A composite polysaccharide, a composite ginger polysaccharide, and a preparation method and use thereof

Abstract

This invention discloses a complex polysaccharide, a complex ginger polysaccharide, its preparation method, and its uses. The monosaccharide composition of the complex polysaccharide includes galactose, xylose, and arabinose; the complex polysaccharide includes a first polysaccharide and a second polysaccharide; the main chain of the first polysaccharide is β-(1-4)-xylan, and the side chains include α-(1-3)-arabinogalactan and arabinogalactan, linked at the C-2 or C-3 position of the xylose residue in the main chain; the main chain of the second polysaccharide is β-(1-3)-galactan, and the side chains include terminal arabinose and β-(1-6)-galactan, linked at the C-6 position of the galactose residue in the main chain. The complex ginger polysaccharide provided in this application has the effect of regulating individual food intake, improving individual weight and fat volume, and improving obesity.

Description

Technical Field

[0001] This invention belongs to the field of polysaccharide development and preparation, specifically relating to a complex polysaccharide, a complex ginger polysaccharide, a complex ginger polysaccharide containing the aforementioned complex polysaccharide, and its preparation method and uses. Background Technology

[0002] Obesity is a nutritional and metabolic disorder and a contributing factor to many chronic diseases. How to prevent and control obesity is a global research hotspot. Soluble polysaccharides and other dietary fibers have effects such as improving blood lipids, aiding weight loss, and promoting bowel movements.

[0003] Ginger is one of the most widely used spices in the world and is a plant resource that can be used for both medicinal and culinary purposes. Ginger contains functional components such as ginger essential oil, essential resins, gingerol, and dietary fiber, which have effects such as lowering blood lipids, anti-oxidation, and enhancing immunity. However, due to the low content of effective substances and its poor taste, ginger's high medicinal effects cannot be fully realized.

[0004] How to separate and identify the effective substances in ginger is a technical problem that needs to be solved in this field. Summary of the Invention

[0005] To address the shortcomings of existing technologies, one objective of this invention is to provide a complex polysaccharide, wherein the monosaccharide composition of the polysaccharide includes galactose, xylose, and arabinose.

[0006] The main chain of the first polysaccharide is β-(1-4)-xylan, and the side chains include α-(1-3)-arabinan and arabinogalactan, which are linked at the C-2 or C-3 position of the xylose residues in the main chain.

[0007] The second polysaccharide backbone is β-(1-3)-galactan, and the side chains include terminal arabinose and β-(1-6)-galactan, which are linked at the C-6 position of the galactose residues in the backbone.

[0008] It should be noted that the main components of the complex polysaccharide defined in this application include galactose, xylose and arabinose, as well as a small amount of other polysaccharides, including glucosamine, rhamnose, glucuronic acid and galacturonic acid. However, since the content of the other polysaccharides is small and there are no obvious linkage signals in the two-dimensional heteronuclear multi-bond correlation spectrum HMBC and the two-dimensional homonuclear correlation spectrum NOESY, it is considered that there are few or no linkages.

[0009] Based on the correlation spectral signals of the two-dimensional heteronuclear multi-bond correlation spectrum HMBC and the two-dimensional homonuclear correlation spectrum NOESY, galactose, xylose, and arabinose may be linked in the following manner:

[0010] The complex polysaccharide includes a first polysaccharide and a second polysaccharide; the main chain of the first polysaccharide is xylan linked by β-(1-4)-glycosidic bonds, and the side chain branches include arabinogalactan or arabinogalactan linked by α-(1-3)-glycosidic bonds, with the linkage point at the C-2 or C-3 position of the main chain; the main chain of the second polysaccharide is galactan linked by β-(1-3)-glycosidic bonds, and the side chain mainly includes galactan linked by β-(1-6)-glycosidic bonds, with the terminal sugar residue of the side chain branch being an arabinose residue, and the linkage point at the C-6 position of the main chain.

[0011] In the complex polysaccharide provided in this application, the monosaccharide composition can be determined by pre-column derivatization of the complex polysaccharide by methylation, followed by qualitative and quantitative analysis by reversed-phase high-performance liquid chromatography.

[0012] The complex polysaccharide provided in this application can be identified using two-dimensional heteronuclear multi-bond correlation spectroscopy (HMBC) and two-dimensional homonuclear correlation spectroscopy (NOESY). The complex polysaccharide described in this application exhibits a correlation signal peak in the HMBC spectrum of β-(1-4)-xylan self-linking, with the linking site located at position C-4, and side chain branches at positions C-3 and C-2; it also exhibits a correlation signal peak of α-(1-3)-arabinose self-linking and a correlation signal peak of galactarabinan, with the linking site located at position C-2 or C-3 of β-(1-4)-xylan. Furthermore, the composite polysaccharide described in this application also exhibits a related signal peak in the two-dimensional homonuclear correlation spectrum NOESY, which shows the self-linking of β-(1-3)-galactose with the linking site located at C-3 and a side chain branch at C-6; there is also a self-linking of β-(1-6)-galactose with the linking site located at C-6 of β-(1-3)-galactose; and there is a related signal peak showing the linking of terminal arabinose residues to galactan side chains with the linking site located at C-6 of β-(1-6)-galactose.

[0013] This application provides a complex polysaccharide that differs from existing polysaccharides, possessing the effect of regulating food intake and improving obesity. Specifically, the complex polysaccharide described in this application can significantly improve diet-induced individual weight and fat volume.

[0014] Preferably, the monosaccharide composition of the polysaccharide, in molar percentage, mainly includes: 35.0-40.0% galactose, 15.0-20.0% xylose, and 35.0-40.0% arabinose.

[0015] Preferably, the molecular weight of the complex polysaccharide is 105-115 kDa, such as 107 kDa, 109 kDa, 110 kDa, 113 kDa, etc.

[0016] Preferably, the monosaccharide composition further includes any one or at least two of glucosamine, rhamnose, glucuronic acid, and galacturonic acid.

[0017] Preferably, the monosaccharide composition of the polysaccharide, by molar percentage, further includes: 0.5-1.0% glucosamine, 2.0-4.0% rhamnose, 2.0-4.0% glucuronic acid, and 1.0-3.0% galacturonic acid.

[0018] As an optional specific embodiment, the monosaccharide composition of the polysaccharide, by molar percentage, is as follows: glucosamine 0.50-0.60%, rhamnose 3.00-3.40%, glucuronic acid 2.50-3.00%, galacturonic acid 1.10-1.50%, galactose 37.00-38.50%, xylose 18.00-18.40%, and arabinose 36.00-36.40%.

[0019] The second objective of this application is to provide a method for preparing a compound ginger polysaccharide, comprising the following steps:

[0020] (1) Using young ginger as raw material, remove its pigments and lipids to obtain ginger powder;

[0021] (2) The ginger powder is enzymatically hydrolyzed and then extracted with water to obtain a ginger polysaccharide extract;

[0022] (3) The ginger polysaccharide extract is subjected to alcohol precipitation, water redissolved, and small molecule impurities are removed. After water removal, a compound ginger polysaccharide is obtained, wherein the compound ginger polysaccharide contains one of the compound polysaccharides described in the present invention.

[0023] The term "young ginger" in this application refers to the tender shoots at the tip of the rhizome of ginger (Zingiber officinale), a plant in the ginger family.

[0024] As a preferred option, the young ginger described in this application is selected from the small yellow ginger of Weiyuan, Sichuan.

[0025] The compound ginger polysaccharide obtained by the preparation method of this application also has the effects of regulating food intake, improving individual weight and fat volume, and improving obesity.

[0026] Preferably, the content of the compound ginger polysaccharide in the compound ginger polysaccharide is 70wt% to 80%.

[0027] Preferably, the starch content of the young ginger described in this application is ≤0.65wt%.

[0028] More preferably, the young ginger comprises 0.20-0.35 wt% crude fat, 0.70-1.23 wt% crude fiber, 0.90-1.58 wt% crude protein, 0.98-1.72 wt% total sugar, 0.64-1.12 wt% starch, and the balance being moisture.

[0029] Using young ginger as raw material to extract the complex polysaccharide described in this application has the advantages of short growth cycle of raw material (young ginger), high content of complex polysaccharide, and good functional effects.

[0030] Preferably, the enzyme used in step (2) for enzymatic hydrolysis is a complex enzyme composed of mesophilic amylase, neutral cellulase and neutral protease.

[0031] Preferably, in the complex enzyme, the mesophilic amylase is 35wt%–45wt%, the neutral cellulase is 25wt%–35wt%, and the neutral protease is 25wt%–35wt%; the sum of all components of the complex enzyme is 100wt%.

[0032] The compound enzyme used in this application is a combination of medium-temperature amylase, neutral cellulase and neutral protease, which can decompose starch polysaccharides to remove impurities, destroy the fiber structure and promote the dissolution of heteropolysaccharides; and the appropriate ratio can increase the content of the compound polysaccharide mentioned in one of the objectives in the compound ginger polysaccharide to a greater extent.

[0033] For example, the activity of the mesophilic amylase is 1000 U / g to 2000 U / g, preferably 1000 U / g to 1500 U / g; the activity of the neutral cellulase is 5000 U / g to 10000 U / g, preferably 5000 to 6000 U / g; and the activity of the neutral protease is 5000 U / g to 10000 U / g, preferably 5000 to 6000 U / g.

[0034] As a further preferred embodiment, the mass ratio of mesophilic amylase, neutral cellulase and neutral protease in the complex enzyme is 4:3:3.

[0035] Preferably, in the enzymatic hydrolysis process described in step (2), the amount of compound enzyme added is 1.5 to 2 g of compound enzyme per gram of ginger powder.

[0036] Excessive use of compound enzymes will reduce the content of compound polysaccharides in compound ginger polysaccharides, while insufficient use will lead to increased starch impurities and reduced compound polysaccharide content.

[0037] Preferably, the enzymatic hydrolysis process in step (2) involves first dispersing ginger powder in water, then adding the compound enzyme, and performing the enzymatic hydrolysis reaction under ultrasound. After inactivation, the supernatant of the enzymatic hydrolysate is the ginger polysaccharide extract.

[0038] Preferably, the ultrasonic power is 300-350W, the temperature is 55-65℃, and the time is 55-65min.

[0039] The ultrasound described in this application can promote a more thorough enzymatic hydrolysis reaction. However, if the ultrasound power is too high, the temperature is too high, or the time is too long, the content of complex polysaccharides in the complex ginger polysaccharide will decrease. This may be because high-power ultrasound causes problems such as enzyme inactivation. Similarly, if the ultrasound power is too low, it will also lead to a decrease in the extraction rate of complex polysaccharides.

[0040] Preferably, the solvent for alcohol precipitation is ethanol with a purity of 95% or higher.

[0041] Preferably, during the alcohol precipitation process, the volume ratio of the ginger polysaccharide extract to the alcohol precipitation solvent is 1:2.8 to 1:3.2, for example, 1:2.9, 1:3.0, 1:3.1, etc.

[0042] Preferably, the alcohol precipitation process is as follows: after mixing the ginger polysaccharide extract with the alcohol precipitation solvent, the mixture is magnetically stirred for 25 min to 40 min, then allowed to stand and refrigerated to obtain ginger polysaccharide precipitate.

[0043] Preferably, the method for removing small molecule impurities involves re-dissolving the ginger polysaccharide precipitate in water and then removing the small molecule impurities using an ultrafiltration membrane with a molecular weight cutoff of 3000 Da.

[0044] Preferably, step (4) is performed after step (3): the compound ginger polysaccharide is purified by ion exchange chromatography, and the substance with a molecular weight of 105-115 kDa is retained, which is the compound polysaccharide of one of the purposes.

[0045] Preferably, the ion exchange chromatography purification step is as follows: the compound ginger polysaccharide is dissolved in water, eluted with a DEAE-Sepharose Fast FLow gel column using a 3BV 0.2mol / L NaCl solution, the eluent is precipitated with 80v% ethanol solution, and the precipitate is the first product after sodium chloride is removed.

[0046] Preferably, the method for retaining substances with a molecular weight cutoff of 105-115 kDa includes either gel chromatography purification or ultrafiltration membrane purification.

[0047] The method for determining the molecular weight cutoff can be any known or new technique that is known to those skilled in the art.

[0048] Preferably, the compound ginger polysaccharide contains 86wt% to 93wt% of the compound polysaccharide, and the purity of the obtained compound polysaccharide is 93% to 96%.

[0049] The third objective of this application is to provide a use of the complex polysaccharide as described in the first objective, wherein the complex polysaccharide is used in any one or at least two of the following: foods, pharmaceuticals, or health products for reducing body fat content and / or reducing weight.

[0050] Compared with the prior art, this application has the following beneficial effects:

[0051] The compound polysaccharide provided in this application and the compound ginger polysaccharide prepared by the preparation method have the effects of regulating individual food intake, improving individual weight and fat volume, and improving obesity. Attached Figure Description

[0052] Figure 1 The HMBC pattern of the complex polysaccharide provided in Example 1;

[0053] Figure 2 NOESY spectrum of the complex polysaccharide provided in Example 1;

[0054] Figure 3 The trend of body weight change in mice in different treatment groups over 8 weeks in an experiment on high-fat diet-induced obesity in mice;

[0055] Figure 4 The total weight gain of different treatment groups in the high-fat diet-induced obesity mouse experiment;

[0056] Figure 5 Comparison of animal morphology and white adipose volume in different treatment groups of mice in an experiment induced by a high-fat diet to become obese.

[0057] Figure 6 White adipose tissue and muscle mass in different treatment groups of mice induced to be obese by a high-fat diet.

[0058] Figure 7 Food intake and energy intake in each treatment group of mice in an experiment to induce obesity in mice with a high-fat diet;

[0059] Figure 8 Example images of HE-stained tissue sections from various treatment groups in an experiment on mice with high-fat diet-induced obesity.

[0060] Figure 9 for Figure 8 A statistical chart of adipocyte size in HE-stained tissue pathological sections. Detailed Implementation

[0061] The technical solution of the present invention will be further described in detail below with reference to specific embodiments. The following embodiments are merely illustrative and explanatory of the present invention and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are covered within the scope of protection intended by the present invention.

[0062] Preparation Example 1

[0063] A method for preparing a complex polysaccharide includes the following steps:

[0064] (1) Using young ginger from Weiyuan, Sichuan as raw material, peel and slice the ginger, and dry it at a low temperature of 35℃ in an electric heating drying oven for 24-48 hours; after drying, the ginger slices are ground into powder and passed through a 60-mesh sieve to obtain crude ginger; the crude ginger is dissolved in 95% ethanol solution (V:V = 1:5), and refluxed twice in a water bath at 70℃ for 2 hours each time to remove pigments and lipids from the ginger; centrifuged at 10000r / min for 10min, the precipitate is taken and freeze-dried to obtain ginger powder;

[0065] The composition of the young ginger of Weiyuan small yellow ginger in Sichuan is as follows: crude fat 0.20wt%, crude fiber 0.70wt%, crude protein 0.90wt%, total sugar 0.98wt%, starch 0.64wt%, and moisture 95.90wt%.

[0066] (2) Weigh 1.0g of ginger sample and dissolve it in 35mL of pure water. After mixing well, add 1.8g of mixed enzyme (the mass ratio of medium-temperature amylase, neutral cellulase and neutral protease is 2:1.5:1.5). The mixture is reacted in an ultrasonic cleaner at 60℃ and 320W for 60min. After the reaction, it is transferred to a water bath at 90℃ to inactivate the enzyme for 20min. Centrifuge at 10000r / min for 30min. The supernatant is the ginger polysaccharide extract.

[0067] (3) The ginger polysaccharide extract was slowly mixed with 95% ethanol (1:3, v / v), magnetically stirred for 30 min, stored at 4℃, and centrifuged (8000 rpm / min, 20 min) after 12 hours to collect the polysaccharide precipitate; then the precipitate was redissolved in deionized water, and dialysis was performed through an ultrafiltration membrane with a molecular weight cutoff of 3000 Da to remove small molecule impurities. The precipitate after the removal of small molecules was then concentrated and freeze-dried to obtain the compound ginger polysaccharide; the extraction rate of the compound ginger polysaccharide was calculated to be approximately 5.18%;

[0068] (4) The compound ginger polysaccharide (100 mg) was dissolved in 15 mL of distilled water and passed through a DEAE-Sepharose FastFLow gel column (2.5 × 12 cm). Elution was performed using 0.2 mol / L NaCl solution (3 BV). The eluent was collected, and ethanol was added to the eluent until the ethanol concentration reached 80%. After stirring and mixing, the mixture was stored at 4°C for 12 h to obtain a polysaccharide precipitate. The precipitate was collected by centrifugation (10000 rpm / min, 20 min), and dialyzed (molecular weight cutoff 1000 Da) to remove low molecular weight substances. The precipitate was lyophilized to obtain the preliminarily purified compound polysaccharide. Then, Chromdex was used to precipitate the polysaccharide. The preliminarily purified complex polysaccharide was further purified by 200PG gel filtration chromatography. Purification conditions included dissolving the polysaccharide sample in an appropriate amount of dH2O, centrifuging, and filtering through a 0.45μm microporous membrane. The filtrate was loaded onto a pre-equilibrated gel filtration column at 1% of the column volume. After all the sample solution had entered the column, ultrapure water was used for elution at a flow rate of 1.8 ml / min. An automatic fraction collector was used to collect 4.5 ml of eluent per tube, eluting one column volume. Online detection was used to collect eluent from the same elution peak region with high peak height and good symmetry. This process was repeated multiple times to enrich the purified polysaccharide. The combined eluent fractions were concentrated under reduced pressure and freeze-dried to obtain the polysaccharide purified by gel filtration chromatography, which was the complex polysaccharide. Calculations showed that the complex ginger polysaccharide contained 93% complex polysaccharide.

[0069] High-performance gel permeation chromatography (HPGPC) was used for detection. The detection conditions were as follows: high-performance liquid chromatograph with differential detector; polymer matrix water-soluble SEC (GFC) columns (OHpak SB-803HQ, OHpak SB-804HQ, OHpak SB-805HQ, 8×300mm) in tandem; mobile phase: 0.05M NaCl solution; flow rate: 0.65 mL / min; column temperature: 40℃; injection volume: 30 μL; peak time: 34.41 min. The molecular weight of the complex polysaccharide was calculated to be 109.24 kDa based on the dextran molecular weight standard, and the purity was 96%.

[0070] Preparation of Comparative Example 1

[0071] The only difference from Preparation Example 1 is that the raw material in step (1) is old ginger from Weiyuan, Sichuan. The composition of the old ginger is: crude fat 0.40wt%, crude fiber 1.20wt%, crude protein 1.42wt%, total sugar 1.18wt%, starch 0.70wt%, and moisture 88.50wt%.

[0072] The results showed that the monosaccharide composition of the compound old ginger polysaccharide was 38.2% mannose and 61.8% glucose, with a molecular weight of 42.5 kDa. It can be seen that the polysaccharide obtained from old ginger has a completely different structure from that obtained from young ginger.

[0073] Preparation Example 2

[0074] The only difference from Preparation Example 1 is that the mixing ratio of the medium-temperature amylase, neutral cellulase and neutral protease of the complex enzyme used in step (2) is 0.35:0.30:0.35.

[0075] The test results showed that the compound ginger polysaccharide contained 87.2% compound polysaccharide, with a molecular weight of 98.50 kDa and a purity of 95%.

[0076] Preparation Example 3

[0077] The only difference from Preparation Example 1 is that the mixing ratio of the medium-temperature amylase, neutral cellulase and neutral protease of the complex enzyme used in step (2) is 0.45:0.25:0.30.

[0078] Tests showed that the compound ginger polysaccharide contained 85% compound polysaccharide, with a molecular weight of 100.30 kDa and a purity of 85%.

[0079] Preparation Example 4

[0080] The only difference from Preparation Example 1 is that the amount of the complex enzyme used in step (2) is 2g.

[0081] The test results showed that the compound ginger polysaccharide contained 90.6% compound polysaccharide, with a molecular weight of 102.40 kDa and a purity of 92%.

[0082] Preparation Example 5

[0083] The only difference from Preparation Example 1 is that the amount of the complex enzyme used in step (2) is 1.5g.

[0084] The test results showed that the compound ginger polysaccharide contained 91.30% compound polysaccharide, with a molecular weight of 100.90 kDa and a purity of 93%.

[0085] Example of detection:

[0086] (1) Monosaccharide composition:

[0087] The complex polysaccharides obtained in the preparation example were hydrolyzed into a mixture of monosaccharides, and the specific operations were as follows:

[0088] 2 mg of sample was hydrolyzed with 2 mol / L trifluoroacetic acid (TFA) at 110 °C for 8 h in a closed system. After hydrolysis, the sample was transferred to an EP tube and methanol was added for co-distillation (4 times) to remove excess TFA.

[0089] The monosaccharide mixture was then derivatized pre-column. The dried sample hydrolysate and the mixed standard of monosaccharides were dissolved in 200 μL of sodium hydroxide solution (0.3 mol / L), and then 240 μL of PMP methanol solution (0.5 mol / L) was added at 70 °C and reacted for 1 h. After cooling, 240 μL of HCl solution (0.3 mol / L) was added to neutralize the reaction system, and then 1 mL of dichloromethane was added for extraction three times to remove excess PMP. Finally, the mixture was centrifuged at 8000 rpm / min for 5 min, and the supernatant was filtered through a 0.22 μm nylon filter membrane to obtain the derivatized monosaccharide mixture.

[0090] The monosaccharide mixtures in the derivatized samples were analyzed using reversed-phase high-performance liquid chromatography (RP-HPLC) under the following conditions: an Agilent XDB-C18 column (4.6 mm × 250 mm) and a UV detector. A phosphate buffer (pH 6.7) and acetonitrile mixture (83:17, V:V) was used as the mobile phase at a flow rate of 1.0 mL / min, with an injection volume of 20 μL. The column temperature was 35 °C, and the absorbance was set to 245 nm. Data analysis was performed using LC solution software (Shimadzu Scientific Instruments).

[0091] The test results are as follows:

[0092] In the complex polysaccharide prepared in Example 1, the molar ratio of glucosamine, rhamnose, glucuronic acid, galacturonic acid, galactose, xylose, and arabinose was 0.535:3.184:2.715:1.360:37.739:18.182:36.284.

[0093] The complex polysaccharides prepared in Examples 2-5 have similar monosaccharide content to those prepared in Example 1, and in molar ratio include 0.50-0.60% glucosamine, 3.00-3.40% rhamnose, 2.50-3.00% glucuronic acid, 1.10-1.50% galacturonic acid, 37.00-38.50% galactose, 18.00-18.40% xylose, and 36.00-36.40% arabinose.

[0094] (2) Polysaccharide molecular structure

[0095] The composite polysaccharide provided in Preparation Example 1 was dissolved in 500 μL of D2O (99.6%) at a concentration of 10 mg and then lyophilized three times. Two-dimensional heteronuclear multi-bond correlation spectroscopy was performed using a Bruker 600 MHz NMR spectrometer. The chemical shifts of all carbon and hydrogen atoms in the composite polysaccharide, combined with the 1D and 2D NMR spectra, are shown in Table 1.

[0096] Table 1. Chemical shift assignments of the complex polysaccharides.

[0097]

[0098]

[0099] Note: — indicates no signal was detected.

[0100] Figure 1 The HMBC pattern of the complex polysaccharide provided in Preparation Example 1 is given.

[0101] Figure 2 The NOESY spectrum of the complex polysaccharide provided in Preparation Example 1 is given.

[0102] It should be noted that, Figure 1 and Figure 2 The main information is in the middle part. The peaks in the middle and on both sides do not affect the reading of the data, so some peaks are not fully captured.

[0103] From Table 1, Figure 1 and Figure 2 It can be inferred that the complex polysaccharide obtained in Preparation Example 1 is a mixture of two polysaccharides, most likely specifically:

[0104] The structure of the first polysaccharide is:

[0105]

[0106] The structure of the second polysaccharide is as follows:

[0107]

[0108] The composite polysaccharides provided in Preparation Examples 2 to 5 were tested according to the testing steps for polysaccharide molecular structure. Combined with the spectrum, a signal distribution similar to that of the composite polysaccharide in Preparation Example 1 was obtained, and the same structure as the composite polysaccharide in Preparation Example 1 was inferred.

[0109] (3) High-fat diet-induced obesity in mice

[0110] The complex polysaccharides obtained from Preparation Example 1 (young ginger YGP) and Preparation Comparative Example 1 (old ginger OGP) were used to conduct experiments on high-fat diet-induced obesity in mice.

[0111] Male C57BL / 6 mice (7 weeks old) were randomly divided into four groups (4 mice per cage, n=12 per group) after one week of acclimatization: a standard diet group (NCD), a high-fat diet group (HFD), a high-fat diet group administered with a combination of ginger polysaccharides via gavage (HFD+YGP), and a high-fat diet group administered with a combination of old ginger polysaccharides via gavage (HFD+OGP) (for testing white adipose tissue in the groin only). Food intake and body weight were monitored weekly. After 8 weeks, the mice were sacrificed under isoflurane anesthesia, and white adipose tissue (WAT) was collected.

[0112] like Figure 3 ( Figure 3 As shown in the trend of body weight change in mice in different treatment groups over 8 weeks in the high-fat diet-induced obesity experiment, the body weight of mice in the HFD group was significantly higher than that in the NCD group starting from week 2, while the body weight of mice in the GP treatment group began to increase significantly after week 4.

[0113] At the end of the experiment, such as Figure 4 ( Figure 4 The total weight gain of different treatment groups in the high-fat diet-induced obesity mouse experiment is shown to be more than twice that of the NCD control group.

[0114] like Figure 5 ( Figure 5 (Comparison of animal morphology and white adipose volume in different treatment groups in the high-fat diet-induced obesity mouse experiment) As shown, after 8 weeks, the obesity morphology of the animals in the HFD+GP treatment group was significantly improved, and the weight gain was 120% and 80% of that in the NCD group and the HFD group, respectively.

[0115] like Figure 6 ( Figure 6 In experiments on high-fat diet-induced obesity in mice, the addition of compound ginger polysaccharide (GP) improved the increase in white adipose tissue (WAT) mass induced by the high-fat diet in each treatment group, while there was no significant difference in skeletal muscle mass among the groups, and no significant difference in skeletal muscle weight.

[0116] like Figure 7 ( Figure 7 The figures show the food intake and energy intake of each treatment group in the high-fat diet-induced obesity experiment in mice. Compared with the HFD group, the HFD+GP treatment group had reduced food intake and energy intake, while the energy intake was the same as that of the NCD group. HFD+GP treatment can improve the obesity status of mice by regulating food intake.

[0117] The compound ginger polysaccharide provided in this application has a significant effect on improving the body weight and fat volume of diet-induced obese mice.

[0118] like Figure 8 ( Figure 8(Example images of HE-stained histopathological sections from different treatment groups in an experiment to induce obesity in mice using a high-fat diet) Figure 9 ( Figure 9 As shown in the statistical chart of adipocyte size in HE-stained tissue pathological sections, the YGP group was significantly lower than that of HFD and OGP, and close to the NCD level. It can be inferred that compared with old ginger polysaccharide, the compound ginger polysaccharide obtained from young ginger can improve the enlargement of adipocytes caused by a high-fat diet.

[0119] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A complex polysaccharide, characterized in that, The monosaccharide composition of the polysaccharide includes: galactose, xylose, and arabinose; The complex polysaccharide includes a first polysaccharide and a second polysaccharide; The main chain of the first polysaccharide is β-(1-4)-xylan, and the side chains include α-(1-3)-arabinan and arabinogalactan, which are linked at the C-2 or C-3 position of the xylose residues in the main chain. The second polysaccharide backbone is β-(1-3)-galactan, and the side chains include terminal arabinose and β-(1-6)-galactan, which are linked at the C-6 position of the galactose residues in the backbone. According to molar percentage, the monosaccharide composition of the polysaccharide mainly includes: galactose 35.0~40.0%, xylose 15.0~20.0%, and arabinose 35.0~40.0%. The molecular weight of the complex polysaccharide is 105~115 kDa; The monosaccharide composition also includes glucosamine, rhamnose, glucuronic acid, and galacturonic acid; According to molar percentage, the monosaccharide composition of the polysaccharide further includes: glucosamine 0.5~1.0%, rhamnose 2.0~4.0%, glucuronic acid 2.0~4.0%, and galacturonic acid 1.0~3.0%.

2. A method for preparing a complex ginger polysaccharide, characterized in that, The preparation method includes the following steps: (1) Using young ginger from Weiyuan, Sichuan as raw material, remove its pigments and lipids to obtain ginger powder; (2) The ginger powder is enzymatically hydrolyzed and then extracted with water to obtain a ginger polysaccharide extract; (3) The ginger polysaccharide extract is subjected to alcohol precipitation, water redissolved, and small molecule impurities are removed. After water removal, a compound ginger polysaccharide is obtained, which contains the compound polysaccharide described in claim 1. (4) The compound ginger polysaccharide is purified by ion exchange chromatography, and the molecular weight cutoff is 105~115kDa, which is the compound polysaccharide of claim 1; the ion exchange chromatography purification steps are as follows: the compound ginger polysaccharide is dissolved in water, and eluted with 0.2mol / L NaCl solution at 3BV using a DEAE-Sepharose Fast FLow gel column. After the eluent is precipitated with 80v% ethanol solution, the precipitate is the first product after sodium chloride is removed.

3. The production method according to claim 2, wherein In the compound ginger polysaccharide, the content of the compound polysaccharide according to claim 1 is 70wt%~80wt%.

4. The production method according to claim 2, wherein The starch content of the young ginger is ≤0.65wt%.

5. The production method according to claim 2, wherein The young ginger is composed of 0.20-0.35 wt% crude fat, 0.70-1.23 wt% crude fiber, 0.90-1.58 wt% crude protein, 0.98-1.72 wt% total sugar, 0.64-1.12 wt% starch, and the balance being moisture.

6. The production method according to claim 2, wherein The enzyme used in step (2) is a complex enzyme composed of medium-temperature amylase, neutral cellulase and neutral protease.

7. The production method according to claim 6, wherein In the complex enzyme, the amount of mesophilic amylase is 35wt%~45wt%, the amount of neutral cellulase is 25wt%~35wt%, and the amount of neutral protease is 25wt%~35wt%.

8. The production method according to claim 6, wherein In step (2), the amount of compound enzyme added is 1.5~2g per gram of ginger powder.

9. The production method according to claim 6, wherein The enzymatic hydrolysis process in step (2) involves first dispersing ginger powder in water, then adding the compound enzyme, and performing the enzymatic hydrolysis reaction under ultrasound. After inactivation, the supernatant of the enzymatic hydrolysate is the ginger polysaccharide extract.

10. The production method according to claim 9, wherein The ultrasonic power is 300~350W, the temperature is 55~65℃, and the time is 55~65min.

11. The production method according to claim 2, wherein The solvent for the alcohol precipitation is ethanol with a purity of 95% or higher.

12. The production method according to claim 2, wherein During the alcohol precipitation process, the volume ratio of the ginger polysaccharide extract to the alcohol precipitation solvent is 1:2.8 to 1:3.

2.

13. The production method according to claim 2, wherein The alcohol precipitation process is as follows: the ginger polysaccharide extract is mixed with the alcohol precipitation solvent, magnetically stirred for 25 min to 40 min, allowed to stand, and then refrigerated to obtain ginger polysaccharide precipitate.

14. The production method according to claim 2, wherein The method for removing small molecule impurities involves re-dissolving the ginger polysaccharide precipitate in water and then removing the small molecule impurities using an ultrafiltration membrane with a molecular weight cutoff of 3000 Da.

15. The production method according to claim 2, wherein The method for retaining substances with a molecular weight cutoff of 105-115 kDa includes either gel chromatography purification or ultrafiltration membrane purification.

16. Use of the complex polysaccharide according to claim 1 or the complex ginger polysaccharide prepared by the method according to any one of claims 2-15, characterized in that, The complex polysaccharide and / or complex ginger polysaccharide are used to prepare any one or at least two of the following: food, medicine or health products that reduce body fat content and / or reduce weight.

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