CN113908182A

CN113908182A - Fermented sea-buckthorn juice and preparation method thereof

Info

Publication number: CN113908182A
Application number: CN202111451294.6A
Authority: CN
Inventors: 李昕阳; 朱宝军
Original assignee: Zhejiang Huibao Biotechnology Co ltd
Current assignee: Zhejiang Huibao Biotechnology Co ltd
Priority date: 2021-12-01
Filing date: 2021-12-01
Publication date: 2022-01-11
Anticipated expiration: 2041-12-01
Also published as: CN113908182B

Abstract

The invention discloses a preparation method of fermented sea-buckthorn juice for regulating metabolic syndrome, which selects sea-buckthorn raw juice and sequentially carries out the following steps: uniformly mixing the raw seabuckthorn pulp, isomaltooligosaccharide and water to obtain a starting material; adding beer yeast into the initial material, and fermenting at normal temperature; adding lactobacillus plantarum into the obtained yeast fermentation product, and fermenting at normal temperature; adding acetic acid bacteria into the obtained lactobacillus fermentation product, and fermenting at normal temperature to obtain fermented fructus Hippophae juice. Under the specific three-time fermentation process scheme, the fermented sea buckthorn juice has the effects of relieving weight gain, reducing blood fat, reducing blood sugar and resisting inflammation, and has the potential of regulating metabolic syndrome.

Description

Fermented sea-buckthorn juice and preparation method thereof

Technical Field

The invention relates to fermented sea buckthorn juice for regulating metabolic syndrome and a preparation method thereof.

Background

Seabuckthorn (Hippophae rhamnoides Linn.) is deciduous shrub, has the characteristics of drought resistance, sand resistance and survival on saline-alkali soil, and is widely used for water and soil conservation, and the area of seabuckthorn forest lands in China also occupies the top of the world and accounts for more than 90% of the total area of the seabuckthorn forest lands in the world. The sea buckthorn is a medicinal and edible plant, the fruits of the sea buckthorn are rich in vitamins, organic acids, phenolic acids, saccharides, fatty acids, amino acids, carotenoids, procyanidine alkaloids, flavones, tannins, triterpenes, steroids and the like, and the sea buckthorn has biological effects of resisting inflammation, resisting oxidation, reducing blood sugar, resisting tumors, preventing radiation, delaying photoaging, resisting atherosclerosis, preventing liver injury, preventing thrombosis and the like.

The sea-buckthorn has good application prospect in many fields of national economy such as food, medicine, light industry, agriculture, animal husbandry, fishery and the like. The more popular seabuckthorn products in the market comprise seabuckthorn fruit juice, seabuckthorn tea, seabuckthorn calcium tablets, seabuckthorn capsules, seabuckthorn fruit oil, seabuckthorn seed oil and the like. A new research direction of sea-buckthorn is to prepare sea-buckthorn ferment through fermentation, and although sea-buckthorn has a plurality of efficacies, the research and patents on the fermentation of sea-buckthorn are focused on the functional activities of resisting oxidation and improving intestinal immunity, and the research on other functional activities is very rare.

The invention of CN202010728224.X discloses a seabuckthorn fermentation stock solution and a production method thereof: the fermentation raw materials comprise fructus Hippophae, fructus Citri Limoniae, fructus Pruni Pseudocerasi, fructus Ananadis Comosi, brown granulated sugar and fructo-oligosaccharide, and Lactobacillus plantarum is used first, and Lactobacillus acidophilus and Bifidobacterium lactis are used later; the main advantage of fermentation is that the original sea-buckthorn juice with sour and astringent taste and poor taste has better taste. And the intestinal flora is adjusted, and the beneficial components of each component are fully released.

201510024785.0 discloses a seabuckthorn fruit fermented product and a preparation method thereof, comprising the following steps: 1) squeezing fructus Hippophae to obtain juice; 2) adding culture medium components into the juice, inoculating leuconostoc mesenteroides for first fermentation to prepare a first fermentation solution; 3) adding pectinase into the fruit and vegetable liquid with the pH value of 4.5-6 for enzymolysis to prepare an enzymolysis fruit and vegetable liquid; 4) adding culture medium components into the enzymolysis fruit and vegetable liquid, and inoculating compound lactobacillus for secondary fermentation to obtain a second fermentation liquid; the compound lactobacillus comprises lactobacillus plantarum; 5) adding culture medium components into the second fermentation liquid, and inoculating composite yeast to perform third fermentation to obtain third fermentation liquid, wherein the composite yeast comprises beer yeast; 6) and (3) uniformly mixing the first fermentation liquid and the third fermentation liquid, centrifuging, homogenizing the centrifuged supernatant, and sterilizing to obtain the seabuckthorn fermentation product. The fermented product has good taste and unique flavor, has good functions of immunoregulation (increasing mouse T lymphocyte activity) and oxidation resistance, and is suitable for a wide range of people.

201811142677.3A fermented fructus Hippophae pudding is prepared by mixing Bacillus coagulans and high temperature resistant lactic acid bacteria, heating, fermenting, and adding edible gum and sweetener to obtain the final product with unique flavor and rich nutrition.

CN 202010239722.8A sea buckthorn enzyme process formula: the fermentation raw materials comprise apple juice, sea buckthorn raw juice, quinoa wheat juice and fructo-oligosaccharide, and the zymophyte is fermented by using the Isaria japonica ferment bacteria, so that the zymophyte has high activity and strong inoxidizability after fermentation.

CN 201711364309.9A preparation method of fermented sea buckthorn composition for improving intestinal immunity comprises: the fermentation raw materials are folium Hippophae and fructus Hippophae, and the fermented materials are mixed with probiotic bacteria and spray dried to obtain the final product with antioxidant activity and capable of improving intestinal tract autoimmunity.

The fermented product of the conventional sea buckthorn is single in functional activity and only limited in oxidation resistance and intestinal immunity.

Disclosure of Invention

The invention aims to provide a preparation method of fermented sea buckthorn juice for regulating metabolic syndrome.

In order to solve the technical problems, the invention provides a preparation method of fermented sea-buckthorn juice for regulating metabolic syndrome, which selects sea-buckthorn raw juice and comprises the following raw materials in parts by weight: 900 parts of seabuckthorn raw stock, (300 +/-45) parts of malto-oligosaccharide, (300 +/-15) parts of water, (2.1 +/-0.1) parts of beer yeast, (2.1 +/-0.1) parts of lactobacillus plantarum and (2.1 +/-0.1) parts of acetic acid bacteria;

the water is conventional drinking water;

the following steps are carried out in sequence:

1) mixing materials:

uniformly mixing the raw hippophae rhamnoides pulp, isomaltooligosaccharide and drinking water to obtain a starting material;

2) and primary fermentation:

adding beer yeast (the beer yeast is activated in advance) into the initial material obtained in the step 1), and fermenting at the normal temperature of 28-30 ℃ for 15 +/-1 days; obtaining yeast fermentation product (liquid state);

3) and (3) secondary fermentation:

adding lactobacillus plantarum into the yeast fermentation product obtained in the step 2), and fermenting at the normal temperature of 28-30 ℃ for 20 +/-1 days; obtaining lactobacillus fermentation product (liquid state);

4) and carrying out third fermentation:

adding acetic acid bacteria into the lactobacillus fermentation product obtained in the step 3), and fermenting at the normal temperature of 28-30 ℃ to obtain fermented sea buckthorn juice.

The improvement of the preparation method of the fermented sea buckthorn juice for regulating the metabolic syndrome of the invention comprises the following steps:

in the step 4), when 3 conditions of acidity less than 3%, pH less than 4 and ethanol content less than 0.5% are met, the fermentation is stopped to obtain the fermented sea buckthorn juice.

The preparation method of the fermented sea buckthorn juice for regulating the metabolic syndrome is further improved as follows:

the lactobacillus plantarum consists of T17 lactobacillus plantarum and HN9 lactobacillus plantarum, wherein T17 lactobacillus plantarum: HN9 lactobacillus plantarum ═ (2 ± 0.2): 1 (viable cell ratio).

Namely, the yeast fermentation product obtained in the step 2) is added with the Lactobacillus plantarum T17 and the Lactobacillus plantarum HN9 for fermentation.

the viable count of the T17 Lactobacillus plantarum is more than or equal to 10⁹CFU/g；

The viable count of HN9 Lactobacillus plantarum is more than or equal to 10⁹CFU/g；

The viable count of acetic acid bacteria is not less than 10⁹CFU/g。

The invention also provides fermented sea buckthorn juice for regulating metabolic syndrome, which is prepared by any one of the methods.

The Lactobacillus plantarum T17 is prepared from Lactobacillus plantarum ZJUF T17(CCTCC NO: M2017342). 201711340889.8/201810724365.7, telling that the bacterium can significantly reduce the cholesterol content in serum and optimize the ratio of high density lipoprotein cholesterol to low density lipoprotein cholesterol; the lactobacillus plantarum can obviously reduce the content of TNF-alpha in serum and has the function of immunoregulation.

The HN9 Lactobacillus plantarum is prepared from Lactobacillus plantarum ZJUF HN9(CCTCC NO: M2017341). 201711404582.X, the plant indicator lactobacillus ZJUF HN9 has the capacity of tolerating high-content bile salt, and is beneficial to survival and propagation in intestinal tract; the compound has higher survival rate and stronger gastrointestinal adhesion in the gastrointestinal tract environment, and can obviously inhibit the propagation of gastrointestinal tract pathogenic bacteria, particularly salmonella typhimurium; in addition, the lactobacillus plantarum has the potential to produce fermented milk.

According to the invention, the sea buckthorn rich in various functional components is used as a fermentation substrate, probiotics with functional activity potential is used, and under the specific three-fermentation process scheme, the fermented sea buckthorn juice has the effects of relieving weight gain, reducing blood fat, reducing blood sugar and resisting inflammation, and has the potential of regulating metabolic syndrome. Moreover, the fermented product enables the original sea-buckthorn juice with sour and astringent taste and poor mouthfeel to have better mouthfeel.

Drawings

The following describes embodiments of the present invention in further detail with reference to the accompanying drawings.

FIG. 1 is a graph comparing the effect of probiotic fermented hippophae rhamnoides juice on body weight, drinking water and energy intake of HFD fed mice;

in fig. 1: a, percent change in weight gain at 16 weeks; b, total weight gain at week 16; c, changing drinking water; d, average daily energy intake.

FIG. 2 is a graph showing the effect of probiotic fermented hippophae rhamnoides juice on the histomorphology of HFD fed mice;

in fig. 2: a, H & E staining of liver; b, epididymis fat H & E staining; c, epididymal adipocyte area.

FIG. 3 is a graph of the effect of probiotic fermented Hippophae rhamnoides on sugar homeostasis in HFD fed mice;

in fig. 3: a, 14 weeks glucose tolerance test blood glucose change; b, 14 week sugar tolerance experiment AUC; c, 15 week insulin resistance test blood glucose change; d, 15 week insulin resistance test AUC; e, 14 week fasting plasma glucose; f, insulin resistance index HOMA-IR.

FIG. 4 shows the effect of probiotic fermented sea buckthorn on HFD fed mouse blood lipid level four;

in fig. 4: a, HDL-C; b, LDL-C; c, TG; d, T-CHO.

FIG. 5 shows the effect of probiotic fermented Hippophae rhamnoides on HFD fed mouse inflammatory factors;

in fig. 5: a, IL-6; b, IL-10; c, TNF-alpha; and D, LPS.

FIG. 6 shows the effect of probiotic fermented hippophae rhamnoides on the oxidative stress of the liver of HFD fed mice;

in fig. 6: a, T-SOD; b, MDA; c, CAT.

Detailed Description

The invention will be further described with reference to specific examples, but the scope of the invention is not limited thereto:

the sea-buckthorn primary pulp is as follows: fructus Hippophae protoplasm is purchased from Gansu Aikang fructus Hippophae products Co.

The viable count of the T17 lactobacillus plantarum is 1-5 multiplied by 10⁹CFU/g, the source is Lactobacillus plantarum ZJUF T17(CCTCC NO: M2017342);

the viable count of HN9 Lactobacillus plantarum is 1-5 multiplied by 10⁹CFU/g, the source is Lactobacillus plantarum ZJUF HN9(CCTCC NO: M2017341);

the viable count of the acetic acid bacteria is 1-5 multiplied by 10⁹CFU/g; it is composed of Shanghai brewing 1.01 aceticus and AS1.41 aceticus (AS 1.41 Acetobacter rankings bred by Beijing microbial research institute of Chinese academy of sciences) with equal viable bacterial count.

The beer yeast is high-activity beer yeast of Yinggao brand produced by Jiangsu YingshengZhishi limited company, and the activation mode is that purified water is activated for 15-20 minutes. The activation of brewers yeast is a conventional technique, such as specifically: selecting purified water as an activation medium, putting the beer yeast into the purified water, and activating for 15-20 minutes at 22-30 ℃ (preferably 28 ℃), so as to obtain activated beer yeast;

example 1, a method for preparing fermented sea buckthorn juice, using the following raw materials: 900kg of raw seabuckthorn pulp, 300kg of malto-oligosaccharide, 300kg of drinking water, 2.1kg of beer yeast, 1.4kg of T17 lactobacillus plantarum, 0.7kg of HN9 lactobacillus plantarum and 2.1kg of acetic acid bacteria.

The viable count of T17 Lactobacillus plantarum is 3 × 10⁹CFU/g, HN9 viable count of Lactobacillus plantarum 3X 10⁹CFU/g, viable count of acetic acid bacteria 3 × 10⁹CFU/g。

The preparation method comprises the following steps in sequence:

the process flow comprises the following steps:

1) mixing materials: adding the raw hippophae rhamnoides pulp, isomaltooligosaccharide and drinking water into a 3-ton fermentation tank, and uniformly mixing to obtain a starting material;

2) and primary fermentation:

adding activated beer yeast into the initial material obtained in the step 1), and fermenting at normal temperature: the temperature of the fermentation tank is kept at 28-30 ℃, and fermentation is carried out for 15 days; obtaining yeast fermentation product (liquid state);

3) and (3) secondary fermentation:

adding T17 lactobacillus plantarum and HN9 lactobacillus plantarum into the yeast fermentation product obtained in the step 2), and fermenting at normal temperature: maintaining the temperature of the fermentation tank at 28-30 deg.C, and fermenting for 20 days; obtaining lactobacillus fermentation product (liquid state);

4) and carrying out third fermentation:

adding acetic acid bacteria into the lactobacillus fermentation product obtained in the step 3), and fermenting at normal temperature: maintaining the temperature of the fermentation tank at 28-30 deg.C, taking a certain amount of fermentation broth every 10 days, and detecting acidity, pH and ethanol content; fermenting for 40 days, wherein the acidity is 2.8%, the pH value is 2.9, and the ethanol content is 0.23%, thereby obtaining the fermented sea buckthorn juice.

Experiment 1, evaluation of the functional activity of fermented sea buckthorn juice: functional evaluation with mouse as model animal

1. Animal feeding and grouping

Male C57BL/6 mice, 6 weeks old, were housed in an animal house and fed normal diet for 7 days to acclimatize to the experimental setting (12h D/N). Randomly divided into 5 groups (12 per group) according to the quality of constitution:

(1) common feed group (NCD): feeding common feed and drinking water;

(2) high fat control group (HFD): feeding high-fat feed with fat function ratio of 45% and drinking water;

(3) unfermented hippophae rhamnoides group (BHJ): feeding high-fat feed with fat function ratio of 45%, and replacing drinking water with diluent which is obtained by diluting mixed raw materials by 20 times before fermentation;

the diluent after the mixed raw materials are diluted by 20 times before fermentation is as follows: namely, a diluent obtained by adding 19 times by mass of drinking water into the initial material obtained in the step 1) of the embodiment 1;

(4) probiotic fermented hippophae rhamnoides group (without sugar supplementation) (FHJ): feeding high-fat feed with fat function ratio of 45%, and replacing drinking water with diluent obtained by diluting 20 times of fermented fructus Hippophae juice;

the diluent after the fermented sea-buckthorn juice is diluted by 20 times is as follows: a diluent obtained by adding 19 times by mass of drinking water to the fermented sea buckthorn juice obtained in example 1;

(5) probiotic fermented hippophae rhamnoides group (top-up sugars) (FHJ + CHO): feeding high-fat feed with fat function ratio of 45%, fermenting, and diluting with sugar (0.32 mg/g glucose, 3.4mg/g fructose, 0.54mg/g sucrose, and 6.92mg/g lactose) to 20 times to obtain diluted solution for replacing drinking water;

after the fermented sea-buckthorn juice is complemented with sugar and diluted by 20 times, the diluent is as follows: 1g of the fermented sea buckthorn juice obtained in example 1 was added with 0.32mg of glucose, 3.4mg of fructose, 0.54mg of sucrose, 6.92mg of lactose, and then 19g of drinking water to obtain a diluted solution.

The NCD group was fed with normal diet, and the other four groups were fed with high-fat diet with fat function ratio of 45%. NCD and HFD mice normally drink distilled water, and mice of BHJ, FHJ + CHO and FHJ + PRO groups respectively drink corresponding sea buckthorn juice diluents. Mice were treated with 12 mice per group, 4 mice per cage, ambient (22 + -2 deg.C), humidity 30-70%, mice were allowed to eat and drink water freely, the test took 16 weeks, body weight was recorded every 7 days, and the amount of food intake and fluid intake was recorded every 3 days. After 16 weeks, the mice are fasted for 24 hours, weighed, then blood is taken from the orbit, cervical vertebra is dislocated and killed, fat and brown fat of the abdominal testis are taken, heart, liver and spleen tissues are quickly taken and quick frozen by liquid nitrogen, transported by dry ice and stored in a refrigerator at-80 ℃ for later use. Serum was obtained by centrifugation at 3000rpm for 10 min.

2 Effect of fermented Hippophae rhamnoides on weight, drinking water and energy intake of mice fed on high-fat diet

As can be seen from FIG. 1, there was no significant difference in body weight (p >0.05) between the groups at the start of the experiment (week 0). After 8 weeks, the percentage body weight gain was lower in NCD, BHJ, FHJ and FHJ + CHO mice than in HFD mice, indicating that both hippophae rhamnoides and fermented hippophae rhamnoides were able to reduce the weight gain caused by high fat diet. At the end of the experiment (16 weeks), the total body weight gain of the mice fed FHJ, FHJ + CHO was significantly lower than that of the HFD group (p < 0.05; fig. 1A, B), and the total body weight gain of the FHJ, FHJ + CHO mice was less than that of the BHJ group, indicating that fermented hippophae rhamnoides contributed to the weight loss of the mice on a high fat diet. As seen from fig. 1C and 1D, the liquid intake and energy intake of NCD group mice were significantly higher than those of other high lipid groups. It is likely that the appetite of the high-fat group will be lower than that of the general diet group, but the modeling of the high-fat mice is successful in view of the change of blood sugar and blood fat later and the like.

3 Effect of fermented Hippophae rhamnoides on organ index and histomorphology of mice fed on high-fat diet

Mouse liver and epididymal fat in 10% formalin were paraffin-sectioned and then stained with hematoxylin-eosin (H & E). H & E stained sections of each epididymal fat were randomly selected for 3 clear fields under a × 400 × microscope and the area of adipocytes in each picture was calculated with Image Pro Plus software. The results of the experiment are shown in FIG. 2.

After 16 weeks of induction with HFD, the epididymal adipocytes area was significantly increased in HFD group mice compared to NCD group mice, while those in BHJ and FHJ groups were significantly smaller (p <0.05) (fig. 2B, C). In order to further observe the influence of BHJ and FHJ on the lipid metabolism of mice fed with high-fat diet, H & E staining is carried out on liver tissues of the mice to find that the number of lipid droplets in the liver tissues of the HFD group is obviously more and more than that of the liver tissues of the NCD group, which indicates that the liver lipid accumulation of the mice of the HFD group is increased. Compared to the HFD group mice, the fat deposition of the liver was significantly reduced in the BHJ and FHJ group mice, especially in the BHJ and FHJ groups (fig. 2A). Overall, BHJ and FHJ have a weaker inhibitory effect on mouse fat accumulation than HFD.

4 Effect of fermented Hippophae rhamnoides on sugar homeostasis in mice fed on high-fat diet

And (3) carrying out a glucose tolerance experiment at 14 weeks of intervention, injecting glucose 2g/kg body weight into the abdominal cavity, measuring blood glucose values of the mice at 0min, 30 min, 60 min, 90 min and 120min respectively, drawing an IGTT (glucose-glucose response time) curve, and calculating the area under the drug absorption curve (AUC) according to the blood glucose values and time. Performing insulin resistance experiment at 15 weeks, injecting insulin into abdominal cavity at 0.75IU/kg body weight, measuring blood glucose values of 0, 30, 60, 90 and 120min mice respectively, drawing blood glucose variation curve (ITT), and calculating area under drug absorption curve (AUC) according to blood glucose value and time. Blood glucose and insulin in mouse serum were detected separately according to the procedure of the commercial kit.

Insulin resistance evaluation index (HOMA-IR) calculation formula:

HOMA-IR FI × FBG/22.5 (formula 3-1)

Note: fasting insulin (FI, mU/L) and Fasting blood glucose (FBG, mmol/L)

The results of the experiment are shown in FIG. 3.

In terms of IGTT and ITT, the results indicate that HFD induced impaired glucose tolerance and insulin resistance in mice (fig. 3A, C). BHJ, FHJ and FHJ + CHO groups improved the glucose tolerance of HFD-fed mice compared to HFD group mice, and FHJ, FHJ + CHO groups had a significant improvement in glucose tolerance (p <0.001) compared to HFD group (fig. 3B). As can be seen from fig. 3E, the fasting plasma glucose was also significantly lower in FHJ group mice than in HFD group mice (p < 0.05). In terms of insulin resistance, the HOMA-IR results showed significant improvement in insulin sensitivity in BHJ, FHJ and FHJ + CHO groups compared to the HFD group (p <0.05, fig. 3F).

5 Effect of fermented Hippophae rhamnoides on blood lipid level of mice fed on high-fat diet

The contents of Triglyceride (TG), total cholesterol (T-CHO), high density lipoprotein (HDL-C) and low density lipoprotein (LDL-C) in the serum of mice were measured according to the procedures of the commercial kit, and the results of the experiment are shown in FIG. 4.

The four terms of blood fat are important indexes reflecting the steady state of the lipid metabolism of the organism. LDL-C and T-CHO are inducers of atherosclerosis development, are closely associated with diabetes and metabolic syndrome, and BHJ, FHJ and FHJ + CHO intervention groups all reduced serum LDL-C, T-CHO levels in mice compared to HFD group mice, and FHJ group showed significant reduction (p <0.05, FIG. 4B, D). The serum HDL-C concentrations were elevated in the BHJ, FHJ and FHJ + CHO groups compared to the HFD group (p <0.05, FIG. 4A). The improvement in FHJ over the HFD group in three indices LDL-C, T-CHO and HDL-C indicates that fermented hippophae rhamnoides juice is helpful in regulating lipid metabolism. However, TG levels were elevated in BHJ, FBJ and FHJ + CHO group mice compared to NCD and HFD groups (p <0.05, fig. 4C), suggesting that ingestion of hippophae rhamnoides and fermentation of hippophae rhamnoides juice may not completely restore lipid metabolism disorders.

6 Effect of fermented sea buckthorn on inflammatory factors of mice fed on high-fat diet

The results of the measurement of inflammatory factors (TNF-. alpha., IL6, IL10, LPS) in serum using the double sandwich ELISA kit according to the instructions are shown in FIG. 5.

Lipopolysaccharide (LPS) is one of the components of gram-negative bacterial cell wall, and changes in serum LPS level are associated with changes in intestinal permeability and inflammatory reactions of the body system. LPS can be transported in the intestinal tract in the form of chylomicrons, and a high fat diet with excessive fat intake will lead to an increase in chylomicrons in the small intestine, thereby facilitating the penetration of LPS into the circulation. It has been shown that obesity, insulin resistance and type II diabetes are associated with elevated blood LPS levels, and that this adverse change in circulating LPS levels is also associated with elevated expression levels of the proinflammatory cytokine TNF- α in adipocytes. Compared with the mice in the HFD group, the mice in the BHJ, FHJ and FHJ + CHO groups have reduced LPS concentration and TNF-alpha level, and the mice in the FHJ and FHJ + CHO groups have significant reduction (p is less than 0.05, and figure 5C, D), and the BHJ, FHJ and HFD groups have no significant difference (p is more than 0.05 and figure 5A, B) in the proinflammatory factor IL-6 and the inflammation inhibiting factor IL-10 indexes.

7 Effect of fermented Hippophae rhamnoides on liver oxidation level of mice fed on high-fat diet

Accurately weighing 0.1g of mouse liver sample, adding 0.9mL of precooled 0.01mol/L PBS buffer solution, holding a homogenizer on an ice box to homogenize for 1min, then centrifuging for 20min at 12000rpm and 4 ℃, taking supernatant to obtain 10% liver tissue homogenate, and measuring the levels of MDA, CAT and T-SOD in the liver according to the kit operation instruction, wherein the experimental result is shown in figure 6.

In the aspect of mouse liver oxidative stress state, MDA of each group has no significant difference (p > 0.05; fig. 6B), BHJ and FHJ have certain influence on liver CAT activity content, but have no significant difference (p > 0.05; fig. 6C). However, compared with HFD mice, the liver SOD activity was significantly improved in FHJ group (p < 0.05; FIG. 6A).

Comparative example 1-1, "1.4 kg of T17 Lactobacillus plantarum, 0.7kg of HN9 Lactobacillus plantarum" was changed to 2.1kgT17 Lactobacillus plantarum, and the remainder was identical to example 1.

Comparative examples 1-2, the same procedures as in example 1 were repeated except that "1.4 kg of Lactobacillus plantarum T17 and 0.7kg of Lactobacillus plantarum HN 9" were changed to "0.7 kg of Lactobacillus plantarum T17 and 1.4kg of Lactobacillus plantarum HN 9".

Comparative example 2-1, the fermentation sequence of step 2), step 3) was changed with respect to example 1, i.e. in particular:

2) and primary fermentation:

adding T17 lactobacillus plantarum and HN9 lactobacillus plantarum into the initial material obtained in the step 1), and fermenting at normal temperature: maintaining the temperature of the fermentation tank at 28-30 deg.C, and fermenting for 20 days; obtaining a lactobacillus fermentation product;

3) and (3) secondary fermentation:

adding activated beer yeast into the lactobacillus fermentation product obtained in the step 2), and fermenting at normal temperature: the temperature of the fermentation tank is kept at 28-30 ℃, and fermentation is carried out for 15 days; obtaining yeast fermentation products;

carrying out the three-time fermentation of the subsequent step 4) on the yeast fermentation product;

the rest is equivalent to embodiment 1.

Comparative examples 2-2, the fermentation sequence of step 3), step 4) was changed with respect to example 1, i.e. in particular:

3) and (3) secondary fermentation:

adding acetic acid bacteria into the yeast fermentation product obtained in the step 2), and fermenting at normal temperature: the temperature of the fermentation tank is kept at 28-30 ℃, and the fermentation time is 40 days;

4) and carrying out third fermentation:

adding T17 lactobacillus plantarum and HN9 lactobacillus plantarum into the product obtained in the secondary fermentation in the step 3) to carry out normal-temperature fermentation: maintaining the temperature of the fermentation tank at 28-30 deg.C, and fermenting for 20 days; obtaining the fermented sea-buckthorn juice.

The rest is equivalent to embodiment 1.

Comparative example 3, with respect to example 1, the following changes were made:

the use of acetic acid bacteria was eliminated, i.e., step 4 of example 1 was eliminated); directly taking the lactobacillus fermentation product obtained by the secondary fermentation in the step 3) as fermented sea buckthorn juice; the rest is the same as example 1.

The above 5 comparative examples were tested according to the above test method and the results were compared with those of example 1, as shown in the following table,

comparative examples 1-1, 1-2 in comparison with comparative example 1, the fermented hippophae rhamnoides juice obtained was weaker than example 1 in improving metabolic syndrome. In comparative examples 2-1 and 2-2, compared with example 1, the results showed significant differences in 16-week total weight gain, fasting plasma glucose, HOMA-IR, LDL-C and T-CHO indices, and it was found that fermented Hippophae rhamnoides juice produced by a process comprising fermentation with Saccharomyces cerevisiae and fermentation with Lactobacillus plantarum was more advantageous for improving metabolic syndrome. Comparative example 3 compared with example 1, the acetic acid bacteria were eliminated, and the HOMA-IR and T-CHO were significantly improved after the elimination of the strain, which indicates that the elimination of the acetic acid bacteria fermentation process is not beneficial to the improvement of the glycolipid metabolism of the final product.

Finally, it is also noted that the above-mentioned lists merely illustrate a few specific embodiments of the invention. It is obvious that the invention is not limited to the above embodiments, but that many variations are possible. All modifications which can be derived or suggested by a person skilled in the art from the disclosure of the present invention are to be considered within the scope of the invention.

Claims

1. A preparation method of fermented seabuckthorn juice for regulating metabolic syndrome selects seabuckthorn raw juice, and is characterized by sequentially carrying out the following steps:

1) mixing materials:

uniformly mixing the raw seabuckthorn pulp, isomaltooligosaccharide and water to obtain a starting material;

2) and primary fermentation:

adding beer yeast into the initial material obtained in the step 1), and fermenting at the normal temperature of 28-30 ℃ for 15 +/-1 days; obtaining yeast fermentation products;

3) and (3) secondary fermentation:

adding lactobacillus plantarum into the yeast fermentation product obtained in the step 2), and fermenting at the normal temperature of 28-30 ℃ for 20 +/-1 days; obtaining a lactobacillus fermentation product;

4) and carrying out third fermentation:

2. The method for preparing fermented hippophae rhamnoides juice for regulating metabolic syndrome according to claim 1, characterized in that the raw materials comprise the following components in parts by weight: 900 parts of raw seabuckthorn pulp, (300 +/-45) parts of maltooligosaccharide, (300 +/-15) parts of water, (2.1 +/-0.1) parts of beer yeast, (2.1 +/-0.1) parts of lactobacillus plantarum and (2.1 +/-0.1) parts of acetic acid bacteria.

3. The method for preparing fermented hippophae rhamnoides juice for regulating metabolic syndrome according to claim 2, wherein:

4. The method for preparing fermented hippophae rhamnoides juice for regulating metabolic syndrome according to claim 3, wherein:

the lactobacillus plantarum consists of T17 lactobacillus plantarum and HN9 lactobacillus plantarum, wherein T17 lactobacillus plantarum: HN9 lactobacillus plantarum ═ (2 ± 0.2): 1 in terms of the amount of bacteria.

5. The method for preparing fermented hippophae rhamnoides juice for regulating metabolic syndrome according to claim 4, wherein:

The viable count of acetic acid bacteria is not less than 10⁹CFU/g。

6. A fermented juice of Hippophae rhamnoides for regulating metabolic syndrome, prepared by the method of any one of claims 1 to 5.