Extracellular Polysaccharide Research Articles

Terrestrial algae: pioneer organisms of carbonate rock solutional weathering in South China karst.

The formation of soil in karst ecosystem has always been a scientific problem of great concern to human beings. Algae can grow on the exposed and non-nutrition carbonate surface, inducing and accelerating weathering of rock substrates, thus promoting soil formation. Yet the actual contribution of algae to solutional weathering intensity remains unclear. In this study, we performed weathering simulation experiment on two algae species (Klebsormidium dissectum (F.Gay) H.Ettl & G.Gärtner and Chlorella vulgaris Beijerinck), which were screened from carbonated rock surfaces from a typical karst region in South China. The results showed: (1) both algae have solutional weathering effect on carbonate rock, (2) there is no difference of solutional intensity observed, yet the solutional modes are different, suggesting different ecological adaptative strategies, (3) algae on carbonate rocks have higher carbonic anhydrase activity (CAA) and secrete more extracellular polysaccharide (EPS), accelerating rock weathering. (4) The absolute dissolution amount of carbonate rock with algae participation is 3 times of that of without algae. These results indicate the significant impact of terrestrial algae on carbonate rock solutional weathering and provides quantitative evidence that terrestrial algae are pioneer species. It also contributes to our further understanding of soil formation in karst ecosystems in South China.

L-lysine as a Potential Agent for Controlling Biofilm Formation Using Fusobacterium nucleatum and Porphyromonas gingivalis

Background Periodontitis, a globally-pervasive chronic inflammatory condition, is precipitated by forming microbial biofilms. A key player in this process is Fusobacterium nucleatum (F. nucleatum), an anaerobic bacterium exhibiting invasive and pro-inflammatory characteristics primarily associated with the onset of periodontitis. Consequently, developing novel biofilm control strategies specifically targeting F. nucleatum is essential. The objective of this research is to examine the suppressive impact of L-lysine on biofilm formation using F. nucleatum. Methods Fusobacterium nucleatum(F. nucleatum) and Porphyromonas gingivalis (P. gingivalis) were cultured in a liquid medium containing L-lysine of different concentrations. The antimicrobial activity of L-lysine was evaluated using the broth microdilution method. The effect of L-lysine on the mono-species and dual-species F. nucleatum biofilms was assessed using techniques such as crystal violet staining. The synthesis of extracellular polysaccharide substances (EPSs) from biofilms was measured using phenol-sulfuric acid. The expression level of virulence factors (FN1420, FN0669, FN0634, FN1162) and the biofilm-formation-related gene radD were quantified using real-time PCR. Results L-lysine exhibited the antibacterial and antibiofilm activity of F. nucleatum and F. nucleatum/ P. gingivalis double strains, which was concentration-dependent. The MIC and MBC values were 10 mg/ml and 20 mg/ml in F. nucleatum single strains, while those in F. nucleatum/ P. gingivalis double strains were 20 mg/ml and 40 mg/ml. L-lysine with a concentration ≥20 mg/ml could reduce the EPS production in F. nucleatum and F. nucleatum/ P. gingivalis dual-species biofilms. L-lysine also inhibited the expression of genes encoding the F. nucleatum virulence factor (FN1420, FN0669, FN0634, FN1162) and the adhesive factor radD. Conclusion These findings corroborate the potential of L-lysine as an agent impeding the development of both mono-species biofilms of F. nucleatum and dual-species biofilms of F. nucleatum and P. gingivalis.

Enhancing Pullulan Production in Aureobasidium pullulans through UV Mutagenesis Breeding and High-Throughput Screening System

This study addresses the production enhancement of pullulan, an extracellular polysaccharide with various applications. Pullulan is primarily produced by Aureobasidium pullulans (A. pullulans), and genetic modification is commonly used to increase its yield. However, there is a need for a more efficient and safer method. To achieve this, we designed a high-throughput screening system utilizing a unique fluorescent protein specific to pullulan. Ultraviolet (UV) mutagenesis was applied to create a pool of mutant strains, and flow cytometry allowed for single-cell screening. Our approach yielded strain M1-B3, which exhibited a substantial increase in pullulan production from 26.5 g/L to 76.88 g/L. Additionally, the molecular weight of the produced pullulan significantly increased, expanding its potential commercial application. This study demonstrates an efficient and safe method to enhance pullulan production in A. pullulans. The UV mutagenesis and flow cytometry based on screening not only increased yield but also improved pullulan’s molecular weight. The adaptability of this method to other polysaccharides and its potential for genomic analysis and broader applications make it a valuable tool in bioproduction.

Antibacterial Activity and Mechanism of Three Root Exudates from Mulberry Seedlings against Ralstonia pseudosolanacearum.

Bacterial wilt is a significant soil-borne disease that poses a threat to mulberry production yield and quality of agricultural production worldwide. However, the disease resistance mechanisms dependent on root exudates are not well understood. In this present study, we investigated the antibacterial mechanisms of the main active substances (erucamide, oleamide, and camphor bromide) present in mulberry root exudates (MRE) against Ralstonia pseudosolanacearum (Rp), the causal agent of bacterial wilt. Our findings revealed that these three active substances inhibited the growth activity of Rp by affecting the cell morphology and extracellular polysaccharide content, as well as triggering a burst of reactive oxygen species. The active substances induced oxidative stress, leading to a decrease in Rp growth. Additionally, the expression levels of key genes in the hrp gene cluster (hrpB, hrpX, and hrpF) and other virulence-related genes (such as ripAW, ripAE, Rs5-4819, Rs5-4374, ace, egl3, and pehB) were significantly reduced upon treatment with the active substances. Further pathogenicity experiments demonstrated that root exudates (at a concentration of 1.5 mg·mL-1) delayed or slowed down the occurrence of bacterial wilt in mulberry. These findings provide valuable insight into the antimicrobial mechanisms of MRE against Rp and lay a theoretical foundation for the development and application of biocontrol agents to control mulberry bacterial wilt.

Prodigiosin as an Antibiofilm Agent against the Bacterial Biofilm-Associated Infection of Pseudomonas aeruginosa.

Pseudomonas aeruginosa is known to generate bacterial biofilms that increase antibiotic resistance. With the increase of multi-drug resistance in recent years, the formulation of a new therapeutic strategy has seemed urgent. Preliminary findings show that Prodigiosin (PG), derived from chromium-resistant Serratia marcescens, exhibited efficient anti-biofilm activity against Staphylococcus aureus. However, its anti-biofilm activity against P. aeruginosa remains largely unexplored. The anti-biofilm activity of PG against three clinical single drug-resistant P. aeruginosa was evaluated using crystal violet staining, and the viability of biofilms and planktonic cells were also assessed. A model of chronic lung infection was constructed to test the in vivo antibiofilm activity of PG. The results showed that PG inhibited biofilm formation and effectively inhibited the production of pyocyanin and extracellular polysaccharides in vitro, as well as moderated the expression of interleukins (IL-1β, IL-6, IL-10) and tumor necrosis factor (TNF-α) in vivo, which might be attributed to the downregulation of biofilm-related genes such as algA, pelA, and pslM. These findings suggest that PG could be a potential treatment for drug-resistant P aeruginosa and chronic biofilm infections.

BolA promotes the generation of multicellular behavior in S. typhimurium by regulating the c-di-GMP pathway genes yeaJ and yhjH

The generation of multicellular behavior enhances the stress adaptability, antibiotic resistance, and pathogenic potential of Salmonella enterica serovar Typhimurium (S. typhimurium), which is challenging for its prevention and control. Therefore, determination of the mechanism of multicellular behavior development is urgently required. Accordingly, this study investigated BolA, a transcription factor that promotes bacterial survival under different stresses. We found that BolA promoted the generation of multicellular behavior. Furthermore, transcriptome analysis revealed that BolA affected the expression of numerous genes, including biofilm formation and motility-related genes. In terms of biofilm formation, compared with the wild-type strain, bolA overexpression (269BolA+) increased the extracellular matrix content (extracellular polysaccharide, extracellular protein, and extracellular DNA (eDNA) by upregulating gene expression, ultimately increasing the biofilm formation ability by 2.56 times. For motility, bolA overexpression inhibited the expression of flagella synthesis genes, resulting in a 91.15 % decrease in motility compared with the wild-type (6 h). Further mechanistic analysis demonstrated that BolA affected the expression of the C-di-GMP pathway genes yeaJ and yhjH, which influenced the generation of multicellular behavior. In terms of biofilms, the extracellular polysaccharide content of 269BolA + ∆Yeaj (bolA overexpression and yeaJ deletion) was reduced by 89.91 % compared with 269BolA+, resulting in a 71.1 % reduction in biofilm forming ability. The motility of the 269∆BolA∆Yhjh (bolA/yhjH double deletion) strain was significantly decreased compared with that of 269∆BolA. Finally, the LacZ gene reporting showed that BolA promoted and inhibited the expression of yeaJ and yhjH, respectively. In conclusion, BolA mainly improves the content of extracellular polysaccharide by promoting the expression of yeaJ, thus enhancing the formation of biofilms. BolA also restricts flagellar synthesis by inhibiting yhjH expression, therefore reducing motility, ultimately promoting multicellular behavior arises. These findings lay a theoretical foundation for the prevention and control of S. typhimurium.

Photoregulation of the biosynthetic activity of the edible medicinal mushroom Lentinula edodes in vitro.

The findings of the study demonstrate the impact of low-intensity laser and quasi-monochromatic light on the biosynthetic activity of the edible medicinal fungus L. edodes during submerged cultivation. An artificial lighting installation based on matrices of light-emitting diodes (LED) emitting light at 470nm (blue), 530nm (green), 650nm (red), and argon gas laser (488nm) was used. Irradiation with blue and red LED and laser led to a shortening of the lag phase by 2days and an increase in the mycelial mass. Irradiation with laser light resulted in the highest mycelial mass yield (14.1g/L) on the 8th day of cultivation. Irradiation in all used wavelength ranges caused an increase in the synthesis of both extracellular and intracellular polysaccharides. Laser light at 488nm and LED at 470nm proved to be the most effective. Irradiation with red, green, and blue laser light caused an increase in the total amount of fatty acids in the mycelial mass compared to the control. A significant distinction in qualitative composition was observed: short-chain acids C6‒C12 compounds were produced under red light irradiation, whereas long-chain C20‒C24 were formed under green light irradiation. The most significant changes in the aromatic profile of the mycelial mass and culture liquid were recorded upon irradiation with green light. The content of aromatic components increased 24.6 times in the mycelial mass and 38.5 times in the culture liquid. The results suggest the possibility of using low-intensity quasi-monochromatic light for targeted regulation of L. edodes biosynthetic activity.

Extraction and Chemical Composition Analyses of Intracellular and Extracellular Polysaccharides from Trametes lactinea Liquid Fermentation

Trametes lactinea is a macrofungus commonly found on broadleaf tree felling piles with rich bioactivity, and its main active substances are polysaccharides, which have a certain medicinal value. Few studies have been conducted on the simultaneous study of intracellular and extracellular polysaccharides of fermented mycelia in liquid culture. In this study, T. lactinea collected from Eucalyptus stumps was subjected to liquid fermentation culturing and the extraction of intracellular and extracellular polysaccharides from the mycelium was optimized using unidirectional and orthogonal test methods. The intracellular and extracellular crude polysaccharides were isolated and purified, and polysaccharide fractions were obtained and assayed for their chemical composition. The main findings of this study are as follows: (1) The optimal conditions for the extraction of intracellular polysaccharides from T. lactinea mycelium were a material–liquid ratio of 1:40, an extraction temperature of 100 °C, a time of 5 h, and four repeats, wherein the polysaccharide extraction rate reached 5.1%. The optimal extraction conditions for the extracellular polysaccharides were a concentration ratio of 5:1, a concentration temperature of 70 °C, an ethanol concentration of 100.0%, and an alcohol settling time of 12 h. The polysaccharides could thus be extracted up to 0.63 mg/mL. (2) The intracellular and extracellular crude polysaccharides of T. lactinea mycelium were successfully isolated and purified, resulting in three homogeneous fractions of intracellular polysaccharides (IP-1, IP-2, and IP-3) and two homogeneous fractions of extracellular polysaccharides (EP-1 and EP-2). (3) The polysaccharide chemical composition resulted in the highest total sugar content of EP-1. IP-3 had the highest uronic acid content. The results of the monosaccharide composition analysis showed that the mycelial intracellular polysaccharides IP-1, IP-2, and IP-3 were all highest in glucose. Both IP-2 and IP-3 contained guluronic acid and glucuronic acid. Mannose was the most abundant extracellular polysaccharide in both EP-1 and EP-2. This study provides theoretical and technical bases for the comprehensive development and utilization of T. lactinea polysaccharides.

Transcriptome Reveals Regulation of Quorum Sensing of Hafnia alvei H4 on the Coculture System of Hafnia alvei H4 and Pseudomonas fluorescens ATCC13525.

In the food industry, foodborne spoilage bacteria often live in mixed species and attach to each other, leading to changes in spoilage characteristics. Quorum sensing (QS) has been reported to be a regulating mechanism for food spoiling by certain kinds of bacteria. Here, the contents of biofilm, extracellular polysaccharides, and biogenic amines in the coculture system of Hafnia alvei H4 and Pseudomonas fluorescens ATCC13525 were significantly reduced when the QS element of H. alvei H4 was deleted, confirming that QS of H. alvei H4 is involved in the dual-species interactions. Then, transcriptomics was used to explore the regulatory mechanism at the mRNA molecular level. The deletion of the QS element decreased the transcript levels of genes related to chemotaxis, flagellar assembly, and the two-component system pathway of H. alvei H4 in the coculture system. Furthermore, a total of 732 DEGs of P. fluorescens ATCC13525 were regulated in the dual species, which were primarily concerned with biofilm formation, ATP-binding cassette transporters, and amino acid metabolism. Taken together, the absence of the QS element of H. alvei H4 weakened the mutual cooperation of the two bacteria in the coculture system, making it a good target for managing infection with H. alvei and P. fluorescens.

A trinuclear Zn (II) schiff base dicyanamide complex attenuates bacterial biofilm formation by ROS generation and membrane damage and exhibits anticancer activity

A trinuclear Zn (II) complex, [(ZnL{N(CN)2})2Zn], termed complex 1 has been synthesized by the reaction of an aqueous solution of sodium dicyanamide to the methanolic solution of Zn (CH3COO)2, 2H2O and corresponding Schiff base (H2L) which is derived from 1:2 condensation of 1, 4 butane diamine with 3-ethoxy salicylaldehyde. Complex 1 is characterized by elemental analysis, IR, UV and Single X-ray diffraction study. Drug resistance is a growing global public health concern that has prompted researchers to look into advanced alternative treatment modalities. In this context, complex 1 has shown promising antibacterial and antibiofilm efficacy against gram-positive Staphylococcus aureus and Methicillin-resistant Staphylococcus aureus strains. Complex 1 attenuated Staphylococcal biofilm formation by reducing several virulence factors including the formation of extracellular polysaccharide matrix, slime, haemolysin, staphyloxanthin, auto-aggregation, cell surface hydrophobicity, and motility. Notably, complex 1 mechanistically potentiated Reactive Oxygen Species (ROS) generation within the bacterial cells, leading to the damage of bacterial cell membrane followed by DNA leakage and thereby impeding the growth of Staphylococcus aureus. Furthermore, complex 1 significantly exhibited anticancer activity by reducing the growth of prostate adenocarcinoma cells. It obstructed the migration of cancer cells by potentiating apoptosis and arresting the cell cycle at the G2/M phase. In summary, complex 1 could act as a potent candidate for the generation of novel antibacterial, antibiofilm as well as anticancer treatment regimens for the management of drug-resistant biofilm-mediated Staphylococcus aureus infection and lethal prostate malignancy.

Physiological regulation of microalgae under cadmium stress and response mechanisms of time-series analysis using metabolomics

The investigation of heavy metal wastewater treatment utilizing microalgae adsorption has been extensively demonstrated. However, the response mechanism based on metabolomics to analyze the time-series changes of microalgae under Cd stress has not been described in detail. In this study, SEM/TEM demonstrated that Cd accumulated on the cell surface of microalgae and was bioconcentrated in the cytoplasm, vesicles, and chloroplasts. Carbonyl/quinone/ketone/carboxyl groups (OCO), membrane polysaccharides (OH), and phospholipids (PO) were involved in the interaction of Cd ions, and the chlorophyll content underwent a process of decreasing in the early stage (1.62 mg/g at 48 h) and recovering to the normal level in the late stage, and the contents of MDA, GSH, and SOD were all increased (29.7 nmol/g, 0.23 mg/g, and 30.01 u/106 cells) and then gradually returned to the steady state. The results of EPS content and fluorescent labeling showed that Cd induced the overexpression and synthesis of extracellular polysaccharides and proteins, which is one of the defense mechanisms participating in the reduction of cellular damage by complexed Cd. Metabolomics results indicated that the malate synthesis pathway was activated after Cd-20 h, and the microalgal cells began to shift the metabolic pathway to storage lipid or polysaccharide biosynthesis. In the Calvin cycle, the expression of D-Sedoheptulose 7-phosphate in Cd-20 h_vs_ck and Cd-72 h_vs_Cd-20 h firstly declined and then increased, and the photosynthesis system was suppressed at the beginning, and then gradually returned to normal to maintain the successful development of the dark reaction. The results of time series analysis revealed that the response of microalgae to Cd was categorized into fast response and slow response to regulate cell adsorption and growth metabolism.

Improvement of extracellular polysaccharides production from Cordyceps militaris immobilized alginate beads in repeated-batch fermentation

Cordyceps militaris (C. militaris) is a unique medicinal fungus of the genus Cordyceps. It has high economic value due to various biological functions from the extracellular polysaccharides (EPSs) it produces. The aim of this study was to establish a mycelium-immobilization system for EPS production in a bubble column system. First, EPS production was optimized by 90.2% (from 0.72 to 1.37 g/L) using response surface methodology under an alginate-immobilized mycelium system with 6 days of cultivation. The highest EPS production was obtained (5.3 g/L) in a flask system through elongation to 15 days of cultivation. EPS production subsequently further increased to 10.42 g/L utilizing a bubble column reactor. Furthermore, the system can be reused for at least three rounds. In the EPS characteristics analysis, the produced EPSs were mainly composed of glucose and galactose. They presented strong antibacterial activity against Staphylococcus aureus. In terms of antioxidant activity, respective the EPS concentration that caused 50% scavenging of DPPH and ABTS free radicals values (IC50) were 4.65 and 7.92 mg/mL. In summary, these findings should be helpful for establishing a suitable industrial fermentation system for C. militaris and can be studied in the food and medical industries in future work.

Inhibition of Streptococcus mutans growth and biofilm formation through protein acetylation.

Numerous cellular processes are regulated in response to the metabolic state of the cell, and one such regulatory mechanism involves lysine acetylation. Lysine acetylation has been proven to play an important role in the virulence of Streptococcus mutans, a major cariogenic bacterial species. S. mutans' glucosyltransferases (Gtfs) are responsible for synthesizing extracellular polysaccharides (EPS) and contributing to biofilm formation. One of the most common nonsteroidal anti-inflammatory drugs is acetylsalicylic acid (ASA), which can acetylate proteins through a nonenzymatic transacetylation reaction. Herein, we investigated the inhibitory effects of ASA on S. mutans. ASA treatment was observed to impede the growth of S. mutans, leading to a reduction in the production of water-insoluble EPS and the formation of biofilm. Moreover, ASA decreased the enzyme activity of Gtfs while increasing the protein acetylation level. The in vivo anticaries efficacy of ASA has further been proved using the rat caries model. In conclusion, ASA as an acetylation agent attenuated the cariogenic virulence of S. mutans, suggesting the potential value of protein acetylation on antimicrobial and anti-biofilm applications to S. mutans.

New insights into rare earth element-induced microalgae lipid accumulation: Implication for biodiesel production and adsorption mechanism

A coupling technology for lipid production and adsorption of rare earth elements (REEs) using microalgae was studied in this work. The microalgae cell growth, lipid production, biochemical parameters and lipid profiles were investigated under different REEs (Ce3+, Gd3+and La3+). The results showed that the maximum lipid production was achieved at different concentrations of REEs, with lipid productivities of 300.44, 386.84 and 292.19 mg L−1 d−1 under treatment conditions of 100 μg L−1 Ce3+, 250 μg L−1 Gd3+ and 1 mg L−1 La3+, respectively. Moreover, the adsorption efficiency of Ce3+, Gd3+ and La3+exceeded 96.58 %, 93.06 % and 91.3 % at concentrations of 25–1000 μg L−1, 100–500 μg L−1 and 0.25–1 mg L−1, respectively. In addition, algal cells were able to adsorb 66.2 % of 100 μg L−1 Ce3+, 48.4 % of 250 μg L−1 Gd3+ and 59.9 % of 1 mg L−1 La3+. The combination of extracellular polysaccharide and algal cell wall could adsorb 25.2 % of 100 μg L−1 Ce3+, 44.5 % of 250 μg L−1 Gd3+ and 30.5 % of 1 mg L−1 La3+, respectively. These findings indicated that microalgae predominantly adsorbed REEs through the intracellular pathway. This study elucidates the mechanism of effective lipid accumulation and adsorption of REEs by microalgae under REEs stress conditions. It establishes a theoretical foundation for the efficient microalgae lipid production and REEs recovery from wastewater or waste residues containing REEs.

Phytofabricated bimetallic synthesis of silver-copper nanoparticles using Aerva lanata extract to evaluate their potential cytotoxic and antimicrobial activities

In this study, we demonstrate the green synthesis of bimetallic silver-copper nanoparticles (Ag–Cu NPs) using Aerva lanata plant extract. These NPs possess diverse biological properties, including in vitro antioxidant, antibiofilm, and cytotoxic activities. The synthesis involves the reduction of silver nitrate and copper oxide salts mediated by the plant extract, resulting in the formation of crystalline Ag–Cu NPs with a face-centered cubic structure. Characterization techniques confirm the presence of functional groups from the plant extract, acting as stabilizing and reducing agents. The synthesized NPs exhibit uniform-sized spherical morphology ranging from 7 to 12 nm. They demonstrate significant antibacterial activity against Staphylococcus aureus and Pseudomonas aeruginosa, inhibiting extracellular polysaccharide secretion in a dose-dependent manner. The Ag–Cu NPs also exhibit potent cytotoxic activity against cancerous HeLa cell lines, with an inhibitory concentration (IC50) of 17.63 µg mL−1. Additionally, they demonstrate strong antioxidant potential, including reducing capability and H2O2 radical scavenging activity, particularly at high concentrations (240 µg mL−1). Overall, these results emphasize the potential of A. lanata plant metabolite-driven NPs as effective agents against infectious diseases and cancer.

Effect of a novel pH-responsive tertiary amine monomer dodecylmethylaminoethyl methacrylate modified resin adhesive on biofilm formation of Streptococcus mutans and Lactobacillus casei in vitro

Objective: To explore the application prospect of a new pH-responsive tertiary amine monomer dodecylmethylaminoethyl methacrylate (DMAEM) modified resin adhesive (DMAEM@RA) in the prevention and treatment of secondary caries. Methods: Five percents DMAEM was added to the resin adhesive to synthesize DMAEM@RA for modifying. Streptococcus mutans (Sm) and Lactobacillus casei (Lc) biofilms were cultured on resin adhesive and DMAEM@RA, respectively. The culture systems were set up at pH=7.4, 6.0, 5.5, and 5.0. The antimicrobial activity of DMAEM@RA was evaluated by quantitative PCR. The effects of DMAEM@RA on biofilm thickness, bacterial amount, and extracellular polysaccharides were studied by scanning electron microscope (SEM) and extracellular polysaccharide staining. Real-time fluorescence quantitative PCR was used to study the effect of DMAEM@RA on the expression levels of cariogenic genes in Sm. Results: DMAEM@RA could significantly reduce the amount of Sm and Lc under acidic conditions, especially Lc. At pH=5.0, the logarithm value of co-cultured Sm bacteria [lg (CFU/ml)] in DMAEM@RA group (7.58±0.01) was significantly lower than that in control group (7.87±0.03) (t=14.32, P<0.001), and the logarithm value of Lc bacteria [lg (CFU/ml)] (7.29±0.04) was also significantly lower than that in control group (7.93±0.15) (t=6.93, P=0.002). SEM observed that the bacteria decreased and the cell fragments appeared in DMAEM@RA group. In addition, DMAEM@RA significantly reduced the biomass of extracellular polysaccharides in the dual-species biofilm under acidic conditions. At pH=5.0, the biomass of extracellular polysaccharides in DMAEM@RA group [(25.13±3.14) mm3/mm2] was significantly lower than that in the control group [(42.66±7.46) mm3/mm2] (t=3.75, P=0.020). DMAEM@RA could significantly up-regulate the expressions of gtfB and gtfC genes in Sm under acidic conditions. At pH=5.0, gtfB and gtfC genes were significantly up-regulated by (14.64± 0.44) times and (2.99±0.20) times, respectively (t=-42.74, P<0.001; t=-13.55, P<0.001). Conclusions: The DMAEM@RA has a good antibacterial effect under acidic conditions, demonstrating that it has a good potential to prevent the occurrence and development of secondary caries.

Production with Fermentation Culture and Antioxidant Activity of Polysaccharides from Morchella esculenta

Morchella esculenta is a precious edible and medicinal fungus rich in protein, polysaccharides, polyphenols, amino acids, triterpenes, and other active components. In this study, MS-1 was isolated from the fruiting body of M. esculenta. Through conducting single-factor experiments and the response surface analysis of the culture conditions, the optimal culture components of an M. esculenta fermentation broth for extracellular polysaccharide production were determined, namely, 3.7% glucose, 2% yeast extract, and 0.15% sodium chloride. The polysaccharides MSF and MSL were extracted from the fruiting body of M. esculenta and the fermentation broth, respectively, and analyzed with gel permeation chromatography (GPC), monosaccharide composition, scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA), and in vivo and in vitro antioxidant and anti-inflammatory activity analyses. The research results show that the calculated MW of MSF is 2.34 × 105 Da, and the calculated MW of MSL is 1.40 × 105 Da. MSF is composed of three monosaccharides: D-galactose, D-glucose, and D-mannose (molar ratio of 4.34:90.22:5.45). MSL consists of five monosaccharides: D-arabinose, D-galactose, D-glucose, D-mannose, and glucuronic acid (molar ratio of 0.31:14.71:13.03:71.43:0.53). The in vitro antioxidant test results show that MSF and MSL both have significant antioxidant activities. Activity experiments on MSF and MSL in zebrafish showed that MSF and MSL have significant repair effects on the oxidative damage caused by metronidazole in zebrafish embryos, and there were significant changes in the transcriptional activity levels of the oxidative stress-related genes SOD, Keap1, and Nrf2. Therefore, the polysaccharides MSF and MSL from MS-1 can be used as important raw materials for functional foods and drugs.

Structural properties and biological activities of the extracellular polysaccharide of Bacillus subtilis LZ13-4

The remarkable functional characteristics of Bacillus subtilis extracellular polysaccharides (BSPS) are of great interest. Therefore, in the present study, BSPS was isolated and characterized to obtain two fractions, BSPS-1 and BSPS-2, respectively, and to investigate their biological activities. BSPS-1 contained fructose, glucose, and galactose (molar ratio: 25.27:43.37:31.36), while BSPS-2 contained fructose with only trace amounts of glucose, galactose, and mannose (molar ratio: 55.08:19.03:19.21:6.68), and their respective average molecular weights were 16.9 kDa and 202.67 kDa. With a 93.55 % clearance of ABTS•+ at a concentration of 2 mg/mL of BSPS-1, the antioxidant activity revealed that BSPS-1 had greater antioxidant activity than BSPS-2 and that both were concentration-dependent. The inhibitory effect on HepG2 cells demonstrated that BSPS-1 and BSPS-2 significantly inhibited the proliferation of HepG2 and increased the expression of apoptotic proteins, causing apoptosis. The inhibition rate on HepG2 cells was dose-dependent and reached 52.7 % and 40.3 % after 48 h of action. BSPS-2 and 800 μg/mL BSPS-1 growth was inhibited in the G1/G0 phase, while 200 and 400 μg/mL BSPS-1 growth was inhibited in the S phase. In conclusion, the study of the BSPS's structure and properties can offer a theoretical foundation for real-world industrial applications.

Digestive characteristics of extracellular polysaccharide from Lactiplantibacillus plantarum T1 and its regulation of intestinal microbiota

This study assessed the potential prebiotic characteristics of the previously reported Lactiplantibacillus plantarum extracellular polysaccharide (EPS-T1) with immunological activity. EPS-T1 was a novel heteropolysaccharide composed of glucose and galactose (1.00:1.21), with a molecular weight of 1.41 × 106 Da. The monosaccharide composition, molecular weight, fourier transform infrared, and 1H NMR analysis showed that EPS-T1 was well tolerated in the simulated oral cavity, gastric fluid, and small intestinal fluid environments, and was not easily degraded. Meanwhile, EPS-T1 could effectively be used as a carbon source to promote the growth of beneficial Lactobacillus species (Lactobacillus delbrueckii ssp. Bulgaricus, Streptococcus thermophilus, Lacticaseibacillus rhamnose GG, Lactiplantibacillus plantarum, Lacticaseibacillus paracasei and Lactobacillus reuteri). After 24 h of fecal fermentation, EPS-T1(5 mg/mL) effectively reduced the relative abundance of harmful bacteria such as the Escherichia-Shigella, Citrobacter, Fusobacterium, Parasutterella, and Lachnoclostridium. While, the level content of beneficial flora (Bacteroides, Blautia, Phascolarctobacterium, Bifidobacterium, Parabacteroides, and Subdoligranulum) were significantly increased. In addition, EPS-T1 was able to significantly promote the enrichment of short-chain fatty acids such as acetic acid, propionic acid and butyric acid. These results provide some basis for the functional application of EPS-T1 as a potential prebiotic.

Polysaccharides Produced by Plant Growth-Promoting Rhizobacteria Strain Burkholderia sp. BK01 Enhance Salt Stress Tolerance to Arabidopsis thaliana.

Salt stress is one of the most serious abiotic stresses leading to reduced agricultural productivity. Polysaccharides from seaweed have been used as biostimulants to promote crop growth and improve plant resistance to abiotic stress. In this study, PGPR strain Burkholderia sp. BK01 was isolated from the rhizosphere of wheat, and it was characterized for phosphorus (Pi) dissolution, indole-3-acetic acid (IAA) production, ammonia (NH3) and exopolysaccharides (EPS). In particular, strain BK01 can efficiently produce extracellular polysaccharide with a yield of 12.86 g/L, using sorbitol as carbon source. BK01 EPS was identified as an heteropolysaccharide with Mw 3.559 × 106 Da, composed of (D)-galactose (75.3%), (D)-glucose (5.5%), (L)-rhamnose (5.5%), (D)-galactouronic acid (4.9%) and (D)-glucuronic acid (8.8%). The present work aims to highlight the effect of the BK01 EPS on growth and biochemical changes in Arabidopsis thaliana under salt stress (100 mM). The purified BK01 EPS at a concentration of 100 mg/L efficiently promoted the growth of plants in pot assays, improved the chlorophyll content, enhanced the activities of SOD, POD and CAT, and decreased the content of MDA. This results suggested that the polysaccharides produced by PGPR strain Burkholderia sp. BK01 can be used as biostimulants to promote plant growth and improve plant resistance to salt stress.