Sugar Side Chains Research Articles

Physicochemical Properties of Low-Molecular-Weight Homogalacturonan Pectin from Enzyme-Hydrolyzed Red Okra.

In this study, we focused on reducing the molecular weight of purified red okra pectin using various hydrolytic enzymes and evaluating its physicochemical properties or characterization. The enzyme treatments targeted both the main pectin chain and the side-chain sugars, resulting in a reduction in the molecular weight by approximately 10% (from 647 kDa) to 60% (down to 252 kDa). Both the purified red okra pectin and enzyme-treated pectins exhibited a homogalacturonan (HG)-type backbone. Fourier transform infrared (FT-IR) spectroscopy revealed a decrease in the absorbance peak for the pectin backbone (1200-1000 cm-1) in the low-molecular-weight (LMW) pectin. The most significant decrease was observed at 3300 cm-1 in pectin treated with both RGH+RGAE enzymes, indicating reduced sugar bonds. These results demonstrate the physicochemical changes in LMW red okra pectin following enzyme treatment and confirm its potential applications due to its unique characteristics.

Arabinan branches in the RG-I region of citrus pectin aid acid-induced gelation

Gelation is a critical property of citrus pectin. However, the roles played by neutral sugar side-chains on acid-induced pectin gelation remain poorly understood. Herein, galactan- or/and arabinan-eliminated pectins (P-G, P-A, and P-AG) were used to investigate the effects of side-chains on gelation. The gel hardness values of citrus pectin, P-G, P-A, and P-AG were 42.6, 39.9, 5.3, and 2.1 g, respectively, suggesting that arabinan contributed more to gelation than galactan. We next found that arabinan branches promoted pectin chain entanglement more effectively than arabinan backbones. Destabilizer addition experiments showed that hydrogen bonding, electrostatic interaction, and hydrophobic interaction were the main forces affecting pectin gel networks and strength, which was further validated by molecular dynamic simulations. The total number of hydrogen bonds between the arabinan branches and galactan/HG (65.7) was significantly higher than that between the arabinan backbones and galactan/HG (39.1), indicating that arabinan branches predominated in terms of such interactions. This study thus elucidated the roles played by neutral-sugar side-chains, especially the arabinan branches of acid-induced pectin gels, in term of enhancing high-methoxyl pectin gelation, and offers novel insights into the structure-gelling relationships of citrus pectin.

Structure of a truncated human GlcNAc-1-phosphotransferase variant reveals the basis for its hyperactivity

Mutations that cause loss of function of GlcNAc-1-phosphotransferase (PTase) lead to the lysosomal storage disorder mucolipidosis II. PTase is the key enzyme of the mannose 6-phosphate (M6P) targeting system that is responsible for tagging lysosomal hydrolases with the M6P moiety for their delivery to the lysosome. We had previously generated a truncated hyperactive form of PTase termed S1S3 which was shown to notably increase the phosphorylation level of secreted lysosomal enzymes and enhance their uptake by cells. Here, we report the 3.4Å cryo-EM structure of soluble S1S3 lacking both transmembrane domains and cytosolic tails. The structure reveals a high degree of conservation of the catalytic core to full-length PTase. In this dimeric structure, the EF-hand of one protomer is observed interacting with the conserved region four of the other. In addition, we present a high-quality EM 3D map of the UDP-GlcNAc bound form of the full-length soluble protein showing the key molecular interactions between the nucleotide sugar donor and side chain amino acids of the protein. Finally, although the domain organization of S1S3 is very similar to that of the Drosophila melanogaster (fruit fly) PTase homolog, we establish that the latter does not act on lysosomal hydrolases.

RG-I-containing sugar domains from Centella Asiatica bind strongly to galectin-3 to inhibit cell–cell interactions

BackgroundCentella Asiatica has been shown to have beneficial value for the treatment of tumors. However, its active ingredients and molecular mechanisms of action have not been fully elucidated. Pectic polysaccharides are the primary active components from medicinal plants. Moreover, these polysaccharides are regarded as potential inhibitors of galectins, such as galectin-1, -3, -7, that generally promote tumor growth. Nevertheless, detailed structural analysis of pectic polysaccharides from Centella Asiatica is sorely lacking, as is knowledge of their interactions with galectins.MethodsWater-soluble pectic polysaccharides (WCAP) isolated from Centella Asiatica were purified into two homogeneous fractions (WCAP-A2b and WCAP-A5b) by a combination of anion-exchange and gel-permeation chromatography. Monosaccharide composition, FT-IR, NMR and enzymatic analyses were used to characterize their structural features. Furthermore, the interactions between galectin-1, -3, -7 and a series of these polysaccharides, including two pectin fractions and their structural domains produced by enzymatic hydrolysis, were evaluated by using hemagglutination and biolayer interferometry.ResultsWCAP-A2b and WCAP-A5b have weight averaged molecular weights of 30.0 kDa and 34.0 kDa, respectively, and both polysaccharides consist of rhamnogalacturonan I (RG-I), rhamnogalacturonan II (RG-II) and homogalacturonan (HG) domains, with mass ratios of 1.3: 1.0: 1.4 and 1.1: 1.0: 2.4, respectively. Their RG-I domains contain arabinan, galactan, and/or arabinogalactan, along with neutral sugar side chains that are more prevalent in WCAP-A2b than in WCAP-A5b. Hemagglutination and biolayer interferometry binding assays indicate that galectin-3 vis-à-vis galectin-1 and -7, binds strongly to the RG-I domain (likely via its neutral side chains) in WCAP-A5b, thereby inhibiting galectin-3-mediated cell–cell interactions.ConclusionsOur study provides structural information on pectin polysaccharides from Centella Asiatica. Results suggest that RG-I domains from WCAP-A5b and WCAP-A2b may be developed as potential inhibitors of galectin-3-mediated cell–cell adhesion and tumor growth.Graphical

Preparation, physicochemical and functional characterization of pectic polysaccharides from fresh passion fruit peel by magnetic-induced electric field-assisted three-phase partitioning

In this study, we explored the use of magnetic-induced electric field (MIEF) treatment at various excitation voltages to assist three-phase partitioning (TPP) methods in extracting and separating pectic polysaccharides (PFPs) from the peel of fresh passion fruit (Passiflora edulis f. flavicarpa). The findings indicated that applying MIEF treatment prior to TPP separation significantly reduced the total sugar and galacturonic acid contents, as well as the degree of esterification of PFPs, with increasing excitation voltage. This treatment notably enhanced their yields and molecular weights. Although the MIEF treatment did not alter the monosaccharide composition of PFPs, it significantly modified their molar ratios of monosaccharides, thereby altering the backbone structure and neutral sugar side chains, which resulted in varied microstructures. Furthermore, especially at higher excitation voltages (≥300 V), the MIEF treatment improved the thermal stability, rheological, and emulsifying properties of PFPs. Consequently, our data suggest that the MIEF-assisted TPP method is a promising approach for the efficient and environmentally friendly preparation of pectic polysaccharides from plant materials.

Requirement for Toll-like Receptor 4 (TLR4) in LPS-induced lung inflammation caused by atypical Cyanobacteria Lipopolysaccharide

Abstract Cyanobacterial blooms found in freshwater sources cause multiple human diseases, including lung inflammation. Cyanobacteria produce atypical lipopolysaccharides (LPS) that can act as either weak TLR4 agonists or even antagonists in in vitro and in vivo models. However, our laboratory demonstrated that LPS from the cyanobacteria Geitlerinema sp. activates immune cells, despite containing altered LPS sugar side chains and inducing modified TLR4 signaling. Additionally, Geitlerinema sp. LPS induced lung inflammation in vivo based on significant neutrophil infiltration 24 hours post LPS exposure. In the current study, we extend our initial findings and asked whether the atypical LPS from an additional cyanobacterial strain, Brasilonema sp., also induces lung inflammation and if TLR4 is required. Our data demonstrate that in vitro, Brasilonema sp. LPS stimulates both Interleukin (IL)-6 and IL-8 production by immortalized airway epithelial cells that is TLR4-dependent, since the addition of the SPA-4 peptide blocks LPS-mediated production of these cytokines. More importantly, Brasilonema sp. LPS induced lung inflammation in wildtype mice but did not induce lung inflammation in TLR4-deficient mice, suggesting that the atypical LPS from cyanobacteria binds to TLR4 to induce lung inflammation. These findings help us to understand the impact of cyanobacteria LPS in neutrophil-dependent airway inflammation in individuals living near freshwater sources contaminated with cyanobacteria.

Comparative Analysis of Physicochemical and Functional Properties of Pectin from Extracted Dragon Fruit Waste by Different Techniques.

Dragon fruit peel, often discarded, is a valuable source of commercial pectin. This study investigates different extraction methods, including cold-water (CW), hot-water (HW), ultrasound (US), and novel enzyme extraction (xylanase: EZX), to extract pectins from dragon fruit peel and compare their characteristics. The pectin yield ranged from 10.93% to 20.22%, with significant variations in physicochemical properties across methods (p < 0.05). FTIR analysis revealed that extraction methods did not alter the primary structural configuration of the pectins. However, molecular weights (Mws) varied significantly, from 0.84 to 1.21 × 103 kDa, and the degree of esterification varied from 46.82% to 51.79% (p < 0.05). Monosaccharide analysis identified both homogalacturonan (HG) and rhamnogalacturonan-I (RG-I) pectic configurations in all pectins, predominantly comprising galacturonic acid (77.21-83.12 %mol) and rhamnose (8.11-9.51 %mol), alongside minor side-chain sugars. These properties significantly influenced pectin functionalities. In the aqueous state, a higher Mw impacted viscosity and emulsification performance, while a lower Mw enhanced antioxidant activities and promoted the prebiotic function of pectin (Lactis brevies growth). This study highlights the impact of extraction methods on dragon fruit peel pectin functionalities and their structure-function relationship, providing valuable insights into predicting dragon fruit peel's potential as a food-grade ingredient in various products.

Effect of enzymatic de-esterification and RG-I degradation of high methoxyl pectin (HMP) on sugar-acid gel properties

The influence of RG-I domains on high methoxyl pectin (HMP) sugar-acid gel properties has rarely been reported. In our work, HMP was modified by enzymatic de-esterification and degradation of RG-I domains to compare and analyze the relationship between the structure and final sugar-acid gel properties. The results showed that the degree of esterification (DE) of REP (pectin degraded by rhamnosidase) and GEP (pectin debranched by galactosidase) was the same as that of untreated HMP, whereas the DE of PMEP (pectin de-esterified by pectin methyl esterase) decreased from 59.63 % to 54.69 %. The monosaccharide composition suggested no significant changes in the HG and RG-I structural domains of PMEP. In contrast, the percentage of RG-I structural domains of REP and GEP dropped from 37 % to about 28 %, accompanied by a reduction in the proportion of the RG-I backbones and side chains. The rheological characterization of sugar-acid gels demonstrated an enhanced gel grade for PMEP and a weakened one for REP and GEP. Moreover, we constructed a correlation relationship between the fine structure of pectin and the properties of the sugar-acid gels, confirming the critical contribution of the RG-I region (especially the neutral sugar side chains) to the HMP sugar-acid gels.

Understanding structure/rheology relationships of amidated low-methoxyl citrus and apple pectin gels: Implications of sucrose, pectin types and characteristics

In this study, the relationships between structure and rheology (at small and large deformations) of amidated low-methoxyl (LMA) citrus and apple pectin gels were investigated considering how does these relationships can be modulated by soluble solids (sucrose), pectin types and characteristics. For that purpose, various LMA pectins with varying degrees of methyl esterification (DE) and amidation (DA), processed from citrus peels and apple pomace, were used. Scaling models of biopolymer gels have been critically discussed in respect to pectin critical gelling concentration (C0) and their structural variabilities. It was demonstrated that, whatever the types of LMA pectins and their structural variabilities, scalar percolation and quadratic models can be efficiently used for the prediction of the storage modulus G′ and C0 as other biopolymer gels. Fractal approach reveals that the origin of the LMA pectin gel elasticity is more in harmony with an enthalpic network of rod-like filaments with bending rigidity ruling the network's elasticity. The addition of sucrose induced an anticipation of the gelation temperature and an important improvement of the viscoelastic properties of the resulting gels. It was also shown that the most tough gels are obtained for citrus pectins due to the higher level of neutral sugar side chains in apple pectins that can hinder preponderant ca-bridging sites unlike citrus pectins. Finally, it was shown that unlike the firmness and hardness of the gels which appear to be more driven by the amide groups, the deformability of the gels is rather governed by the methyl ester groups.

Effect of ultrasonic modification on the binding ability of pectin to anthocyanin.

Pectin was considered as a potential candidate to improve the thermal stability of anthocyanins, and the binding ability of pectin to anthocyanins was influenced by its structure. In this study, sunflower pectins, modified by ultrasound (40 kHz) for different periods of time, were prepared and used to bind with anthocyanins, extracted from purple sweet potato. Characterization and thermal stability of pectin-anthocyanin complexes were investigated. The ultrasonic modification of pectin resulted in many changes in pectin chemical structure, including degradation of neutral sugar side chains, breakage of methoxyl groups, and increased molecular flexibility. Extension of ultrasonic modification time led to greater changes in pectin chemical structure. Analysis of the binding ability, as determined by Fourier transform infrared spectroscopy and molecular dynamics simulations, revealed that the interaction between pectin and anthocyanins was driven by hydrogen bonding, electrostatic interaction, and hydrophobic interaction. Pectins with different ultrasonic modification times bound with anthocyanins to different extents, mainly resulting from an increase in the number of hydrogen bonds. According to high-performance liquid chromatographic analysis, during heating at 90 °C the stronger the binding ability of pectin and anthocyanin complex, the better was its thermal stability. Ultrasonic modification of pectin could effectively enhance its binding ability to anthocyanin. © 2023 Society of Chemical Industry.

Extraction of citrus pectin using pressurized carbon dioxide and production of its oligosaccharides

In this study, we utilized pressurized carbon dioxide (pCO2) for pectin extraction from citrus residues because of its potential to produce acid without chemicals, that is non-toxic, green technology for humans and the environment. The pCO2 was performed at 2.5 MPa and 90 °C for 90 min, while the conventional method (CM) for pectin extraction using HCl was performed under atmospheric pressure at 90 °C for 90 min. Pectin extraction yields with pCO2 depended on the type of citrus fruit and were lower than those obtained with the CM. However, pCO2-extracted pectin (pCO2-pectin) provided a higher degree of esterification (DE), neutral sugar side-chain substitution on the pectate backbone, and greater molecular size than those of pectin extracted using the conventional method (CM-pectin). Pectin oligosaccharide (POS) was prepared from CM-pectin and pCO2-pectin using viscozyme L hydrolysis under optimum conditions. These POSs were then designated as CM-POS and pCO2-POS, respectively. Both pCO2-POS and CM-POS stimulated the growth of Lactobacillus and Bifidobacterium species, and induced them to produce short-chain fatty acids, and slightly suppressed immunoglobulin E production in U266 cells. Furthermore, pCO2-POS had higher angiotensin-converting enzyme inhibitory activity compared to CM-POS. This study suggests that pCO2 is a novel method for extracting pectin with unique molecular properties affecting POS functionality in vitro.

Pectic polysaccharides from Lilium brownii and Polygonatum odoratum exhibit significant antioxidant effects in vitro

Two pectic polysaccharides (WLBP-A3-c and WPOP-A-c) were isolated from traditional Chinese medicines Lilium brownii and Polygonatum odoratum, respectively. Monosaccharide composition, FT-IR, NMR and enzymatic analyses indicated that both WLBP-A3-c (59 kDa) and WPOP-A-c (33 kDa) contained homogalacturonan (HG), rhamnogalacturonan I (RG-I), and rhamnogalacturonan II (RG-II) domains, with mass ratios of 76.0: 17.2:6.8 and 76.8:10.6:12.6, respectively. Two RG-I domains WLBP-A3-c-DE1 and WPOP-A-c-DE1, correspondingly obtained from WLBP-A3-c and WPOP-A-c by enzymatic hydrolysis, were composed of repeating units of [→2)-α-L-Rhap-(1 → 4)-α-D-GalpA-(1→] with highly branched neutral sugar side chains at the O-4 position of Rhap, which contained arabinan, galactan, arabinogalactan I and II (AG-I and AG-II) side chains in different proportions. By comparison, WPOP-A-c exhibited higher scavenging effects against DPPH, ABTS and hydroxy radicals than WLBP-A3-c, probably because WPOP-A-c had higher contents of GalA residues and HG domains and lower molecular weight. Among three domains of WPOP-A-c, HG domain possessed the strongest activity in decreasing ROS production and promoting SOD activity, resulting in the effective protection of HepG2 cells against H2O2-induced oxidative stress. Our study provides evidence that pectins rich in HG domains from Lilium brownii and Polygonatum odoratum exhibit significant antioxidant effects, which hold potential for the application in the field of healthcare products.

An Acid-Free Alternative to Pectin Production from the Cell Walls of Olive Oil Waste and Different Fruits Using Choline Chloride.

The pectin from the cell walls of olive waste (alperujo) and apple, orange and strawberry fruits was extracted using choline chloride (ChCl) and the yield and chemical and structural compositions were compared to pectin extracted using citric acid (CA) and ammonium oxalate/oxalic acid (AOOA). According to the results, the alperujo pectin extracted using ChCl from alcohol-insoluble residue (AIR) showed a higher yield (2.20-2.88% on the basis of dry weight of AIR) than using CA (0.65-1.22%) but lower than using AOOA (3.92-5.42%). For fruit pectin, the highest yield was obtained using CA (8.81-16%), followed by AOOA (5.4-6.63%), although for apple pectin, ChCl gave a similar yield (5.36%) to AOOA. The uronic acid contents in all ChCl pectins (45.9-70.6% dry basis AIR) were higher or similar to that of the other extracting agents (30.6-65.2%), although a lower level of neutral sugar side chains was detected, with a lower degree of branching and degree of methylation. The NMR and FT-IR spectroscopy of the pectin isolated using ChCl confirmed its slightly different structural composition with respect to CA and AOOA pectin. Therefore, depending on the source material and functionality, pectin isolated using ChCl could be an acid-free alternative to pectin production.

The Equilibria of Triterpene Sapogenins-Phosphatidylcholine in Monolayers at the Air/Water Interface.

Sapogenins are the non-sugar parts of saponins (aglycones), high-molecular-weight glycosides linked to one or more sugar side chains. This group of compounds presents many properties, e.g., the potent properties of reducing surface tension and foaming properties, as evidenced by the amphipathic nature of these substances. They are used in the cosmetics industry, the washing and detergent industry, and the food industry. In addition, they have many healing properties. They lower blood cholesterol but are also used to synthesize steroid drugs or hormones. As reported in the literature, saponins also show antitumor activity, leading to cell cycle inhibition and apoptosis of various neoplastic cells. In this study, the influence of two sapogenins: asiatic acid (AA) and oleanolic acid (OA), on the properties of monolayers made of phosphatidylcholine (DPPC) was investigated. The method used in these studies was the Langmuir method with Brewster angle microscopy. The interactions between the tested compounds in mixed monolayers were described. Using mathematical equations, we established that oleanolic acid and asiatic acid formed complexes with DPPC at 1:1 ratios, characterized by high stability constants. We derived the parameters characterizing the formed complexes and described the phase transitions that occur during the formation of pure and mixed monolayers.

Discovery of novel benzimidazole acyclic C-nucleoside DNA intercalators halting breast cancer growth.

Breast cancer continues to be the most frequent cancer worldwide. In practice, successful clinical outcomes were achieved via targeting DNA. Along with the advances in introducing new DNA-targeting agents, the "sugar approach" design was employed herein to develop new intercalators bearing pharmacophoric motifs tethered to carbohydrate appendages. Accordingly, new benzimidazole acyclic C-nucleosides were rationally designed, synthesized and assayed via MTT(3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay to evaluate their cytotoxicity against MCF-7 and MDA-MB-231 breast cancer cells compared to normal fibroblasts (Wi-38), compared to doxorubicin. (1S,2R,3S,4R)-2-(1,2,3,4,5-Pentahydroxy)pentyl-1H-5,6-dichlorobenzimidazole 7 and (1S,2R,3S,4R)-2-(1,2,3,4,5-pentahydroxy)pentyl-1H-naphthimidazole 13 were the most potent and selective derivatives against MCF-7 (half-maximal inhibitory concentration[IC50 ] = 0.060 and 0.080 µM, selectivity index [SI] = 9.68 and 8.27, respectively) and MDA-MB-231 cells (IC50 = 0.299 and 0.166 µM, SI = 1.94 and 3.98, respectively). Thus, they were identified as the study hits for mechanistic studies. Both derivatives induced DNA damage at 0.24 and 0.29 μM, respectively. The DNA damage kinetics were studied compared to doxorubicin, where they both induced faster damage than doxorubicin. This indicated that 7 and 13 showed a more potent DNA-damagingeffect than doxorubicin. Docking simulations within the DNA double strands highlighted the role of both the heterocyclic core and the sugar side chain in exhibiting key H-bond interactions with DNA bases.

Structure characterization and calcium ions induced emulsion gel properties of pectic polysaccharide from Fructus aurantii

A novel pectic polysaccharide was extracted from Fructus aurantii using ammonium oxalate extraction, whose primary structure and emulsification properties were investigated for the first time. Fructus aurantii ammonium oxalate-extracted pectin (FAOP) was a highly methyl-esterified pectin with a 60.36% methyl-esterification degree and a relative weight-average molecular weight (Mw) of 1.30 × 106 Da. The backbone of FAOP was composed of a 2:3 molar ratio of →4)-α-GalpA-(1→ and →4)-α-GalpA-6-O-methyl-(1→ linkage, which contained an ∼57.04% linear homogalacturonan (HG) structural domain. In addition, neutral sugar side chains, such as arabinan and galactan belonging to the rhamnogalacturonan I (RG-I) region, were identified. Subsequently, the effects of Ca2+ on the properties of oil-in-water emulsions were investigated by droplet size, zeta potential, rheological properties and emulsion stability. Results indicated that FAOP formed a solid-like gel emulsion and exhibited excellent emulsion stability when the Ca2+ concentration was 5 mM. At this concentration, the droplet size was the smallest, the backscatter signal (BS) was almost unchanged; the Turbiscan stability index (TSI) value was the lowest, and tanδ (G"/G′) was less than 1. These findings indicated that the improved emulsion stability of FAOP by the introduction of Ca2+ was related to the emulsion gel network structure. These results indicated the potential application of Fructus aurantii pectin as a food stabiliser and thickener.

Impact of non-thermal modifications on the physicochemical properties and functionality of litchi pomace dietary fibre

Pomace is frequently underestimated as a complex by-product. High pressure homogenisation (HPH), enzymatic hydrolysis (EH) and high hydrostatic pressure (HHP) modifications are applied to improve the functional properties of litchi pomace insoluble dietary fibre (LPDF). The relationships between physicochemical properties and structural alterations and digestibility of LPDF are studied. The results showed that HPH, EH and HHP changed the LPDF's microstructure, causing it to develop more porous structures. Meanwhile, HPH modification exhibited a remarkable effect on LPDF's water holding (15.6 g/g), water swelling (26.1 mL/g) and oil retention (5.4 g/g) capabilities, resulting in the change of the complexity of the neutral sugar side chain. Notably, the highest total phenol content (3.4 GAE mg/g) and antioxidant activities (DPPH 153.6, ABTS 188.9, FRAP 195.1 μmol Trolox/g) were also observed in LPDF modified with HPH. Moreover, the diversity of structural features and high affinity of modified LPDFs are conducive to its functional properties, i.e., inhibition of starch digestion. The remarkable potential of LPDF modified with HPH was attributed to the beneficial effects of dietary fibre on human health. As a result, litchi pomace could be a useful and effective source of functional fibres. The incorporation of litchi pomace into the human diet is still underexplored but is a promising alternative.

Analysis of steroidal glycoalkaloids and their metabolites in Solanum nigrum fruits based on liquid chromatography-tandem mass spectrometry and molecular networking.

Solanum nigrum fruit is like a treasure house for anticancer drugs because of its steroidal alkaloids. However, the clinical treatment of cancer mainly uses immature fruits, which can cause a toxic reaction if eaten directly, while mature fruits are eaten as fruit. In order to clarify the reasons for the differences in pharmacodynamics and toxicity between them, we studied the composition and metabolism of steroidal alkaloids in fruits of different maturities based on liquid chromatography-tandem mass spectrometry and molecular networking. As a result, 114 steroidal glycoalkaloids were identified. During fruit ripening, the aglycones of steroidal alkaloids mainly undergo hydroxylation and carboxylation, and the sugar side chains mainly undergo acylation and glycosylation reactions. Furthermore, 219 steroidal alkaloids were identified in a metabolism experiment in rats. Metabolic processes include deglycosylation, redox, sulfuric acid binding, acetyl binding, and glucuronic acid-binding. Steroidal alkaloids in mature fruits have high molecular weight and polarity, which are difficult to absorb, and most of them are excreted through feces and urine, which may be the reason for their poor efficacy. This study lays a foundation for research on the biosynthesis of steroidal alkaloids and provides potential candidates for the discovery of new steroidal alkaloid anticancer drugs.

Cytotoxic effects of recombinant proteins enhanced by momordin Ic are dependent on cholesterol and ganglioside GM1

Plant-derived triterpenoid saponins have been shown to play a powerful role in enhancing the cytotoxic activity of protein therapeutics. However, the mechanism of how saponins are acting is not clearly understood. In this study, momordin Ic (MIC), a triterpenoid saponin derived from Kochia scoparia (L.) Schrad., specifically enhance the antiproliferative effect of recombinant MAP30 (a type I ribosome inactivating protein, RIP) in breast cancer cells. Subsequently, the possible mechanism of how MIC enhanced the cytotoxicity of MAP30 was analyzed in detail. We observed the level of intracellular labeled MAP30 using fluorescence microscopy and flow cytometry. And a reporter protein, GAL9, was used to monitor the role of MIC in promoting endosomal escape. We found endosomal escape does not play a role for the enhancer effect of MIC while the effect of MIC on MAP30 is cholesterol dependent and that ganglioside GM1, a lipid raft marker, can competitively inhibit cytotoxicity of MAP30 enhanced by MIC. Finally, we provided some insights into the correlation between the sugar side chain of MIC and its role in enhancing of RIP cytotoxicity and altering of drug cell tropism.

Structural characteristics and gel properties of pectin from citrus physiological premature fruit drop

This study is the first to extract and characterize pectin from citrus physiological premature fruit drop. The extraction yield of pectin reached 4.4 % by acid hydrolysis method. The degree of methoxy-esterification (DM) of citrus physiological premature fruit drop pectin (CPDP) was 15.27 %, indicating it was low-methoxylated pectin (LMP). The monosaccharide composition and molar mass test results showed CPDP was a highly branched macromolecular polysaccharide (β: 0.02, Mw: 2.006 × 105 g/mol) with rich rhamnogalacturonan I domain (50.40 %) and long arabinose and galactose side chain (32.02 %). Based on the fact that CPDP is LMP, Ca2+ was used to induce CPDP to form gels. Textural and rheological tests showed that the gel strength and storage modulus of CPDP were higher than commercial citrus pectin (CP) used in this paper due to the lower DM and rich neutral sugar side chains of CPDP. Scanning electron microscope (SEM) results showed CPDP had stable gel network structure.