CN117089580A - Lactobacillus casei SF-L-12 fermented product and preparation process and application thereof - Google Patents
Lactobacillus casei SF-L-12 fermented product and preparation process and application thereof Download PDFInfo
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- CN117089580A CN117089580A CN202311354341.4A CN202311354341A CN117089580A CN 117089580 A CN117089580 A CN 117089580A CN 202311354341 A CN202311354341 A CN 202311354341A CN 117089580 A CN117089580 A CN 117089580A
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
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Abstract
The invention relates to the technical field of microorganism application, in particular to lactobacillus casei SF-L-12 fermented product, and a preparation process and application thereof. Lactobacillus casei @Lactobacillus casei) SF-L-12 is preserved in China general microbiological culture Collection center (CGMCC) at the date of 10 and 21 in 2020, and the preservation number is CGMCC No.20927, and the substrate to be fermented comprises 64-82 parts of purified water, 1-3 parts of radix codonopsis pilosulae extract, 1-4 parts of radix angelicae sinensis extract, 1-4 parts of fructus lycii extract, 2-5 parts of fructus schisandrae extract, 2-3 parts of dandelion extract, 2-3 parts of poria cocos extract, 2-5 parts of fructus jujubae powder and 2 parts of semen coicis powder4 parts of apple juice and 4-5 parts of Chinese medicinal materials with homology of medicine and food, and the Chinese medicinal materials are reasonably compatible, safe and harmless. The lactobacillus casei SF-L-12 fermented product prepared by the biological fermentation process can obviously improve the content of IgG, igA, igM and other immune globulins in blood, and effectively improve the immunity of organisms.
Description
Technical Field
The invention relates to the technical field of microorganism application, in particular to lactobacillus casei SF-L-12 fermented product, and a preparation process and application thereof.
Background
Immunoglobulin (immunoglobulin) refers to an animal protein with antibody activity, mainly found in plasma, as well as in other body fluids, tissues and some secretions. Immunoglobulins in human plasma are mostly found in gamma-globulin (gamma-globulin). Immunoglobulins can be classified into five classes IgG, igA, igM, igD, igE. Severe deficiency of immunoglobulins can cause immunodeficiency and can clinically induce a variety of diseases such as infection, tumor, allergy, autoimmune disease, etc. In general, people with low immunoglobulin levels are prone to develop immunodeficiency diseases, and the incidence of malignant tumors in such people is higher than that of normal people.
Patent CN106999588A discloses an immunoglobulin-containing pharmaceutical product with increased stability comprising a polyclonal immunoglobulin solution in a pre-filled polymer syringe in a second package containing an oxygen scavenger, for the treatment of immunodeficiency-type diseases by intravenous or subcutaneous injection of IgG. However, this type of therapy is costly and has significant side effects. The patent CN113598361A discloses a herbal ferment for improving the immunity of human bodies and a preparation method thereof, wherein the main raw materials of the herbal ferment are medicinal and edible traditional Chinese medicinal materials, including ginseng, poria cocos, chinese date, mulberry, rhizoma polygonati, medlar, liquorice, raspberry, pollen pini, honey and the like, and the herbal ferment is finally prepared by extracting the traditional Chinese medicinal materials, adding honey and brown granulated sugar, and sequentially carrying out lactic acid fermentation, alcohol fermentation and acetic acid fermentation. However, according to the description, the IgG, igA, igM content of the subjects in the test group had an ascending trend compared with that before the test after 90 days of taking the herbal ferment product, but no statistical difference (P > 0.05); compared with the control group, the content of IgG, igA, igM in the test food group has an ascending trend, but no statistical difference (P is more than 0.05), so that the preparation process of the herbal ferment is complex, and the effect of increasing the secretion amount of the immunoglobulin is not obvious.
Disclosure of Invention
Aiming at the defects of high production cost, large side effect, insignificant therapeutic effect and slow effect of the preparation for increasing the immune globulin in the prior art, the invention provides the lactobacillus casei SF-L-12 fermented product, which is prepared by adopting traditional Chinese medicines with homology of medicine and food as raw materials and applying a biological fermentation technology, and has safety, no harm and remarkable effect.
In a first aspect, the invention provides a lactobacillus casei SF-L-12 fermented product, which is prepared by fermenting a substrate to be fermented through lactobacillus casei SF-L-12, wherein the lactobacillus casei is [ ]Lactobacillus casei) SF-L-12 is preserved in China general microbiological culture Collection center (CGMCC) at the date of 21 and 10 months in 2020, with the preservation number of CGMCC No.20927 and the preservation address of North Chen Xilu No. 1 and 3 in the Chaoyang area of Beijing city; the fermented substrate comprises the following components in parts by weight: 64-82 parts of purified water, 1-3 parts of radix codonopsis extract, 1-4 parts of Chinese angelica extract, 1-4 parts of medlar extract, 2-5 parts of shizandra berry extract, 2-3 parts of dandelion extract, 2-3 parts of poria cocos extract, 2-5 parts of jujube powder, 2-4 parts of coix seed powder and 4-5 parts of apple juice.
Further, the substrate to be fermented comprises the following components in parts by weight: 74 parts of purified water, 2 parts of radix codonopsis extract, 2 parts of Chinese angelica extract, 2 parts of medlar extract, 3 parts of schisandra chinensis extract, 3 parts of dandelion extract, 3 parts of poria cocos extract, 3 parts of jujube powder, 3 parts of coix seed powder and 5 parts of apple juice.
The fermented substrate of the fermented beverage has the following effects:
radix codonopsis pilosulae: strengthening spleen, benefiting lung, nourishing blood and promoting fluid production;
chinese angelica root: replenishing blood, promoting blood circulation, regulating menstruation and relieving pain;
wolfberry fruit: nourishing liver and kidney, replenishing essence and improving eyesight.
Chinese magnoliavine fruit: astringing, inducing astringency, invigorating qi, promoting fluid production, invigorating kidney, and calming heart;
dandelion: clearing heat and detoxicating, detumescence and resolving hard mass, inducing diuresis for treating stranguria;
poria cocos: induce diuresis and excrete dampness, invigorate spleen and calm heart;
jujube: spleen and stomach tonifying, qi invigorating and body fluid production promoting;
semen Coicis: induce diuresis and excrete dampness, invigorate spleen and check diarrhea.
The formula takes radix codonopsis as a monarch drug, strengthens the middle warmer and Qi, takes Chinese angelica as a ministerial drug, enriches the blood and invigorates the blood, takes Chinese magnoliavine fruit as an adjuvant drug, calms the lung and calms the heart, and is matched with medlar, dandelion, poria cocos, jujube and coix seed, so as to clear away heat and toxic materials, and invigorate the spleen and tonify the kidney.
In a second aspect, the invention provides a preparation process of the lactobacillus casei SF-L-12 fermented product, which specifically comprises the following steps:
(1) Firstly, mixing the components of a substrate to be fermented to obtain mixed slurry;
(2) Delivering the mixed slurry into a sterilization container, sterilizing under high pressure, and cooling to obtain a sterilization liquid;
(3) Inoculating lactobacillus casei SF-L-12 into an MRS liquid culture medium, and culturing to obtain a lactobacillus casei SF-L-12 inoculant;
(4) Inoculating lactobacillus casei SF-L-12 inoculant into the sterilizing liquid, and fermenting to obtain fermentation liquid;
(5) Conveying the fermentation liquor to a high-pressure spraying wall breaking machine for wall breaking treatment to obtain wall breaking fermentation liquor;
(6) Pasteurizing the wall-broken fermentation broth to obtain lactobacillus casei SF-L-12 fermented product.
Further, in the step (2), the autoclave temperature was 120℃for 30 minutes, and then cooled to 35 ℃.
Further, in the step (3), the inoculation amount of the lactobacillus casei SF-L-12 is 1%, the culture temperature is 37 ℃, and the culture time is 18 hours.
Further, in the step (4), the inoculum size of the inoculant was 1%, the fermentation temperature was 37℃and the fermentation time was 24 hours.
Further, in the step (5), the pressure of the high-pressure spraying wall breaking machine is set to be 60Mpa.
In a third aspect, the invention provides an application of the lactobacillus casei SF-L-12 fermented product in preparing a product for increasing the secretion amount of immunoglobulin.
The invention has the beneficial effects that:
the substrate to be fermented provided by the invention basically adopts the traditional Chinese medicine with homology of medicine and food, is reasonable in compatibility, safe and nontoxic, and simultaneously uses the lactobacillus casei SF-L-12 for fermentation, so that the absorption of the medicine is improved, the synergy is achieved, and finally the lactobacillus casei SF-L-12 fermented product is obtained, and animal experiments prove that the content of the immunoglobulin in blood IgG, igA, igM and the like can be obviously improved, the immunity of an organism is effectively improved, and the substrate has no side effect and quick response.
Detailed Description
In order to better understand the technical solutions of the present invention, the following description will clearly and completely describe the technical solutions of the embodiments of the present invention, and it is obvious that the described embodiments are only some embodiments of the present invention, not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the present invention without making any inventive effort, shall fall within the scope of the present invention.
The MRS solid plate culture medium used in the invention comprises the following components: 10g of peptone, 5g of beef powder, 4g of yeast powder and K 2 HPO 4 ·7H 2 O2 g, triammonium citrate 2g, glucose 20g and CH 3 COONa·3H 2 O 5g、MgSO 4 ·7H 2 O 0.2g、MnSO 4 ·4H 2 O0.05 g, tween 80 1mL, agar 15.0g, distilled water 1000mL。
The MRS liquid culture medium used in the invention comprises the following components: 10g of peptone, 5g of beef powder, 4g of yeast powder and K 2 HPO 4 ·7H 2 O2 g, triammonium citrate 2g, glucose 20g and CH 3 COONa·3H 2 O 5g、MgSO 4 ·7H 2 O 0.2g、MnSO 4 ·4H 2 O0.05 g, tween 80 1mL, distilled water 1000mL.
The preparation process of the codonopsis pilosula extract, the angelica extract, the medlar extract, the shizandra berry extract, the dandelion extract and the poria cocos extract used in the invention comprises the following steps: cleaning raw materials, crushing into coarse particles, decocting with 10 times of water for 2 times for 3h and 1h, mixing the two filtrates, standing for 2h, and filtering to obtain extractive solution. Concentrating the extractive solution under vacuum degree of-0.06-0.08 Mpa to relative density of 1.10 (70deg.C), spray drying, and pulverizing into 80 mesh fine powder to obtain extract.
The preparation method of the jujube powder and the coix seed powder comprises the following steps: drying fructus Jujubae and Coicis semen without core until the water content is below 5%, and grinding into 80 mesh powder for use.
The preparation method of the apple juice comprises the following steps: pre-boiling fructus Mali Pumilae for 5min, pulping, extracting juice from the pulp by horizontal screw machine, concentrating the material to 40% by two-effect vacuum evaporator, sterilizing at high temperature, and cooling to obtain fructus Mali Pumilae juice.
Lactobacillus casei used in the inventionLactobacillus casei) SF-L-12 is obtained by self-collection and is preserved in China general microbiological culture Collection center (CGMCC) at 21 months in 2020, the preservation number is CGMCC No.20927, the preservation address is North Chen Xilu No. 1 and 3 in the Chaoyang area of Beijing city, and other related information is disclosed in China patent CN113444675A.
EXAMPLE 1 Lactobacillus casei SF-L-12 fermentation
Firstly, preparing a substrate to be fermented, wherein 74 parts of purified water, 2 parts of radix codonopsis pilosulae extract, 2 parts of Chinese angelica extract, 2 parts of medlar extract, 3 parts of shizandra berry extract, 3 parts of dandelion extract, 3 parts of poria cocos extract, 3 parts of jujube powder, 3 parts of coix seed powder and 5 parts of apple juice are calculated according to parts by weight.
The preparation method comprises the following steps: (1) Firstly, mixing the components of a substrate to be fermented to obtain mixed slurry; (2) Delivering the mixed slurry into a sterilization container, sterilizing at 120 ℃ for 30min under high pressure, and cooling to 35 ℃ to obtain a sterilization liquid; (3) Inoculating lactobacillus casei SF-L-12 into MRS liquid culture medium with an inoculum size of 1%, and culturing at 37deg.C for 18 hr to obtain lactobacillus casei SF-L-12 inoculant; (4) Inoculating lactobacillus casei SF-L-12 inoculant into sterilizing liquid with an inoculum size of 1%, and fermenting at 37deg.C for 24 hr to obtain fermentation liquid; (5) Conveying the fermentation liquor to a high-pressure spraying wall breaking machine, setting the pressure to 60MPa, and carrying out wall breaking treatment to obtain wall breaking fermentation liquor; and (6) performing pasteurization on the wall-broken fermentation broth to obtain a fermentation product.
EXAMPLE 2 Lactobacillus casei SF-L-12 fermentation
Firstly, preparing a substrate to be fermented, wherein the substrate to be fermented comprises, by weight, 64 parts of purified water, 3 parts of codonopsis pilosula extract, 4 parts of Chinese angelica extract, 4 parts of medlar extract, 5 parts of schisandra chinensis extract, 3 parts of dandelion extract, 3 parts of poria cocos extract, 5 parts of jujube powder, 4 parts of coix seed powder and 4 parts of apple juice.
A fermentation was prepared according to the procedure of example 1.
EXAMPLE 3 Lactobacillus casei SF-L-12 fermentation
Firstly, preparing a substrate to be fermented, wherein the substrate to be fermented comprises 82 parts of purified water, 1 part of radix codonopsis pilosulae extract, 1 part of Chinese angelica extract, 1 part of medlar extract, 2 parts of Chinese magnoliavine fruit extract, 2 parts of dandelion extract, 2 parts of poria cocos extract, 2 parts of jujube powder, 2 parts of coix seed powder and 5 parts of apple juice in parts by weight.
A fermentation was prepared according to the procedure of example 1.
Comparative example 1 Lactobacillus casei CICC 20251 fermented product
First, the same substrate to be fermented as in example 1 was prepared.
The preparation method comprises the following steps: (1) Firstly, mixing the components of a substrate to be fermented to obtain mixed slurry; (2) Delivering the mixed slurry into a sterilization container, sterilizing at 120 ℃ for 30min under high pressure, and cooling to 35 ℃ to obtain a sterilization liquid; (3) Inoculating lactobacillus paracasei CICC 20251 into MRS liquid culture medium with an inoculum size of 1%, and culturing at 37deg.C for 18 hr to obtain lactobacillus paracasei CICC 20251 inoculant; (4) Inoculating lactobacillus paracasei CICC 20251 inoculant into the sterilizing liquid, fermenting at 37 ℃ for 24 hours to obtain fermentation liquid; (5) Conveying the fermentation liquor to a high-pressure spraying wall breaking machine, setting the pressure to 60MPa, and carrying out wall breaking treatment to obtain wall breaking fermentation liquor; and (6) performing pasteurization on the wall-broken fermentation broth to obtain a fermentation product.
Comparative example 2 Lactobacillus casei ACCC 10640 ferment
First, the same substrate to be fermented as in example 1 was prepared.
The preparation method comprises the following steps: (1) Firstly, mixing the components of a substrate to be fermented to obtain mixed slurry; (2) Delivering the mixed slurry into a sterilization container, sterilizing at 120 ℃ for 30min under high pressure, and cooling to 35 ℃ to obtain a sterilization liquid; (3) Inoculating lactobacillus casei ACCC 10640 into MRS liquid culture medium with an inoculum size of 1%, and culturing at 37deg.C for 18 hr to obtain lactobacillus casei ACCC 10640 inoculant; (4) inoculating lactobacillus casei ACCC 10640; inoculating into sterilized solution with inoculum size of 1%, fermenting at 37deg.C for 24 hr to obtain fermentation liquid; (5) Conveying the fermentation liquor to a high-pressure spraying wall breaking machine, setting the pressure to 60MPa, and carrying out wall breaking treatment to obtain wall breaking fermentation liquor; and (6) performing pasteurization on the wall-broken fermentation broth to obtain a fermentation product.
Test case Effect of Lactobacillus casei SF-L-12 ferments on mouse serum immunoglobulin
Selecting 50 clean-grade male Kunming mice, keeping the weight of the mice at 18-22 g, carrying out 3-day adaptive feeding, taking 48 mice with similar weight, randomly dividing the mice into 6 groups, 8 mice in each group, respectively filling the fermented products prepared in the example 1, the example 2, the example 3, the comparative example 1 and the comparative example 2 with the purified water, filling the stomach with 2ml of the control group once daily with normal diet drinking water, continuously filling the stomach for 10 days, further feeding for 1 day, taking the eyeballs for the second day after feeding, placing the eyeballs in a low-temperature refrigerator for overnight, centrifuging for 15min at 3000r/min, separating the supernatant, and measuring the content of IgA, igM, igG by adopting an indirect enzyme-linked immunosorbent assay. Statistical analysis was performed using SPSS17.0 and the results are shown in table 1.
TABLE 1 Effect of groups of ferments on mouse serum immunoglobulins
The following is noted: * Indicating p < 0.05.
Immunoglobulins are the main antibodies of humoral immunity, and the high and low serum antibody content reflects the resistance of the body to diseases to a certain extent. Serum immunoglobulin assays are a common method for examining humoral immunity levels, and typically detecting class 3 immunoglobulins of IgG, igA, igM reflects the overall serum immunoglobulin levels. As can be seen from the above table, administration of the Lactobacillus casei SF-L-12 fermented product increased the IgG, igA and IgM contents in the serum of the mice, indicating that the Lactobacillus casei SF-L-12 fermented product increased the immunity of the organism, wherein the mice of group 1 had the highest IgG and IgA contents in the serum, and the IgM contents were significantly increased but not statistically significant differences compared to the control group.
Although the present invention has been described in detail by way of preferred embodiments, the present invention is not limited thereto. Various equivalent modifications and substitutions may be made in the embodiments of the present invention by those skilled in the art without departing from the spirit and scope of the present invention, and it is intended that all such modifications and substitutions be within the scope of the present invention/be within the scope of the present invention as defined by the appended claims.
Claims (8)
1. A lactobacillus casei SF-L-12 ferment is characterized in that the ferment is prepared by fermenting a ferment substrate through lactobacillus casei SF-L-12, and the lactobacillus casei is @Lactobacillus casei) SF-L-12 is preserved in China general microbiological culture Collection center (CGMCC) at the date of 21 and 10 months in 2020, with the preservation number of CGMCC No.20927 and the preservation address of North Chen Xilu No. 1 and 3 in the Chaoyang area of Beijing city; the fermented substrate comprises the following components in parts by weight: 64-82 parts of purified water, 1-3 parts of radix codonopsis extract and radix angelicae sinensis extract1-4 parts of an extract, 1-4 parts of a wolfberry extract, 2-5 parts of a schisandra chinensis extract, 2-3 parts of a dandelion extract, 2-3 parts of a poria cocos extract, 2-5 parts of jujube powder, 2-4 parts of coix seed powder and 4-5 parts of apple juice.
2. Lactobacillus casei SF-L-12 ferment according to claim 1, characterized in that the substrate to be fermented comprises the following components in parts by weight: 74 parts of purified water, 2 parts of radix codonopsis extract, 2 parts of Chinese angelica extract, 2 parts of medlar extract, 3 parts of schisandra chinensis extract, 3 parts of dandelion extract, 3 parts of poria cocos extract, 3 parts of jujube powder, 3 parts of coix seed powder and 5 parts of apple juice.
3. A process for preparing lactobacillus casei SF-L-12 ferment as claimed in claim 1, comprising the steps of:
(1) Firstly, mixing the components of a substrate to be fermented to obtain mixed slurry;
(2) Delivering the mixed slurry into a sterilization container, sterilizing under high pressure, and cooling to obtain a sterilization liquid;
(3) Inoculating lactobacillus casei SF-L-12 into an MRS liquid culture medium, and culturing to obtain a lactobacillus casei SF-L-12 inoculant;
(4) Inoculating lactobacillus casei SF-L-12 inoculant into the sterilizing liquid, and fermenting to obtain fermentation liquid;
(5) Conveying the fermentation liquor to a high-pressure spraying wall breaking machine for wall breaking treatment to obtain wall breaking fermentation liquor;
(6) Pasteurizing the wall-broken fermentation broth to obtain lactobacillus casei SF-L-12 fermented product.
4. A process according to claim 3, wherein in step (2) the autoclave is at 120 ℃ for 30 minutes and then cooled to 35 ℃.
5. The process according to claim 3, wherein the lactobacillus casei SF-L-12 is inoculated in an amount of 1% in step (3), the culturing temperature is 37℃and the culturing time is 18 hours.
6. The process according to claim 3, wherein the inoculum size in step (4) is 1%, the fermentation temperature is 37℃and the fermentation time is 24 hours.
7. The process according to claim 3, wherein in the step (5), the high-pressure spray wall breaker pressure is set to 60Mpa.
8. Use of a lactobacillus casei SF-L-12 ferment as claimed in claim 1 in the manufacture of a product for increasing the secretion of immunoglobulins.
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