A method for preparing coconut oligosaccharides for adjunctive treatment of hyperglycemia and hyperlipidemia.

Abstract

This invention provides a method for preparing coconut oligosaccharides for adjuvant treatment of hyperglycemia and hyperlipidemia, comprising the following steps: (1) cutting coconut meat into pieces, pulping, homogenizing under high pressure to obtain a homogenate, and then cooling and filtering to obtain a primary filtrate; (2) adding a compound enzyme preparation to the primary filtrate for enzymatic hydrolysis, centrifuging and collecting the supernatant to obtain a secondary filtrate; (3) adding an ionic liquid to the secondary filtrate for fractional extraction, collecting the extract, evaporating, and vacuum concentrating to obtain a concentrated solution; (4) decolorizing the concentrated solution by adsorption through a macroporous adsorption resin column, eluting with deionized water, collecting the sugar-containing components, and freeze-drying to obtain the target coconut oligosaccharide. This invention achieves a high yield of coconut oligosaccharides through the synergistic effect of the compound enzyme preparation and the ionic liquid. This coconut oligosaccharide can inhibit the activity of α-glucosidase, while also exhibiting good cholesterol-binding ability and a good inhibition rate against lipase, thus providing a good adjuvant treatment effect for hyperglycemia and hyperlipidemia.

Description

Technical Field

[0001] This invention relates to the field of oligosaccharide preparation technology, and in particular to a method for preparing coconut oligosaccharides for adjuvant treatment of hyperglycemia and hyperlipidemia. Background Technology

[0002] Hyperglycemia and hyperlipidemia are common metabolic diseases in modern society, seriously affecting people's health and potentially leading to various complications such as cardiovascular and cerebrovascular diseases and diabetes. Currently, although there are many types of drugs available for treating hyperglycemia and hyperlipidemia, long-term use may cause certain side effects. Therefore, finding safe and effective adjunctive treatment methods is of significant practical importance.

[0003] Oligosaccharides, also known as oligosaccharides or low-glucose compounds, refer to compounds composed of 2-10 glycosidic bonds. They are a new type of functional sugar source with various physiological functions, such as regulating intestinal flora, assisting in the regulation of blood sugar and blood lipids, promoting nutrient metabolism, and helping mineral absorption. Oligosaccharides integrate nutrition, health care, and dietary therapy, and are widely used in food, health products, beverages, medicine, feed additives and other fields.

[0004] Coconut is a common tropical fruit, and its flesh is rich in carbohydrates. Currently, the main processed products of coconut are coconut oil, coconut powder, coconut sugar, and coconut water. However, there are relatively few methods for extracting coconut oligosaccharides, and the extraction process suffers from drawbacks such as poor dissolution of oligosaccharides and low extraction rates. Therefore, finding a suitable extraction method for coconut oligosaccharides can further expand the deep processing and utilization of coconut. Summary of the Invention

[0005] In view of this, the present invention proposes a method for preparing coconut oligosaccharides for adjuvant treatment of hyperglycemia and hyperlipidemia.

[0006] The technical solution of this invention is implemented as follows:

[0007] A method for preparing coconut oligosaccharides for adjuvant treatment of hyperglycemia and hyperlipidemia includes the following steps:

[0008] (1) After cutting the coconut meat into pieces, mix it with water and then beat it to obtain a pulp. The pulp is then subjected to high-pressure homogenization to obtain a homogenate. The homogenate is then cooled at 2-4℃ for 6-8 hours and filtered to obtain a primary filtrate.

[0009] (2) Add a compound enzyme preparation to the primary filtrate and shake to hydrolyze it. Centrifuge and collect the supernatant to obtain the secondary filtrate.

[0010] (3) Add the secondary filtrate to an ionic liquid for fractional extraction, collect the extract, evaporate the extract, and concentrate it under vacuum to obtain a concentrated solution;

[0011] (4) The concentrated liquid was adsorbed and decolorized by a macroporous adsorption resin column, eluted with deionized water, and the sugar-containing components were collected and freeze-dried to obtain coconut oligosaccharides for adjuvant treatment of hyperglycemia and hyperlipidemia.

[0012] Furthermore, the mass ratio of coconut meat to water is 3 to 5:1; the high-pressure homogenization is carried out 2 to 3 times under conditions of 30 to 50 MPa.

[0013] Furthermore, the amount of the compound enzyme preparation added is 2% to 4% of the mass of the first filtrate.

[0014] Furthermore, the complex enzymes in the complex enzyme preparation include cellulase, papain, pectinase, and glucanase in a mass ratio of 7-10:2-3:2-4:3-5.

[0015] Furthermore, the preparation method of the compound enzyme preparation is as follows: take the compound enzyme and add it to the phosphate buffer solution and stir to mix well. Then, while stirring, add polyethylene glycol 400 to prepare a suspension. Add 1,8-bismaleimide-diethylene glycol to the suspension and stir and crosslink in an ice-water bath for 2-4 hours. Centrifuge to collect the precipitate and wash the precipitate 3-4 times to obtain the compound enzyme preparation.

[0016] Furthermore, the concentration of the phosphate buffer solution is 18–20 mg / mL; the mass-to-volume ratio of the complex enzyme, phosphate buffer, and polyethylene glycol 400 is 1–2 mg: 6–8 mL: 20–30 mL.

[0017] Furthermore, the 1,8-bismaleimide-diethylene glycol added is 2% to 3% of the mass of the suspension.

[0018] Furthermore, the centrifugation conditions are: rotation speed 3500-4000 r / min, time 20-25 min.

[0019] Furthermore, the volume ratio of the secondary filtrate to the ionic liquid is 5-7:1-2.

[0020] Furthermore, the ionic liquid is prepared by mixing 1-ethyl-3-methylimidazolium lactate with an ethanol solution; the mass concentration of the ethanol solution is 70% to 80%; and the volume ratio of 1-ethyl-3-methylimidazolium lactate to the ethanol solution is 3 to 5: 1 to 2.

[0021] Furthermore, the oscillating enzymatic hydrolysis conditions are as follows: temperature 45–55℃, pH 6.0–7.5, oscillation speed 100–150 r / min, and oscillation enzymatic hydrolysis time 60–300 min.

[0022] Furthermore, the pore size of the ultrafiltration membrane is 3000–5000 Da.

[0023] Compared with the prior art, the beneficial effects of the present invention are:

[0024] This invention first uses low-temperature cooling to solidify and separate lipids during the extraction of coconut oligosaccharides. Then, the synergistic effect of a compound enzyme preparation and ionic liquid effectively extracts the coconut oligosaccharides, resulting in a high yield. Furthermore, the coconut oligosaccharides prepared by this invention are more easily absorbed, inhibit α-glucosidase activity, reduce glucose absorption, and possess good cholesterol-binding capacity. They also exhibit good inhibition of lipase, thus reducing the body's absorption of fat and providing good adjunctive treatment for hyperglycemia and hyperlipidemia.

[0025] In this invention, the compound enzyme preparation exhibits high stability and enhances the activity of the compound enzyme after precipitation and cross-linking. Furthermore, through the synergistic effect of the compound enzyme preparation and ionic liquid, it can enhance the destruction of coconut cell walls, accelerate the rate of sugar dissolution from cells, and accelerate the degradation of macromolecular polysaccharides into oligosaccharides. It can also remove impurities such as cellulose, protein, and pectin, which is beneficial for the subsequent separation and purification of coconut oligosaccharides, effectively shortens the preparation time, maintains the activity of oligosaccharides, and improves the yield of coconut oligosaccharides. Detailed Implementation

[0026] To better understand the technical content of this invention, specific embodiments are provided below to further illustrate the invention.

[0027] Unless otherwise specified, the experimental methods used in the embodiments of this invention are all conventional methods.

[0028] Unless otherwise specified, all materials and reagents used in the embodiments of this invention are commercially available.

[0029] The cellulase used in this invention is manufactured by Nanning Pangbo Biotechnology Co., Ltd., and its specification is 20,000 u / g.

[0030] The papain of this invention is manufactured by Nanning Pangbo Biotechnology Co., Ltd., and the specification is 20,000 u / g.

[0031] The pectinase used in this invention is manufactured by Nanning Pangbo Bioengineering Co., Ltd., and the specification is 30,000 u / g.

[0032] The glucanase used in this invention is manufactured by Nanning Pangbo Biotechnology Co., Ltd., and the specification is 30,000 u / g.

[0033] Example 1

[0034] The compound enzymes in the compound enzyme preparation include cellulase, papain, pectinase and glucanase in a mass ratio of 7:3:4:3;

[0035] The preparation method of the compound enzyme preparation is as follows: The compound enzyme is added to a phosphate buffer solution with a concentration of 19 mg / mL and stirred until homogeneous. Then, while stirring, polyethylene glycol 400 is added to form a suspension. 1,8-bismaleimide-diethylene glycol is added to the suspension and stirred in an ice-water bath for 2 hours to crosslink. The precipitate is collected by centrifugation. The precipitate is washed four times with a phosphate buffer solution with a concentration of 19 mg / mL to obtain the compound enzyme preparation. The mass-to-volume ratio of the compound enzyme, phosphate buffer, and polyethylene glycol 400 is 1 mg:8 mL:26 mL; the mass of 1,8-bismaleimide-diethylene glycol added is 3% of the mass of the suspension.

[0036] The ionic liquid is prepared by mixing 1-ethyl-3-methylimidazolium lactate with an ethanol solution; the ethanol solution has a mass concentration of 75%; and the volume ratio of 1-ethyl-3-methylimidazolium lactate to the ethanol solution is 3:2.

[0037] A method for preparing coconut oligosaccharides for adjuvant treatment of hyperglycemia and hyperlipidemia includes the following steps:

[0038] (1) The mass ratio of coconut meat to water is 4:1. After cutting the coconut meat into pieces, it is mixed with water and then pulped to obtain a slurry. The slurry is homogenized three times under 50MPa to obtain a homogenate. Then, the homogenate is cooled at 4℃ for 8 hours and filtered to obtain a first filtrate.

[0039] (2) Add the compound enzyme preparation to the primary filtrate and perform enzymatic hydrolysis for 120 min at a temperature of 55℃, pH of 7.5 and a shaking speed of 100 r / min. Then, centrifuge at a speed of 3800 r / min for 20 min and collect the supernatant to obtain the secondary filtrate. The amount of compound enzyme preparation added is 4% of the mass of the primary filtrate.

[0040] (3) The secondary filtrate is added to the ionic liquid for fractional extraction according to the volume ratio of secondary filtrate to ionic liquid is 6:2. The extract is collected, evaporated, and concentrated under vacuum to obtain the concentrate.

[0041] (4) The concentrate was adsorbed and decolorized by a macroporous adsorption resin column, eluted with deionized water, and the sugar-containing components were collected. Then, it was filtered through an ultrafiltration membrane with a pore size of 5000 Da to obtain the permeate, which was then freeze-dried to obtain coconut oligosaccharides.

[0042] Example 2

[0043] The compound enzymes in the compound enzyme preparation include cellulase, papain, pectinase and glucanase in a mass ratio of 9:2:2:4.

[0044] The preparation method of the compound enzyme preparation is as follows: The compound enzyme is added to a phosphate buffer solution with a concentration of 18 mg / mL and stirred until homogeneous. Then, while stirring, polyethylene glycol 400 is added to form a suspension. 1,8-bismaleimide-diethylene glycol is added to the suspension and stirred in an ice-water bath for 3 hours to crosslink the mixture. The precipitate is collected by centrifugation. The precipitate is washed three times with a phosphate buffer solution with a concentration of 18 mg / mL to obtain the compound enzyme preparation. The mass-to-volume ratio of the compound enzyme, phosphate buffer, and polyethylene glycol 400 is 2 mg:6 mL:30 mL; the mass of 1,8-bismaleimide-diethylene glycol added is 2% of the mass of the suspension.

[0045] The ionic liquid is prepared by mixing 1-ethyl-3-methylimidazolium lactate with an ethanol solution; the ethanol solution has a mass concentration of 70%; and the volume ratio of 1-ethyl-3-methylimidazolium lactate to the ethanol solution is 5:1.

[0046] A method for preparing coconut oligosaccharides for adjuvant treatment of hyperglycemia and hyperlipidemia includes the following steps:

[0047] (1) The mass ratio of coconut meat to water is 5:1. After the coconut meat is cut into pieces, it is mixed with water and then pulped to obtain a slurry. The slurry is homogenized twice under 30MPa to obtain a homogenate. Then the homogenate is cooled at 3℃ for 6 hours and filtered to obtain a first filtrate.

[0048] (2) Add the compound enzyme preparation to the primary filtrate and perform enzymatic hydrolysis for 150 min at a temperature of 45℃, pH of 6.8 and a shaking speed of 120 r / min. Then, centrifuge at a speed of 3500 r / min for 25 min and collect the supernatant to obtain the secondary filtrate. The amount of compound enzyme preparation added is 2% of the mass of the primary filtrate.

[0049] (3) The secondary filtrate is added to the ionic liquid for fractional extraction according to the volume ratio of secondary filtrate to ionic liquid of 7:1. The extract is collected, evaporated, and concentrated under vacuum to obtain the concentrate.

[0050] (4) The concentrate was adsorbed and decolorized by a macroporous adsorption resin column, eluted with deionized water, and the sugar-containing components were collected. Then, it was filtered through an ultrafiltration membrane with a pore size of 5000 Da to obtain the permeate, which was then freeze-dried to obtain coconut oligosaccharides.

[0051] Example 3

[0052] The compound enzymes in the compound enzyme preparation include cellulase, papain, pectinase and glucanase in a mass ratio of 10:3:3:5;

[0053] The preparation method of the compound enzyme preparation is as follows: The compound enzyme is added to a 20 mg / mL phosphate buffer solution and stirred until homogeneous. Then, while stirring, polyethylene glycol 400 is added to form a suspension. 1,8-bismaleimide-diethylene glycol is added to the suspension and stirred in an ice-water bath for 3 hours to crosslink. The precipitate is collected by centrifugation. The precipitate is washed four times with a 20 mg / mL phosphate buffer solution to obtain the compound enzyme preparation. The mass-to-volume ratio of the compound enzyme, phosphate buffer, and polyethylene glycol 400 is 2 mg:7 mL:20 mL; the mass of 1,8-bismaleimide-diethylene glycol added is 2.5% of the suspension mass.

[0054] The ionic liquid is prepared by mixing 1-ethyl-3-methylimidazolium lactate with an ethanol solution; the ethanol solution has a mass concentration of 80%; and the volume ratio of 1-ethyl-3-methylimidazolium lactate to the ethanol solution is 4:1.

[0055] A method for preparing coconut oligosaccharides for adjuvant treatment of hyperglycemia and hyperlipidemia includes the following steps:

[0056] (1) The mass ratio of coconut meat to water is 3:1. After cutting the coconut meat into pieces, it is mixed with water and then pulped to obtain a pulp. The pulp is homogenized three times under 40MPa to obtain a homogenate. Then, the homogenate is cooled at 2℃ for 7h and filtered to obtain a first filtrate.

[0057] (2) Add the compound enzyme preparation to the primary filtrate and perform enzymatic hydrolysis at a temperature of 50℃, pH of 7.2 and a shaking speed of 150 r / min for 120 min. Then, centrifuge at a speed of 4000 r / min for 20 min and collect the supernatant to obtain the secondary filtrate. The amount of compound enzyme preparation added is 3% of the mass of the primary filtrate.

[0058] (3) According to the volume ratio of secondary filtrate to ionic liquid of 5:2, the secondary filtrate is added to the ionic liquid for fractional extraction, the extract is collected, the extract is evaporated, and the extract is concentrated under vacuum to obtain the concentrate.

[0059] (4) The concentrate was adsorbed and decolorized by a macroporous adsorption resin column, eluted with deionized water, and the sugar-containing components were collected. Then, it was filtered through an ultrafiltration membrane with a pore size of 5000 Da to obtain the permeate, which was then freeze-dried to obtain coconut oligosaccharides.

[0060] Comparative Example 1

[0061] The difference between this comparative example and Example 3 is that no compound enzyme preparation was added for oscillating enzymatic hydrolysis, while the rest is the same as Example 3.

[0062] The comparative method for preparing coconut oligosaccharides for adjuvant treatment of hyperglycemia and hyperlipidemia includes the following steps:

[0063] (1) The mass ratio of coconut meat to water is 3:1. After cutting the coconut meat into pieces, it is mixed with water and then pulped to obtain a pulp. The pulp is homogenized three times under 40MPa to obtain a homogenate. Then, the homogenate is cooled at 2℃ for 7h and filtered to obtain a first filtrate.

[0064] (2) The primary filtrate is added to the ionic liquid at a volume ratio of 5:2 to the primary filtrate and then fractionally extracted. The extract is collected, evaporated, and concentrated under vacuum to obtain the concentrate.

[0065] (3) The concentrate was decolorized by adsorption through a macroporous adsorption resin column, eluted with deionized water, and the sugar-containing components were collected. Then, it was filtered through an ultrafiltration membrane with a pore size of 5000 Da to obtain the permeate, which was then freeze-dried to obtain coconut oligosaccharides.

[0066] Comparative Example 2

[0067] The difference between this comparative example and Example 3 is that no ionic liquid was added for fractional extraction, while the rest is the same as Example 3.

[0068] The comparative example describes a method for preparing coconut oligosaccharides for the adjunctive treatment of hyperglycemia and hyperlipidemia, comprising the following steps:

[0069] (1) The mass ratio of coconut meat to water is 3:1. After cutting the coconut meat into pieces, it is mixed with water and then pulped to obtain a pulp. The pulp is homogenized three times under 40MPa to obtain a homogenate. Then, the homogenate is cooled at 2℃ for 7h and filtered to obtain a first filtrate.

[0070] (2) Add the compound enzyme preparation to the primary filtrate and perform enzymatic hydrolysis at 50℃, pH 7.2 and shaking speed of 150 r / min for 120 min. Then, centrifuge at 4000 r / min for 20 min and collect the supernatant to obtain the secondary filtrate. The amount of compound enzyme preparation added is 3% of the mass of the primary filtrate. Evaporate the supernatant and concentrate it under vacuum to obtain the concentrated solution.

[0071] (3) The concentrate was decolorized by adsorption through a macroporous adsorption resin column, eluted with deionized water, and the sugar-containing components were collected. Then, it was filtered through an ultrafiltration membrane with a pore size of 5000 Da to obtain the permeate, which was then freeze-dried to obtain coconut oligosaccharides.

[0072] Comparative Example 3

[0073] The difference between this comparative example and Example 3 is that the preparation method of the compound enzyme preparation of the present invention is different from that of Example 3. That is, the compound enzyme in this comparative example is simply mixed evenly without precipitation and cross-linking, while the rest is consistent with Example 3.

[0074] The preparation method of the compound enzyme preparation in this comparative example is as follows: cellulase, papain, pectinase and glucanase are mixed evenly in a mass ratio of 10:3:3:5.

[0075] Test case

[0076] The properties of the coconut oligosaccharides prepared by the methods of Examples 1-3 and Comparative Examples 1-3 were determined.

[0077] (1) Yield of coconut oligosaccharides

[0078] The coconut oligosaccharides prepared according to the preparation methods of Examples 1-3 and Comparative Examples 1-3 were then used as glucose as a standard, and the yield (%) of the coconut oligosaccharides was determined according to the phenol-sulfuric acid method.

[0079] (2) α-glucosidase inhibitory ability

[0080] The coconut oligosaccharides obtained by the preparation methods of Examples 1-3 and Comparative Examples 1-3 were prepared into sample solutions with a concentration of 4 mg / mL using 67 mmol / L potassium phosphate buffer (pH 6.8). The inhibition rate of α-glucosidase activity was then determined colorimetrically. 1.0 mL of the above sample solution was thoroughly mixed with 2.0 mL of α-glucosidase at an enzyme activity concentration of 2 U / mL. After incubation at 37°C for 20 min, 4.5 mL of p-NPG at a concentration of 20 mmol / L was added. The mixture was incubated at 37°C for another 30 min, and finally, 7.0 mL of sodium carbonate solution at a concentration of 2 mol / L was added to terminate the reaction. In the control tube, the same amount of potassium phosphate buffer was used as a blank group instead of the sample group, with the sample group without α-glucosidase as the background group. Acarbose was used as the positive control. The absorbance at 405 nm was measured to determine the α-glucosidase inhibition ability. The formula for calculating the α-glucosidase inhibition rate is as follows:

[0081] α-glucosidase inhibition rate (%) = [1 - (A1 - A2) / A0] × 100

[0082] In the formula: A0: blank group test value; A1: sample group test value; A2: background group test value.

[0083] (3) Cholesterol binding capacity

[0084] Fresh egg yolks were mixed with 9 times their volume of distilled water and stirred to form a homogeneous emulsion. 0.2 g of coconut oligosaccharides prepared in Examples 1-3 and Comparative Examples 1-3 were weighed and added to 25 mL of the emulsion. The mixture was shaken in a 37°C water bath for 2 hours, then centrifuged at 4000 rpm for 20 minutes. The supernatant was collected, and the absorbance at 550 nm was measured using the o-phthalaldehyde method. The formula for calculating cholesterol binding capacity is as follows:

[0085] Cholesterol binding capacity (mg / g) = (M1 - M2) / M

[0086] Where: M: constant weight sample mass, g; M1: cholesterol content in the supernatant after centrifugation, mg; M2: cholesterol content in the egg yolk emulsion before adsorption, mg.

[0087] (4) Lipase inhibition ability

[0088] The coconut oligosaccharides obtained in Examples 1-3 and Comparative Examples 1-3 were prepared into sample solutions with a concentration of 4 mg / mL using distilled water. 40 μL of a 100 mg / mL lipase solution was mixed thoroughly with 100 μL of the sample solution. 3.0 mL of a 100 mmol / L, pH 7.0 potassium phosphate buffer solution was added, and the mixture was reacted at 30°C for 3 min. Then, 20 μL of p-nitrobenzene acetate-dimethyl sulfoxide solution was added, and the reaction was carried out for 10 min. The absorbance was measured at 405 nm. A control group was prepared using the same concentration and volume of phosphate buffer solution, and its absorbance was measured at 405 nm. The lipase inhibition rate was then calculated using the following formula:

[0089] Lipase inhibition rate (%) = (A1-A2) / A1×100

[0090] In the formula: A1: test value of the control group; A2: test value of the sample group.

[0091] The results of the above measurements are shown in Table 1 below:

[0092] Table 1

[0093]

[0094] The above results indicate that the combination of the complex enzyme preparation obtained by the specific method and the ionic liquid in Examples 1-3 of this invention, when preparing coconut oligosaccharides, can improve the yield of coconut oligosaccharides. This demonstrates that the complex enzyme preparation of this invention exhibits high stability after precipitation cross-linking and can enhance the activity of the complex enzyme. Furthermore, the synergistic effect of the complex enzyme preparation and the ionic liquid can enhance the disruption of coconut cell walls, accelerate the rate of sugar dissolution from cells, and accelerate the degradation of macromolecular polysaccharides into oligosaccharides. It can also remove impurities such as cellulose, protein, and pectin, which is beneficial for the subsequent separation and purification of coconut oligosaccharides, effectively shortening the preparation time and increasing the yield.

[0095] Furthermore, the coconut oligosaccharides prepared in Examples 1-3 of this invention exhibit better inhibitory effects on α-glucosidase than those in Comparative Examples 1-3, effectively reducing glucose absorption and thus assisting in lowering blood sugar levels. Regarding cholesterol binding capacity and lipase inhibition rate, Examples 1-3 of this invention can bind more cholesterol compared to Comparative Examples 1-3. Simultaneously, by inhibiting lipase activity, they can reduce the body's absorption of fat, thereby achieving the goal of assisting in lowering blood lipids.

[0096] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A method for preparing coconut oligosaccharides for adjuvant treatment of hyperglycemia and hyperlipidemia, characterized in that, Includes the following steps: (1) After cutting the coconut meat into pieces, mix it with water and then beat it to obtain a pulp. The pulp is subjected to high pressure homogenization to obtain a homogenate. Then, the homogenate is cooled at 2~4℃ for 6~8h and filtered to obtain a primary filtrate. (2) Add the compound enzyme preparation to the primary filtrate and shake to hydrolyze it. Centrifuge and collect the supernatant to obtain the secondary filtrate. The amount of the compound enzyme preparation added is 2% to 4% of the mass of the primary filtrate. The preparation method of the compound enzyme preparation is as follows: take a compound enzyme composed of cellulase, papain, pectinase and glucanase in a mass ratio of 7 to 10: 2 to 3: 2 to 4: 3 to 5 and add it to a phosphate buffer solution and stir to mix. Then, while stirring, add polyethylene glycol 400 to prepare a suspension. Add 1,8-bismaleimide-diethylene glycol to the suspension and stir and crosslink in an ice water bath for 2 to 4 hours. Centrifuge and collect the precipitate. Wash the precipitate 3 to 4 times to obtain the compound enzyme preparation. (3) The secondary filtrate is added to the ionic liquid for fractional extraction, the extract is collected, the extract is evaporated, and the extract is concentrated under vacuum to obtain the concentrate; the ionic liquid is prepared by mixing 1-ethyl-3-methylimidazolium lactate with an ethanol solution. (4) The concentrated liquid was adsorbed and decolorized by a macroporous adsorption resin column, eluted with deionized water, and the sugar-containing components were collected. Then, it was filtered through an ultrafiltration membrane to obtain the permeate, which was then freeze-dried to obtain coconut oligosaccharides for adjuvant treatment of hyperglycemia and hyperlipidemia.

2. The method for preparing coconut oligosaccharides for adjuvant treatment of hyperglycemia and hyperlipidemia according to claim 1, characterized in that, The mass ratio of coconut meat to water is 3~5:1; the high-pressure homogenization is carried out 2~3 times under 30~50MPa conditions.

3. The method for preparing coconut oligosaccharides for adjuvant treatment of hyperglycemia and hyperlipidemia according to claim 1, characterized in that, The concentration of the phosphate buffer solution is 18-20 mg / mL; the mass-to-volume ratio of the complex enzyme, phosphate buffer solution, and polyethylene glycol 400 is 1-2 mg: 6-8 mL: 20-30 mL; and the 1,8-bismaleimide-diethylene glycol added is 2%-3% of the mass of the suspension.

4. The method for preparing coconut oligosaccharides for adjuvant treatment of hyperglycemia and hyperlipidemia according to claim 1, characterized in that, The centrifugation conditions for collecting the supernatant are: rotation speed 3500-4000 r / min, time 20-25 min.

5. The method for preparing coconut oligosaccharides for adjuvant treatment of hyperglycemia and hyperlipidemia according to claim 1, characterized in that, The volume ratio of the secondary filtrate to the ionic liquid is 5~7:1~2.

6. The method for preparing coconut oligosaccharides for adjuvant treatment of hyperglycemia and hyperlipidemia according to claim 1, characterized in that, The mass concentration of the ethanol solution is 70%~80%; the volume ratio of 1-ethyl-3-methylimidazolium lactate to the ethanol solution is 3~5:1~2.

7. The method for preparing coconut oligosaccharides for adjuvant treatment of hyperglycemia and hyperlipidemia according to claim 1, characterized in that, The pore size of the ultrafiltration membrane is 3000~5000 Da.

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