The The Effect of Concentration of Citric Acid Solution on Extraction of Pectin from Watermelon Albedo

Watermelon albedo or the white flesh of watermelon rind contains pectin with high enough content. In this study was conducted the extraction of pectin from watermelon albedo by the liquid-solid extraction method. The citric acid solution was used as a solvent with concentrations varied by 4%, 7%, and 10%. The research steps include the pretreatment of raw material, the extraction, the posttreatment of extraction, and the analysis of pectin. The pretreatment aimed to reduce water content and reduce the size of raw material. The extraction process was the main process in which pectin dissolution occurred in the citric acid solvent. Furthermore, the posttreatment of extraction aimed to obtain pectin solid with the addition of ethanol. The final step was the analysis, including pectin yield, moisture content, methoxyl content, galacturonic acid content, and pectin functional group. Based on research obtained, an increase in the concentration of citric acid increased pectin yield, methoxyl content, and galacturonic acid content. The moisture content of pectin decreased with the increasing concentration of citric acid. Pectin yield, moisture content, methoxyl content, galacturonic acid content resulted from 10% concentration of citric acid solvent are 8.356%, 19.748%, 7.029%, and 69.048%, respectively. Based on FTIR analysis, the functional groups contained in pectin are hydroxyl group, methyl group, carbonyl group, and ether group. This functional groups are main constituents of pectin structure.

Methylation Profiling of Biosynthetic Genes Reveals The Role of D-Galacturonic Acid Reductase In Ascorbic Acid Accumulation In Tomato Fruit

Ascorbic acid (AsA) is an important nutrient component contributing to major flavor value of tomato fruit and human health. Although transcription regulation of AsA biosynthetic genes have been well demonstrated, epigenetic modification underlying AsA accumulation remains unclear. In this study, we exposed immature tomato fruits to a methyltransferase inhibitor (5-azacytidine) and detected the impacts on AsA accumulation. Inhibition of DNA methylation enhanced AsA accumulation in tomato leaves and fruits. We further isolated a AsA biosynthetic gene, SlGalUR5, which encodes a D-galacturonic acid reductase. SlGalUR5 showed reduced DNA methylation levels and higher transcription levels in Slmet1 mutant while have converse pattern in Sldml2 mutant. 5-azacytidine treatment significantly decreased DNA methylation levels of SlGalUR5 in fruits. Conversely, transcription profiles of SlGalUR5 and enzyme activity of GalUR were enhanced in 5-azacytidine–treated fruits. Our finding revealed a new insight into epigenome modification of SlGalUR5 involved in ascorbic acid accumulation and provide a potential means of increasing AsA levels for tomato breeding.

A Novel Polysaccharide Isolated From Fresh Longan (Dimocarpus longan Lour.) Activates Macrophage via TLR2/4-Mediated PI3/AKT and MyD88/TRAF6 Pathways

A novel immunomodulatory polysaccharide (LP4) with a molecular weight 6.31 × 104 g/mol was purified from fresh longan pulp. It was composed of mannose, glucose, glucuronic acid, galactose, xylose, arabinose, galacturonic acid, fucose, and rhamnose in a molar percentage of 36:31:10:7:4:4:3:2:2, and mainly linked by (1→6)-β-Man, (1→4)-β-Glc and (1→6)-α-Glc. LP4 can obviously enhance the phagocytosis of macrophages and promote the proliferation of lymphocytes. After treating macrophages with LP4 (12.5–50 μg/ml), the production of IL-1β and TNF-α was significantly increased. These increases of cytokines were suppressed when the TLR2/TLR4 receptors were inhibited by anti-TLR2 and/or anti-TLR4 antibodies. Moreover, the mRNA expression of INOS, AKT, PI3K, TRAF6 and MyD88 was significantly suppressed by TLR2/TLR4 antibodies. These results indicated that LP4 induced macrophage activation mainly via the TLR2 and TLR4-induced PI3K/AKT and MyD88/TRAF6 pathways.

Ascorbic Acid Content and Transcriptional Profiling of Genes Involved in Its Metabolism during Development of Petals, Leaves, and Fruits of Orange (Citrus sinensis cv. Valencia Late)

Citrus fruit is one of the most important contributors to the ascorbic acid (AsA) intake in humans. Here, we report a comparative analysis of AsA content and transcriptional changes of genes related to its metabolism during development of petals, leaves and fruits of Valencia Late oranges (Citrus sinensis). Petals of close flowers and at anthesis contained the highest concentration of AsA. In fruits, AsA content in the flavedo reached a maximum at color break, whereas the pulp accumulated lower levels and experienced minor fluctuations during development. AsA levels in leaves were similar to those in the flavedo at breaker stage. The transcriptional profiling of AsA biosynthetic, degradation, and recycling genes revealed a complex and specific interplay of the different pathways for each tissue. The D-galacturonic acid pathway appeared to be relevant in petals, whereas in leaves the L-galactose pathway (GGP and GME) also contributed to AsA accumulation. In the flavedo, AsA content was positively correlated with the expression of GGP of the L-galactose pathway and negatively with DHAR1 gene of the recycling pathway. In the pulp, AsA appeared to be mainly controlled by the coordination among the D-galacturonic acid pathway and the MIOX and GalDH genes. Analysis of the promoters of AsA metabolism genes revealed a number of cis-acting elements related to developmental signals, but their functionalities remain to be investigated.

Comparison and Optimization of Quantification Methods for Shigella flexneri Serotype 6 O-antigen Containing Galacturonic Acid and Methyl-Pentose

Shigella is a leading diarrheal cause of morbidity and mortality worldwide, especially in low- and middle-income countries and in children under five years of age. Increasing levels of antimicrobial resistance make vaccine development an even higher global health priority. S. flexneri serotype 6 is one of the targets of many multicomponent vaccines in development to ensure broad protection against Shigella. The O-antigen (OAg) is a key active ingredient and its content is a critical quality attribute for vaccine release in order to monitor their stability and to ensure appropriate immune response. Here, the optimization of two methods to quantify S. flexneri 6 OAg is reported together with the characterization of their performances. The optimized Dische colorimetric method allows a tenfold increment of the sensitivity with respect to the original method and is useful for fast analysis detecting selectively methyl-pentoses, as rhamnose in S. flexneri 6 OAg. Also, a more specific HPAEC-PAD method was developed, detecting the dimer galacturonic acid-galactosamine (GalA-GalN) coming from S. flexneri 6 OAg acid hydrolysis. These methods will facilitate characterization of S. flexneri 6 OAg based vaccines. The colorimetric method can be used for quantification of other polysaccharide containing methyl-pentoses, and the HPAEC-PAD could be extended to other polysaccharides containing uronic acids.

Identification of the Aldo-Keto Reductase Responsible for d-galacturonic Acid Conversion to l-galactonate in Saccharomyces cerevisiae

d-galacturonic acid (d-GalUA) is the main constituent of pectin, a complex polysaccharide abundant in several agro-industrial by-products such as sugar beet pulp or citrus peel. During several attempts to valorise d-GalUA by engineering the popular cell factory Saccharomyces cerevisiae, it became obvious that d-GalUA is, to a certain degree, converted to l-galactonate (l-GalA) by an endogenous enzymatic activity. The goal of the current work was to clarify the identity of the responsible enzyme(s). A protein homology search identified three NADPH-dependent unspecific aldo-keto reductases in baker’s yeast (encoded by GCY1, YPR1 and GRE3) that show sequence similarities to known d-GalUA reductases from filamentous fungi. Characterization of the respective deletion mutants and an in vitro enzyme assay with a Gcy1 overproducing strain verified that Gcy1 is mainly responsible for the detectable reduction of d-GalUA to l-GalA.

Structural Characteristics, Rheological Properties, and Antioxidant Activity of Novel Polysaccharides from “Deer Tripe Mushroom”

The polysaccharide was extracted by water extraction and alcohol precipitation method from “deer tripe mushroom,” which was domesticated and bred from wild Auricularia delicata in Zambia. The structure of the deer tripe mushroom polysaccharide (DTMP) was characterized by gas chromatography-mass spectrometry (GC-MS), Fourier transform infrared spectroscopy (FT-IR), nuclear magnetic resonance spectroscopy (NMR), scanning electron microscope (SEM), and other analytical techniques. And the rheological and gel properties and antioxidant capacities of the polysaccharide were studied. The results showed that DTMP was mainly composed of mannose, fructose, glucose, and galacturonic acid, of which the galacturonic acid was the main component. By calculation, the molar ratio of mannose : fructose : glucose : galactoacid in DTMP is 0.8 : 14.8 : 1.0 : 26.32. The polysaccharide was mainly composed by α-1,4-glycosidic bond. DTMP solution showed a shear-thinning (pseudoplastic) behaviour, and its pseudoplasticity was more obvious at a concentration of 2%. The power law model was used to evaluate the viscosity curves of DTMP, and its viscosity and consistency indices both increased as the concentration increased, whereas both indices decreased as the concentration decreased. The viscosity of the polysaccharide solution changed as the pH changed: the polysaccharide solutions had a higher viscosity at pH = 10. DTMP showed gel-like behaviour (G′ ＞ G″), and the gel strength enhanced with the increase of concentration (2%–10%). In addition, the antioxidant experiment of DTMP showed that it had good antioxidant activity, and there was a significant dose-effect relationship between its activity and concentration in the low concentration range.

Production of Pectin from Citrus Residues: Process Alternatives and Insights on Its Integration under the Biorefinery Concept

This chapter describes the pectin production process from citrus residues. It discusses the importance of essential oils removal before processing through steam distillation, hydrodistillation, or solvent extraction. Also, it presents different extraction methods (acid hydrolysis, microwave-assisted acid hydrolysis, and hydrodistillation) that have been employed and different solvents that can be used for its purification. Since all these processing parameters can affect the final pectin yield and quality, a discussion is made on which processing options and conditions could be used based on recently reported data. The best operational conditions based on the percentages of pectin recovery and their relationship with quality parameters, such as the galacturonic acid content and degree of esterification are presented. Finally, a discussion is made regarding the opportunities for its integration under the biorefinery concept that could help to enhance several economic and environmental aspects of the process.