Cassava cell wall delivery vehicle and method of making same

Abstract

The application discloses a cassava cell wall delivery carrier and a preparation method thereof, and belongs to the field of special diet ingredient development. The application provides a preparation method of a natural cassava cell wall delivery carrier. The method is characterized in that: the whole cassava cell is separated by using pectinase, the cell wall permeability is regulated by hydrothermal treatment, the amylase is ensured to pass through the cell wall, the damage to the cell wall is minimized, the intracellular starch is enzymatically hydrolyzed by using digestive enzyme, and then a hollow cell wall with an intact cell wall structure is obtained. The obtained product can load guest molecules with biological activity, enhance the stability of the guest molecules, and slow down the release of the guest molecules in the stomach and intestines, so that the bioavailability of the guest molecules is improved.

Description

Technical Field

[0001] This invention relates to a cassava cell wall delivery carrier and its preparation method, belonging to the field of special dietary ingredient development. Background Technology

[0002] Cassava tubers are composed of neatly stacked, intact cells. Starch is tightly wrapped in the cell walls, which are made up of cellulose, hemicellulose, and pectin. These cells provide rigidity and strength to the cell structure. Furthermore, the small pores in the natural cell walls act as a physical barrier, inhibiting the digestive enzymes from hydrolyzing intracellular starch.

[0003] Currently, existing technologies utilize carriers such as gels, emulsions, and microcapsules to bind with active guest molecules (phenolic substances, antioxidants, drugs, dietary supplements, etc.) using physical or chemical methods to achieve the delivery and sustained release of guest molecules, thereby enhancing their bioavailability in vivo. However, this system has low drug loading capacity and requires a large amount of wall material to encapsulate the guest substance. At the same time, the digestion and decomposition of the wall material during the sustained release process in vivo increases the burden on the gastrointestinal tract.

[0004] Therefore, designing an embedding wall material for natural small molecule guest substances can enable the guest molecules to be released slowly in the gastrointestinal tract while reducing the burden on the gastrointestinal tract, which is of great significance for improving the bioavailability of guest molecules. Summary of the Invention

[0005] To address the aforementioned issues, the inventors previously discovered that the cell wall surface possesses small pores, allowing small molecules to pass through and be retained within the cell wall cavity, thus possessing the potential to serve as a delivery carrier. This invention provides a method for preparing a natural cassava cell wall delivery carrier. Intact cassava whole cells are separated using pectinase, and cell wall permeability is controlled through hydrothermal treatment to ensure amylase can pass through the cell wall while minimizing cell wall damage. Subsequently, intracellular starch is enzymatically hydrolyzed using digestive enzymes, resulting in a hollow cell wall with an intact cell wall structure. The resulting product can be loaded with bioactive guest molecules, enhancing the stability of the guest substances and their sustained-release effect in the gastrointestinal tract, thereby improving their bioavailability.

[0006] The first objective of this invention is to provide a cassava cell wall delivery carrier, the preparation method of which includes:

[0007] (1) Whole cell isolation of cassava: Cassava blocks were enzymatically hydrolyzed using pectinase hydrolysate. The ratio of cassava blocks to pectinase hydrolysate was 1 kg: 8-10 L. Cassava samples after enzymatic hydrolysis were prepared.

[0008] (2) Whole cell sieving: After enzymatic hydrolysis and separation, cassava samples were sieved and whole cell samples of cassava were collected.

[0009] (3) Regulation of cell wall permeability: Cassava whole cell samples were incubated at 70-75℃ for 1-5 min to prepare cassava whole cell samples with regulated cell walls;

[0010] (4) Preparation of hollow cell walls: Cassava whole cell samples with cell walls regulated by amylase were used to prepare cassava cell wall delivery vectors.

[0011] Unless otherwise specified, the unit for the feed-to-liquid ratio in this application is kg:L; the unit for w / v is kg / L.

[0012] In one embodiment, the pectinase hydrolysate in step (1) is prepared by dissolving pectinase in citrate buffer to a final concentration of 0.5–5 U / mL; adding 3–5% w / v ascorbic acid and activating at 45–50°C for 0.5–1 h to obtain the pectinase hydrolysate; the hydrolysis conditions are 45–50°C for 1–2 h.

[0013] In one embodiment, the citrate buffer solution has a concentration of 0.1 M and a pH of 3.5.

[0014] In one embodiment, the enzymatic hydrolysis reaction is accompanied by stirring at a speed of 80–100 r / min.

[0015] In one embodiment, the sieving in step (2) involves using two layers of sieves to sieve the cells. The first layer of sieve has a mesh size of 200-250 μm, and the second layer of sieve has a mesh size of 50-75 μm. The cells are rinsed with running water 5-8 times, and the cassava sample on the second layer of sieve is collected to obtain a whole cassava cell sample with a water content of 70-80%.

[0016] In one embodiment, in step (3), the cassava whole cells are incubated at 70°C for 3 minutes. Under these reaction conditions, amylase can pass through the cell wall while minimizing damage to the cell wall.

[0017] In one embodiment, the cassava whole cell sample after cell wall regulation in step (4) is dispersed in acetate buffer; wherein the volume ratio of the incubated cassava whole cell solution to the acetate buffer is 1:5 to 10.

[0018] In one embodiment, the enzymatic hydrolysis conditions in step (4) are 5-8 U / mL of amylase and 10-30 min of hydrolysis time.

[0019] In one embodiment, the amylase in step (4) includes: α-amylase, β-amylase, glucoamylase, and amylase.

[0020] In one embodiment, after the cassava whole cell sample with cell wall regulation is digested with amylase in step (4), the reaction is terminated by adding 90% ethanol, and the cassava cell wall delivery carrier is obtained by washing and centrifugation.

[0021] In one embodiment, the centrifugation speed in step (4) is 5000 rpm and the centrifugation time is 10 to 15 min.

[0022] A second object of the present invention is to provide a product that uses any of the above-described cassava cell wall delivery carriers; the method for preparing the product includes:

[0023] The guest molecule is mixed with any of the above-mentioned cassava cell wall delivery carriers at a mass ratio of 1:1 to 5, and the mixture is prepared by reacting for 0.5 to 1 h under the conditions of 1 to 3 MPa pressure, 20 to 25 °C temperature, and 80 to 100 rpm rotation speed.

[0024] In one embodiment, the guest molecule is dissolved in a solvent to prepare a guest molecule solution with a concentration of 10-20% w / v; the cassava cell wall delivery carrier is resuspended in water to prepare a cassava cell wall delivery carrier dispersion with a concentration of 10-20% w / v.

[0025] In one embodiment, the product includes food, health products, or medicine.

[0026] In one embodiment, the guest molecule includes: an antioxidant, a nutrient, a flavoring agent, or a drug.

[0027] Optionally, the antioxidants include, but are not limited to, one or more of anthocyanins, quercetin, ferulic acid, curcumin, and catechins.

[0028] Optionally, the nutrients include, but are not limited to, one or more of the following: vitamins, carotene, amino acids, minerals, unsaturated fatty acids, starch, and peptides.

[0029] Optionally, the flavor substances include, but are not limited to, one or more of the following: alcohols, esters, sugars, phenols, aldehydes, and glycosides.

[0030] A third object of the present invention is to provide the use of any of the above-described cassava cell wall delivery carriers in the preparation of products, including food, health products, or pharmaceuticals.

[0031] Optionally, the food includes, but is not limited to: starch, bread, noodles, steamed buns, dumplings, biscuits, and cakes; optionally, the additives include, but are not limited to, thickeners, emulsifiers, stabilizers, or preservatives; optionally, the medicine includes, but is not limited to, fillers, binders, disintegrants, flow aids, or drug carriers; optionally, the health products include, but are not limited to, resistant starch, pregelatinized starch, slow-digesting starch, and modified starch.

[0032] A fourth objective of this invention is to provide a method for reducing the release rate of guest molecules in the gastrointestinal tract, using a cassava cell wall delivery carrier to load the guest molecules. The preparation method includes: mixing the guest molecules with any of the above-mentioned cassava cell wall delivery carriers, and reacting for 0.5 to 1 hour under conditions of a pressure of 1 to 3 MPa, a temperature of 20 to 25°C, and a rotation speed of 80 to 100 rpm to obtain a cassava cell wall delivery carrier loaded with guest molecules.

[0033] In one embodiment, the guest molecule includes: antioxidants, nutrients, flavor compounds, or drugs; optionally, the antioxidants include one or more of anthocyanins, quercetin, ferulic acid, curcumin, and catechins; optionally, the nutrients include one or more of vitamins, carotene, amino acids, minerals, unsaturated fatty acids, starch, and peptides; optionally, the flavor compounds include one or more of alcohols, esters, sugars, phenols, aldehydes, and glycosides.

[0034] A fifth object of the present invention is to provide the application of any of the above-mentioned cassava cell wall delivery carriers in vivo for the delivery of drugs, food ingredients, and antioxidants, wherein the food ingredients include: medical food ingredients and dietary supplement ingredients; and the antioxidants include: phenolic active substances.

[0035] The beneficial effects of this invention are:

[0036] This invention provides a method for preparing a natural cassava cell wall delivery carrier. The method involves separating intact cassava cells using pectinase and regulating cell wall permeability through hydrothermal treatment to ensure amylase can pass through the cell wall while minimizing cell wall damage. Then, digestive enzymes are used to enzymatically hydrolyze intracellular starch, resulting in a hollow cell wall with an intact cell wall structure. The resulting product can be loaded with bioactive guest molecules, enhancing the stability of the guest substances and their sustained-release effect in the gastrointestinal tract, thereby improving their bioavailability.

[0037] (1) This invention uses enzymatic separation of cassava whole cells, regulates cell wall permeability through heat treatment, and uses digestive enzymes to hydrolyze intracellular starch to obtain cassava cell walls with relatively complete cell wall structures. This product is a natural, safe, non-toxic, and biodegradable wall material.

[0038] (2) The cassava cell wall material prepared by the present invention has rigidity and strength and strong acid and alkali resistance due to the presence of polysaccharides such as cellulose, hemicellulose and pectin in the cell wall, which can enhance the stability of guest molecules.

[0039] (3) The cassava cell wall material prepared by the present invention can encapsulate guest molecules in the hollow structure of the cell wall and enhance the encapsulation rate by using high pressure reaction, thereby increasing its drug loading capacity and reducing the burden of the wall material on gastrointestinal digestion.

[0040] (4) In this invention, due to the physical barrier effect of the cell wall, the guest molecules can have a slow release effect during the gastrointestinal digestion process, which improves their bioavailability. Moreover, the cell wall components are degraded by microorganisms in the colon as dietary fiber, which regulates the intestinal microorganisms and promotes intestinal health. Attached Figure Description

[0041] Figure 1 Microscopic images of whole cassava cells and their cell walls.

[0042] Figure 2 Simulate the release curve in gastric juice for cassava cell wall delivery carrier;

[0043] Figure 3 Release curves in small intestinal fluid were simulated for cassava cell wall delivery carriers. Detailed Implementation

[0044] The following describes preferred embodiments of the present invention. It should be understood that the embodiments are for better explanation of the present invention and are not intended to limit the present invention. In the following embodiments and comparative examples, curcumin is selected as the representative guest molecule to illustrate the advantages of the present invention.

[0045] 1. Observation of the whole cell and cell wall microstructure of cassava

[0046] The starch crystal structure of cassava whole cells after hydrothermal repair was observed under polarized light using an optical microscope, and the hollow cell walls were observed by staining the cells with 0.1% Congo red.

[0047] 2. Determination of drug loading and encapsulation efficiency

[0048] Prepare a series of curcumin-ethanol standard solutions with concentrations ranging from 2 to 6 μg / mL. Use anhydrous ethanol as a blank control and measure the absorbance at 425 nm. Plot a standard curve with concentration on the x-axis and absorbance on the y-axis.

[0049] Weigh 0.5 mg of sample into 10 mL of anhydrous ethanol, sonicate for 5 min, centrifuge at 3500 r / min for 10 min, combine the supernatants after two centrifugations, and measure the absorbance at 425 nm wavelength. Calculate the drug loading and encapsulation efficiency of the inclusion complex based on the standard curve.

[0050]

[0051] 3. Simulated curcumin release curves in gastric and intestinal fluids

[0052] 2.00 g NaCl and 0.26 g pepsin were dissolved in water, and the pH was adjusted to 2 with 1 mol / L HCl, bringing the volume to 1 L to obtain simulated gastric juice. 6.80 g KH₂PO₄ was dissolved in 500 mL of water, and then trypsin (10 g / L) was dissolved in the above solution. The volume was brought to 1 L with water, and the pH was adjusted to 6.5 with 0.1 mol / L NaOH to obtain simulated intestinal juice. 5.00 mg of sample was dispersed in 5 mL of gastric and intestinal juices, placed in a dialysis bag, and immersed in 200 mL of dialysis fluid. The curcumin release was measured every 0.5 h. The curcumin release was determined by measuring the absorbance of 2 mL of the supernatant at 425 nm using a UV spectrophotometer.

[0053] Raw materials used in the examples:

[0054] Cassava was purchased from Guangxi Kangyi Ecological Agriculture Development Co., Ltd.

[0055] Example 1: Preparation of cassava cell wall delivery carrier

[0056] The steps for preparing the cassava cell wall delivery vector are as follows:

[0057] (1) Whole-cell isolation of cassava: Peel and wash the cassava, cut it into pieces (3mm×3mm), and soak it in water; dissolve pectinase in citrate buffer (0.1M pH=3.5) to prepare 1U / mL enzymatic hydrolysate, and add 5% w / v ascorbic acid, activate at 50℃ for 0.5h to prepare preheated activated enzymatic hydrolysate; add cassava pieces to the preheated activated enzymatic hydrolysate, with a material-to-liquid ratio of 1kg:8L, and stir continuously at 50℃ (100rpm) for 2h to prepare enzymatically isolated cassava samples;

[0058] (2) Cassava whole cell sieving: The cassava sample prepared in step (1) after enzymatic hydrolysis and separation was rinsed with running water and the cells were sieved through a double-layer sieve. The first layer of the sieve had a pore size of 250 μm and the second layer of the sieve had a pore size of 75 μm. The sample was rinsed with running water 5 times and the cassava whole cells separated on the second layer of the sieve were collected. The cassava whole cell sample had a water content of 75%.

[0059] (3) Cell wall permeability regulation: The water-containing cassava whole cell sample obtained in step (2) was incubated at 70℃ for 3 min to regulate cell permeability and ensure that the amylase could enzymatically hydrolyze intracellular starch through the cell wall to prepare a cassava whole cell sample after cell wall regulation (because the water content of the cells was high before incubation, the cassava whole cell sample after incubation was in solution form).

[0060] (4) Preparation of hollow cell walls: The cassava whole cell samples with cell walls regulated in step (3) were dispersed in sodium acetate (0.1M pH=5.2) buffer at a volume ratio of 1:5, and amyloglucosidase (final enzyme concentration of 5U / mL) was added. The mixture was incubated at 37℃ for 30 min to enzymatically hydrolyze intracellular starch, and 90% ethanol was added to terminate the reaction.

[0061] (5) Centrifugation and dehydration: Centrifuge the product obtained in step (4) at 5000 rpm for 15 min, discard the supernatant, and wash it three times with distilled water to prepare the cell wall;

[0062] (6) Curcumin loading: The cell wall prepared in step (5) was dispersed in water to prepare a 10% w / v cell wall suspension; curcumin was dissolved in anhydrous ethanol to prepare a 10% w / v curcumin-ethanol solution, which was added to the uniformly dispersed cell wall suspension, wherein the ratio of curcumin to cell wall was 1:1 (dry matter weight ratio), and placed in a high-pressure reactor to accelerate the encapsulation efficiency of the cell wall on the guest substance. The reaction pressure was 1 MPa, the temperature was 20℃, the reaction speed was 80 rpm, and the reaction time was 0.5 h; after the reaction was completed, the supernatant was removed by centrifugation (to remove liquid and free curcumin), and the precipitate was freeze-dried to prepare a cassava cell wall loaded with curcumin (curcumin cassava cell wall delivery carrier).

[0063] Microscopic examination of the cell walls of whole cassava cells and those after enzymatic starch hydrolysis yielded the following results: Figure 1 As shown.

[0064] The results showed that hollow cell walls with intact cell wall structures could be obtained by sieving cassava tubers with pectinase, regulating cell wall permeability through hydrothermal treatment, and hydrolyzing intracellular starch with digestive enzymes.

[0065] Example 2: Preparation of cassava cell wall delivery carrier

[0066] Based on Example 1, step (1) was modified as follows: pectinase was dissolved in citrate buffer (0.1M pH=3.5) to prepare a 5U / mL enzymatic hydrolysate, and 3% w / v ascorbic acid was added. The hydrolysate was activated at 45℃ for 1h to obtain a preheated and activated enzymatic hydrolysate. Cassava chunks were added to the preheated and activated enzymatic hydrolysate at a material-to-liquid ratio of 1kg:10L. The hydrolysate was stirred continuously at 50℃ (100rpm) for 2h to obtain an enzymatically separated cassava sample. The remaining steps were the same as in Example 1 to obtain a cassava cell wall loaded with curcumin.

[0067] Example 3: Preparation of cassava cell wall delivery carrier

[0068] Based on Example 1, step (3) is changed to: the whole cassava cell sample obtained in step (2) is incubated at 75°C for 2 min, and the remaining steps are the same as in Example 1, so as to prepare cassava cell wall loaded with curcumin.

[0069] Example 4: Preparation of cassava cell wall delivery carrier

[0070] Based on Example 1, step (4) is changed as follows: the whole cassava cell sample with cell wall regulation prepared in step (3) is dispersed in sodium acetate (0.1M pH=5.2) buffer at a volume ratio of 1:8, amyloglucosidase (final concentration of enzyme is 8U / mL) is added, and the mixture is incubated at 37℃ for 15 min to enzymatically hydrolyze intracellular starch. 90% ethanol is added to terminate the reaction.

[0071] Example 5: Preparation of cassava cell wall delivery carrier

[0072] Based on Example 1, step (6) was modified as follows: the cell wall prepared in step (5) was dispersed in water to prepare a 15% w / v cell wall suspension; curcumin was dissolved in anhydrous ethanol to prepare a 15% w / v curcumin-ethanol solution, which was added to the uniformly dispersed cell wall suspension, wherein the ratio of curcumin to cell wall was 1:1 (dry weight ratio). The mixture was placed in a high-pressure reactor to accelerate the encapsulation efficiency of the cell wall on the guest substance. The reaction pressure was 3 MPa, the temperature was 25°C, the reaction speed was 100 rpm, and the reaction time was 1 h. After the reaction, the supernatant was removed by centrifugation (to remove liquid and free curcumin), and the precipitate was freeze-dried to prepare cassava cell walls loaded with curcumin (curcumin cassava cell wall delivery carrier). Comparative Example 1: Preparation of curcumin microcapsules by conventional method

[0073] Curcumin microcapsules were prepared using a common spray drying method, with starch as a carrier for encapsulating curcumin. The steps are as follows:

[0074] Prepare a 10% w / v starch solution, boil it in a water bath for 10 min and cool it for later use. Prepare a 30% w / v curcumin-ethanol solution. Mix the curcumin-ethanol solution with the starch solution, wherein the mass ratio of curcumin to starch is 1:1 (dry matter weight ratio). Stir continuously at room temperature for 2 h. The uniformly mixed solution is spray-dried to obtain curcumin microcapsules.

[0075] When analyzing drug loading and encapsulation efficiency, the samples were washed with anhydrous ethanol to remove free curcumin from the surface.

[0076] Comparative Example 2: Crushed whole-cell cassava

[0077] Based on Example 1, the whole cassava cell sample collected in step (2) was broken in a cell wall breaker for 20 seconds to obtain broken whole cassava cells. The remaining steps were the same as in Example 1 to prepare broken cassava cell walls loaded with curcumin.

[0078] Comparative Example 3: Adjusting cell wall permeability to regulate treatment temperature

[0079] Based on Example 1, the treatment conditions for step (3) were: incubation at 90°C for 20 min, and the remaining steps were the same as in Example 1, to prepare cassava cell walls loaded with curcumin.

[0080] Tests showed that the starch granules in the whole cassava cells prepared in this comparative example completely absorbed water and swelled. The cell wall pores enlarged under the expansion pressure, which increased the cell wall permeability. The loading efficiency and sustained-release effect of curcumin were analyzed.

[0081] Comparative Example 4: Omission of Cell Wall Permeability Regulation

[0082] Based on Example 1, step (3) is omitted, and the remaining steps are the same as in Example 1, to prepare complete cassava whole cells loaded with curcumin.

[0083] Comparative Example 5: Changing the conditions of starch enzymatic hydrolysis

[0084] Based on Example 1, step (4) was modified as follows: the whole cassava cell sample with cell wall regulation prepared in step (3) was dispersed in sodium acetate (0.1M pH=5.2) buffer at a volume ratio of 1:10, amyloglucosidase (final enzyme concentration of 0.5U / mL) was added, and the mixture was incubated at 37℃ for 5 min to enzymatically hydrolyze intracellular starch. 90% ethanol was added to terminate the reaction. The remaining steps were the same as in Example 1, and starch-containing cassava cell walls loaded with curcumin were prepared.

[0085] Comparative Example 6: No high-voltage auxiliary power used

[0086] Based on Example 1, step (6) is modified as follows: the cell wall prepared in step (5) is dispersed in water to prepare a 10% w / v cell wall suspension; curcumin is dissolved in anhydrous ethanol to prepare a 10% w / v curcumin-ethanol solution, which is added to the uniformly dispersed cell wall suspension, wherein the ratio of curcumin to cell wall is 1:1 (dry matter weight ratio), magnetically stirred for 1 h, centrifuged to remove the supernatant (to remove liquid and free curcumin), and the precipitate is freeze-dried to prepare a cassava cell wall loaded with curcumin (curcumin cassava cell wall delivery carrier).

[0087] Example 6: Detection of the release curve of curcumin cassava cell wall delivery carrier

[0088] Using commercially available curcumin raw material (Comparative Example 7) as a control, the drug loading and encapsulation efficiency of cassava cell walls loaded with curcumin prepared in Examples 1-5 and Comparative Examples 1-7, as well as the release curves in simulated intestinal and gastric fluids, were determined.

[0089] Table 1 Results of drug loading and encapsulation efficiency determination

[0090]

[0091]

[0092] The results of drug loading and encapsulation efficiency measurements are shown in Table 1. Comparative Example 6 and Example 1 show that high pressure can increase the drug loading of the cell wall. Comparative Example 4 and Example 1 show that cell wall permeability regulation is crucial for subsequent enzyme entry and starch hydrolysis within the cells. Comparative Example 2 and Example 1 show that broken cassava cells have poor drug encapsulation effects. Comparative Examples 5, 3, and the Example show that parameters used in cell wall preparation also affect the final drug encapsulation. Comparative Example 1 and Example 1 show that compared to conventional starch-based carriers, the drug loading of the cell wall increased from 16.57% to 30%–40%, significantly improving the drug loading. The commercially available raw material is pure curcumin (Comparative Example 7), which releases too quickly in the gastrointestinal tract and fails to achieve a sustained-release effect.

[0093] The results of the simulated gastric and small intestinal fluid release curves are as follows: Figure 2 , Figure 3 As shown in Tables 2 and 3, the results indicate that, as demonstrated by Comparative Example 6 and Example 1, high pressure assistance can achieve the loading of phenolic substances, delay their release in the gastrointestinal tract, and improve their bioavailability in vivo. The drug loading in Examples 1 to 5 is significantly increased, and excessively low cell permeability (Comparative Example 4), excessively high cell permeability (Comparative Example 3), and cell wall disruption (Comparative Example 2) all lead to a decrease in drug loading and loading rate.

[0094] Compared with the original curcumin, the curcumin loaded on the cassava cell wall has a significant sustained-release effect in simulated gastric and intestinal juices, and can continue to release curcumin even after 6 hours of reaction. In addition, the product of this invention avoids the use of chemical reagents and has the advantages of natural environmental protection. This product can be used as an ingredient in special dietary foods and can be widely used in special dietary foods with antioxidant, anti-inflammatory and antibacterial properties and regulation of glucose and lipid metabolism.

[0095] Table 2 Gastric juice sustained-release data

[0096] Grouping 0.5h 1h 2h 3h 4h 6h Example 1 11.8％ 15.4％ 20.6％ 24.4％ 26.2％ 30.3％ Example 2 10.7％ 14.4％ 19.3％ 25.4％ 28.1％ 35.7％ Example 3 14.8％ 19.3％ 24.3％ 32.8％ 36.5％ 39.5％ Example 4 12.3％ 17.5％ 22.5％ 25.3％ 28.8％ 33.7％ Example 5 13.1％ 18.2％ 23.3％ 28.3％ 31.4％ 37.4％ Comparative Example 1 28.4％ 33.1％ 37.1％ 44.1％ 49.4％ 51.3％ Comparative Example 2 27.2％ 35.5％ 42.5％ 48.2％ 52.3％ 54.6％ Comparative Example 3 25.1％ 30.5％ 34.6％ 41.1％ 45.5％ 48.3％ Comparative Example 4 34.3％ 41.3％ 53.4％ 62.2％ 65.5％ 69.3％ Comparative Example 5 20.5％ 24.7％ 32.2％ 36.5％ 40.3％ 43.2％ Comparative Example 6 24.3％ 27.9％ 34.5％ 39.4％ 44.8％ 45.1％ Comparative Example 7 41.7％ 52.3％ 66.5％ 78.2％ 81.4％ 83.7％

[0097] Table 3. Small Intestinal Fluid Sustained-Release Data

[0098]

[0099]

[0100] Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Anyone skilled in the art can make various modifications and alterations without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be determined by the claims.

Claims

1. A product, characterized in that, The product uses a cassava cell wall delivery carrier; The product includes food or medicine; the preparation method of the product includes: The guest molecule was mixed with a cassava cell wall delivery carrier at a mass ratio of 1:1 to 5, and the mixture was reacted for 0.5 to 1 h under the conditions of 1 to 3 MPa pressure, 20 to 25 °C temperature, and 80 to 100 rpm rotation speed. The method for preparing the cassava cell wall delivery carrier includes: (1) Whole cell isolation of cassava: Cassava blocks were enzymatically hydrolyzed using pectinase hydrolysate. The ratio of cassava blocks to pectinase hydrolysate was 1 kg: 8~10 L. Cassava samples after enzymatic hydrolysis were prepared. (2) Whole cell sieving: After enzymatic hydrolysis and separation, cassava samples were sieved and whole cell samples of cassava were collected. (3) Regulation of cell wall permeability: Cassava whole cell samples were incubated at 70-75℃ for 1-5 min to prepare cassava whole cell samples with regulated cell walls; (4) Preparation of hollow cell walls: Cassava whole cell samples with cell walls regulated by amylase were used to prepare cassava cell wall delivery vectors; The preparation method of pectinase hydrolysate in step (1) is as follows: dissolve pectinase in citrate buffer to make the final concentration of pectinase 0.5~5 U / mL; add 3~5% w / v ascorbic acid, activate at 45~50℃ for 0.5~1 h to prepare pectinase hydrolysate; the hydrolysis conditions are 45~50℃ for 1~2 h. In step (2), the cells are sieved using two layers of sieves. The first layer of sieves has a pore size of 200-250 μm, and the second layer of sieves has a pore size of 50-75 μm. The cassava sample on the second layer of sieves is collected to obtain whole cassava cell powder. The amylase in step (4) includes α-amylase, β-amylase or glucoamylase; the enzymatic hydrolysis conditions are an amylase concentration of 5~8 U / mL and a hydrolysis time of 10~30 min. The guest molecules include: antioxidants, nutrients, flavorings, or drugs.

2. The product according to claim 1, characterized in that, The antioxidants include one or more of the following: anthocyanins, quercetin, ferulic acid, curcumin, and catechins.

3. The product according to claim 1, characterized in that, The nutrients include one or more of the following: vitamins, carotene, amino acids, minerals, unsaturated fatty acids, starch, and peptides.

4. The product according to claim 1, characterized in that, The flavor compounds include one or more of the following: alcohols, esters, sugars, phenols, aldehydes, and glycosides.

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