Isolation and characterization of a nephroprotective polysaccharide from Dendrobium chrysotoxum Lindl against LPS-induced acute kidney injury mice.

The structure and bioactivity of a novel polysaccharide from Dendrobium Chrysotoxum Lindl (DCP-1) were investigated. The crude polysaccharides of Dendrobium Chrysotoxum Lindl (DCP) were extracted by hot water extraction, and the protein was removed by enzymatic hydrolysis and Sevage. After purification, the chemical structure of polysaccharides was identified by infrared spectroscopy, methylation analysis and nuclear magnetic resonance spectroscopy. Then, a mouse model of acute kidney injury (AKI) was constructed using lipopolysaccharide (LPS), and pretreated with DCP. Structure characterization demonstrated that the number-average molecular weight and mass average molar mass of DCP-1 were 28.43 kDa and 15.00 kDa, respectively. DCP-1 mainly consisted of mannose (37.8 %) and glucose (55.6 %). The main linkage types of DCP-1 were contained 1,4-Linked Manp and 1,4-Linked Glcp. And DCP-1 was demonstrated to be an O-acetylglucomannan with ß-ᴅ-configuration in pyranoid form. Besides, the bioactivity of DCP was further investigated. The results showed that DCP exhibited notable anti-inflammatory activity in LPS-induced AKI mice. After treated with DCP, the creatinine (CREA) and urea nitrogen (BUN) in serum were successfully down-regulated in AKI mice. DCP treatment prevented the characteristic morphological changes of LPS-induced renal tubular injury. The results showed that DCP treatment significantly reduced the concentration of oxidative damage indicators (MDA, SOD) and the expression of inflammatory indices (TNF-α, IL-6, MCP-1, COX-2). In general, the newly extracted polysaccharide DCP showed excellent nephroprotective effect, which enabled it to be an ideal natural medicine for kidney diseases therapy.

Comparative Analysis of Water-Soluble Polysaccharides from Dendrobium Second Love 'Tokimeki' and Dendrobium nobile in Structure, Antioxidant, and Anti-Tumor Activity In Vitro.

With potential anti-tumor and antioxidant properties, the polysaccharide content of D. nobile is relatively lower than that of the other medicinal Dendrobium. To find high-content polysaccharide resources, the polysaccharide (DHPP-Ⅰs) was prepared from D. Second Love 'Tokimeki' (a D. nobile hybrid) and compared with DNPP-Ⅰs from D. nobile. DHPP-Is (Mn 31.09 kDa) and DNPP-Is (Mn 46.65 kDa) were found to be O-acetylated glucomannans (-Glcp-(1,4) and O-acetylated-D-Manp-(1,4) backbones), analogous to other Dendrobium polysaccharides. DHPP-Ⅰs had higher glucose content (31.1%) and a lower degree (0.16) of acetylation than DNPP-Ⅰs (15.8%, 0.28). Meanwhile, DHPP-Ⅰs and DNPP-Ⅰs had the same ability in the radical scavenging assay, which was milder than the control of Vc. Both DHPP-Is and DNPP-Is inhibited SPC-A-1 cell proliferation in vitro, with obvious differences in dose concentrations (0.5-2.0 mg/mL) and treatment times (24-72 h). Therefore, the antioxidant activity of DHPP-Ⅰs and DNPP-Ⅰs is not associated with distinction in anti-proliferative activity. As a glucomannan derived from non-medicinal Dendrobium, DHPP-Ⅰs has similar bioactivity to other medicinal Dendrobium, and this could serve as a starting point for studying the conformational-bioactivity relationship of Dendrobium polysaccharides.

High light triggers flavonoid and polysaccharide synthesis through DoHY5-dependent signaling in Dendrobium officinale.

Dendrobium officinale is edible and has medicinal and ornamental functions. Polysaccharides and flavonoids, including anthocyanins, are important components of D. officinale that largely determine the nutritional quality and consumer appeal. There is a need to study the molecular mechanisms regulating anthocyanin and polysaccharide biosynthesis to enhance D. officinale quality and its market value. Here, we report that high light (HL) induced the accumulation of polysaccharides, particularly mannose, as well as anthocyanin accumulation, resulting in red stems. Metabolome and transcriptome analyses revealed that most of the flavonoids showed large changes in abundance, and flavonoid and polysaccharide biosynthesis was significantly activated under HL treatment. Interestingly, DoHY5 expression was also highly induced. Biochemical analyses demonstrated that DoHY5 directly binds to the promoters of DoF3H1 (involved in anthocyanin biosynthesis), DoGMPP2, and DoPMT28 (involved in polysaccharide biosynthesis) to activate their expression, thereby promoting anthocyanin and polysaccharide accumulation in D. officinale stems. DoHY5 silencing decreased flavonoid- and polysaccharide-related gene expression and reduced anthocyanin and polysaccharide accumulation, whereas DoHY5 overexpression had the opposite effects. Notably, naturally occurring red-stemmed D. officinale plants similarly have high levels of anthocyanin and polysaccharide accumulation and biosynthesis gene expression. Our results reveal a previously undiscovered role of DoHY5 in co-regulating anthocyanin and polysaccharide biosynthesis under HL conditions, improving our understanding of the mechanisms regulating stem color and determining nutritional quality in D. officinale. Collectively, our results propose a robust and simple strategy for significantly increasing anthocyanin and polysaccharide levels and subsequently improving the nutritional quality of D. officinale.

RID serve as a more appropriate measure than phenol sulfuric acid method for natural water-soluble polysaccharides quantification.

With structural diversity of water-soluble polysaccharides, their precise quantitative analysis by phenol­sulfuric acid method becomes more difficult and challenging. In this study, the quantification analysis of dextran and glucose in phenol sulfuric acid method was compared in this paper. When the concentration is below 90 µg/mL, the quantification of glucose is close to theoretical value, however, glucose derivatives have significantly different absorption. Later, quantitative factors of water-soluble polysaccharide in RID measurement were investigated. The optimum temperature was 40 °C and linear range was 0.3125-10.0 mg/mL in RID for dextrans (1.0-500 kDa) and glucose derivatives. Method validation studies of the RID method were further performed and compared to conventional phenol sulfuric acid method, which demonstrated that RID measurement is more reliable and satisfactory method. The intervention of water-soluble impurity in RID response should be well control below 6% (w/w). By comparison, the RID measurement could well alleviate drawbacks in phenol­sulfuric acid method.

Structural characterization and antitumor activity of a polysaccharide from Dendrobium wardianum.

Through hot water extraction, protein removal and chromatographic purification, DWPP-Is was found to be the major polysaccharide present in the stem of D. wardianum. The Mn and Mw of DWPP-Is were 29.0 kDa and 98.6 kDa, respectively. Furthermore, mannose and glucose were found to be the most abundant monosaccharides in DWPP-Is. Their backbones consist of (1 â†’ 4)-ß-d-Glcp and O-acetylated (1 â†’ 4)-ß-d-Manp, which are similar to the structures of other anti-tumour Dendrobium polysaccharides. The inhibition rate of DWPP-Is treatment on SPC-A-1 cells (2 mg/mL, 72 h) reached 56.0%. Intragastric administration of DWPP-Is on A549 tumour-bearing KM mice (10 mg/mL, 0.2 mL) exhibited similar inhibition ratios to that of erlotinib hydrochloride (2 mg/mL). Moreover, the highest inhibition was observed in P-CK treatment combined with DWPP-Is, reaching an inhibition rate of 23.4%. These results suggest that DWPP-Is has the potential to be a functional agent for lung cancer prevention.

Complete chloroplast genomes of three important species, Abelmoschus moschatus, A. manihot and A. sagittifolius: Genome structures, mutational hotspots, comparative and phylogenetic analysis in Malvaceae.

Abelmoschus is an economically and phylogenetically valuable genus in the family Malvaceae. Owing to coexistence of wild and cultivated form and interspecific hybridization, this genus is controversial in systematics and taxonomy and requires detailed investigation. Here, we present whole chloroplast genome sequences and annotation of three important species: A. moschatus, A. manihot and A. sagittifolius, and compared with A. esculentus published previously. These chloroplast genome sequences ranged from 163121 bp to 163453 bp in length and contained 132 genes with 87 protein-coding genes, 37 transfer RNA and 8 ribosomal RNA genes. Comparative analyses revealed that amino acid frequency and codon usage had similarity among four species, while the number of repeat sequences in A. esculentus were much lower than other three species. Six categories of simple sequence repeats (SSRs) were detected, but A. moschatus and A. manihot did not contain hexanucleotide SSRs. Single nucleotide polymorphisms (SNPs) of A/T, T/A and C/T were the largest number type, and the ratio of transition to transversion was from 0.37 to 0.55. Abelmoschus species showed relatively independent inverted-repeats (IR) boundary traits with different boundary genes compared with the other related Malvaceae species. The intergenic spacer regions had more polymorphic than protein-coding regions and intronic regions, and thirty mutational hotpots (≥200 bp) were identified in Abelmoschus, such as start-psbA, atpB-rbcL, petD-exon2-rpoA, clpP-intron1 and clpP-exon2.These mutational hotpots could be used as polymorphic markers to resolve taxonomic discrepancies and biogeographical origin in genus Abelmoschus. Moreover, phylogenetic analysis of 33 Malvaceae species indicated that they were well divided into six subfamilies, and genus Abelmoschus was a well-supported clade within genus Hibiscus.

Enhanced co-generation of cellulosic ethanol and methane with the starch/sugar-rich waste mixtures and Tween 80 in fed-batch mode.

BACKGROUND: The mixed-feedstock fermentation is a promising approach to enhancing the co-generation of cellulosic ethanol and methane from sugarcane bagasse (SCB) and molasses. However, the unmatched supply of the SCB and molasses remains a main obstacle built upon binary feedstock. Here, we propose a cellulose-starch-sugar ternary waste combinatory approach to overcome this bottleneck by integrating the starch-rich waste of Dioscorea composita Hemls. extracted residue (DER) in mixed fermentation. RESULTS: The substrates of the pretreated SCB, DER and molasses with varying ratios were conducted at a relatively low solids loading of 12%, and the optimal mixture ratio of 1:0.5:0.5 for the pretreated SCB/DER/molasses was determined by evaluating the ethanol concentration and yield. Nevertheless, it was found that the ethanol yield decreased from 79.19 ± 0.20 to 62.31 ± 0.61% when the solids loading increased from 12 to 44% in batch modes, regardless of the fact that the co-fermentation of three-component feedstock was performed under the optimal condition defined above. Hence, different fermentation processes such as fed-batch and fed-batch + Tween 80 were implemented to further improve the ethanol concentration and yield at higher solids loading ranging between 36 and 44%. The highest ethanol concentration of 91.82 ± 0.86 g/L (69.33 ± 0.46% of theoretical yield) was obtained with fed-batch + Tween 80 mode during the simultaneous saccharification and fermentation at a high solids loading of 44%. Moreover, after the ethanol recovery, the remaining stillage was digested for biomethane production and finally yielded 320.72 ± 6.98 mL/g of volatile solids. CONCLUSIONS: Integrated DER into the combination of SCB and molasses would be beneficial for ethanol production. The co-generation of bioethanol and biomethane by mixed cellulose-starch-sugar waste turns out to be a sustainable solution to improve the overall efficacy in biorefinery.

Integrating sugarcane molasses into sequential cellulosic biofuel production based on SSF process of high solid loading.

BACKGROUND: Sugarcane bagasse (SCB) is one of the most promising lignocellulosic biomasses for use in the production of biofuels. However, bioethanol production from pure SCB fermentation is still limited by its high process cost and low fermentation efficiency. Sugarcane molasses, as a carbohydrate-rich biomass, can provide fermentable sugars for ethanol production. Herein, to reduce high processing costs, molasses was integrated into lignocellulosic ethanol production in batch modes to improve the fermentation system and to boost the final ethanol concentration and yield. RESULTS: The co-fermentation of pretreated SCB and molasses at ratios of 3:1 (mixture A) and 1:1 (mixture B) were conducted at solid loadings of 12% to 32%, and the fermentation of pretreated SCB alone at the same solid loading was also compared. At a solid loading of 32%, the ethanol concentrations of 64.10 g/L, 74.69 g/L, and 75.64 g/L were obtained from pure SCB, mixture A, and mixture B, respectively. To further boost the ethanol concentration, the fermentation of mixture B (1:1), with higher solid loading from 36 to 48%, was also implemented. The highest ethanol concentration of 94.20 g/L was generated at a high solid loading of 44%, with an ethanol yield of 72.37%. In addition, after evaporation, the wastewater could be converted to biogas by anaerobic digestion. The final methane production of 312.14 mL/g volatile solids (VS) was obtained, and the final chemical oxygen demand removal and VS degradation efficiency was 85.9% and 95.9%, respectively. CONCLUSIONS: Molasses could provide a good environment for the growth of yeast and inoculum. Integrating sugarcane molasses into sequential cellulosic biofuel production could improve the utilization of biomass resources.

Ethanol production from mixtures of sugarcane bagasse and Dioscorea composita extracted residue with high solid loading.

Various mixing ratios of alkali pretreated sugarcane bagasse and starch-rich waste Dioscorea composita hemls extracted residue (DER) were evaluated via simultaneous saccharification and fermentation (SSF) with 12% (w/w) solid loading, and the mixture ratio of 1:1 achieved the highest ethanol concentration and yield. When the solid loading was increased from 12% to 32%, the ethanol concentration was increased to 72.04 g/L, whereas the ethanol yield was reduced from 84.40% to 73.71%. With batch feeding and the addition of 0.1% (w/v) Tween 80, the final ethanol concentration and yield of SSF at 34% loading were 82.83 g/L and 77.22%, respectively. Due to the integration with existing starch-based ethanol industry, the co-fermentation is expected to be a competitive alternative form for cellulosic ethanol production.

Enhanced enzymatic hydrolysis of sugarcane bagasse with ferric chloride pretreatment and surfactant.

A FeCl3 pretreatment methodology was developed to convert raw sugarcane bagasse to highly digestible pretreated solid and selectively extract up to ∼100% of the hemicellulose from lignocellulosic biomass. FeCl3 pretreated solids yielded a quite high fermentable sugar yield compared to the native material. In addition, characterization of raw material and pretreated solid by X-ray diffraction (XRD), scanning electron microscopy (SEM), and thermogravimetric (TG) analysis was carried out to better understand how hemicellulose removal affected subsequent enzymatic hydrolysis. Furthermore, the addition of surfactants during enzymatic hydrolysis achieved higher glucose yields. 82.3% of glucose could be obtained with addition of BSA, combined with that generated during pretreatment process, the total glucose yield reached 42.2g/100g raw material, representing 93.8% of glucose in the raw sugarcane bagasse. The FeCl3 process offered the potential to co-produce xylose-derived and glucose-derived chemicals in the bio-refinery.