Process for synergistically extracting rape pollen polysaccharide through compound enzyme

Abstract

The invention relates to the technical field of plant polysaccharide extraction, in particular to a process for synergistically extracting rape pollen polysaccharide by a compound enzyme, which comprises the following steps: crushing rape pollen to 200-300 meshes by pulse airflow, performing microwave-ultrasonic degreasing by using a 85% ethanol solution, mixing with an acetic acid-sodium acetate buffer solution with the pH value of 4.8, and performing ultrasonic extraction to obtain a crude extract; the method comprises the following steps: adding a compound enzyme system which is immobilized by Fe3O4 (at) SiO2 nanoparticles by cellulase, pectinase, protease and beta-glucosidase according to a mass ratio of 2: 1: 1: 0.5, carrying out enzymolysis for 4 hours in a 50 DEG C constant-temperature water bath and a 15V / cm pulsed electric field, carrying out enzyme deactivation on enzymatic hydrolysate, carrying out gradient ethanol precipitation, carrying out ultrafiltration deproteinization, and carrying out vacuum pulsation freeze drying to obtain the polysaccharide. According to the method disclosed by the invention, the cell wall breakage rate and the enzymolysis efficiency are improved through pulsed airflow crushing, magnetic nano immobilized enzyme and pulsed electric field assistance, the polysaccharide yield reaches 7.32%, the purity is 91%, the beta-glucosidic bond proportion and the antioxidant activity are remarkably improved, the enzyme can be recycled and reused, the drying time is shortened, and efficient and low-consumption extraction is realized.

Description

Technical Field

[0001] The invention relates to the technical field of plant polysaccharide extraction, in particular to a process for extracting rapeseed pollen polysaccharides in a coordinated manner using a complex enzyme. Background Art

[0002] Rapeseed pollen is a natural resource rich in active ingredients such as polysaccharides, proteins, and flavonoids. Its polysaccharide components have attracted much attention due to their biological activities such as immunomodulation, antioxidant, and anti-tumor. However, the traditional rapeseed pollen polysaccharide extraction process has significant technical bottlenecks. The early hot water extraction method relied on high temperature to destroy the cell wall, which not only led to the denaturation of the polysaccharide structure, but also the yield was generally less than 4%, which was difficult to meet the needs of industrial production. Although the subsequent enzymatic hydrolysis method uses cellulase, pectinase, etc. to destroy the cell wall, its free enzyme system has the defects of low catalytic efficiency and a single enzyme-substrate action site. Especially when faced with the complex cell wall structure of rapeseed pollen, the polysaccharide release is insufficient and the enzyme cannot be recycled and reused, resulting in high production costs.

[0003] With the increasing requirements for natural product extraction efficiency and environmental protection, although the existing methods such as ultrasound-assisted and microwave-assisted can enhance mass transfer, the destruction of cell walls by simple physical means lacks selectivity, is often accompanied by degradation of effective ingredients, and significantly increases energy consumption costs. For example, although traditional ultrasonic extraction can shorten the extraction time, the purity of polysaccharides is easily interfered by impurities such as lipids and proteins, and the subsequent purification process is complicated. In addition, the existing complex enzyme systems are mostly limited to the simple combination of cellulase and pectinase, and are not designed for the synergistic degradation of proteins and glycosidic bonds in the cell wall of rapeseed pollen, resulting in a low ratio of β-glycosidic bonds and difficulty in fully exerting the biological activity of polysaccharides.

[0004] In current industrial production, rapeseed pollen polysaccharide extraction faces multiple challenges: low yield, poor purity, weak activity, and high cost. There is an urgent need to develop an extraction process that combines efficient cell wall disruption, selective enzymatic hydrolysis, low energy consumption, and recyclability to overcome the bottlenecks of traditional technologies in polysaccharide yield, structure preservation, and industrial application. Summary of the Invention

[0005] (1) Technical problems solved In view of the shortcomings of the existing technology, the present invention provides a process for extracting rapeseed pollen polysaccharides in a coordinated manner using a complex enzyme.

[0006] (2) Technical solution A process for extracting rapeseed pollen polysaccharides using a complex enzyme, comprising the following steps: S1. Rapeseed pollen was pulverized to 200-300 mesh using pulsed airflow. During the pulverization process, nitrogen gas at -5°C was introduced to control the temperature to ≤30°C. Microwave-ultrasound synergistic degreasing was then performed using an 85% ethanol solution at a microwave power of 400W and an ultrasonic frequency of 45kHz for 25 minutes. The precipitate was collected after centrifugation. S2. The precipitate was mixed with an acetic acid-sodium acetate buffer solution at pH 4.8, and a magnetic nano-immobilized complex enzyme system was added. The enzyme system consisted of cellulase, pectinase, protease, and β-glucosidase immobilized by Fe3O4@SiO2 nanoparticles. The enzyme addition amount was 2% of the pollen mass. The enzyme was hydrolyzed in a 50°C constant temperature water bath combined with a pulsed electric field. During the enzymatic hydrolysis process, 0.1 M acetic acid was automatically added dropwise by an online pH sensor to maintain the stability of the system. S3. After the enzyme was inactivated at 85°C, the hydrolyzate was subjected to a gradient ethanol precipitation method: 2 volumes of 95% ethanol were added and allowed to stand for 8 hours. After centrifugation, the precipitate was precipitated again with 4 volumes of 70% ethanol, allowed to stand at 4°C for 12 hours, and the precipitate was collected by centrifugation. S4. The precipitate was dynamically deproteinized using a hollow fiber ultrafiltration membrane at an operating pressure of 0.15 MPa and a flow rate of 20 L / h. The precipitate was then pre-frozen at -40°C for 2 hours and vacuum pulse freeze-dried for 30 hours to obtain rapeseed pollen polysaccharide.

[0007] Preferably, during the microwave-ultrasound synergistic degreasing, 0.5% wt citric acid is added to the ethanol solution as a penetration enhancer.

[0008] Preferably, the magnetic nano-immobilized complex enzyme system realizes enzyme recovery and reuse through an external magnet with a magnetic field strength of 0.8T.

[0009] Preferably, during the pulsed electric field enzymolysis process, a pulsed electric field is applied for 5 minutes at intervals of 30 minutes.

[0010] Preferably, in the gradient ethanol precipitation method, the first precipitation temperature is controlled at 25°C, and the second precipitation temperature is lowered to 4°C.

[0011] Preferably, when the hollow fiber ultrafiltration membrane is used for deproteinization, a 0.05%wt chitosan solution is introduced as a filter aid.

[0012] Preferably, the pulsating pressure range of the vacuum pulsating freeze drying is 5-10 Pa, the drying time is shortened by 20% compared with traditional freeze drying, and the polysaccharide activity retention rate is ≥95%.

[0013] Preferably, a near-infrared spectroscopy online monitoring system is integrated into the process to measure the polysaccharide concentration in the enzymatic hydrolysate in real time, and the enzyme inactivation program is automatically triggered when the concentration reaches 12 mg / mL.

[0014] Preferably, the pulse airflow pulverization pressure is 0.6 MPa, and the pulverization times are 3 times.

[0015] Preferably, the yield of rapeseed pollen polysaccharide extracted by this process is ≥7.1%, the purity is ≥91%, and the β-glycosidic bond ratio is increased by 35% compared with the traditional enzymatic hydrolysis method.

[0016] (3) Beneficial technical effects Compared with the existing technology, the beneficial effects of the present invention are: 1. The combination of pulsed airflow pulverization and microwave-ultrasound synergistic degreasing technology achieves a pollen cell wall fragmentation rate of over 95%, improving degreasing efficiency and creating sufficient contact conditions for subsequent enzymatic hydrolysis. Compared to traditional pulverization and degreasing processes, this process increases polysaccharide yield, reaching a purity of over 92%. The magnetic nano-immobilized complex enzyme system, immobilized by Fe3O4@SiO2 nanoparticles, retains over 80% enzyme activity even after five reuses, reducing costs compared to free enzyme systems. Furthermore, the pulsed electric field-assisted enzymatic hydrolysis increases cell wall porosity, increases the proportion of β-glycosidic bonds, and significantly enhances the antioxidant activity of polysaccharides, with a DPPH scavenging rate of over 86%.

[0017] 2. The integration of gradient ethanol precipitation and vacuum pulse freeze-drying solves the problems of polysaccharide loss and activity degradation during traditional precipitation. Compared to single-concentration precipitation, this method improves yield, shortens drying time, and maintains over 95% polysaccharide activity. This process dynamically regulates the enzymatic hydrolysis process through online near-infrared spectroscopy monitoring. When the polysaccharide concentration reaches 12 mg / mL, the enzyme inactivation process is automatically triggered, ensuring process stability and repeatability. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 This is a process flow chart of the invention's proposed method of extracting rapeseed pollen polysaccharides using a composite enzyme. Figure 2 This is a comparison chart of polysaccharide yield and purity between the examples and comparative examples; Figure 3 DPPH clearance rate comparison chart of Example and Comparative Example; Figure 4 It is a radar comparison chart made by unifying the dimensions of the comparative data of the embodiment and the comparative example. DETAILED DESCRIPTION

[0019] according to Figures 1 to 4 , the specific implementation of the present invention is as follows: 1. Process principle and system composition This process uses pulsed airflow to disrupt rapeseed pollen cell walls, combined with microwave-ultrasound synergistic degreasing to remove lipid interference. A magnetic nanoparticle-immobilized complex enzyme system (cellulase, pectinase, protease, and β-glucosidase) is then used to release polysaccharides through enzymatic hydrolysis assisted by a pulsed electric field. This process then proceeds through gradient ethanol precipitation, ultrafiltration to deproteinize, and vacuum pulse freeze-drying to achieve high-purity rapeseed pollen polysaccharides. The Fe3O4@SiO2 nanoparticle-immobilized enzyme is recovered using a magnetic field.

[0020] 2. Example 1: Standard process parameter extraction S1. Raw material pretreatment 100g of rapeseed pollen at peak flowering stage was placed in a pulsed airflow mill at a pulverization pressure of 0.6 MPa, with nitrogen gas flowing through at -5°C for temperature control. The milled pollen was pulverized three times to a 200-mesh size, achieving a cell wall fragmentation rate of 95%. 1000mL of 85% ethanol solution and 0.5% wt citric acid were added to the pulverized pollen. The milled pollen was then defatted using a microwave at 400W and an ultrasonic frequency of 45kHz for 25 minutes. The milled pollen was then incubated in a 45°C waterbath and centrifuged at 4000 rpm for 15 minutes. The supernatant was discarded and the pellet was collected.

[0021] S2. Magnetic Nanoparticle Immobilization Enzyme Hydrolysis Prepare 1200 mL of acetic acid-sodium acetate buffer at pH 4.8 and add the precipitate to the buffer. Prepare a Fe₃O₄@SiO₂ nanoparticle-immobilized enzyme system: 2 g of cellulase, 1 g of pectinase, 1 g of protease, and 0.5 g of β-glucosidase were mixed with 100 mg of Fe₃O₄@SiO₂ nanoparticles in a mass ratio of 2:1:1:0.5. The mixture was then immobilized using a crosslinker. The immobilized enzyme system was added to the reaction system at an enzyme concentration of 2% of the pollen mass. In a 50°C water bath, a pulsed electric field with an electric field strength of 15 V / cm and a pulse frequency of 10 Hz was applied. Enzymatic hydrolysis was carried out for 4 hours with stirring at 200 rpm. An online pH sensor was used for real-time monitoring, and 0.1 M acetic acid was added dropwise to maintain a stable pH.

[0022] S3. Gradient Ethanol Precipitation The enzymatic hydrolysate was inactivated at 85°C for 18 minutes and centrifuged at 4000 rpm for 15 minutes to obtain the supernatant. Two volumes of 95% ethanol were added to the supernatant, and the mixture was allowed to stand at 25°C for 8 hours. The precipitate was then collected by centrifugation. Four volumes of 70% ethanol were then added to the precipitate, and the mixture was allowed to stand at 4°C for 12 hours. The polysaccharide precipitate was then collected by centrifugation at 6000 rpm for 10 minutes.

[0023] S4. Purification and Drying The precipitate was deproteinized by passing it through a hollow fiber ultrafiltration membrane with a molecular weight cutoff of 5000 Da at an operating pressure of 0.15 MPa and a flow rate of 20 L / h. A 0.05% wt chitosan solution was simultaneously introduced. The ultrafiltration solution was pre-frozen at -40°C for 2 hours and then dried in a vacuum pulsation freeze dryer at a vacuum of 5 Pa and a pulsation frequency of 0.5 Hz for 30 hours to yield 7.32 g of rapeseed pollen polysaccharide.

[0024] III. Example 2: Pulsed Electric Field Parameter Optimization S1. Raw material pretreatment 100g of rapeseed pollen at peak flowering stage was placed in a pulsed airflow mill at a pulverization pressure of 0.6 MPa, with nitrogen gas flowing through at -5°C for temperature control. The milled pollen was pulverized three times to a 200-mesh size, achieving a cell wall fragmentation rate of 95%. 1000mL of 85% ethanol solution and 0.5% wt citric acid were added to the pulverized pollen. The milled pollen was then defatted using a microwave at 400W and an ultrasonic frequency of 45kHz for 25 minutes. The milled pollen was then incubated in a 45°C waterbath and centrifuged at 4000 rpm for 15 minutes. The supernatant was discarded and the pellet was collected.

[0025] S2. Magnetic Nanoparticle Immobilization Enzyme Hydrolysis Prepare 1200 mL of acetic acid-sodium acetate buffer at pH 4.8 and add the precipitate to the buffer. Prepare the Fe₃O₄@SiO₂ nanoparticle-immobilized complex enzyme system: 2 g of cellulase, 1 g of pectinase, 1 g of protease, and 0.5 g of β-glucosidase were weighed and mixed with 100 mg of Fe₃O₄@SiO₂ nanoparticles in a mass ratio of cellulase:pectinase:protease:β-glucosidase = 2:1:1:0.5. The enzymes were then immobilized using a crosslinker. The immobilized enzyme system was added to the reaction system at an enzyme concentration of 2% of the pollen mass. In a 50°C water bath, a pulsed electric field with an electric field strength of 20 V / cm and a pulse frequency of 15 Hz was applied for 5 minutes at intervals of 20 minutes. Enzymatic digestion was carried out for 4 hours with stirring at 200 rpm. An online pH sensor was used for real-time monitoring, and 0.1 M acetic acid was added dropwise to maintain a stable pH.

[0026] S3. Gradient Ethanol Precipitation The enzymatic hydrolysate was inactivated at 85°C for 18 minutes and centrifuged at 4000 rpm for 15 minutes to obtain the supernatant. Two volumes of 95% ethanol were added to the supernatant, and the mixture was allowed to stand at 25°C for 8 hours. The precipitate was then collected by centrifugation. Four volumes of 70% ethanol were then added to the precipitate, and the mixture was allowed to stand at 4°C for 12 hours. The polysaccharide precipitate was then collected by centrifugation at 6000 rpm for 10 minutes.

[0027] S4. Purification and Drying The precipitate was deproteinized by passing it through a hollow fiber ultrafiltration membrane with a molecular weight cutoff of 5000 Da at an operating pressure of 0.15 MPa and a flow rate of 20 L / h. A 0.05% wt chitosan solution was simultaneously introduced. The ultrafiltration solution was pre-frozen at -40°C for 2 hours and then dried in a vacuum pulsation freeze dryer at a vacuum of 8 Pa and a pulsation frequency of 0.5 Hz for 28 hours to yield 7.51 g of rapeseed pollen polysaccharide.

[0028] Example 3: Adjustment of the ratio of complex enzymes S1. Raw material pretreatment 100g of rapeseed pollen at peak flowering stage was placed in a pulsed airflow mill at a pulverization pressure of 0.6 MPa, with nitrogen gas flowing through at -5°C for temperature control. The milled pollen was pulverized three times to a 200-mesh size, achieving a cell wall fragmentation rate of 95%. 1000mL of 85% ethanol solution and 0.5% wt citric acid were added to the pulverized pollen. The milled pollen was then defatted using a microwave at 400W and an ultrasonic frequency of 45kHz for 25 minutes. The milled pollen was then incubated in a 45°C waterbath and centrifuged at 4000 rpm for 15 minutes. The supernatant was discarded and the pellet was collected.

[0029] S2. Magnetic Nanoparticle Immobilization Enzyme Hydrolysis Prepare 1200 mL of acetic acid-sodium acetate buffer at pH 4.8 and add the precipitate to the buffer. Prepare a Fe₃O₄@SiO₂ nanoparticle-immobilized enzyme system: 1.5 g of cellulase, 1 g of pectinase, 1 g of protease, and 0.5 g of β-glucosidase were mixed with 100 mg of Fe₃O₄@SiO₂ nanoparticles in a mass ratio of 1.5:1:1:0.5. The mixture was then immobilized using a crosslinker. The immobilized enzyme system was added to the reaction system at an enzyme concentration of 2% of the pollen mass. In a 50°C water bath, a pulsed electric field with an electric field strength of 15 V / cm and a pulse frequency of 10 Hz was applied. Enzymatic hydrolysis was carried out for 4 hours with stirring at 200 rpm. An online pH sensor was used for real-time monitoring, and 0.1 M acetic acid was added dropwise to maintain a stable pH.

[0030] S3. Gradient Ethanol Precipitation The enzymatic hydrolysate was inactivated at 85°C for 18 minutes and centrifuged at 4000 rpm for 15 minutes to obtain the supernatant. Two volumes of 95% ethanol were added to the supernatant, and the mixture was allowed to stand at 25°C for 8 hours. The precipitate was then collected by centrifugation. Four volumes of 70% ethanol were then added to the precipitate, and the mixture was allowed to stand at 4°C for 12 hours. The polysaccharide precipitate was then collected by centrifugation at 6000 rpm for 10 minutes.

[0031] S4. Purification and Drying The precipitate was deproteinized by passing it through a hollow fiber ultrafiltration membrane with a molecular weight cutoff of 5000 Da at an operating pressure of 0.15 MPa and a flow rate of 20 L / h. A 0.05% wt chitosan solution was simultaneously introduced. The ultrafiltration solution was pre-frozen at -40°C for 2 hours and then dried in a vacuum pulsation freeze dryer at a vacuum of 5 Pa and a pulsation frequency of 0.5 Hz for 30 hours to yield 7.15 g of rapeseed pollen polysaccharide.

[0032] 5. Comparative Example: Traditional Hot Water Extraction Method S1. Preprocessing 100 g of rapeseed pollen was crushed to 80 mesh with a common grinder, defatted with 85% ethanol solution at room temperature for 30 minutes under stirring, and the precipitate was collected by centrifugation.

[0033] S2. Hot water extraction The precipitate was mixed with water at a solid-liquid ratio of 1:20, extracted in a 90°C water bath for 3 hours, and the supernatant was filtered.

[0034] S3. Precipitation and Purification The supernatant was directly added with 4 times the volume of 95% ethanol for precipitation, allowed to stand for 24 hours, centrifuged and deproteinized 5 times by Sevag method, dialyzed for 48 hours, and 3.21 g of polysaccharide was obtained by ordinary freeze-drying.

[0035] The effects of the embodiment and the comparative example are compared in the following table: Table 1 Group Example 1 Example 2 Example 3 Comparative Example Polysaccharide yield (%) 7.32 7.51 7.15 3.21 purity(%) 92.5 93.2 91.8 68.3 β-glycosidic bond ratio (%) 48.7 51.2 47.3 32.5 DPPH clearance rate (%) 86.3 88.5 85.1 52.1 Enzyme activity retention rate (after 5 reuses) 82% 80% 81% 0 The yields of Examples 1-3 were 128%-134% higher than those of the control group, indicating that the composite enzyme synergized with the pulsed electric field significantly improved the efficiency of polysaccharide release. In Example 2, by optimizing the pulsed electric field parameters, the β-glycosidic bond ratio was increased to 51.2%, confirming the promoting effect of electric field intensity and frequency on cell wall porosity. After 5 reuses, the enzyme activity retention rate of the magnetic nano-immobilized enzyme was ≥80%, which was significantly improved compared to the free enzyme, reducing costs. Vacuum pulse freeze drying shortened the time by 20% compared to traditional drying, and the polysaccharide activity retention rate reached 95%, while the DPPH clearance rate of the control group was only 52.1%, verifying the protective effect of the process on the active ingredients.

[0036] The response comparison of electric field intensity and pulse frequency to gain is shown in the following table: Table 2 Electric field strength (V / cm) Pulse frequency (Hz) Polysaccharide yield (%) 15 10 7.32 15 15 7.45 20 10 7.48 20 15 7.51 The pulsed electric field parameters significantly affected the polysaccharide yield. When the electric field intensity was 20 V / cm and the pulse frequency was 15 Hz, the yield reached a maximum of 7.51%, an increase of 2.6% compared with the basic parameters (15 V / cm, 10 Hz), indicating that there is an optimal range for the synergistic effect of the electric field.

[0037] While embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions, and variations may be made to these embodiments without departing from the principles and spirit of the invention, and that the scope of the invention is defined by the appended claims and their equivalents.

Claims

1. A process for extracting rapeseed pollen polysaccharides by using a complex enzyme, characterized in that: The following steps are involved: S1. Rapeseed pollen was pulverized to 200-300 mesh using pulsed airflow. During the pulverization process, nitrogen gas at -5°C was introduced to control the temperature to ≤30°C. Microwave-ultrasound synergistic degreasing was then performed using an 85% ethanol solution at a microwave power of 400W and an ultrasonic frequency of 45kHz for 25 minutes. The precipitate was collected after centrifugation. S2. The precipitate was mixed with an acetic acid-sodium acetate buffer solution at pH 4.8, and a magnetic nano-immobilized complex enzyme system was added. The enzyme system consisted of cellulase, pectinase, protease, and β-glucosidase immobilized by Fe3O4@SiO2 nanoparticles. The enzyme addition amount was 2% of the pollen mass. The enzyme was hydrolyzed in a 50°C constant temperature water bath combined with a pulsed electric field. During the enzymatic hydrolysis process, 0.1 M acetic acid was automatically added dropwise by an online pH sensor to maintain the stability of the system. S3. After the enzyme was inactivated at 85°C, the hydrolyzate was subjected to a gradient ethanol precipitation method: 2 volumes of 95% ethanol were added and allowed to stand for 8 hours. After centrifugation, the precipitate was precipitated again with 4 volumes of 70% ethanol, allowed to stand at 4°C for 12 hours, and the precipitate was collected by centrifugation. S4. The precipitate was dynamically deproteinized using a hollow fiber ultrafiltration membrane at an operating pressure of 0.15 MPa and a flow rate of 20 L / h. The precipitate was then pre-frozen at -40°C for 2 hours and vacuum pulse freeze-dried for 30 hours to obtain rapeseed pollen polysaccharide.

2. The process for extracting rapeseed pollen polysaccharides by complex enzyme synergistically according to claim 1, characterized in that: During the microwave-ultrasound synergistic degreasing, 0.5% wt citric acid is added to the ethanol solution as a penetration enhancer.

3. The process for extracting rapeseed pollen polysaccharides by complex enzyme synergistically according to claim 1, characterized in that: The magnetic nano-immobilized complex enzyme system realizes enzyme recovery and reuse through an external magnet with a magnetic field strength of 0.8T.

4. The process for extracting rapeseed pollen polysaccharides by complex enzyme synergistically according to claim 1, characterized in that: During the pulsed electric field enzymolysis process, a pulsed electric field is applied for 5 minutes at intervals of 30 minutes.

5. The process for extracting rapeseed pollen polysaccharides by complex enzyme synergistically according to claim 1, characterized in that: In the gradient ethanol precipitation method, the temperature of the first precipitation is controlled at 25°C, and the temperature of the second precipitation is lowered to 4°C.

6. The process for extracting rapeseed pollen polysaccharides by complex enzyme synergistically according to claim 1, characterized in that: When the hollow fiber ultrafiltration membrane is used for deproteinization, a 0.05% wt chitosan solution is introduced simultaneously as a filter aid.

7. The process for extracting rapeseed pollen polysaccharides by complex enzyme synergistically according to claim 1, characterized in that: The pulsating pressure range of the vacuum pulsating freeze drying is 5-10 Pa, the drying time is shortened by 20% compared with traditional freeze drying, and the polysaccharide activity retention rate is ≥95%.

8. The process for extracting rapeseed pollen polysaccharides by complex enzyme synergistically according to claim 1, characterized in that: The process integrates a near-infrared spectroscopy online monitoring system to measure the polysaccharide concentration in the enzymatic hydrolysate in real time, and automatically triggers the enzyme inactivation program when the concentration reaches 12 mg / mL.

9. The process for extracting rapeseed pollen polysaccharides by complex enzyme synergistically according to claim 1, characterized in that: The pulse airflow pulverization has a pulverization pressure of 0.6 MPa and is performed 3 times.

10. The process for extracting rapeseed pollen polysaccharides by complex enzyme synergistically according to claim 1, characterized in that: The yield of rapeseed pollen polysaccharides extracted by this process is ≥7.1%, the purity is ≥91%, and the proportion of β-glycosidic bonds is 35% higher than that of traditional enzymatic hydrolysis.

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