A near-rose-colored *Syngonium spp.* polysaccharide and its application in adsorbing zearalenone.

Abstract

This invention discloses a polysaccharide from *Synthia spp.* and its application in the adsorption of zearalenone, belonging to the field of biotechnology. *Synthia spp.* is inoculated into a liquid culture medium, cultured, and the supernatant is removed. The mixture is washed with sterile water and inactivated. Crude polysaccharide is extracted using an alkaline extraction method. Proteins are removed, and the mixture is dialyzed. The polysaccharide is redissolved in pure water, centrifuged, and the supernatant is purified using a weak anion exchange column. The polysaccharide is then separated and purified using an AKTA purification system at a flow rate of 4 mL / min. Elution with pure water yields SPYP 1, and elution with 0.1 M NaCl followed by dialyzing yields SPYP 2. SPYP 2 can adsorb zearalenone, with stronger adsorption capacity in the presence of glucose or mannose.

Description

Technical Field

[0001] This invention belongs to the field of biotechnology, specifically relating to a polysaccharide of *Syngonium spp.* and its application in the adsorption of zearalenone. Background Technology

[0002] Mycotoxins are toxic secondary metabolites produced by molds after food contamination, posing serious health risks with long-term intake. Zearalenone (ZEN), a highly stable and significantly toxic mycotoxin, not only accumulates and is transferred through the "corn-feed-livestock product" chain, posing a systemic biosafety risk to the entire industry chain, but also causes reproductive, liver, and kidney damage, endangering human health. Therefore, reducing the hazards of ZEN has become an urgent problem to be solved. Among existing ZEN treatment methods, microbial adsorption has significant advantages such as low cost and environmental friendliness, especially suitable for treating low-concentration pollutants and resource recovery, avoiding the secondary pollution problems of chemical and physical methods, and is a sustainable toxin treatment method. Microbial adsorbents mainly include beneficial microorganisms such as yeast and lactobacillus; however, these strains generally suffer from unsatisfactory adsorption effects and relatively limited species diversity. Therefore, there is an urgent need to develop new microbial resources to address ZEN contamination. Yeast-based adsorbents, due to their unique cell wall structure, exhibit high adsorption capacity and selectivity for ZEN pollution. Furthermore, their large cell size and good settling properties facilitate solid-liquid separation and recovery, making them an ideal choice for efficient, low-consumption treatment and resource recovery. In recent years, in addition to the yeast adsorbent itself, its active components—especially yeast polysaccharides—have received widespread attention as a highly promising biosorbent material. They not only possess strong toxin adsorption capabilities but also exhibit high biocompatibility, providing efficient and diversified solutions for ZEN pollution control.

[0003] This study screened *Sporobolomyces pararoseus* from food to identify it as having good ZEN adsorption performance and explored its adsorption mechanism in depth. Given the crucial role of yeast polysaccharides in this adsorption process, the polysaccharides of *Sporobolomyces pararoseus* were further isolated, purified, and structurally identified. The in vitro adsorption effect and in vivo attenuation effect of these polysaccharides on ZEN were investigated, providing a theoretical basis and research ideas for the development of novel ZEN adsorbents. Summary of the Invention

[0004] The purpose of this invention is to solve the above-mentioned technical problems by providing a polysaccharide SPYP2 of *Syngonium rosenbergii* and its application in the adsorption of zearalenone.

[0005] A near-rose-colored polysaccharide SPYP 2 from *Syngonium spp.* is prepared by the following method:

[0006] 1) Inoculate nearly 1000 rosédacna into liquid culture medium, remove the supernatant, wash with sterile water, and inactivate.

[0007] 2) Crude polysaccharides of *Syngonium spp.* near-rose color were extracted using the alkaline extraction method;

[0008] 3) Remove protein, then dialyze in a dialysis bag;

[0009] 4) Isolation and purification of polysaccharides: Redissolve in pure water, centrifuge and collect the supernatant, purify with a weak anion exchange column; use an AKTA purification system for separation and purification at a flow rate of 4 mL / min; elute with pure water to obtain SPYP 1, elute with 0.1 M NaCl, and dialyze to obtain SPYP 2;

[0010] Step 4) describes a weak anion exchange column as DEAE seplife FF.

[0011] Step 2) describes the alkaline extraction method, in which the inactivated *Synthia sacchariformis* was dissolved in 2 M NaOH solution and stirred at room temperature for 2 h. After centrifugation, the supernatant was obtained, and anhydrous ethanol was added at a volume ratio of 1:4 to precipitate the supernatant overnight. After centrifugation, the precipitate was redissolved, and the protein was removed using the Sevag method. The supernatant was then freeze-dried to obtain crude yeast polysaccharide.

[0012] Step 3) describes the removal of protein using the Sevag method, followed by dialysis in an 8000–14000 kDa dialysis bag.

[0013] Another objective of this invention is the application of the aforementioned polysaccharide SPYP2 from *Saccharomyces cerevisiae* as a zearalenone adsorbent;

[0014] The application described involves the adsorption of zearalenone in the presence of glucose or mannose.

[0015] This invention provides a method for screening the extraction of yeast polysaccharides from a strain of Sporobolomyces pararoseus that efficiently adsorbs zearalenone. In particular, the method of alkaline extraction after high-temperature inactivation improves the yield of yeast polysaccharides and achieves the best adsorption performance. It is proposed that the high-temperature assisted alkaline extraction method can significantly improve the adsorption characteristics of yeast polysaccharides.

[0016] This invention provides a novel molecular structure for yeast-based polysaccharides, particularly highlighting that increased glucose and mannose content enhances their ability to adsorb zearalenone.

[0017] Based on this, the yeast-based polysaccharide provided by the present invention has adsorption characteristics for zearalenone and can reduce the content of zearalenone in vivo, thereby reducing damage to the body. The extraction method of the yeast polysaccharide of the present invention combines high-temperature and high-pressure extraction and alkaline extraction, which can effectively improve the extraction efficiency of active ingredients and ensure that the extract has a strong adsorption effect.

[0018] The beneficial effects of this invention are:

[0019] This invention provides a screening method for yeast polysaccharide extraction, identifying an alkaline-extracted polysaccharide from *Syngonium rosenbergii* that exhibits good adsorption performance for zearalenone. This polysaccharide is a natural biological macromolecule, non-toxic, and does not produce secondary pollution, making it safe for use as a feed or food additive. The preparation process is simple and cost-effective; the polysaccharide extraction process is straightforward, requiring no complex chemical modification or loading steps, facilitating industrial production and demonstrating good economic feasibility. It also exhibits good stability and wide applicability, suitable for various scenarios such as feed processing, grain storage, and food raw material pretreatment. Attached Figure Description

[0020] Figure 1 Comparison of ZEN adsorption capacity and purity of yeast polysaccharides extracted using different methods. (A, B) Adsorption capacity and ZEN removal rate of polysaccharides extracted using different methods; (C) Comparison of polysaccharide purity and extraction rate extracted using different methods. (Note: Identical letters indicate no significant difference, P<0.05);

[0021] Figure 2 Elution curve of AE-P;

[0022] Figure 3 SEM observations of SPYP 1 and SPYP 2;

[0023] Figure 4 AFM observations of SPYP 1 and SPYP 2. (A) SPYP 1; (B) SPYP 2;

[0024] Figure 5 TEM morphology observation results of SPYP-1 and SPYP-2;

[0025] Figure 6 FT-IR results for SPYP 1 and SPYP 2;

[0026] Figure 7 Adsorption screening of ZEN by SPYP 1 and SPYP 2 and purification results of SPYP 2. (A) Component screening; (B) SPYP 2 purification. (* indicates P<0.05, indicating significant difference);

[0027] Figure 8Monosaccharide composition and molecular weight determination results of SPYP2. (A) Monosaccharide composition; (B) Light scattering (LS) and refractive index (RI) signals; (C) Absolute molecular weight; (D) Molecular configuration diagram;

[0028] Figure 9 SPYP 2's 1H spectrum, 13C spectrum, DEPT-135, and TOCSY. (A) 1H spectrum; (B) 13C spectrum; (C) DEPT-135; (D) TOCSY;

[0029] Figure 10 SPYP 2 two-dimensional nuclear magnetic resonance spectrum. (A) COSY; (B) HSQC; (C) HMBC; (D) NOESY;

[0030] Figure 11 Chemical formula and structural formula of SPYP 2. (A) Chemical formula of SPYP 2; (B) Structural formula of SPYP 2;

[0031] Figure 12 Kinetics, internal diffusion, and thermodynamic models of SPYP2 adsorption of ZEN. (AB) Kinetic model; (CD) Thermodynamic model;

[0032] Figure 13 Mechanism of ZEN adsorption by SPYP 2. (A) FT-IR; (B) XPS; (CE) C1s, O1s and N1s spectra of SPYP 2; (FH) C1s, O1s and N1s spectra of SPYP 2 after ZEN adsorption;

[0033] Figure 14 Molecular dynamics simulations reveal the self-assembly mechanism of SPYP 2-ZEN. (A) Snapshot of the aggregation of SPYP 2 (blue) and ZEN (red) clusters in water. (B) SASA changes of SPYP 2 during the simulation. (C) Time evolution of the interaction energy between SPYP 2 and ZEN (van der Waals forces and electrostatic interactions). (D) Hydrogen bond count between SPYP 2 and ZEN throughout the simulation.

[0034] Figure 15 DFT calculations. (A–C) Different molecular surfaces of mannose and glucose and their surface extrema; (D) HOMO-LUMO orbitals after adsorption and E gap (E) Scatter plot of Sign(λ2)p colored IGM; (F) Distribution of interactions in real space, where red spheres represent oxygen atoms, cyan spheres represent carbon atoms, and white spheres represent hydrogen atoms. Detailed Implementation

[0035] Example 1: Extraction and identification of polysaccharides from *Syndrome affinis*

[0036] 1.1. Extraction of polysaccharides from *Syngonium spp.*

[0037] The inactivated *Synthia spp.* was dissolved in 2 M NaOH solution and stirred at room temperature for 2 h. After centrifugation, the supernatant was obtained. Anhydrous ethanol was added at a ratio of 1:4 (v / v) to precipitate the supernatant overnight. After centrifugation, the precipitate was redissolved, and the protein was removed using the Sevag method. The supernatant was then freeze-dried to obtain crude polysaccharide extracted from the yeast.

[0038] The aforementioned *Sporobolomyces pararoseus* strain used in our laboratory was isolated from *Persimmon arborescens* and identified by Kumei Biotechnology Co., Ltd. as belonging to the genus *Sporobolomyces pararoseus*. Comparison with the standard strain confirmed that no genetic variation had occurred.

[0039] 1.2 Isolation and purification of polysaccharides from *Syngonium spp.*

[0040] Approximately 5 g of yeast polysaccharide sample was dissolved in 200 mL of pure water and centrifuged at 10000 r / min for 10 min. The supernatant was collected and purified using a DEAE Seplife FF weak anion exchange column and an AKTA purification system at a flow rate of 4 mL / min. SPYP 1 was obtained by elution with pure water, SPYP 2 by elution with 0.1 M NaCl, and SPYP 3 by elution with 0.2 M NaCl. Each fraction was collected in 15 mL tubes and dialyzed separately. The total sugar content of the eluent in each collection tube was determined using the sulfuric acid-phenol method. Specifically, 100 μL of diluted polysaccharide supernatant was added to 600 μL of sulfuric acid-phenol reagent (5% phenol solution: concentrated sulfuric acid = 1:5 (v / v)), mixed well, and allowed to stand in the dark for 10 min. The absorbance was measured at 490 nm. An ion purification elution curve was plotted. Since no sample was available after dialysis with SPYP 3, SPYP 1 and SPYP 2 were used for adsorption capacity comparison. SPYP 2 showed better adsorption than SPYP 1, therefore SPYP 2 was selected for subsequent experiments. Approximately 1 g of purified SPYP 2 was added to 20 mL of pure water and centrifuged at 10000 r / min for 10 min. The supernatant was then passed through a Sephacryl S-400HR gel chromatography column and purified using the AKTA system at a flow rate of 1 mL / min. Elution was performed with 1.5 column volumes of pure water, collecting 10 mL of eluent in each tube.

[0041] 1.3 Determination of monosaccharide composition of crude polysaccharide from *Syndrome affinis*

[0042] 1) Preparation of Standards

[0043] Accurately weigh fucose, rhamnose, arabinose, galactose, glucose, xylose, mannose, fructose, ribose, galacturonic acid, glucuronic acid, mannuronic acid, and guluronic acid respectively, and dilute with water to prepare a 10 mg / mL monosaccharide standard stock solution. Then, take appropriate amounts of each monosaccharide stock solution and mix them to prepare mixed standard solutions with the highest index concentration of 60 μg / mL, 50 μg / mL, or 40 μg / mL. Then, dilute stepwise according to the corresponding concentration gradient to prepare a series of standard solutions for instrumental analysis.

[0044] 2) Sample preparation

[0045] Take a clean chromatographic vial, weigh an appropriate amount of polysaccharide sample, add 1 mL of 2 M trifluoroacetic acid (TFA) solution, and heat at 121℃ for 2 h. Purge with nitrogen, dry, wash with 99.99% methanol, dry again, and repeat the methanol washing 2–3 times. Dissolve in sterile water, transfer to a chromatographic vial for analysis.

[0046] 3) Chromatographic determination

[0047] The chromatographic system used was the Thermo ICS 5000+ ion chromatography system, which employed an electrochemical detector to analyze and detect monosaccharide components. A Dionex™ CarboPac™ PA20 (150*3.0 mm, 10 μm) liquid chromatography column was used; the injection volume was 5 μL. Mobile phase A (H2O), mobile phase B (0.1 M NaOH), mobile phase C (0.1 M NaOH, 0.2 M NaAc), flow rate 0.5 mL / min; column temperature 30℃; elution gradient: 0 min A / B / C (95:5:0, V / V), 26 min A / B / C (85:5:10, V / V), 42 min A / B / C (85:5:10, V / V), 42.1 min A / B / C (60:0:40, V / V), 52 min A / B / C (60:40:0, V / V), 52.1 min A / B / C (95:5:0, V / V), 60 min A / B / C (95:5:0, V / V).

[0048] 1.4 Determination of molecular weight of crude polysaccharide from *Syndrome affinis*

[0049] The sample was dissolved in 0.1 M NaNO3 aqueous solution (containing 0.02% NaN3, w / w) to a final concentration of 1 mg / mL, and filtered through a 0.45 μm filter before analysis. The chromatographic system used was a gel permeation chromatography-differential chromatography-multi-angle laser light scattering system. The liquid chromatography system was a U3000 (Thermo, USA), the differential detector was an Optilab T-rEX (Wyatt technology, CA), and the laser light scattering detector was a DAWN HELEOS II (Wyatt technology, CA). Specific column and elution conditions were as follows: Ohpak SB-805 HQ (300 × 8 mm) and Ohpak SB-803HQ (300 × 8 mm) gel size exclusion columns were used in series. The column temperature was 45℃, the injection volume was 100 μL, the mobile phase was A (0.02% NaN3, 0.1 M NaNO3), the flow rate was 0.6 mL / min, isocratic elution was used, and the elution time was 75 min.

[0050] 1.5 Isolation, purification, and component screening of crude polysaccharides from *Syndrome affinis*

[0051] Approximately 5 g of yeast polysaccharide sample was dissolved in 200 mL of pure water and centrifuged at 10000 r / min for 10 min. The supernatant was collected and purified using a DEAE Seplife FF weak anion exchange column and an AKTA purification system at a flow rate of 4 mL / min. A gradient elution was performed sequentially with pure water, 0.1 M, 0.2 M, and 0.3 M NaCl solutions, collecting 15 mL of each eluent tube. The total sugar content of the eluent in each collection tube was determined using the sulfuric acid-phenol method. Specifically, 100 μL of the diluted polysaccharide supernatant was added to 600 μL of sulfuric acid-phenol reagent (5% phenol solution: concentrated sulfuric acid = 1:5 (v / v)), mixed well, and allowed to stand in the dark for 10 min. The absorbance was measured at 490 nm. An ion purification elution curve was plotted. The eluents from the collection tubes corresponding to the same elution peak were combined, rotary evaporated, dialyzed, and lyophilized to obtain two fractions, which were named SPYP 1 and SPYP 2, respectively.

[0052] 1.6 Structural characterization of polysaccharide (SPYP) components

[0053] 1.6.1 Scanning Electron Microscopy (SEM) Measurement

[0054] SPYP 1 and SPYP 2 were observed using SEM. The purified polysaccharide samples were placed on the sample stage, evenly spread on a strip, sputtered with gold, and then examined using SEM. SEM images of the two polysaccharides were obtained at magnifications of 500×, 2500×, and 5000×.

[0055] 1.6.2 Atomic Force Microscopy (AFM) Measurement

[0056] SPYP 1 and SPYP 2 were observed using AFM. A 5.0 μg / mL polysaccharide aqueous solution was heated at 80℃ for 40 min and then sonicated for 10 min to break up the large aggregates and obtain a fully dissolved polysaccharide solution. The solution was then dropped onto fresh mica sheets while hot and air-dried at 35℃ before observation.

[0057] 1.6.3 Transmission Electron Microscopy (TEM) Measurement

[0058] Yeast polysaccharides (SPYP 1 and SPYP 2) were observed using TEM.

[0059] 1.6.4 FT-IR Measurement

[0060] Take a dried polysaccharide sample and mix it with spectroscopically pure potassium bromide at a ratio of 1:100 and grind it. Press it into a transparent thin film and scan it with an FT-IR spectrometer in the range of 4000–400 cm⁻¹ (resolution 4 cm⁻¹, 64 scans).

[0061] 1.6.5 Thermal stability determination

[0062] The polysaccharide samples were analyzed using a thermogravimetric analyzer. The temperature was increased from room temperature (25°C) to 600°C at a linear rate of 10°C / min, and the entire process was carried out under a nitrogen atmosphere.

[0063] 1.7 Gel purification and structural identification of polysaccharides

[0064] Take about 1 g of the ion-purified polysaccharide sample and add it to 20 mL of pure water. Centrifuge at 10000 r / min for 10 min. Take the supernatant and pass it through a Sephacryl S-400HR gel chromatography column. Use the AKTA system for separation and purification at a flow rate of 1 mL / min. Elute with 1.5 column volumes of pure water, collecting one tube for every 10 mL. Collect all the eluent.

[0065] 1.7.1 Molecular weight determination

[0066] The sample was dissolved at a concentration of 1 mg / mL in a 0.1 M NaNO3 aqueous solution containing 0.02% NaN3 and filtered through a 0.45 μm pore size membrane. The homogeneity and molecular weight of the polysaccharide were determined using a technique combining size exclusion chromatography with differential detection and multi-angle laser light scattering (SEC-MALLS-RI).

[0067] 1.7.2 Monosaccharide Composition

[0068] Take approximately 5 mg of sample and hydrolyze it in a sealed tube with 2 M trifluoroacetic acid at 121 °C for 2 h. Dry the sample under nitrogen. Wash with methanol, dry, and repeat the process 2–3 times. Resolve the residue in deionized water and filter it through a 0.22 μm microporous membrane before analysis. Instrument settings are the same as in Method 3.3.3. Data were acquired using ICS5000+ ion chromatography and processed using a Chromeleon 7.2 CDS (Thermo Scientific).

[0069] 1.7.3 Polysaccharide Methylation

[0070] The polysaccharide sample was dissolved in dimethyl sulfoxide (DMSO). This solution was methylated with CH3I in a DMSO / NaOH system. After methylation, it was hydrolyzed with 2 mol / L TFA at 121 °C for 1.5 h, followed by reduction with NaBD4 and acetylation with acetic anhydride at 100 °C for 2.5 h. Gas chromatography-mass spectrometry (GC-MS) conditions were as follows: acetate dissolved in chloroform, equipped with an Agilent BPX70 column (30 m × 0.25 mm × 0.25 µm), and high-purity helium (split ratio 10:1) as the carrier gas, with an injection volume of 1 μL. Mass spectrometry analysis was performed at an initial temperature of 140 °C for 2 min, followed by a temperature ramp to 230 °C at a rate of 3 °C / min for 3 min. The scan mode was SCAN, with a mass range (m / z) of 50–350.

[0071] 1.7.4 Polysaccharide NMR Analysis

[0072] The purified sample was dissolved in 0.5 mL of D2O to a final concentration of 40 mg / mL. 1D-NMR (¹H-NMR, ¹³C-NMR, DEPT135) and two-dimensional NMR (COSY, NOESY, HMBC, HSQC, and TOCSY) measurements were performed at 25 °C using an NMR spectroscopy system at a frequency of 500 MHz.

[0073] 2. Investigation into the adsorption mechanism of yeast polysaccharides

[0074] 2.1 Kinetic experiment of polysaccharide adsorption of ZEN

[0075] 15 mg of SPYP 2 was added to each of the 15 mL Erlenmeyer flasks containing ZEN solutions of different concentrations (4, 8, 12, 16, 20 μg / mL). The Erlenmeyer flasks were then sealed and placed in a constant temperature shaker (150 r / min, 25℃). Samples were taken at preset time intervals, and the ZEN concentration in the solution at different time points was determined by HPLC. The adsorption capacity q of the sample for ZEN at different time points was calculated according to formula (1).e (mg / g), the adsorption process was analyzed using kinetic models (including pseudo-first-order model (2), pseudo-second-order model (3) and intraparticle diffusion model (4)), as follows.

[0076] 2.2 Isotherms and Thermodynamic Experiments of ZEN Adsorption by SPYP 2

[0077] A certain amount of adsorbent (15 mg) was added to conical flasks containing 15 mL of ZEN solutions of different concentrations (2–20 μg / mL). The flasks were then sealed and placed in constant temperature shakers at different temperatures (298, 303, 308 K). After adsorption reached equilibrium, the ZEN concentration in the liquid was determined by HPLC. The adsorption capacity of polysaccharides for ZEN at different times was calculated according to formula (1), and the experimental adsorption data were fitted using Langmuir (5) and Freundlich (6) models.

[0078] Thermodynamic parameters such as ∆G, ∆H, and ∆S can be calculated using adsorption experimental data under different temperature conditions. The values ​​of ∆G, ∆H, and ∆S are calculated using formula (7-10).

[0079] Adsorption capacity:

[0080] ;

[0081] First-order dynamics:

[0082] ;

[0083] Second-order dynamics:

[0084] ;

[0085] Internal diffusion:

[0086] ;

[0087] Where, k1 (min -1 Kp (mg·(g·min)) and k2 (g / (mg⋅min)) are the adsorption kinetic rate constants; 1 / 2 ) -1 ) -1 ) is the rate constant of the intraparticle diffusion model; C (mg·g −1 ) is a constant in the particle diffusion model.

[0088] Adsorption isotherm

[0089] Langmuir

[0090] ;

[0091] Freundlich

[0092] ;

[0093] Where qmax is the maximum adsorption capacity, and a, b, and k are... f And n are model constants.

[0094] ;

[0095] Where Madsorbate (g / mol) is the molar mass of the adsorbate, Cadsorbate is the standard concentration of the adsorbate, and γ is the activity coefficient.

[0096] ;

[0097] ;

[0098] ;

[0099] Where T(K) is the temperature in Kelvin, R is Avogadro's constant, and Ka is the equilibrium constant.

[0100] 2.3 Investigation into the mechanism of polysaccharide adsorption of ZEN

[0101] XPS and FT-IR were used to determine the changes in functional groups of polysaccharides before and after ZEN adsorption.

[0102] 2.4 Molecular dynamics and density functional theory (DFT) calculations of polysaccharide adsorption of ZEN

[0103] Materials Studio (2022) software was used to construct the molecular configurations of polysaccharides and ZEN, respectively. The AC module was used to model amorphous molecular chains, obtaining a mixed aqueous solution system of polysaccharides and ZEN. The smartminimized method was used for optimization to obtain stable chain segments. Molecular dynamics simulations were performed using the forcite module, with the COMPASSII force field selected to describe the interactions between molecular chains. Snapshots of the system were obtained at different times after the molecules reacted over time. Andersen's and Berenden's functions were used to control temperature and pressure (set to 298.15 K, 101325 Pa), with a cutoff radius of 14.5 Å (angstroms) and a maximum duration of 200 ns.

[0104] To better explore potential interaction sites between molecules, we downloaded the structure of ZEN from the PubChem database and converted it to 3D format using the Open Babel program. Due to the relatively large molecular weight of polysaccharides and the presence of multi-scale structures within the particles, the entire polysaccharide structure system is difficult to simulate using DFT. Therefore, we chose to use computational models of monomeric polysaccharide molecules (glucose, mannose). These models were constructed using the website (http: / / www.glycosciences.de / / ). First, the geometric structures of the polysaccharide and ZEN monomeric molecules were optimized using the B3LYP / 6-31G(d) method. Subsequently, the electrostatic potential (ESP) distribution was plotted using the Multiwfn program. Random images of 100 complexes were generated using the Genmer program and optimized using a semi-empirical algorithm in the Molclus program. With ZPE and BSSE corrections, the initial complex structure was further optimized using the same DFT basis set to obtain the lowest energy. The Independent Gradient Model (IGM) method was employed in the Multiwfn software to analyze and visualize non-covalent interactions between molecules.

[0105] This invention obtains a highly efficient extraction method by screening different methods to assess the yield, purity, and adsorption characteristics of yeast polysaccharides, such as... Figure 1 As shown, different extraction methods for polysaccharides were compared to determine their impact on ZEN adsorption and to optimize the method. Hot water extraction, enzymatic extraction, and alkaline extraction were compared, and the results are as follows. Figure 3 As shown in Figure 2 (AB), the alkaline extraction method achieved the highest removal rate and qe value, at 89.64% and 3.49 mg / g, respectively. Furthermore, the extraction rate and purity of the polysaccharides were as follows: Figure 3 As shown in .2(C), the purity of all samples is around 76.00%, while the extraction rate of AE-P is the highest at 7.20%. Therefore, based on the extraction rate, alkaline extraction was chosen for the subsequent extraction of polysaccharides with ZEN adsorption function.

[0106] This invention obtains yeast polysaccharides through separation and purification. The dialyzed SPYP was further purified using a DEAE-52 cellulose column (2.6 × 25 cm). Figure 2 As shown, the polysaccharides were collected and eluted with 0, 0.1, 0.2, and 0.3 M NaCl solutions to obtain three fractions, named SPYP 1, SPYP 2, and SPYP 3, respectively. However, since the recovery rate of SPYP 3 during dialysis was not ideal, the structures of SPYP 1 and SPYP 2 polysaccharides were selected for structural analysis, with purities of 83.4% and 82.0%, respectively.

[0107] This invention characterizes the isolated and purified yeast polysaccharide to obtain its morphological features. The surface structure of the polysaccharide in the two separated components (SPYP 1 and SPYP 2) was observed using SEM, such as... Figure 3 As shown, SPYP 1 exhibits a spherical and network structure with an increased number of surface pores, likely due to extensive branching interactions within the polysaccharide. In contrast, SPYP 2 exhibits a rough, dense, spherical and network structure with fewer but larger surface pores, exhibiting an overall granular shape with irregular aggregation; this structure contributes to enhanced water solubility. Their morphological and conformational characteristics were observed using AFM, such as... Figure 4 As shown, SPYP 1 exhibits a rod-like structure with a height ranging from 6.62 to 9.39 nm and an average surface roughness (Ra) of 1.33 nm, displaying a certain degree of heterogeneity in its overall morphology. However, SPYP 2 shows lower aggregation and height compared to SPYP 1. This study found that both polysaccharides exhibit a certain degree of roughness, which is likely related to the interactions between their molecular chains and the resulting aggregation behavior. To clarify the microstructure of the single molecular chains of SPYP 1 and SPYP 2, higher resolution TEM was used for observation. Figure 5 As shown, SPYP 1 mainly consists of large, clustered particles with cross-linking between them, and a size of approximately 20 nm. In contrast, SPYP 2 exhibits significantly reduced aggregation, with the polysaccharide chains unfolding. Observations revealed that these polysaccharide chains are composed of relatively uniform spherical particles, ranging in size from 10 nm to 50 nm. Furthermore, the morphology of both polysaccharides closely resembles that exhibited by AFM.

[0108] This invention performs FT-IR analysis on the polysaccharides after ion separation and purification to obtain the major functional group composition of the polysaccharides, such as... Figure 6 As shown, both polysaccharides exhibit broad -OH stretching vibration peaks in the 3000–3600 cm⁻¹ range, similar to the structure of the previously studied crude polysaccharide. CH stretching vibration peaks are observed in the 2800–3000 cm⁻¹ range, with weak peaks near 2920 cm⁻¹ and 2879 cm⁻¹ attributed to the asymmetric and symmetric CH stretching vibrations of the methylene (-CH₂-) and methine (-CH-) groups, characteristic absorptions of the CH bonds on the sugar ring. SPYP₂ shows peaks in the 1500–1700 cm⁻¹ range. -1 It has a characteristic absorption peak at 1643 cm⁻¹. -1 The amide I bands on the left and right sides mainly originate from the stretching vibrations of C=O in protein or polypeptide components. Therefore, it can be inferred that SPYP 2 still contains protein impurities or mannose-protein complexes. Similarly, both polysaccharides exhibit the characteristic "fingerprint region" of sugar rings (1200–950 cm⁻¹). -1SPYP 2 was also found at 810 cm⁻¹ in FT-IR. -1 The relatively large peak values ​​of the mannose characteristic peaks around the left and right indicate that the mannose content of SPYP 2 polysaccharide is relatively high.

[0109] This invention screened the ability of two polysaccharides to adsorb ZEN and found that SPYP 2 had the best ZEN removal ability. Figure 7 A) FT-IR analysis revealed a high mannose peak content in SPYP 2, which may explain its good adsorption effect. Therefore, SPYP 2 underwent further gel column purification, resulting in a purity of 92.50% (A). Figure 7 B).

[0110] This invention determines the monosaccharide composition and molecular weight of SPYP2 after isolation and purification to obtain the basic composition of this polysaccharide. The results are as follows: Figure 8 (A) and Table 1 are shown. This polysaccharide is mainly composed of Man (58.11%), and also contains Glc (37.11%) and Gal (4.78%). Furthermore, the molecular weight distribution of SPYP 2 is as follows: Figure 8 As shown in (B–C) and Table 2, Mw is 134.110 kDa, Mn is 121.294 kDa, and the polydispersity index is 1.106, indicating that SPYP 2 has high homogeneity. Molecular configuration diagram ( Figure 8 D) shows that the slope value of SPYP 2 is 0.17 ± 0.01, indicating that the polysaccharide has a compact, uniform and spherical conformation in 0.1 M NaNO3 solution.

[0111] .

[0112] .

[0113] This invention determined the glycosidic bonds and molar percentages of sugar residues in SPYP 2 through methylation analysis. The results are summarized in Table 3. The relative contents of mannose, glucose, and galactose in SPYP 2 are 61.98%, 32.57%, and 5.46%, respectively, which are close to the monosaccharide composition results in Table 1, indicating that the methylation reaction of this polysaccharide is relatively complete. Furthermore, the dominant glycosidic bond types in SPYP 2 include t-Man(p)4-Man(p), 6-Man(p), and 3,6-Man(p), with percentages of 8.84%, 34.13%, 12.58%, and 6.43%, respectively.

[0114] .

[0115] This invention determines the molecular structure of the polysaccharide SPYP 2 by performing 1D and 2D NMR (COSY, HSQC, NOESY, DEPT-135, TOCSY, HMBC) measurements. The sample's proton NMR signal is mainly concentrated between δ 3.0–5.5 ppm. Multiple coupled signal peaks were identified in the anodic signal region of δ 4.3–5.4 ppm, indicating the presence of various sugar residues. The corresponding chemical shifts of the anodic hydrogens are δ 4.48, 4.67, 4.69, 4.79, 4.92, and 5.03 ppm, respectively. Non-anodic hydrogen signals are mainly concentrated in the δ 3.1–4.2 ppm region. Some signals, due to severe overlap, require further analysis using TOCSY spectroscopy (…). Figure 9 D) and COSY Figure 10 A) The H2–H6 chemical shifts of each sugar residue were assigned. Multiple signal peaks were identified in the anomeric carbon region of the sample, combined with... 13 C NMR spectrum and HSQC spectrum Figure 9 The cross-peaks in the anodic regions A and B identified the anodic signals present in the sample as: δ 4.79 / 96.67, 4.48 / 102.68, 4.92 / 97.68, 5.03 / 97.81, 4.67 / 100.07, and 4.69 / 102.48 ppm, which were denoted as sugar residues A, B, C, D, E, and F, respectively. Combined with DEPT-135 ( Figure 9 C) and 13C NMR ( Figure 9 B) The spectrum can yield signals for methylene (δ 60.52, 60.99, 65.51, 62.72, 65.57, 60.7 ppm).

[0116] The NMR signal assignment process for the major sugar residues is as follows: The anodic region signal of sugar residue A (δ 4.79 / 96.67ppm, H1 / C1) indicates that it is α-mannose. The COSY spectrum (…) Figure 10 A) The hydrogen signals were determined to be: H2 (4.06 ppm), H3 (3.78 ppm), H4 (3.78 ppm), H5 (3.45 ppm), and H6 (3.84, 3.68 ppm). HSQC spectrum ( Figure 10 B) The carbon signals assigned are: C1 (96.67 ppm), C2 (70.17 ppm), C3 (71.55 ppm), C4 (76.86 ppm), C5 (69.5 ppm), and C6 (60.52 ppm). C1 and C4 show significant low-field shifts, indicating substitution at the O-1 and O-4 positions. Based on methylation and literature results, this residue is inferred to be →4)-α-D-Manp-(1→).

[0117] The anodic signal of sugar residue B (δ 4.48 / 102.68 ppm, H1 / C1) indicates that it is β-glucose. COSY spectrum ( Figure 10 A) The hydrogen signals are assigned as follows: H2 (3.27 ppm), H3 (3.43 ppm), H4 (3.65 ppm), H5 (3.84 ppm), and H6 (3.87 ppm). HSQC spectrum ( Figure 10 B) The corresponding carbon signals are: C1 (102.68 ppm), C2 (73.15 ppm), C3 (75.95 ppm), C4 (73.38 ppm), C5 (70.08 ppm), and C6 (60.99 ppm). The low-field shifts of C1 and C3 suggest that the O-1 and O-3 positions are substituted. Based on methylation and literature data, it is speculated that the residue is →3)-β-D-Glcp-(1→).

[0118] The anomeric region signal of sugar residue C was δ 4.92 / 97.68 ppm (H1 / C1), suggesting it is α-mannose. COSY spectroscopy (…) Figure 10 A) The hydrogen signals were assigned sequentially as follows: H2 (3.52 ppm), H3 (3.65 ppm), H4 (3.86 ppm), H5 (3.82 ppm), and H6 (3.92, 3.67 ppm). Based on HSQC spectra ( Figure 10 B) Further carbon signal assignments are: C1 (97.68 ppm), C2 (71.38 ppm), C3 (65.06 ppm), C4 (70.11 ppm), C5 (70.3 ppm), and C6 (65.51 ppm). C1 and C6 show significant low-field shifts, indicating substitution at the O-1 and O-6 positions. Based on methylation and literature results, this residue is inferred to be →6)-α-D-Manp-(1→).

[0119] The anodic signal δ of sugar residue D (5.03 / 97.81 ppm, H1 / C1) indicates that residue D may be an α-configuration mannose residue, as shown in the COSY spectrum ( Figure 10 In A), the hydrogen signals of residue D were determined based on the cross-peak δ 5.03 / 3.73 ppm: H2 (3.73 ppm), H3 (3.59 ppm), H4 (3.77 ppm), H5 (3.9 ppm), H6 (3.63 ppm), thus allowing attribution to the chemical shifts of hydrogen atoms on the sugar ring. Then, the HSQC signal ( Figure 10B) The chemical shifts of C on the sugar ring are: C1 (97.81 ppm), C2 (67.82 ppm), C3 (74.87 ppm), C4 (69.77 ppm), C5 (70.31 ppm), and C6 (62.72 ppm). The chemical shift of C1 is shifted to a lower field, indicating that the residue has been substituted at the O-1 position of the sugar ring. Based on the methylation analysis results and literature reports, it is inferred that sugar residue D may be α-D-Manp-(1→).

[0120] The anodic signal δ of sugar residue E (4.67 / 100.07 ppm, H1 / C1) indicates that residue E may be a β-configuration mannose residue, as shown in the COSY spectrum ( Figure 10 In A), the H signals of residue E were determined based on the cross-peak δ 4.67 / 4.2 ppm: H2 (4.2 ppm), H3 (3.89 ppm), H4 (3.64 ppm), H5 (3.78 ppm), H6 (3.7 ppm), thus allowing attribution to the chemical shifts of hydrogen atoms on the sugar ring. Then, the HSQC signal ( Figure 10 B) Assigning the chemical shifts of C on the sugar ring, we know that C1 (100.07 ppm), C2 (67.45 ppm), C3 (78.82 ppm), C4 (64.87 ppm), C5 (71.80 ppm), and C6 (65.57 ppm). Among them, the chemical shifts of C1, C3, and C6 are shifted to the lower field, indicating that the residue has been substituted at the O-1, O-3, and O-6 positions of the sugar ring. Based on the methylation analysis results and literature reports, we infer that the sugar residue E may be →3,6)-β-D-Manp-(1→).

[0121] The anodic signal δ of sugar residue F, 4.69 / 102.48 ppm (H1 / C1), indicates that residue F may be a β-configuration galactose residue, as shown in the COSY spectrum ( Figure 10 In A), the H signals of residue F were determined based on the cross-peak δ 4.69 / 3.31 ppm: H2 (3.31 ppm), H3 (3.46 ppm), H4 (3.64 ppm), H5 (3.7 ppm), H6 (3.82 ppm), thus allowing attribution to the chemical shifts of hydrogen atoms on the sugar ring. Then, the HSQC signal ( Figure 10B) Attributing the chemical shifts of C on the sugar ring, the chemical shifts of residue F are as follows: C1 chemical shift δ 102.48 ppm, C2 chemical shift δ 69.66 ppm, C3 chemical shift δ 75.5 ppm, C4 chemical shift δ 71.37 ppm, C5 chemical shift δ 72.92 ppm, and C6 chemical shift δ 60.7 ppm. Among them, the chemical shifts of C1 and C3 shift to the lower field, indicating that the residue has been substituted at the O-1 and O-3 positions of the sugar ring. Based on the methylation analysis results and literature reports, it is inferred that sugar residue F may be →3)-β-D-Galp-(1→).

[0122] In summary, it is inferred that sugar residue A is →4)-α-D-Manp-(1→), sugar residue B is →3)-β-D-Glcp-(1→), sugar residue C is →6)-α-D-Manp-(1→), sugar residue D is α-D-Manp-(1→), sugar residue E is →3,6)-β-D-Manp-(1→), and sugar residue F is →3)-β-D-Galp-(1→. The assignments of these sugar residue shifts are shown in Table 4.

[0123] Based on each sugar residue in the sample 13 C and 1 The chemical shift of H, combined with HMBC and NOESY spectra ( Figure 9 (C, D) Analyze the structure and linkage mode of this polysaccharide. Based on HMBC spectra ( Figure 9 C), there is a cross-peak between H1 of sugar residue A and C3 of sugar residue E at δ 4.79 / 78.82 ppm; a cross-peak between C1 of sugar residue A and H3 of sugar residue E at δ 96.67 / 3.89 ppm; a cross-peak between C1 of sugar residue B and H3 of sugar residue B at δ 102.68 / 3.43 ppm; a cross-peak between C1 of sugar residue B and H3 of sugar residue F at δ 102.68 / 3.46 ppm; a cross-peak between H1 of sugar residue C and C6 of sugar residue C at δ 4.92 / 65.51 ppm; a cross-peak between C1 of sugar residue C and H6 of sugar residue C at δ 97.68 / 3.92 ppm; a cross-peak between H1 of sugar residue E and C4 of sugar residue A at δ 4.67 / 76.86 ppm; and a cross-peak between C1 of sugar residue E and H4 of sugar residue A at δ 102.68 / 3.43 ppm. 100.07 / 3.78 ppm, with a cross peak δ 102.48 / 3.43 ppm between C1 of sugar residue F and H3 of sugar residue B. According to NOESY spectra ( Figure 9(D) There is a cross-peak between H1 and H4 of sugar residue A (δ 4.79 / 3.78 ppm), a cross-peak between H1 and H3 of sugar residue A (δ 4.79 / 3.43 ppm), a cross-peak between H1 and H6 of sugar residue A (δ 4.79 / 3.67 ppm), a cross-peak between H1 and H4 of sugar residue B (δ 4.48 / 3.78 ppm), a cross-peak between H1 and H3 of sugar residue B (δ 4.48 / 3.43 ppm), a cross-peak between H1 and H3 of sugar residue B (δ 4.48 / 3.46 ppm), and a cross-peak between H1 and H4 of sugar residue C (δ 4.92 / 3.78 ppm). The H1 of sugar residue C and the H6 of sugar residue C have a cross peak at δ 4.92 / 3.92 (3.67) ppm, and the H1 of sugar residue D and the H6 of sugar residue E have a cross peak at δ 5.03 / 3.7 ppm.

[0124] Therefore, based on the analysis of one-dimensional and two-dimensional NMR information and methylation results, it is inferred that this polysaccharide is mainly composed of interconnected main chains such as →4)-α-D-Manp-(1→,→3)-β-D-Glcp-(1→,→6)-α-D-Manp-(1→,→3,6)-β-D-Manp-(1→ and→3)-β-D-Galp-(1→), with side chains consisting of α-D-Manp-(1→) linked to the O-6 positions of the sugar residues →3,6)-β-D-Manp-(1→). Its specific chemical formula and structural formula are as follows: Figure 11 (A, B).

[0125] This invention uses adsorption kinetics to clarify the role of SPYP2 in adsorbing ZEN. For example... Figure 12 As shown in (A–B), the kinetic results indicate that the pseudo-second-order kinetic curve fits the ZEN concentration better than the pseudo-first-order kinetic curve. However, the correlation coefficients of the pseudo-second-order kinetic model, with R² values ​​of 0.97, 0.92, 0.81, 0.92, and 0.97 respectively (Table 5), are relatively low. Furthermore, the qe predicted by the pseudo-second-order kinetic model are 2.36, 5.19, 8.76, 13.33, and 16.79 mg / g, which are closer to the measured values ​​than those predicted by the pseudo-first-order kinetic equation. Based on these results, the pseudo-second-order kinetics can better explain the adsorption of ZEN by SPYP2, suggesting that the adsorption process is mainly chemisorption.

[0126]

[0127] This invention analyzes the adsorption behavior using Langmuir and Freundlich models, determining the adsorption saturation capacity of the adsorbent under equilibrium conditions. Figure 12As can be seen from (C–D) and Table 6, the adsorption capacity q decreases with increasing temperature. e Increase. R-squared of the Langmuir model fit. 2 The values ​​of 0.99, 0.96, and 0.93 are all higher than those of the Freundlich model, demonstrating that it is more suitable for explaining the adsorption reaction of ZEN by SPYP2, indicating that monolayer adsorption occurred at the binding sites on the SPYP2 surface. Although various biosorbents for ZEN adsorption have been reported, the adsorption of ZEN by SPYP2 in *Syndrome aureomarginata* has not yet been studied. The adsorption thermodynamic results are shown in Table 7. The changes in adsorption free energy, heat, and entropy were calculated. At different temperatures, ΔG < 0 indicates that the adsorption of ZEN is spontaneous.

[0128] .

[0129] .

[0130] This invention clarifies the main functional groups of polysaccharides acting on ZEN using FT-IR and XPS. For example... Figure 13 As shown in (A), the absorption peak is at 3385 cm⁻¹ before and after adsorption. -1 The change in the -OH group indicates that -OH binds to ZEN via hydrogen bonds; similar results were observed in the studies by Zheng et al. Furthermore, a 2942 cm⁻¹ region was also found. -1 and 1031 cm -1 CH and CO at 1645cm -1 The C=O peaks at the adsorption sites also changed, indicating their involvement in ZEN adsorption. In addition to FT-IR analysis of the polysaccharides before and after adsorption, this study also used XPS to further investigate the binding mechanism. Figure 13 B–H), the C1s spectrum of SPYP 2 before ZEN adsorption is as follows Figure 13 As shown in (C and F), the binding energies of 285.95, 284.91, and 283.55 eV belong to the OC=O, COC, and CC groups, respectively. After ZEN adsorption, these energies become 288.05, 286.43, and 284.80 eV, respectively, proving that all of the above groups participated in ZEN adsorption. The O1s spectrum of SPYP 2 before adsorption is shown below. Figure 13As shown in (D, G), the binding energies of 529.54, 531.16, and 532.46 eV belong to the C=O, -OH, and OCO groups, respectively. After adsorption, the binding energies shift to higher levels at 531.16, 532.70, and 533.95 eV, attributed to the interaction between lone heteropairs and the molecule. Analysis of the C1s and O1s energy spectrum peaks confirms that SPYP 2 successfully adsorbed ZEN. This analysis also reveals the contribution of carbon and oxygen-related functional groups to ZEN adsorption.

[0131] This invention explores the self-assembly dynamics of ZEN and SPYP through MD simulation. Figure 14 (A) shows a snapshot of the adsorption and aggregation process of SPYP2 and ZEN in an aquatic environment. Initially, SPYP2 and ZEN molecules are randomly dispersed within the simulation chamber. As the simulation progresses, these molecules begin to coalesce, and by 200 ns, stable clusters of SPYP2 and ZEN appear. Solvent-accessible surface area (SASA) (e.g.) Figure 14 The results of examination (B) showed that the SASA value decreased rapidly during adsorption and stabilized at around 150 ns. The study indicates that the pattern of intermolecular interaction energies closely follows the SASA trend. Figure 14 C). As the intermolecular distance decreases, the interaction energy gradually increases. The dominant forces driving these interactions are van der Waals forces and electrostatic interactions, providing the core impetus for the initial approach and preliminary stabilization of the complex. After reaching equilibrium, the average electrostatic interaction and van der Waals force were calculated to be -590.83 kJ / mol and -1149.55 kJ / mol, respectively, indicating a strong mutual attraction between molecules. Furthermore, since the C=O and -OH groups on the ZEN molecule are potential hydrogen bond acceptors and donors, the formation of hydrogen bonds within the system was evaluated, with results as follows: Figure 14 As shown in (D), the number of hydrogen bonds formed between SPYP 2 and ZEN is parallel to the trend of electrostatic interaction energy, indicating that the intermolecular bonding is not a simple, non-specific hydrophobic stacking or random collision, but rather the entire complex system is stabilized by a polar interaction bonding mode. Furthermore, the formation of hydrogen bonds not only further enhances the binding affinity of the complex, but more importantly, it determines the specific bonding mode and orientation of the complex, improving the specificity of the bonding. Therefore, it can be inferred that the self-assembly behavior of SPYP 2 and ZEN is mainly driven by van der Waals forces, hydrogen bonds, and intermolecular electrostatic interactions, prompting the SPYP 2 and ZEN complex to form a stable conformation in solution.

[0132] This invention clarifies the main forces driving the adsorption of zearalenone by polysaccharides through DFT calculations. Since the structure of SPYP 2 is primarily composed of mannose and glucose, using it as the basic unit allows for a better understanding of the adsorption mechanism. The electrostatic potential distribution (ESP) composite structure after ZEN is shown below. Figure 15 As shown in (A–C), the ESP distribution of the free monomer can be represented by a scale, where deeper red indicates a significantly increased electron cloud density and a greater amount of negative charge in the corresponding region, while deeper blue indicates a lower electron cloud density and a greater amount of positive charge. The maximum and minimum ESP sites appear near the hydroxyl and carboxyl groups of the polysaccharide and ZEN, which may be their main molecular interaction sites. Furthermore, the adsorption energies (Eads) of mannose + ZEN and glucose + ZEN are -37.247 and -33.697 kcal / mol, respectively. FMO assesses the binding reactivity and stability between the polysaccharide and ZEN; a higher absolute value of the highest occupied molecular orbital (HOMO) indicates a stronger electron donor capacity. Conversely, a lower absolute value of the lowest unoccupied molecular orbital (LUMO) indicates a stronger electron acceptor capacity. Moreover, the HOMO-LUMO gap indicates difficulty in electron transitions and molecular configuration stability. Figure 15 As shown in (D), mannose + ZEN (3.15630558 eV) and glucose + ZEN (3.12882348 eV) indicate that mannose has a stronger adsorption capacity for ZEN, which is consistent with the results of Eads analysis.

[0133] This invention also included IGM analysis to reveal the weak interactions between mannose and glucose and ZEN. For example... Figure 15 As shown in (E), the blue pattern represents strong non-covalent interactions, the green pattern represents non-covalent interactions with van der Waals forces, and the red pattern represents strong spatial resistance. It can be seen that the polysaccharide and ZEN complex system exhibit vertical bar-shaped "peaks" in all three colors, indicating the presence of non-covalent interactions in the complex system, such as hydrogen bonds and van der Waals forces, further verifying the results of molecular dynamics. Figure 15 (F) represents the non-covalent interaction between the polysaccharide and the ZEN complex system in real space. The hydrogen bonds between them (blue figure) are mainly distributed between the hydroxyl groups of ZEN and the polysaccharide molecules, which are the main interaction sites for the structural stability of this complex system.

Claims

1. A near-rose-colored *Syngonium spp.* polysaccharide SPYP 2, prepared by the following method: 1) Inoculate nearly 1000 rosé yeast into liquid culture medium, remove the supernatant, wash with sterile water, and inactivate; 2) Crude polysaccharides of *Syngonium spp.* near-rose color were extracted using the alkaline extraction method; 3) Remove protein, then dialyze in a dialysis bag; 4) Isolation and purification of polysaccharides: Redissolve in pure water, centrifuge and collect the supernatant, purify with a weak anion exchange column; use an AKTA purification system for separation and purification at a flow rate of 4 mL / min; elute with pure water to obtain SPYP 1, elute with 0.1 M NaCl, and dialyze to obtain SPYP 2.

2. The near-rose-colored *Syngonium spp.* polysaccharide SPYP 2 according to claim 1, characterized in that: Step 4) describes a weak anion exchange column as DEAE seplife FF.

3. The near-rose-colored *Syngonium spp.* polysaccharide SPYP 2 according to claim 2, characterized in that: Step 2) describes the alkaline extraction method, in which the inactivated *Synthia sacchariformis* was dissolved in 2 M NaOH solution and stirred at room temperature for 2 h. After centrifugation, the supernatant was obtained, and anhydrous ethanol was added at a volume ratio of 1:4 to precipitate the supernatant overnight. After centrifugation, the precipitate was redissolved, and the protein was removed using the Sevag method. The supernatant was then freeze-dried to obtain crude yeast polysaccharide.

4. The near-rose-colored *Syngonium spp.* polysaccharide SPYP 2 according to claim 3, characterized in that: Step 3) describes the removal of protein using the Sevag method, followed by dialysis in an 8000–14000 kDa dialysis bag.

5. The application of the near-rose-colored Cyclospora polysaccharide SPYP 2 as described in claim 1 in the adsorption of zearalenone.

6. The application according to claim 5, characterized in that: It involves the adsorption of zearalenone in the presence of glucose or mannose.

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