synthase (GCS), and promotes glycogen synthesis by regulating the expression of insulin/PI3K/Akt pathway proteins. These results indicated that AeP-P-1 could be developed as a potential ingredient in immunostimulatory agents.

Rheological properties of polysaccharides from Pholiota nameko with different temperature extraction: Concentration, pH, temperature, and saltion.

Cold and hot water extracted polysaccharides (CW-PNPs and HW-PNPs) were isolated from Pholiota nameko. The rheological properties of PNPs were investigated by steady shear and oscillatory rheological measurements. The PNPs exhibited typical non-Newtonian and shear-thinning behavior, which are affected by PNP concentration, temperature, pH value, salt ion, and concentration. Specifically, the apparent viscosity of the two PNPs solutions at concentration of 1% (w/w) was shown as HW-PNPs > CW-PNPs. The apparent viscosity of PNPs decreases under acidic and alkaline conditions and when the temperature rises; K+ and Na+ cause the apparent viscosity of CW-PNPs to decrease, while Ca2+ and Al3+ are opposite. The addition of four different salt ions all caused the apparent viscosity of the HW-PNPs to decrease. The results of dynamic rheological experiments show that G' and G″ showed slightly frequency dependency with G' exceeding G″ throughout the accessible range of frequency for CW-PNPs and HW-PNPs.

Structural characterization and prebiotic potential of an acidic polysaccharide from Imperial Chrysanthemum.

A novel water-soluble polysaccharide, named ICP-1, was isolated and purified by Sephadex G-200 after extracting the crude polysaccharide (ICP) from Imperial Chrysanthemum. The structural characterization of ICP-1 was determined by physical and chemical methods, FT-IR, NMR, SEM, HPGPC, periodate oxidation, Smith degradation, methylation and Congo red test. Then, acid production and proliferation of lactic acid bacteria and the tolerance tests of simulated gastrointestinal fluid were measured to investigate the activity of prebiotic potential. The results showed that ICP-1 was an acidic hetero-polysaccharide with an average molecular weight of 2.98 × 103 kDa and a specific optical rotation of +155°. The glycosyl residues of ICP-1 were composed of (1→), (1→4) and (1→6) glucose, (1→5) arabinose, (1→4) galacturonic acid and (1→3,6) mannose. Besides, ICP-1 can speed up the acid production of lactic acid bacteria and promote the growth and proliferation of lactic acid bacteria effectively.

Structure, antioxidant property and protection on PC12 of a polysaccharide isolated and screened from Abelmoschus esculentus L.Moench (okra).

AeP-P-2, a pectic polysaccharide, was extracted from the fruit pod of okra. It composed of rhamnose (Rha), arabinose (Ara), glucose (Glc), galactose (Gal) and galacturonic acid (GalA) with the ratio of 4.75:2.01:1.00:4.91:7.24. The main structural feature of AeP-P-2 are 1,4-linked galacturonan units (homogalacturonan backbone) and (1 → 2) and (1 → 2,4) linked Rha (rhamnogalacturonan I region). And the other side chains contained →1)-linked Ara, (1 → 5)-linked Ara, (1 → 4)-linked Glc, (1 → 6)-linked Gal, (1 → 4)-linked Rha, (1 → 2,4)-linked Rha, →1)-linked Ara and →1)-linked Gal. When the concentration of AeP-P-2 was 3.2 mg/mL, the scavenging rates on DPPH·, ABTS, O2-· and ·OH reached to 61.88%, 87.10%, 52.17% and 60.32%, respectively. AeP-P-2 also could protect PC12 cells from the damage of H2O2 and reduce apoptosis caused by oxidative damage by decreasing the level of ROS. The findings indicated that okra was a functional vegetable and AeP-P-2 was worth studying and developing into antioxidant component.

Effect of steam explosion pretreatment on the structure and bioactivity of Ampelopsis grossedentata polysaccharides.

Steam explosion (SE) was a friendly environmentally pretreatment method. In this study, the effect of steam explosion (SE) pretreatment on structure and α-glucosidase inhibitory activity of Ampelopsis grossedentata polysaccharides was evaluated. Two novel polysaccharides (AGP and AGP-SE) were extracted, isolated, purified and analyzed by NMR, FT-IR and methylation. The results indicated that AGP mainly consisted of Rha, Xyl, Glc, and Ara with a molecular weight of 2.74 × 103 kDa and AGP-SE mainly consisted of Man, Ara, and Gal with a molecular weight of 2.14 × 103 kDa. Furthermore, the backbone of AGP and AGP-SE were mainly composed of 5)-Araf-(1→, -Glcp-(1→, 6)-Glcp-(1→, 6)-Galp-(1→, 3,6)-Manp-(1→, and 2,3,6)-Glcp-(1→. Finally, we demonstrated that all polysaccharides exhibited obviously α-glucosidase inhibition activity and mixed type inhibition. AGP-SE had better α-glucosidase inhibition activity and the binding affinity KD on α-glucosidase by using Surface Plasmon Resonance (SPR) than AGP. Overall, SE pretreatment is an effective method for extracting polysaccharide and provides a new idea into the improvement of biological activity.

Enzymatic characterization and validation of gene expression of phosphoglucomutase from Cordyceps militaris.

The purification and characterization of PGM (Phosphoglucomutase) from Cordyceps militaris (C. militaris) was investigated. PGM was purified using a combination of ultrafiltration, salting-out and ion exchange chromatography resulting in 4.23-fold enhancement of activity with a recovery of 20.01%. Molecular mass was 50.01 kDa by SDS-PAGE. The optimal activity was achieved at pH 7.5 and 30 °C with NADPH as substrate. The results showed that SDS, DTT Li+, Cu2+, Na+, Mn2+ and Al3+ were effective PGM inhibitors; whereas glycerol, Zn2+, Mg2+, Ca2+, Fe2+ and Fe3+ could enhance the activity of PGM, and the Km and Vmax values were 11.62 mmol/L and 416.67 U/mL, respectively. At the same time, qRT-PCR was used to test the changes of mRNA transcription level of PGM gene encoding under two fermentation conditions: basic medium and optimized medium. The relative quantitative results of PGM target genes resulting in 2.60-fold enhancement than the control group.

Abscisic acid stimulates a calcium-dependent protein kinase in grape berry.

It has been demonstrated that calcium plays a central role in mediating abscisic acid (ABA) signaling, but many of the Ca2+-binding sensory proteins as the components of the ABA-signaling pathway remain to be elucidated. Here we identified, characterized, and purified a 58-kD ABA-stimulated calcium-dependent protein kinase from the mesocarp of grape berries (Vitis vinifera x Vitis labrusca), designated ACPK1 (for ABA-stimulated calcium-dependent protein kinase1). ABA stimulates ACPK1 in a dose-dependent manner, and the ACPK1 expression and enzyme activities alter accordantly with the endogenous ABA concentrations during fruit development. The ABA-induced ACPK1 stimulation appears to be transient with a rapid effect in 15 min but also with a slow and steady state of induction after 60 min. ABA acts on ACPK1 indirectly and dependently on in vivo state of the tissues. Two inactive ABA isomers, (-)-2-cis, 4-trans-ABA and 2-trans, 4-trans-(+/-)-ABA, are ineffective for inducing ACPK1 stimulation, revealing that the ABA-induced effect is stereo specific to physiological active (+)-2-cis, 4-trans-ABA. The other phytohormones such as auxin indoleacetic acid, gibberellic acid, synthetic cytokinin N-benzyl-6-aminopurine, and brassinolide are also ineffective in this ACPK1 stimulation. Based on sequencing of the two-dimensional electrophoresis-purified ACPK1, we cloned the ACPK1 gene. The ACPK1 is expressed specifically in grape berry covering a fleshy portion and seeds, and in a developmental stage-dependent manner. We further showed that ACPK1 is localized in both plasma membranes and chloroplasts/plastids and positively regulates plasma membrane H+-ATPase in vitro, suggesting that ACPK1 may be involved in the ABA-signaling pathway.