The antioxidant and free-radical scavenging activities of extract and fractions from corn silk (Zea mays L.) and related flavone glycosides

This study was designed to evaluate both the antioxidant and free-radical scavenging activities of extract and fractions from corn silk. N-butanol fraction (BF) demonstrated the highest total phenolic content (164.1 ± 9.7 μg GAE/g DCS) and total flavonoids content (69.4 ± 5.1 μg RE/g DCS), accompanied with the highest antioxidant activity compared to other fractions through all antioxidant assays. Two flavone glycosides showing potent antioxidant activity were isolated from BF and identified, by comparing spectral data (UV, FAB-MS and NMR) with literature values, to be isoorientin-2″-O-α-l-rhamnoside and 3′-methoxymaysin. The two isolated flavone glycosides, particularly isoorientin-2″-O-α-l-rhamnoside, demonstrated significant total antioxidant activity, DPPH radical scavenging activity, reducing power and iron-chelating capacity, with EC₅₀ values of 14.24 ± 1.49, 22.69 ± 2.33, 6.58 ± 1.07 and 30.25 ± 3.05 μg/ml, respectively. Results obtained indicated that corn silk extracts can be used potentially as a ready accessible and valuable bioactive source of natural antioxidants.     

Identification of flavonol glycosides in American cranberry fruit

Two flavonol glycosides, quercetin galactoside and quercetin arabinoside, have been identified in American cranberry fruit, as a complementary investigation of our previous study. The analysis processes included separation, hydrolysis and structure elucidation of flavonol glycosides. The separation of flavonol glycosides was carried out by solvent extraction, thin-layer chromatography (TLC) and high performance liquid chromatography (HPLC). After hydrolysis of the obtained flavonol glycosides, flavonol aglycones and sugars were identified by HPLC and gas chromatography–mass spectrometry (GC–MS), respectively.     

Preparation and Characterization of C10–C14 Alkyl Cellulose Ester Sulfate Surfactant

The aim of this work is to synthesize surfactants based on cellulose with different molecular weights. Raw cotton cellulose was tailored into cellulose segments with different molecular weights by a hydrothermal process, then the average degree of polymerization (DP) was determined by viscosimetry and the molecular weight distribution was estimated by gel permeation chromatography. The C₁₀–C₁₄ alkyl cellulose ester sulfate surfactants were prepared by hydrophilic sulfonation and hydrophobic esterification. The surface tension of the surfactants solution was obtained by the Wilhelmy plate method. Results showed that the cellulose segments presented a broader distribution compared with the raw material. The critical micelle concentration (CMC) value decreased from 1.08 to 0.86 wt% as the hydrophobic chain length was increased from 10 to 14. The CMC values of cellulose surfactants with C₁₄-acyl chloride hydrophobization decreased from 1.32 to 0.86 wt% as the DP was decreased from 2,700 to 296.     

GH-16 Type β-1,3-Glucanase from Lysobacter sp. MK9-1 Enhances Antifungal Activity of GH-19 Type Chitinase,and Its Glucan-binding Domain Binds to Fungal Cell-wall

The GH-16 type β-1,3-glucanase (BgluC16MK) gene of Lysobacter sp. MK9-1 was cloned to study its antifungal activities. BgluC16MK displays amino acid sequence similarity with GluC from L. enzymogenes strain N4-7. BgluC16MK includes a signal sequence, a catalytic domain and carbohydrate-binding module family 6-type β-glucan binding domain (B-GBD). The expression of the BgluC16MK gene in Escherichia coli without the signal sequence resulted in antifungal activity at a dose of 0.6-0.8 nmol/disk. However, BgluC16MK displayed antifungal activity at a dose of 0.025 nmol/disk in combination with Chi19MK. Substrate-specific assay revealed that purified BgluC16MK hydrolyzed insoluble curdlan more readily than the soluble substrate. Furthermore, to explore the binding selectivity of B-GBD of BgluC16MK, we constructed a fusion protein (B-GBD-GFP) using the B-GBD and green fluorescent protein. The activity of the fusion protein against various substrates indicates that B-GBD was selective for glucans with β-1,3-linkages. An additional study demonstrated the binding ability of B-GBD-GFP to the cell-wall of living fungi, such as T. reesei and Aspergillus oryzae. These findings suggest that BgluC16MK can be utilized to generate antifungal enzyme preparations and that the fusion protein B-GBD-GFP can be used to identify the fungal cell surface structure using β-glucans.     

Comprehensive Analysis of Copy Number Variations on Glycoside Hydrolase 45 Genes among Different Bursaphelenchus xylophilus Strains

Bursaphelenchus xylophilus is considered the most dangerous quarantine pest in China. It causes enormous economic and ecological losses in many countries from Asia and Europe. The glycoside hydrolase 45 gene family has been demonstrated in early studies to contribute to the cell wall degradation ability of B. xylophilus during its infection. However, the copy number variation (CNV) of the GH45 gene and its association with B. xylophilus pathogenicity were not fully elucidated. In this study, we found that the GH45 gene with two copies is the most predominant type among 259 B. xylophilus strains collected from China and Japan. Additionally, 18 strains are identified as GH45 genes with a single copy, and only two strains are verified to have three copies. Subsequent expression analysis and inoculation test suggest that the copy numbers of the GH45 gene are correlated with gene expression as well as the B. xylophilus pathogenicity. B. xylophilus strains with more copies of the GH45 gene usually exhibit more abundant expression and cause more severe wilt symptoms on pine trees. The aforementioned results indicated the potential regulatory effects of CNV in B. xylophilus and provided novel information to better understand the molecular pathogenesis of this devastating pest.     

A Bacillus licheniformis Glycoside Hydrolase 43 Protein Is Recognized as a MAMP

Glycoside hydrolases from pathogens have often been reported as inducers of immune responses. However, the roles of glycoside hydrolase from plant-growth-promoting rhizobacteria (PGPR) in the resistance of plants against pathogens is not well studied. In this study, we identified a glycoside hydrolase 43 protein, H1AD43, produced by Bacillus licheniformis BL06 that can trigger defense responses, including cell death. Ion-exchange and size-exclusion chromatography were used for separation, and the amino acid sequence was identified by mass spectrometry. The recombinant protein generated by prokaryotic expression was able to elicit a hypersensitive response (HR) in Nicotiana benthamiana and trigger early defense responses, including reactive oxygen species (ROS) burst, callose accumulation, and the induction of defense genes. In addition, the protein could induce resistance in N. benthamiana, in which it inhibited infection by Phytophthora capsici Leonian and tobacco mosaic virus-green fluorescent protein (TMV-GFP) expression. H1AD43 thus represents a microbe-associated molecular pattern (MAMP) of PGPR that induces plant disease resistance and may provide a new method for the biological control of plant disease.     

The Flavonol Quercitrin Hinders GSK3 Activity and Potentiates the Wnt/β-Catenin Signaling Pathway

The Wnt/β-catenin signaling pathway dictates cell proliferation and differentiation during embryonic development and tissue homeostasis. Its deregulation is associated with many pathological conditions, including neurodegenerative disease, frequently downregulated. The lack of efficient treatment for these diseases, including Alzheimer’s disease (AD), makes Wnt signaling an attractive target for therapies. Interestingly, novel Wnt signaling activating compounds are less frequently described than inhibitors, turning the quest for novel positive modulators even more appealing. In that sense, natural compounds are an outstanding source of potential drug leads. Here, we combine different experimental models, cell-based approaches, neuronal culture assays, and rodent behavior tests with Xenopus laevis phenotypic analysis to characterize quercitrin, a natural compound, as a novel Wnt signaling potentiator. We find that quercitrin potentiates the signaling in a concentration-dependent manner and increases the occurrence of the Xenopus secondary axis phenotype mediated by Xwnt8 injection. Using a GSK3 biosensor, we describe that quercitrin impairs GSK3 activity and increases phosphorylated GSK3β S9 levels. Treatment with XAV939, an inhibitor downstream of GSK3, impairs the quercitrin-mediated effect. Next, we show that quercitrin potentiates the Wnt3a-synaptogenic effect in hippocampal neurons in culture, which is blocked by XAV939. Quercitrin treatment also rescues the hippocampal synapse loss induced by intracerebroventricular injection of amyloid-β oligomers (AβO) in mice. Finally, quercitrin rescues AβO-mediated memory impairment, which is prevented by XAV939. Thus, our study uncovers a novel function for quercitrin as a Wnt/β-catenin signaling potentiator, describes its mechanism of action, and opens new avenues for AD treatments.     

大豆异黄酮糖苷酸法水解工艺的研究

通过单因素和正交试验得到了大豆异黄酮糖苷酸法水解为大豆异黄酮苷元的最佳工艺条件为:盐酸乙醇的浓度为3 mol/L,水解温度为80℃,水解时间180 min,酸法水解率为81.31%.     

发酵处理对大豆制品中异黄酮含量与组分的影响

利用发酵大豆食品———豆豉与酸豆乳作为试验对象 ,通过与对照品比较 ,发现发酵处理对大豆食品中的异黄酮总含量的影响不大 ,但对其异黄酮的组分有较大的影响 ,发酵处理使糖苷型大豆异黄酮在 β 葡萄糖苷酶的作用下水解为游离型大豆异黄酮 ,从而使发酵制品中的游离型大豆异黄酮的含量增高 ,而糖苷型大豆异黄酮的含量降低     

高效液相色谱法同时测定柚子中的4种黄酮苷和柠檬苦素

为同时测定柚子中芸香柚皮苷、柚皮苷、橙皮苷、新橙皮苷和柠檬苦素,研究了高效液相色谱的洗脱条件、测定波长等对测定结果的影响。结果表明,优选的测定条件为色谱柱:Bio-Bond 5μm C₁₈(4.6 mm×250 mm);进样量10μL;黄酮苷与柠檬苦素的检测波长分别为283 nm和220 nm;流动相:乙腈(A)和水(B),梯度洗脱:0 min,22%A;8 min,22%A;10 min,95%A;16 min,95%A;18 min,22%A;20 min,22%A,流速1 mL/min。该测定条件下,上述5种成分分离良好;其标准曲线相关系数均大于0.9997,检出限为0.69~1.72 mg/L,定量限为1.51~3.15 mg/L,重复性相对标准偏差小于2%。加样回收率均大于98%,加样回收率相对标准偏差均小于2%。