CN112515077A - Functional lycium ruthenicum fermented drink and preparation method and application thereof - Google Patents

Functional lycium ruthenicum fermented drink and preparation method and application thereof Download PDF

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CN112515077A
CN112515077A CN202011443523.5A CN202011443523A CN112515077A CN 112515077 A CN112515077 A CN 112515077A CN 202011443523 A CN202011443523 A CN 202011443523A CN 112515077 A CN112515077 A CN 112515077A
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lycium ruthenicum
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lactobacillus rhamnosus
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李海权
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Shanghai Zhiyesheng Biotechnology Co ltd
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Abstract

A functional lycium ruthenicum fermented drink and a preparation method and application thereof relate to the field of probiotic fermentation. The functional black wolfberry fermented beverage comprises lactobacillus rhamnosus YL198 bacterial suspension and black wolfberry in a dosage ratio of L to kg by weight of 1: 100-1: 500. Lactobacillus rhamnosus YL198 is preserved in China center for type culture Collection (CCTCC M2020781) at 11 months and 27 days in 2020. According to the invention, the black wolfberry is fermented by using the lactobacillus rhamnosus YL198, and the magnetic field generated by the terahertz ore is matched with the probiotic fermentation and the fermentation conversion of the bioactive components of the black wolfberry, so that the content of the bioactive components in the black wolfberry is obviously improved, a large amount of short-chain fatty acid, vitamins, amino acid and extracellular polysaccharide can be generated by the growth and metabolism of lactobacillus, and new nutritional components are endowed to the fermented black wolfberry. The invention has the functions of improving the intestinal function, bidirectionally regulating constipation and treating diarrhea.

Description

Functional lycium ruthenicum fermented drink and preparation method and application thereof
Technical Field
The invention relates to the technical field of probiotic fermentation, and in particular relates to a functional lycium ruthenicum fermented beverage as well as a preparation method and application thereof.
Background
The probiotics refer to living microorganisms which are beneficial to the health of a human body after being given to the human body with proper dosage, and the main strains are lactobacillus and bifidobacterium. Probiotics have received considerable attention because of their recognized edible safety and unique biological functions, which has led to unprecedented development in the use of probiotic fermentation technology. The method is characterized in that one or more of the optimized probiotic floras are used as strains, the intestinal environment of a human body and the digestive decomposition process of medicinal and edible homologous components in the human body are simulated in vitro, the effective components of the medicinal and edible homologous raw materials are biologically converted, and macromolecular substances in the medicinal and edible homologous raw materials are converted into small molecular components which can be directly absorbed by the intestinal tract of the human body. The probiotic fermentation liquor contains a large amount of active enzyme, and after the probiotic fermentation liquor is quickly absorbed by organisms, the probiotic fermentation liquor can improve intestinal functions, regulate immune functions and the like, so that the biological activity of medicinal and edible raw materials can be better exerted.
Lycium ruthenicum Murr is a functional raw material used as both medicine and food, is rich in various health care components, comprises flavonoid compounds, anthocyanin, procyanidine, phytosterol, phenolic acid, betaine, polysaccharide and the like, has extremely high nutrition and health care values, and can resist oxidation, delay aging of human body cell tissues, protect eyesight, resist fatigue and protect cardiovascular. The lycium ruthenicum and the functional substances thereof have wide development prospects.
Disclosure of Invention
In order to solve the problems in the prior art, the invention provides a functional lycium ruthenicum fermented drink and a preparation method and application thereof.
The technical scheme adopted by the invention for solving the technical problem is as follows:
the functional black wolfberry fermented beverage comprises Lactobacillus rhamnosus YL198 bacterial suspension and black wolfberry, wherein the dosage ratio of the Lactobacillus rhamnosus YL198 bacterial suspension to the black wolfberry is as follows: the weight of the powder is 1L:100 kg-1L: 500 kg;
the Lactobacillus rhamnosus YL198 has been preserved in China center for type culture Collection at 11 month and 27 month in 2020, with the preservation number as follows: CCTCC M2020781.
In a preferred embodiment, the ratio of the Lactobacillus rhamnosus (Lactobacillus rhamnosus) YL198 bacterial suspension to lycium ruthenicum is as follows: the volume and weight are 1L to 300 kg.
The preparation method of the functional lycium ruthenicum fermented drink comprises the following steps:
inoculating Lactobacillus rhamnosus (Lactobacillus rhamnosus) YL198 to a liquid MRS culture medium, standing and culturing at 37-45 ℃ for 16-25 h, centrifuging at 4-10 ℃ and 4000-8000 rpm for 5-10 min, and collecting thallus precipitates; suspending with aqueous solution of fermentation and sterilization culture medium, adjusting viable count to 1.0 × 109～1.0× 1010CFU/ml to obtain YL198 bacterial suspension;
naturally drying fresh lycium ruthenicum, soaking for 2 times by using 0.5-1% of salt solution, cleaning with clear water, putting terahertz ore at the bottom of a fermentation tank, putting the lycium ruthenicum, covering the lycium ruthenicum with the terahertz ore, wherein the total weight of the terahertz ore is 1-2% of the total weight of the lycium ruthenicum, adding YL198 bacterial suspension, covering a plastic film, sealing, keeping the temperature at 30-40 ℃, fermenting for 10-20 days, performing solid-liquid separation after the fermentation is finished, fermenting the lycium ruthenicum again once under the same condition, combining fermentation liquids obtained in two times, filtering, and packaging to obtain the functional lycium ruthenicum fermented drink, wherein the pH value is 3.0-3.5.
As a preferred embodiment, the preparation method of the functional lycium ruthenicum fermented drink comprises the following steps:
inoculating Lactobacillus rhamnosus YL198 into liquid MRS culture medium, standing at 37 deg.C for 16 hr, centrifuging at 4 deg.C and 6000rpm for 8min, and collecting thallus precipitate; suspending with aqueous solution of fermentation and sterilization culture medium, adjusting viable count to 1.0 × 109～1.0×1010CFU/ml to obtain YL198 bacterial suspension;
naturally drying fresh lycium ruthenicum, soaking for 2 times by using 0.75% of salt water, cleaning with clear water, placing terahertz ore at the bottom of a fermentation tank, placing the lycium ruthenicum, covering the terahertz ore on the lycium ruthenicum, wherein the total weight of the terahertz ore is 1.5% of the total weight of the lycium ruthenicum, adding YL198 bacterial suspension, covering a plastic film, sealing, fermenting at the constant temperature of 30 ℃ for 10 days, performing solid-liquid separation after the fermentation is finished, performing repeated fermentation on the lycium ruthenicum once under the same condition, combining fermentation liquor obtained in two times, filtering, and packaging to obtain a functional lycium ruthenicum fermented drink, wherein the pH value is between 3.0 and.
In a preferred embodiment, the aqueous solution of the fermentative sterilization medium is prepared by the following method: adding 30g of corn steep liquor powder and 30g of molasses into each liter of liquid culture medium, and autoclaving at 121 deg.C for 15min to obtain the final product.
The functional lycium ruthenicum fermented drink is applied to preparation of products with intestinal function improvement.
As a preferred embodiment, the product is a pharmaceutical, nutraceutical or food product.
As a more preferred embodiment, the pharmaceutical product is in the form of: aqua, capsule, powder or tablet.
The invention has the beneficial effects that:
the Lactobacillus rhamnosus (Lactobacillus rhamnosus) YL198 is preserved in China center for type culture Collection in 11 months and 27 days of 2020, and the preservation numbers are as follows: CCTCC M2020781.
According to the invention, the functional lycium ruthenicum fermented beverage is prepared by fermenting lycium ruthenicum through Lactobacillus rhamnosus YL198 and simultaneously assisting probiotic fermentation through a magnetic field generated by terahertz ore and assisting fermentation conversion of bioactive components of the lycium ruthenicum, the preparation method is simple and efficient, the content of bioactive components such as polysaccharide, flavone, anthocyanin, polyphenol and procyanidine in the lycium ruthenicum can be obviously improved, and the curative effect is enhanced. Meanwhile, lactic acid bacteria grow and metabolize to generate a large amount of short-chain fatty acids, vitamins, amino acids, exopolysaccharides and the like, and new nutrient components are given to the fermented lycium ruthenicum. In addition, compared with the prior art, the functional lycium ruthenicum fermented beverage has the advantages that the intestinal function is improved, and constipation and diarrhea are regulated in a bidirectional mode.
Detailed Description
The functional black wolfberry fermented beverage mainly comprises Lactobacillus rhamnosus YL198 bacterial suspension and black wolfberry, and the dosage ratio of the Lactobacillus rhamnosus YL198 bacterial suspension to the black wolfberry is as follows: the weight of the powder is 1L:100 kg-1L: 500 kg; preferably, the dosage ratio of the Lactobacillus rhamnosus (Lactobacillus rhamnosus) YL198 bacterial suspension to the lycium ruthenicum is as follows: the volume and weight are 1L to 300 kg.
Wherein, the adopted Lactobacillus rhamnosus YL198 has been preserved in China Center for Type Culture Collection (CCTCC) at 11 month and 27 month in 2020, and the addresses are as follows: wuhan university, the preservation number is: CCTCC NO: m2020781.
The functional black wolfberry fermented drink is prepared by fermenting black wolfberry serving as a raw material with Lactobacillus rhamnosus YL 198. The specific preparation method comprises the following steps:
inoculating Lactobacillus rhamnosus YL198 to a liquid MRS culture medium, performing static culture at 37-45 ℃ for 16-25 h (preferably, 16h at 37 ℃), centrifuging at 4-10 ℃ and 4000-8000 rpm for 5-10 min (preferably, 8min at 4 ℃ and 6000 rpm), and collecting thallus precipitates; suspending with aqueous solution of fermentation and sterilization culture medium, adjusting viable count to 1.0 × 109～1.0×1010CFU/ml to obtain YL198 bacterial suspension;
naturally drying fresh lycium ruthenicum, soaking for 2 times by using 0.5-1% (preferably 0.75%) of salt solution, cleaning with clear water, placing terahertz ore at the bottom of a fermentation tank, placing the lycium ruthenicum, covering the lycium ruthenicum with the terahertz ore, wherein the total weight of the terahertz ore is 1-2% (preferably 1.5%) of the total weight of the lycium ruthenicum, adding YL198 bacterial suspension, covering a plastic film, sealing, keeping the temperature at 30-40 ℃ (preferably, keeping the temperature at 30 ℃), fermenting for 10-20 days (preferably, fermenting for 10 days), performing solid-liquid separation after the fermentation is finished, fermenting the lycium ruthenicum again once under the same condition, combining two fermentation liquids, filtering and packaging to obtain the functional lycium ruthenicum fermented drink, wherein the pH value is 3.0-3.5.
Preferably, the fermentation sterilization culture medium aqueous solution is prepared by the following method: adding 30g of corn steep liquor powder and 30g of molasses into each liter of liquid culture medium, and autoclaving at 121 deg.C for 15min to obtain the final product.
The functional lycium ruthenicum fermented drink is applied to preparation of products with intestinal function improvement. Preferably, the product is a pharmaceutical, nutraceutical or food product, but is not limited thereto. More preferably, the pharmaceutical product is in the form of: aqueous, capsule, powder or tablet, but not limited thereto.
According to the invention, through probiotic fermentation and microbial transformation, the content of bioactive components in the fermentation product is increased, the nutritive value and the functional effect of the fermentation product are improved, and meanwhile, the probiotics and the lycium ruthenicum are used in a combined manner to supplement each other, so that the taste and the flavor of the lycium ruthenicum are improved, and the effects of quality improvement and synergy are also achieved.
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The experimental procedures in the following examples are conventional unless otherwise specified.
The test materials used in the following examples were purchased from a conventional biochemical reagent store unless otherwise specified.
The quantitative tests in the following examples, all set up three replicates and the results averaged.
Example 1 isolation, identification and preservation of strains
First, isolation of the Strain
In 12 months of 2017, a fresh pickled Chinese sauerkraut sample collected from the city of Tonghua of Jilin province is put into a transport culture medium, is quickly placed in an ice box and is sent to a laboratory for strain separation. Grinding and diluting the pickled Chinese cabbage sample in a gradient manner, separating lactobacillus in the sample by using a lactobacillus selective culture medium, culturing for 48h at 37 ℃, selecting typical bacterial colonies (milky white, round, neat edges, convex, opaque and moist and smooth surfaces) on a flat plate, carrying out streak culture on an MRS solid culture medium, and separating to obtain pure bacterial colonies. Inoculating the pure cultured strain to liquid MRS culture medium, culturing, adding 60% glycerol, and storing in refrigerator at-80 deg.C. Of these, 1 strain of Lactobacillus was designated YL 198.
II, identification of the strains
Physiological and biochemical identification results of YL 198: gram staining is positive, catalase test is negative, benzidine test is negative, indole test is negative, and acetyl methyl methanol test is positive; starch is not hydrolyzed, gelatin is not liquefied, hydrogen sulfide is not generated, and acid and gas are not generated by fermenting glucose; immotile bacilli; the growth can be carried out at 15 ℃ and 45 ℃, and the optimal growth temperature is 37-42 ℃; the pH is suitably 5.0 to 7.0; tolerates 6.5% NaCl; YL198 shows uniform turbid growth in liquid MRS medium, and the thallus is white precipitate after long-term storage.
The API 50CH (Merria, France) identification of YL198 is given in the following table:
Figure BDA0002830775460000051
Figure BDA0002830775460000061
in the above table, "+" indicates that the strain is a positive reaction; "-" indicates that the strain was negative.
16S rDNA sequence homology analysis: YL198 genome DNA is extracted, PCR amplification is carried out on the gene by utilizing lactobacillus primers (16 sF: 5 '-AGAGTTTGATCCTGGCTCAG-3'; 16 sR: 5 '-AGAAAGGAGGTGATCCAGC-3'), an amplification product is sent to the company Limited of biological engineering (Shanghai) to carry out sequencing, and the obtained 16s rDNA sequence is shown as a sequence 1 in a sequence table. Homology alignment analysis of the 16s rDNA sequence in GenBank database by BLAST program revealed that YL198 has 99% homology with Lactobacillus rhamnosus CP-134(GenBank: MK 601693.1).
From the above results, the strain YL198 was identified as Lactobacillus rhamnosus (Lactobacillus rhamnosus).
Third, preservation of the Strain
The Lactobacillus rhamnosus (Lactobacillus rhamnosus) YL198 is preserved in China Center for Type Culture Collection (CCTCC) in 11 months and 27 days of 2020, and has the address as follows: wuhan university, the preservation number is: CCTCC M2020781.
Example 2 preparation of a functional Lycium ruthenicum fermented beverage of the present invention
Preparation of zymocyte suspension
Inoculating Lactobacillus rhamnosus YL198 (hereinafter referred to as YL 198) in liquid MRS culture medium, standing at 37 deg.C for 16 hr, centrifuging at 4 deg.C and 6000rpm for 8min, collecting thallus precipitate, suspending with fermentation sterilization culture medium aqueous solution, adjusting viable count to 1.0 × 109CFU/ml to obtain YL198 bacterial suspension; wherein, the aqueous solution of the fermentation and sterilization culture medium is prepared by the following method: adding 30g of corn steep liquor powder and 30g of molasses into each liter of liquid culture medium, and autoclaving at 121 ℃ for 15 min.
Preparation of functional lycium ruthenicum fermented beverage
Naturally drying fresh lycium ruthenicum, soaking for 2 times by using 0.75% of salt water, cleaning with clear water, placing terahertz ore at the bottom of a fermentation tank, placing the lycium ruthenicum, covering the lycium ruthenicum with the terahertz ore, wherein the total weight of the terahertz ore is 1.5% of the total weight of the lycium ruthenicum, adding YL198 bacterial suspension, adding the YL198 bacterial suspension and the lycium ruthenicum according to the volume ratio of 1L to 300kg, compacting, covering a plastic film, sealing, carrying out constant temperature of 30 ℃, carrying out fermentation for 10 days, carrying out solid-liquid separation after the fermentation is finished, carrying out re-fermentation on the lycium ruthenicum once under the same condition, combining two fermentation liquids, filtering and packaging to obtain the functional lycium ruthen.
After the fermentation is finished, the content of the fermentation thalli in the obtained fermentation product is not less than 2.5 hundred million/ml, and the pH value is between 3.0 and 3.5. The lycium ruthenicum probiotic fermentation product, namely the functional lycium ruthenicum fermented drink, obtained in the embodiment can be directly drunk.
Example 3 Effect of the functional Lycium ruthenicum fermented drink of the present invention on improving constipation
1. Experimental animals: balb/c male mice, 6-8 weeks old, 18-22 g in weight, SPF grade, purchased from Vickland hundred million laboratory animals, Inc.
2. Grouping and processing experimental animals. The test is divided into a control group, a model group and an experimental group. The control group and the model group mice are perfused with 0.5mL/d of normal saline; the experimental group drenches the functional lycium ruthenicum fermented drink of the invention with the volume of 0.5 mL/d/piece, once a day, and freely eats during the experimental period.
3. Stool test of mice
After 7d of the experiment, the mice in each group were fasted for 16h without water deprivation. The mice of the model group and the experimental group are administered with 10mg/kg of compound diphenoxylate by gavage, and the mice of the control group are administered with distilled water. After the compound diphenoxylate is given for 0.5h, each group of mice is perfused with 10mL/kg of gastric ink, and the animals are fed in a single cage and normally drink water to eat. Starting from the gavage ink, the first time of defecation of each animal, the number of black stool particles within 5h and the mass were recorded. And drying the collected excrement at 100 ℃ until the quality is constant, and calculating the water content of the excrement. The moisture content (%) of feces was (1-mass of feces after drying/mass of feces after drying) × 100%. The results of the functional lycium ruthenicum fermented drink of the invention on the indexes of defecation of mice of constipation model are shown in table 1.
TABLE 1
Figure BDA0002830775460000081
Indicates significant difference (P <0.05) compared to the model group and indicates significant difference (P <0.01) compared to the model group.
As can be seen from Table 1, after the oral gavage for 7d, the experimental group has very significant difference (P <0.01) compared with the model group in terms of the first defecation time; for the quality of the black feces within 5h, the experimental group has very significant difference (P <0.01) compared with the model group; for the number of the black excrement grains within 5h, the experiment has very obvious difference (P is less than 0.01) compared with a model group, and a live bacterium group is higher than a dead bacterium group; for fecal water content, there was no significant difference between the experimental group and the model group (P > 0.05). Therefore, the functional lycium ruthenicum fermented drink has an obvious effect of improving the defecation quality of experimental mice.
4. Small intestine propulsion experiment
After 15d of the experiment, the mice in each group were fasted for 16h without water deprivation. The mice of the model group and the experimental group are administered 5mg/kg of compound diphenoxylate by gastric lavage, and the control group is administered physiological saline. After the compound diphenoxylate is given for 0.5h, the gastric lavage ink is 0.1mL/kg for each group. After 25min, the cervical vertebrae is taken off immediately to kill the mouse, the abdominal cavity is opened to separate mesentery, the intestinal canal with the upper end from the pylorus, the lower end to the ileocecal part is cut and placed on a tray, the small intestine is slightly pulled into a straight line, the length of the intestinal canal is measured as the total length of the small intestine, and the length from the pylorus to the front edge of ink is measured as the ink propelling length. And calculating the ink propelling rate. The ink propulsion rate is [ ink propulsion length (cm)/total small intestine length (cm) ] × 100. The results of the effect of the functional lycium ruthenicum fermented drink on the intestinal ink propulsion rate of a constipation model mouse are shown in table 2.
TABLE 2
Figure BDA0002830775460000082
Figure BDA0002830775460000091
Indicates significant difference (P <0.05) compared to the model group and indicates significant difference (P <0.01) compared to the model group.
As can be seen from Table 2, the ink propelling rate of the model group is obviously lower than that of the control group (P is less than 0.01), and the ink propelling experiment mice successfully model constipation models. After the lavage is carried out for 15d, the difference of the ink propulsion rate of the experimental group is extremely obvious (P is less than 0.01) compared with that of the model group, and the ink propulsion rate of the experimental group is obviously increased compared with that of the model group. Therefore, the functional lycium ruthenicum fermented drink can reduce the intestinal peristalsis of experimental mice and is beneficial to feces formation.
Example 4 therapeutic effect of the functional Lycium ruthenicum fermented beverage of the present invention on enteritis
1. Experimental animals: 30 male mice, C57BL/6 at 6 weeks of age, were purchased from Vinca hundred million laboratory animals, Inc.
2. Grouping and processing experimental animals. The method comprises the following steps of randomly dividing the black wolfberry into 3 groups, wherein each group comprises 10 control groups, 10 model groups and 10 experimental groups, after adaptive feeding for 1 week, the experimental groups are drenched with 0.5mL/d functional black wolfberry fermented drink for 21 days continuously, the control groups and the model groups are drenched with equal amount of normal saline, the test days are 15-21, and 2.5% of DSS is added into drinking water of the model groups and the experimental groups every day to induce colitis. After the test was completed, the mice were sacrificed by anesthesia, serum was collected, colon tissue was taken, and homogenate was prepared by mashing with a tissue homogenizer. The colon length, spleen weight and number of intestinal flora of the mice were measured. The results are shown in Table 3.
TABLE 3
Group of Colon Length (cm) Spleen weight (g) Lactobacillus count
Control group 9.33±0.13** 0.07±0.22** 30.13±0.13**
Model set 5.09±0.27 0.12±0.11 9.88±0.98
Experimental group 7.98±0.24** 0.06±0.21** 23.30±0.16**
Indicates significant difference (P <0.05) compared to the model group and indicates significant difference (P <0.01) compared to the model group.
During the course of persistent colonic inflammation, the length of the colon in mice was shortened by ulceration. Accompanied by colonic tissue adhesion. As can be seen from Table 3, the colon length was significantly shorter in the model group than in the control group (p < 0.01). Although the colon was shortened and the tissue was adhered due to inflammation and ulcer, the colon length of the mice of the experimental group was significantly recovered (p <0.01) compared to the model group. The spleen weight of the model group mice was also significantly higher than the control group (p < 0.01). The mean spleen weight in the experimental group was significantly lower than in the model group (p < 0.01). The lactobacillus count of the experimental group was significantly higher than that of the model group (p < 0.01). Therefore, the functional lycium ruthenicum fermented drink has an obvious treatment effect on colitis, can obviously improve the number of beneficial microorganisms such as lactobacillus in intestinal tracts and is beneficial to regulating the balance of intestinal flora.
4. Detection of antioxidant enzymes in colonic tissue
The total antioxidant capacity of colon tissue of mice was determined using a commercially available antioxidant assay kit. The activity of glutathione peroxidase (GPx), Glutathione Reductase (GR), Glutathione (GSH), Catalase (CAT) and superoxide dismutase (SOD) is measured by a special kit. Malondialdehyde (MDA) was detected with a commercially available thiobarbituric acid reactant (TBARS) kit. All assay kits were purchased from Nanjing institute of bioengineering and tested according to the manufacturer's instructions. The results of the functional lycium ruthenicum fermented drink of the invention on the total oxidation resistance and GSH redox cycle of DSS treated mouse colon tissues are shown in Table 4.
TABLE 4
Figure BDA0002830775460000101
Indicates significant difference (P <0.05) compared to the model group and indicates significant difference (P <0.01) compared to the model group.
As can be seen from Table 4, the total antioxidant capacity of colon tissue in the model group is significantly lower than that in the control group (p < 0.05). In contrast, the total antioxidant capacity of the experimental group was significantly higher than that of the model group (p < 0.05). The GPx activity of the model group was significantly lower than the control group (p < 0.05). The GPx activity of the experimental group was significantly higher than that of the model group (p < 0.05). The GR activity of model animals is also significantly lower than that of the control group (p <0.05), and the GR activity of the experimental group is significantly enhanced. Therefore, the functional lycium ruthenicum fermented drink can obviously improve the oxidation resistance of the colon of an experimental model mouse and improve the resistance to intestinal diseases.
The results of the functional lycium ruthenicum fermented beverage of the invention on the effects of CAT, SOD and lipid peroxidation products MDA of DSS-treated mouse colon tissues are shown in Table 5.
TABLE 5
Figure BDA0002830775460000111
Indicates significant difference (P <0.05) compared to the model group and indicates significant difference (P <0.01) compared to the model group.
As can be seen from table 5, the CAT activity in the model group was significantly lower than that in the control group, and the difference was significant (P < 0.05). The CAT activity of the experimental group is increased, and the difference is obvious (p < 0.05). The SOD activity of the experimental group is significantly higher than that of the model group (p <0.05), but the SOD activity of the model group is significantly lower than that of the control group (p < 0.05). The model group had significantly higher MDA content than the control group (p < 0.05). The MDA content in the experimental group was significantly lower than in the model group (p <0.05), returning to almost the same level as in the control group animals. Therefore, the functional lycium ruthenicum fermented drink can obviously improve the activity of the antioxidant enzyme of the colon of the mouse.
5. Serum inflammatory factor level assay
All assay kits for Interferon (IFN) -g, Tumor Necrosis Factor (TNF) -a, Interleukin (IL) -6, IL-12 (p70), and IL-10 were purchased from Nanjing institute of bioengineering and tested according to the manufacturer's instructions. The results of the effects of the functional lycium ruthenicum fermented drink on the serum inflammatory factors of mice treated with DSS are shown in table 6.
TABLE 6
Figure BDA0002830775460000112
Indicates significant difference (P <0.05) compared to the model group and indicates significant difference (P <0.01) compared to the model group.
As can be seen from Table 6, TNF- α secretion in the model group was significantly higher than that in the control group (p <0.05), which showed increased inflammation and enhanced host immune response. The experimental group showed a significant reduction of TNF-alpha secretion compared to the model (p < 0.05). Compared with the control group, the IL-6 secretion of the model group is obviously increased (p < 0.05). The IL-6 secretion was significantly reduced in the experimental group compared to the model group (p < 0.05). The IL-12 secretion in the model group was significantly higher than that in the control group (p < 0.05). The IL-12 secretion of the experimental group is obviously lower than that of the model group (p < 0.05. the IL-10 secretion of the model group is obviously lower than that of the control group (p < 0.05). The IL-10 secretion of the experimental group is obviously higher than that of the model group (p < 0.05). The functional lycium ruthenicum fermented drink improves the intestinal inflammation by reducing the expression of serum proinflammatory cytokines and increasing and inhibiting the expression of the inflammatory cytokines.
The invention discloses a functional lycium ruthenicum fermented drink, a preparation method and application thereof, and a person skilled in the art can appropriately improve process parameters by referring to the content. It is expressly intended that all such similar substitutes and modifications which would be obvious to one skilled in the art are deemed to be included in the invention. While the invention has been described in terms of preferred embodiments, it will be apparent to those skilled in the art that the technology can be practiced and applied by modifying or appropriately combining the products described herein without departing from the spirit and scope of the invention.
Sequence listing
<110> Shanghai plant fermentation Living Biotech Co., Ltd
<120> functional black wolfberry fermented beverage, and preparation method and application thereof
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1310
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
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ttggaaacag atgctaatac cgcataaatc caagaaccgc atggttcttg gctgaaagat 60
ggcgtaagct atcgcttttg gatggacccg cggcgtatta gctagttggt gaggtaacgg 120
ctcaccaagg caatgatacg tagccgaact gagaggttga tcggccacat tgggactgag 180
acacggccca aactcctacg ggaggcagca gtagggaatc ttccacaatg gacgcaagtc 240
tgatggagca acgccgcgtg agtgaagaag gctttcgggt cgtaaaactc tgttgttgga 300
gaagaatggt cggcagagta actgttgtcg gcgtgacggt atccaaccag aaagccacgg 360
ctaactacgt gccagcagcc gcggtaatac gtaggtggca agcgttatcc ggatttattg 420
ggcgtaaagc gagcgcaggc ggttttttaa gtctgatgtg aaagccctcg gcttaaccga 480
ggaagtgcat cggaaactgg gaaacttgag tgcagaagag gacagtggaa ctccatgtgt 540
agcggtgaaa tgcgtagata tatggaagaa caccagtggc gaaggcggct gtctggtctg 600
taactgacgc tgaggctcga aagcatgggt agcgaacagg attagatacc ctggtagtcc 660
atgccgtaaa cgatgaatgc taggtgttgg agggtttccg cccttcagtg ccgcagctaa 720
cgcattaagc attccgcctg gggagtacga ccgcaaggtt gaaactcaaa ggaattgacg 780
ggggcccgca caagcggtgg agcatgtggt ttaattcgaa gcaacgcgaa gaaccttacc 840
aggtcttgac atcttttgat cacctgagag atcaggtttc cccttcgggg gcaaaatgac 900
aggtggtgca tggttgtcgt cagctcgtgt cgtgagatgt tgggttaagt cccgcaacga 960
gcgcaaccct tatgactagt tgccagcatt tagttgggca ctctagtaag actgccggtg 1020
acaaaccgga ggaaggtggg gatgacgtca aatcatcatg ccccttatga cctgggctac 1080
acacgtgcta caatggatgg tacaacgagt tgcgagaccg cgaggtcaag ctaatctctt 1140
aaagccattc tcagttcgga ctgtaggctg caacttccgt acatggagtc ggaatcgcta 1200
tagattgcga ccctgcctca gtacgtggta acaaggtagc ggtagagttt gatcgtggct 1260
cagtaagtgg taacaaggta accgtagagt ttgatcatgg ctcagtaagt 1310

Claims (8)

1. The functional black wolfberry fermented beverage is characterized by comprising a Lactobacillus rhamnosus YL198 bacterial suspension and black wolfberry, wherein the dosage ratio of the Lactobacillus rhamnosus YL198 bacterial suspension to the black wolfberry is as follows: the weight of the powder is 1L:100 kg-1L: 500 kg;
the Lactobacillus rhamnosus YL198 has been preserved in China center for type culture Collection at 11 month and 27 month in 2020, with the preservation number as follows: CCTCC M2020781.
2. The functional fermented lycium ruthenicum drink according to claim 1, wherein the ratio of the Lactobacillus rhamnosus YL198 bacterial suspension to the lycium ruthenicum is as follows: the volume and weight are 1L to 300 kg.
3. The method for preparing the functional fermented lycium ruthenicum drink according to claim 1, comprising the following steps:
inoculating Lactobacillus rhamnosus (Lactobacillus rhamnosus) YL198 to a liquid MRS culture medium, standing and culturing at 37-45 ℃ for 16-25 h, centrifuging at 4-10 ℃ and 4000-8000 rpm for 5-10 min, and collecting thallus precipitates; suspending with aqueous solution of fermentation and sterilization culture medium, adjusting viable count to 1.0 × 109～1.0×1010CFU/ml to obtain YL198 bacterial suspension;
naturally drying fresh lycium ruthenicum, soaking for 2 times by using 0.5-1% of salt solution, cleaning with clear water, putting terahertz ore at the bottom of a fermentation tank, putting the lycium ruthenicum, covering the lycium ruthenicum with the terahertz ore, wherein the total weight of the terahertz ore is 1-2% of the total weight of the lycium ruthenicum, adding YL198 bacterial suspension, covering a plastic film, sealing, keeping the temperature at 30-40 ℃, fermenting for 10-20 days, performing solid-liquid separation after the fermentation is finished, fermenting the lycium ruthenicum again once under the same condition, combining fermentation liquids obtained in two times, filtering, and packaging to obtain the functional lycium ruthenicum fermented drink, wherein the pH value is 3.0-3.5.
4. The method of claim 3, comprising the steps of:
inoculating Lactobacillus rhamnosus YL198 into liquid MRS culture medium, standing at 37 deg.C for 16 hr, centrifuging at 4 deg.C and 6000rpm for 8min, and collecting thallus precipitate; suspending with aqueous solution of fermentation and sterilization culture medium, adjusting viable count to 1.0 × 109～1.0×1010CFU/ml to obtain YL198 bacterial suspension;
naturally drying fresh lycium ruthenicum, soaking for 2 times by using 0.75% of salt water, cleaning with clear water, placing terahertz ore at the bottom of a fermentation tank, placing the lycium ruthenicum, covering the terahertz ore on the lycium ruthenicum, wherein the total weight of the terahertz ore is 1.5% of the total weight of the lycium ruthenicum, adding YL198 bacterial suspension, covering a plastic film, sealing, fermenting at the constant temperature of 30 ℃ for 10 days, performing solid-liquid separation after the fermentation is finished, performing repeated fermentation on the lycium ruthenicum once under the same condition, combining fermentation liquor obtained in two times, filtering, and packaging to obtain a functional lycium ruthenicum fermented drink, wherein the pH value is between 3.0 and.
5. The method according to claim 3, wherein the aqueous solution of the fermentative sterilization medium is prepared by the following method: adding 30g of corn steep liquor powder and 30g of molasses into each liter of liquid culture medium, and autoclaving at 121 deg.C for 15min to obtain the final product.
6. The use of the functional fermented lycium ruthenicum drink according to claim 1 or 2 for preparing a product with improved intestinal function.
7. The use according to claim 6, wherein the product is a pharmaceutical, nutraceutical or food product.
8. The use of claim 7, wherein the pharmaceutical product is in a dosage form of: aqua, capsule, powder or tablet.
CN202011443523.5A 2020-12-11 2020-12-11 Functional lycium ruthenicum fermented drink and preparation method and application thereof Pending CN112515077A (en)

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