Luzhou-flavor distiller's grains water-soluble glucose oligosaccharide as well as preparation method and application thereof

Abstract

The invention discloses Luzhou-flavor distiller's grains water-soluble glucose oligosaccharide as well as a preparation method and application thereof, and belongs to the technical field of medicines. The method comprises the following steps: extracting water-soluble glucose oligosaccharide from Luzhou-flavor distiller's grains serving as a raw material; the structure main chain of the Luzhou-flavor distiller's grains water-soluble glucose oligosaccharide is composed of glucose, and the branch chain comprises arabinose and glucose; the arabinose powder is prepared from arabinose, galactose, glucose and xylose, and the molar ratio of the arabinose to the galactose to the glucose to the xylose is 8.81: 2.54: 76.82: 6.90. The water-soluble glucose oligosaccharide prepared by the invention can obviously improve the symptom of alcoholic liver, and has the effects of inhibiting liver injury and improving the activity of liver antioxidant enzyme. In addition, the Luzhou-flavor vinasse is a Baijiu fermentation byproduct, the raw material yield is high, the cost is low, and the resource utilization value of the vinasse can be improved.

Description

Technical Field

[0001] The invention belongs to the technical field of medicines, and in particular relates to a Luzhou-flavor liquor lees water-soluble glucose oligosaccharide and a preparation method and application thereof. Background Art

[0002] Alcoholic liver disease refers to liver damage caused by long-term excessive drinking. The liver is affected by the metabolism of alcohol and shows varying degrees of liver cell damage, which can develop into cirrhosis or liver cancer in severe cases. At present, the treatment of alcoholic liver disease mainly includes the use of silymarin drugs, corticosteroids, immunomodulatory drugs, biologics and small molecule drugs. These drugs can relieve symptoms to a certain extent, but there is a risk of side effects. Therefore, the development of new drugs and auxiliary therapies to relieve alcoholic liver disease is an urgent problem that the current medical community needs to solve.

[0003] Luzhou-flavor liquor lees contain a variety of bioactive ingredients, such as polyphenols, amino acids, peptides, oligosaccharides, etc., which have high potential resource utilization value. Oligosaccharides have the characteristics of small molecular weight, easy absorption by cells, and strong activity, which have attracted the attention of many researchers. However, there are relatively few studies on the extraction of water-soluble glucose oligosaccharides from liquor lees, and the research on its structure and function is not in-depth enough, and there is a lack of research on its ability to alleviate alcoholic liver disease.

[0004] Exploring the activity of water-soluble oligosaccharides in distiller's grains and exploring their potential biological activities will be beneficial to the resource utilization of distiller's grains. Summary of the invention

[0005] In view of the above technical problems, the present invention provides a Luzhou-flavor liquor lees water-soluble glucose oligosaccharide and a preparation method and application thereof.

[0006] The technical solution adopted by the present invention to solve its technical problem is:

[0007] In a first aspect, the present invention provides a water-soluble glucose oligosaccharide from Luzhou-flavor liquor lees, wherein the structural main chain of the glucose oligosaccharide is composed of glucose, and the side chains include arabinose and glucose;

[0008] The sugar chain structure of the glucose oligosaccharide is shown in Formula 1:

[0009]

[0010] The water-soluble glucose oligosaccharide from Luzhou-flavor liquor lees is composed of arabinose, galactose, glucose and xylose, and the molar ratio thereof is 8.81:2.54:76.82:6.90.

[0011] The molecular weight of the water-soluble glucose oligosaccharide from the Luzhou-flavor liquor lees is 2800-3000Da.

[0012] In a second aspect, the present invention provides a method for preparing the above-mentioned Luzhou-flavor liquor lees water-soluble glucose oligosaccharide, comprising the following steps:

[0013] (1) mixing Luzhou-flavor liquor lees with deionized water, performing ultrasonic pretreatment to obtain a mixed solution, then adding amylase and protease to fully react, centrifuging, filtering, and obtaining a supernatant;

[0014] (2) dialyzing the supernatant obtained in step (1) with a dialysis bag, taking the dialyzate, concentrating it by rotary evaporation, and then ultrafiltration and retentate, taking the retentate, concentrating it by rotary evaporation, taking the concentrate and mixing it with ethanol for sufficient reaction, centrifuging, and freeze-drying to obtain crude oligosaccharides;

[0015] (3) The crude oligosaccharides obtained in step (2) are redissolved in water, and eluted with water at a flow rate of 0.5 mL / min using a DEAE-52 cellulose chromatography column; the eluted portion is further separated by a Sephacryl G25 chromatography column, and a large molecular weight portion is collected to obtain Luzhou-flavor liquor lees water-soluble glucose oligosaccharides.

[0016] In step (1) of the above preparation method, the moisture content of the Luzhou-flavor liquor lees is 57-60wt%, the crude protein content is 14.25-16.31wt% dry weight, the starch content is 8.80-10.20wt% dry weight, and the cellulose content is 10.25-12.55wt% dry weight.

[0017] In step (1) of the above preparation method, the solid-liquid ratio of Luzhou-flavor liquor lees to deionized water is 1 g:10 mL; the amount of amylase added is 600-800 U / mL of the mixed solution; and the amount of protease added is 600-1000 U / mL of the mixed solution.

[0018] In step (1) of the above preparation method, the ultrasonic pretreatment condition is 5kW ultrasound for 20 to 35 minutes.

[0019] In step (1) of the above preparation method, the reaction condition is 45° C. for 1 h.

[0020] In step (1) of the above preparation method, the centrifugation conditions are 5000 rpm and 25° C. for 20 min.

[0021] In step (2) of the above preparation method, the retentate is a component with a mass less than 5000Da; the volume of the concentrate is 1 / 4 of the volume of the retentate, and the volume ratio of the concentrate to ethanol is 1:3.

[0022] In step (2) of the above preparation method, the molecular weight cut-off of the dialysis bag is 300 Da and the dialysis time is 48 h.

[0023] In step (2) of the above preparation method, the reaction conditions are alcohol precipitation reaction at 4°C for 12 hours.

[0024] In step (2) of the above preparation method, the centrifugation conditions are 5000 rpm and 4°C for 20 min.

[0025] In step (3) of the above preparation method, the purity of the Luzhou-flavor liquor lees water-soluble glucose oligosaccharide reaches 83% or above.

[0026] In a third aspect, the present invention provides a pharmaceutical composition, comprising the above-mentioned Luzhou-flavor liquor lees water-soluble glucose oligosaccharide, or the Luzhou-flavor liquor lees water-soluble glucose oligosaccharide prepared by the above-mentioned preparation method.

[0027] In a fourth aspect, the present invention provides the use of the above-mentioned Luzhou-flavor liquor lees water-soluble glucose oligosaccharide, or the Luzhou-flavor liquor lees water-soluble glucose oligosaccharide prepared by the above-mentioned preparation method, or the above-mentioned pharmaceutical composition in the preparation of a drug, reagent, diet or nutritional supplement having one or more of the following effects:

[0028] a. Relieve and treat alcoholic liver disease;

[0029] b. Improve the activity of antioxidant enzymes in liver tissue;

[0030] c. Regulate liver lipid metabolism;

[0031] d. Relieve and repair liver cell damage.

[0032] Furthermore, in the above-mentioned effect b, the liver tissue antioxidant enzymes are SOD, GPX and CAT.

[0033] The beneficial effects of the present invention are as follows: the method for preparing water-soluble glucose oligosaccharides from Luzhou-flavor liquor lees provided by the present invention has a high yield of crude oligosaccharides, which is greater than 28%; the purity of the obtained glucose oligosaccharides is high, which is greater than 83%, and the molecular weight is between 2800 and 3000 Da, which is suitable for large-scale production. The water-soluble glucose oligosaccharides from Luzhou-flavor liquor lees prepared by the present invention can significantly alleviate alcoholic liver disease, regulate liver lipid metabolism, and alleviate liver cell damage, and can realize the high-value utilization of liquor brewing byproducts, lees. In addition, the Luzhou-flavor liquor lees in the present invention are byproducts of liquor fermentation, with high raw material yield and low cost. The present invention can improve the resource utilization value of lees. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 It is the nuclear magnetic signal peak of the water-soluble glucose oligosaccharide from the Luzhou-flavor liquor lees in Example 5;

[0035] Figure 2 The effect of the water-soluble glucose oligosaccharide from Luzhou-flavor liquor lees on the body weight (A) and liver index (B) of mice with alcoholic liver disease in Example 6;

[0036] Figure 3Pathological observation of liver injury in Example 6 (×200 and ×400 magnifications);

[0037] Figure 4 The effect of water-soluble glucose oligosaccharides from Luzhou-flavor liquor lees on blood parameters of mice with alcoholic liver disease in Example 6;

[0038] Figure 5 This is the effect of the water-soluble glucose oligosaccharides from the Luzhou-flavor liquor lees in Example 6 on the levels of antioxidant enzymes in mice with alcoholic liver disease. DETAILED DESCRIPTION

[0039] The technical solution of the present invention can be specifically implemented in the following manner.

[0040] The present invention provides a Luzhou-flavor liquor lees water-soluble glucose oligosaccharide (EP), which is an oligosaccharide with glucose constituting a main chain and glucose and arabinose constituting a side chain, and its sugar chain structure is shown in Formula 1:

[0041] Its components are arabinose, galactose, glucose and xylose, with a molar ratio of 8.81:2.54:76.82:6.90; its molecular weight is 2800-3000Da.

[0042] The present invention also provides a method for preparing the above-mentioned Luzhou-flavor liquor lees water-soluble glucose oligosaccharide (EP), comprising the following steps:

[0043] (1) Luzhou-flavor liquor lees and deionized water were mixed at a solid-liquid ratio of 1 g:10 mL, and pretreated with 5 kW ultrasonic waves for 20 to 35 min. Oligosaccharides were then extracted by enzymatic hydrolysis. Amylase (600 to 800 U / mL of the mixed solution) and protease (600 to 1000 U / mL of the mixed solution) were added to the mixed solution at 45°C for 1 h, and then centrifuged at 5000 rpm at 25°C for 20 min, and the supernatant was filtered;

[0044] (2) The supernatant obtained in step (1) was dialyzed with a 300Da dialysis bag for 48 hours, the dialyzate was taken for rotary evaporation, the concentrated solution was taken for ultrafiltration interception, and the components less than 5000Da in the interception solution were concentrated to 1 / 4 of the original volume, anhydrous ethanol was added at a volume ratio of 1:3, and alcohol precipitation was performed at 4°C for 12 hours, followed by centrifugation at 5000rpm, 4°C for 20 minutes to obtain the precipitate, and freeze-dried to obtain crude oligosaccharides;

[0045] (3) The crude oligosaccharides obtained in step (2) are redissolved in water, and eluted sequentially with water at a flow rate of 0.5 mL / min using a DEAE-52 cellulose chromatography column; the eluted portion is further separated by a Sephadex G25 chromatography column to obtain Luzhou-flavor liquor lees water-soluble glucose oligosaccharides.

[0046] The technical solution and effects of the present invention are further illustrated below through practical examples.

[0047] Example

[0048] 1. Preparation of water-soluble glucose oligosaccharides (EP) from Luzhou-flavor liquor lees

[0049] 1. Physical and chemical properties of Luzhou-flavor liquor lees: The types of lees and the range of lees components used in all embodiments and comparative examples of the present invention are shown in Table 1.

[0050] Table 1 Index of Luzhou-flavor liquor lees

[0051]

[0052] *Based on distillers dry weight

[0053] The preparation method of Luzhou-flavor liquor lees water-soluble glucose oligosaccharides (Examples 1 to 3) specifically comprises the following steps:

[0054] (1) Mix the lees with deionized water at a solid-liquid ratio of 1 g:10 mL, pre-treat with 5 kW ultrasonic wave for 20 to 35 min, and then extract oligosaccharides by enzymatic hydrolysis. Add amylase and protease to the mixture, heat at 45 ° C for 1 h, and then centrifuge at 5000 rpm at 25 ° C for 20 min, and filter the supernatant;

[0055] (2) The supernatant obtained in step (1) was dialyzed with a 300Da dialysis bag for 48 hours, the dialyzate was taken for rotary evaporation, the concentrated solution was taken for ultrafiltration interception, and the components less than 5000Da in the interception solution were concentrated to 1 / 4 of the original volume, anhydrous ethanol was added at a volume ratio of 1:3, and alcohol precipitation was performed at 4°C for 12 hours, followed by centrifugation at 5000rpm, 4°C for 20 minutes to obtain the precipitate, and freeze-dried to obtain crude oligosaccharides;

[0056] (3) The crude oligosaccharides obtained in step (2) are redissolved in water, and eluted sequentially with water at a flow rate of 0.5 mL / min using a DEAE-52 cellulose chromatography column; the eluted portion is further separated by a Sephadex G25 chromatography column to obtain vinasse water-soluble glucose oligosaccharides (EP).

[0057] In step (1) of Example 1, ultrasonic pretreatment was performed for 20 min; the amount of amylase added was 600 U / mL of the mixed solution, and the amount of protease added was 600 U / mL of the mixed solution;

[0058] In step (1) of Example 2, ultrasonic pretreatment was performed for 25 min; the amount of amylase added was 700 U / mL of the mixed solution, and the amount of protease added was 800 U / mL of the mixed solution;

[0059] In step (1) of Example 3, ultrasonic pretreatment was performed for 35 min; the amount of amylase added was 800 U / mL of the mixed solution, and the amount of protease added was 1000 U / mL of the mixed solution;

[0060] 2. The glucose oligosaccharides (EP) prepared in Examples 1 to 3 were tested. The results are shown in Table 2.

[0061] Calculation of crude oligosaccharide yield: record the weight of enzymatically hydrolyzed vinasse M1, then weigh the weight of the extracted freeze-dried crude oligosaccharides (M2), and calculate the crude oligosaccharide yield using the formula: crude oligosaccharide yield (%) = M2 / M1*100.

[0062] Purity calculation of water-soluble glucose oligosaccharides from distiller's grains: water-soluble glucose oligosaccharides from Luzhou-flavor liquor lees purified by Sephadex G25 were taken, freeze-dried and then re-dissolved in deionized water, and the purity of the total amount of oligosaccharides was determined by the phenol-sulfuric acid method.

[0063] Table 2 Test results of water-soluble glucose oligosaccharides (EP) from Luzhou-flavor liquor lees

[0064]

[0065] Comparative Examples 1 to 3 show that the crude oligosaccharide yield obtained by the method of the present invention is relatively high, greater than 28%; the obtained Luzhou-flavor liquor lees water-soluble glucose oligosaccharide (EP) has a high purity of greater than 83%, and a molecular weight of 2800 to 3000 Da.

[0066] 2. Physical and Chemical Properties of Water-soluble Glucose Oligosaccharides (EP) from Luzhou-flavor Liquor Grains

[0067] Example 4 Experimental analysis was performed on the monosaccharide composition and methylation of water-soluble glucose oligosaccharides from Luzhou-flavor liquor grains.

[0068] The results of sugar composition analysis are shown in Table 3. The water-soluble glucose oligosaccharides in Luzhou-flavor liquor lees are composed of arabinose, galactose, glucose, and xylose, and the molar ratio is 8.81:2.54:76.82:6.90.

[0069] Table 3 Monosaccharide composition of water-soluble glucose oligosaccharides (EP) from Luzhou-flavor liquor lees

[0070]

[0071] After the water-soluble glucose oligosaccharides from Luzhou-flavor liquor lees were completely methylated through multiple methylation reactions, they were hydrolyzed, reduced and acetylated to generate partially methylated sugar alcohol acetates, which were then detected by GC-MS. According to the total ion chromatogram and each mass spectrum fragment analysis, a total of five glycosidic bonds were detected in the water-soluble glucose oligosaccharides from Luzhou-flavor liquor lees (Table 4), including →4)-Glcp-(1→, t-Glcp-(1→, →4,6)-Glcp-(1→, →3,4)-Glcp-(1→ and t-Araf-(1→), with a molar ratio (mol%) of 44.28:15.11:11.34:8.22:6.88.

[0072] Table 4 Methylation analysis of water-soluble glucose oligosaccharides (EP) from Luzhou-flavor liquor grains

[0073]

[0074]

[0075] Example 5: Nuclear magnetic resonance analysis was performed on the water-soluble glucose oligosaccharide structure of Luzhou-flavor liquor lees.

[0076] Figure 1 The one-dimensional and two-dimensional NMR spectra of water-soluble glucose oligosaccharides from Luzhou-flavor liquor lees are shown in Figure 1. From the one-dimensional NMR spectrum, it can be seen that EP has multiple signal peaks in the anomeric hydrogen and anomeric carbon regions ( Figure 1 A and B), and then through COSY ( Figure 1 C) Analyze the HH related signal peak to exclude impurity peaks, and then HSQC spectrum ( Figure 1 D) Six groups of glycosidic bond H1 / C1 signal peaks were identified, with δ5.36 / 102.85, 4.97 / 101.40, 4.64 / 98.78, 5.23 / 94.96, and 5.09 / 110.39 ppm signals respectively attributed to residue A→4)-α-D-Glcp-(1→, residue Bα-D-Glcp-(1→, residue C→4,6)-β-D-Glcp-(1→, residue D→3,4)-α-D-Glcp-(1→, residue Eα-L-Araf-(1→).

[0077] In the HSQC spectrum ( Figure 1 D), the anomeric signal at 5.36 / 102.85 ppm was assigned to the residue A→4)-α-D-Glcp-(1→. COSY spectrum ( Figure 1C) further shows related signals derived from H-1 signal, including H1 / H2 5.36 / 3.58, H2 / H3 3.58 / 3.88, H3 / H4 3.88 / 3.40, H4 / H5 3.40 / 3.38 and H5 / H6 3.38 / 3.75. Using the chemical shifts of H-2 to H-6, the corresponding chemical shifts of C-2 to C-6 of residue A were determined to be 5.36 / 102.85, 3.58 / 74.3, 3.88 / 73.26, 3.4 / 78.93, 3.38 / 72.47, 3.75 / 63.62 ppm, respectively. Similarly, the hydrogen proton and carbon signals of residues B, C, D, and E were also determined.

[0078] According to HMBC ( Figure 1 E) and NOESY ( Figure 1 F) The cross-peak data in the two spectra, the correlation signals between C-1 of residue A and H-4 of residue A and between C-1 of residue A and H-4 of residue C, can infer the existence of the unit →4)-α-D-Glcp-(1→4,6)-β-D-Glcp-(1→4)-α-D-Glcp-(1→4). This structural feature is further confirmed by the signals at 5.36 / 78.93ppm and 5.36 / 75.69ppm in the HMBC spectrum. The correlation signals between H-1 of residue B and C-6 of residue C The signal peaks (4.97 / 70.22) between the two residues can determine the existence of α-D-Glcp-(1→4,6)-β-D-Glcp-(1→. Similarly, the connection between residue D and A, residue C and residue B, and residue D and residue E were also determined. It was confirmed that EP is composed of a glucose backbone, and the substitution mainly occurs at the C-6 position of the glucose group of →4,6)-β-D-Glcp-(1→ and the C-4 position of →3,4)-α-D-Glcp-(1→, while glucose and arabinose exist in the side chains.

[0079] Example 6 Experimental analysis was conducted on the activity of water-soluble glucose oligosaccharides from Luzhou-flavor liquor grains in alleviating alcoholic liver disease.

[0080] Healthy 8-week-old female mice were selected and randomly divided into 3 groups: normal control group (NC), alcohol group (Alcohol) and Luzhou-flavor liquor lees glucose oligosaccharide (EP) group. There were 10 mice in each group. The Alcohol group and the EP group were given alcohol gavage for 30 days. The alcohol dose increased from 1.5g / kg / day on days 0-5 to 2.7g / kg / day on day 25, and gradually increased by 0.3g / kg every 5 days. WP and EP oligosaccharides were dissolved in alcohol, and water-soluble glucose oligosaccharides (EP) were administered to the mice in the EP group daily by gavage, with a gavage dose of 200mg / kg body weight. The NC group was given the same volume of water by gavage, without alcohol or oligosaccharides. The experimental timeline is as follows. Figure 2As shown in A. After an overnight fasting period, the mice were euthanized. Blood and liver tissue were collected to analyze the effects of EP on mice with alcoholic liver disease.

[0081] The results showed that EP treatment could significantly improve the symptoms of liver injury. There was no significant difference in the initial body weight of mice in each group (p>0.05, Figure 2 A). However, drinking alcohol caused mice to lose weight and increase liver weight. During the 30-day experiment, the weight of mice in the NC group increased by 14.16%, which was significantly higher than that in the alcohol group (5.39%) (p<0.05). The liver index in the alcohol group was significantly higher than that in the NC group (p<0.05). However, compared with the alcohol group, the alcoholic liver mice taking EP had a higher weight and lower liver index ( Figure 2 B). The results show that water-soluble glucose oligosaccharides (EP) from Luzhou-flavor liquor lees can alleviate the symptoms of alcoholic liver disease in mice.

[0082] The liver tissues of mice in each experimental group were examined by histopathological examination ( Figure 3 ). In the NC group, hepatocytes showed normal morphology, characterized by uniform cytoplasm and clear nuclei. In contrast, the alcohol group showed obvious signs of liver damage, including cell swelling, loss of cell borders, and cell degeneration and death. Compared with the alcohol group, the turbidity and swelling of hepatocytes in the EP group were significantly reduced. The results show that water-soluble glucose oligosaccharides from Luzhou-flavor liquor lees can repair hepatocyte damage in mice with alcoholic liver disease.

[0083] Figure 4 The effect of water-soluble glucose oligosaccharides from Luzhou-flavor liquor lees on blood parameters of mice with alcoholic liver disease (Note: different lowercase letters represent significant differences between groups, p<0.05). Alcohol-induced liver damage, early lipid metabolism disorders, further develop into liver fibrosis. Compared with the NC group, the levels of liver tissue metabolism indicators total cholesterol (TC), triglycerides (TG), and low-density lipoprotein cholesterol (LDL-C) in the serum of mice in the alcohol group were increased, while the level of high-density lipoprotein cholesterol (HDL-C) was decreased ( Figure 4 AD). At the same time, compared with the alcohol group, the levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), important indicators of liver damage, in the EP group were reduced by approximately 26.47% and 16.70%, respectively ( Figure 4 EF). The results showed that the administration of water-soluble glucose oligosaccharides (EP) from Luzhou-flavor liquor lees could regulate liver lipid metabolism and alleviate alcohol-induced liver damage.

[0084] Figure 5The effect of water-soluble glucose oligosaccharides from Luzhou-flavor liquor lees on the levels of antioxidant enzymes in mice with alcoholic liver disease (Note: different lowercase letters represent significant differences between groups, p<0.05). Alcohol intake significantly reduced the activities of liver antioxidant enzymes, including SOD, GSH-Px, and CAT, indicating that alcohol injection caused severe oxidative stress in the liver. Administration of EP at a given dose can increase the enzymatic activity levels of antioxidant enzymes. Compared with the alcohol group, water-soluble glucose oligosaccharides (EP) from Luzhou-flavor liquor lees increased the antioxidant enzyme activities of SOD, GSH-Px, and CAT by 12.74%, 31.96%, and 12.58%, respectively.

[0085] In summary, water-soluble glucose oligosaccharides (EP) from Luzhou-flavor liquor lees have the combined effects of alleviating alcoholic liver symptoms, repairing liver tissue cell damage, regulating liver lipid metabolism levels, and increasing the activity level of antioxidant enzymes, ultimately achieving the effect of alleviating alcoholic liver disease.

Claims

1. Luzhou-flavor liquor lees water-soluble glucose oligosaccharide, characterized by: The structural main chain of the glucose oligosaccharide is composed of glucose, and the side chains include arabinose and glucose; The sugar chain structure of the glucose oligosaccharide is shown in Formula 1:

2. The Luzhou-flavor liquor lees water-soluble glucose oligosaccharide according to claim 1, characterized in that: The glucose oligosaccharide is composed of arabinose, galactose, glucose and xylose, and the molar ratio thereof is 8.81:2.54:76.82:6.

90.

3. The Luzhou-flavor liquor lees water-soluble glucose oligosaccharide according to claim 1, characterized in that: The molecular weight of the glucose oligosaccharide is 2800-3000Da.

4. The method for preparing Luzhou-flavor liquor lees water-soluble glucose oligosaccharides according to any one of claims 1 to 3, characterized in that: The steps include: (1) mixing Luzhou-flavor liquor lees with deionized water, performing ultrasonic pretreatment to obtain a mixed solution, then adding amylase and protease to fully react, centrifuging, filtering, and obtaining a supernatant; (2) dialyzing the supernatant obtained in step (1) with a dialysis bag, taking the dialyzate, concentrating it by rotary evaporation, and then ultrafiltration and retentate, taking the retentate, concentrating it by rotary evaporation, taking the concentrate and mixing it with ethanol for sufficient reaction, centrifuging, and freeze-drying to obtain crude oligosaccharides; (3) The crude oligosaccharides obtained in step (2) are redissolved in water, and eluted with water at a flow rate of 0.5 mL / min using a DEAE-52 cellulose chromatography column; the eluted portion is further separated by a Sephacryl G25 chromatography column, and a large molecular weight portion is collected to obtain Luzhou-flavor liquor lees water-soluble glucose oligosaccharides.

5. The method for preparing the Luzhou-flavor liquor lees water-soluble glucose oligosaccharide according to claim 4, characterized in that: In step (1), the moisture content of the Luzhou-flavor liquor lees is 57-60wt%, the crude protein content is 14.25-16.31wt% dry weight, the starch content is 8.80-10.20wt% dry weight, and the cellulose content is 10.25-12.55wt% dry weight.

6. The method for preparing the Luzhou-flavor liquor lees water-soluble glucose oligosaccharide according to claim 4, characterized in that: In step (1), the solid-liquid ratio of Luzhou-flavor liquor lees to deionized water is 1 g:10 mL; the amount of amylase added is 600-800 U / mL of the mixed solution; the amount of protease added is 600-1000 U / mL of the mixed solution; In step (1), at least one of the following is also included: Ultrasonic pretreatment conditions were 5kW ultrasound for 20 to 35 min; The reaction conditions were 45°C for 1 h; The centrifugation conditions were 5000 rpm and 25°C for 20 min.

7. The method for preparing Luzhou-flavor liquor lees water-soluble glucose oligosaccharides according to claim 4, characterized in that: In step (2), the retentate is a component less than 5000Da; the volume ratio of the concentrate to ethanol is 1:3; In step (2), at least one of the following is also included: The molecular weight cut-off of the dialysis bag is 300Da, and the dialysis time is 48h; The reaction conditions were 4°C alcohol precipitation for 12 h; The centrifugation conditions were 5000 rpm and 4°C for 20 min.

8. A pharmaceutical composition, characterized in that: The invention comprises the Luzhou-flavor liquor lees water-soluble glucose oligosaccharide as claimed in any one of claims 1 to 3, or the Luzhou-flavor liquor lees water-soluble glucose oligosaccharide prepared by the preparation method as claimed in any one of claims 4 to 7.

9. Use of the Luzhou-flavor liquor lees water-soluble glucose oligosaccharide according to any one of claims 1 to 3, or the Luzhou-flavor liquor lees water-soluble glucose oligosaccharide prepared by the preparation method according to any one of claims 4 to 7, or the pharmaceutical composition according to claim 8 in the preparation of a drug, reagent, diet or nutritional supplement having one or more of the following effects: a. Relieve and treat alcoholic liver disease; b. Improve the activity of antioxidant enzymes in liver tissue; c. Regulate liver lipid metabolism; d. Relieve and repair liver cell damage.

10. The use according to claim 9, characterized in that: In action b, the liver tissue antioxidant enzymes are SOD, GPX and CAT.

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