CN115838676A - Traditional Chinese medicine fermentation by using novel lactobacillus plantarum strain and application - Google Patents
Traditional Chinese medicine fermentation by using novel lactobacillus plantarum strain and application Download PDFInfo
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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Abstract
The invention relates to the technical field of microorganisms, in particular to traditional Chinese medicine fermentation and application by utilizing a novel lactobacillus plantarum strain. The invention provides lactobacillus plantarum (Lactplantibibacillus plantarum) LP-K0901 with the preservation number of GDMCC No. 63007, which has high biological activity, acid resistance, bile salt resistance, capability of inhibiting harmful bacteria and regulating flora balance, capability of resisting oxidation and removing free radicals, and capability of improving plant fermentation capacity and product functions after fermentation.
Description
Technical Field
The invention relates to the technical field of microorganisms, in particular to traditional Chinese medicine fermentation and application by utilizing a novel lactobacillus plantarum strain.
Background
The lactobacillus plantarum belongs to a gram-positive probiotic which does not produce spores, and is mainly applied to probiotic products and traditional Chinese medicine fermentation. The higher the biological activity of the lactobacillus plantarum is, the higher the yield of industrial production is, the stronger the efficacy effect is, the production cost is further reduced, and the production efficiency is improved. Therefore, screening of high-activity lactobacillus plantarum is an industrial goal.
The lactobacillus plantarum used for developing probiotic products needs to have strong gastric acid and bile salt resistance so as to ensure that the lactobacillus plantarum effectively passes through the stomach and the small intestine to reach the large intestine to play a role. The increase of harmful intestinal bacteria such as Escherichia coli can cause flora imbalance and cause various diseases, and if the lactobacillus plantarum has a good effect of inhibiting the harmful bacteria, the restoration of the intestinal flora balance can be accelerated. Free radicals are harmful compounds generated in oxidation reaction of organisms, can promote aging and diseases of the organisms and cause health problems, remove oxidation resistance such as free radicals and the like, and are beneficial to maintaining the health of the organisms.
The traditional Chinese medicine with homology of medicine and food and the like can be applied to food due to safety, but the effect is relatively not ideal, how to improve the effect of the traditional Chinese medicine with homology of medicine and food becomes an industrial difficulty, the problem of low effect of plants such as the traditional Chinese medicine with homology of medicine and food can be solved through a traditional Chinese medicine fermentation technology, and the key point of traditional Chinese medicine fermentation is to find an excellent strain for fermenting the traditional Chinese medicine.
No reports of lactobacillus plantarum strains with remarkable performance in the fermentation aspect of herbaceous plants such as probiotics, traditional Chinese medicines and the like are found.
The lower the biological activity of the existing lactobacillus plantarum is, the weak gastric acid and bile salt resistance, the weak functions of inhibiting harmful bacteria and regulating flora balance, the weak capabilities of scavenging free radicals and resisting oxidation, and the low fermentation capability and the low functions of fermented products of Chinese medicines and other plants.
Disclosure of Invention
In view of the above, the invention provides traditional Chinese medicine fermentation and application by using a novel lactobacillus plantarum strain, wherein the strain has the functions of high biological activity, acid resistance, bile salt resistance and strong oxidation resistance, and can adjust plant fermentation and flora balance, and improve plant fermentation capacity and functions of fermented products.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides Lactobacillus plantarum (Lactplantibibacillus plantarum) LP-K0901, the preservation number of which is GDMCC No. 63007.
In some embodiments of the present invention, the lactobacillus plantarum LP-K0901 includes one or more of viable bacteria, inactivated bacteria, fermentation broth, exosomes or metabolites thereof.
In some embodiments of the present invention, the Lactobacillus plantarum LP-K0901 has high biological activity, high gastric acid resistance, and high bile salt resistance.
The invention also provides application of the lactobacillus plantarum (Lactplantibibacillus plantarum) LP-K0901 in preparation of any of the following items:
(I) A probiotic preparation, a synbiotic preparation and/or a metabiotic preparation; and/or
(II) products for plant fermentation;
the micro-ecological product, probiotic product, synbiotic product, post-biotic product and/or the product for plant fermentation include, but are not limited to, one or more of a drug, a nutraceutical, a food for special medical use, a functional food, a cosmetic, a feed additive or a pet food; and/or
The dosage form of the microecological product, the probiotic product, the synbiotic product, the metazoan product and/or the product for plant fermentation comprises one or more of but not limited to powder, granule, capsule, tablet, pill, extract, liquid preparation, sugar or jelly; and/or
Including but not limited to fondants and/or tabletted confections; and/or
The plant fermentation includes but is not limited to one or more of traditional Chinese medicine fermentation, fruit and vegetable fermentation or new food raw material fermentation;
the traditional Chinese medicine fermentation includes but is not limited to the fermentation of medicine-food homologous traditional Chinese medicines;
the traditional Chinese medicine comprises but is not limited to one or more of poria cocos, radix puerariae and lucid ganoderma.
The invention also provides a product which comprises the Lactobacillus plantarum LP-K0901;
the article comprises any of:
(I) A probiotic preparation, a synbiotic preparation, and/or a metabiotic preparation; and/or
(II) products for plant fermentation;
the micro-ecological product, probiotic product, synbiotic product, post-biotic product and/or the product for plant fermentation include, but are not limited to, one or more of a drug, a nutraceutical, a food for special medical use, a functional food, a cosmetic, a feed additive or a pet food; and/or
The dosage form of the microecological preparation, probiotic preparation, metazoan preparation and/or the preparation for plant fermentation comprises but is not limited to one or more of powder, granule, capsule, tablet, pill, extract, liquid, sugar or jelly; and/or
Including but not limited to fondants and/or tabletted confections; and/or
The plant fermentation includes but is not limited to one or more of traditional Chinese medicine fermentation, fruit and vegetable fermentation or new food raw material fermentation;
the traditional Chinese medicine fermentation includes but is not limited to the fermentation of medicine-food homologous traditional Chinese medicines;
the traditional Chinese medicine comprises but is not limited to one or more of poria cocos, radix puerariae and lucid ganoderma.
The invention also provides a product which is prepared by fermenting the lactobacillus plantarum (Lactplantibibacillus plantarum) LP-K0901.
In some embodiments of the invention, the above products include, but are not limited to, products of plant fermentation; and/or
The product includes, but is not limited to, one or more of a pharmaceutical product, a nutraceutical product, a food for special medical use, a functional food, a food product, a cosmetic product, a feed additive or a pet food; and/or
The dosage form of the product comprises but is not limited to one or more of powder, granules, capsules, tablets, pills, extractum, liquid preparations, sugar or jelly; and/or
Including but not limited to fondants and/or tabletted confections; and/or
The plant fermentation includes but is not limited to one or more of traditional Chinese medicine fermentation, fruit and vegetable fermentation or new food raw material fermentation;
the traditional Chinese medicine fermentation includes but is not limited to the fermentation of medicine-food homologous traditional Chinese medicines;
the traditional Chinese medicine comprises but is not limited to one or more of poria cocos, radix puerariae and lucid ganoderma.
The invention also provides the Lactobacillus plantarum (Lactplantibibacillus plantarum) LP-K0901; and/or
The above-mentioned articles; and/or
Use of the product in any of the following:
(I) Preparing a product for inhibiting the growth of harmful bacteria; and/or
(II) preparing a product for promoting the growth of beneficial bacteria; and/or
(III) preparing a product for regulating the flora balance; and/or
(IV) preparing an antioxidant product; and/or
(V) improving the product performance of plant fermentation; and/or
(VI) preparing a medicament or formulation for the prevention and/or treatment of disease; and/or
(VII) preparing a health-care product.
In some embodiments of the present invention, in the above applications, the harmful bacteria include, but are not limited to, staphylococcus aureus and/or escherichia coli; and/or
The beneficial bacteria include but are not limited to bifidobacteria and/or lactobacilli; and/or
The flora balance includes, but is not limited to, one or more of intestinal flora, vaginal flora, skin flora, oral flora, gastric flora, or urinary system flora; and/or
The antioxidant includes, but is not limited to scavenging free radicals; preferably, the scavenging of free radicals includes, but is not limited to, scavenging DPPH free radicals; and/or
Improving the product performance of plant fermentation includes but is not limited to improving safety and/or enhancing immunity function; and/or
The prevention and/or treatment of diseases include, but are not limited to, one or more of digestive system diseases, motor system diseases, respiratory system diseases, urinary system diseases, reproductive system diseases, circulatory system diseases, endocrine system diseases, nervous system diseases, or immune system diseases; and/or
The health care includes, but is not limited to, one or more of nutritional health care, maintaining physical health, or beautifying.
In some embodiments of the present invention, in the above-mentioned applications, the digestive system disease includes, but is not limited to, one or more of spleen deficiency, stomach nourishing, diarrhea, irritable bowel syndrome, ulcerative colitis, crohn's disease, antibiotic-associated diarrhea, pseudomembranous enteritis, infantile secondary diarrhea to pneumonia, intestinal cancer, tumor radiotherapy and chemotherapy adjuvant therapy, abdominal distension and dyspepsia, constipation, stool odor, hepatobiliary disease, viral diarrhea, feeding intolerance, necrotizing enterocolitis, jaundice, oral ulcer, thrush, promotion of infantile intestinal development maturation, digestive absorption, nutritional health or growth development or nutritional malabsorption diseases;
the liver and gall diseases comprise but are not limited to liver diseases and/or alcohol dispelling and liver protecting;
preferably, the hepatobiliary disease includes, but is not limited to, one or more of fatty liver, hepatitis, cirrhosis or liver cancer;
preferably, the hepatobiliary disease includes, but is not limited to, alcoholic and/or non-alcoholic liver disease;
preferably, the hepatobiliary disease includes, but is not limited to, one or more of alcoholic fatty liver, alcoholic hepatitis, non-alcoholic fatty liver, or non-alcoholic steatohepatitis; and/or
The motor system disease includes, but is not limited to, one or more of arthritis, osteoporosis, or rickets;
preferably, the motor system disease includes, but is not limited to, one or more of degenerative arthritis, rheumatoid arthritis, or ankylosing spondylitis; and/or
The respiratory disease includes but is not limited to one or more of nourishing lung, moistening lung to arrest cough, pulmonary nodule, pneumonia, novel coronavirus infection, recurrent respiratory tract infection or antiviral infection; and/or
The urinary system diseases include but are not limited to kidney deficiency;
preferably, the urinary system disorder includes, but is not limited to, kidney yin deficiency and/or kidney yang deficiency; and/or
The reproductive system disease includes but is not limited to one or more of vaginitis, uterine fibroids, dysmenorrhea, breast nodules, breast cancer or climacteric syndrome; and/or
The circulatory system diseases include but are not limited to one or more of cardiovascular and cerebrovascular diseases, hyperlipidemia or hypertension; and/or
The endocrine system disease includes but is not limited to one or more of hyperglycemia, dyslipidemia, diabetes, hyperuricemia, gout or thyroid nodule; and/or
The neurological disorder includes, but is not limited to, one or more of insomnia, mood relief, sleep disorder, headache, depression, or anxiety; and/or
The immune system diseases include but are not limited to one or more of allergic diseases, allergy, skin diseases, immunity enhancement, allergic rhinitis; and/or
The maintaining of body health comprises but is not limited to one or more of eliminating dampness, invigorating qi, tonifying yang, replenishing blood, tonifying yin or resisting oxidation; and/or
The beautifying includes but is not limited to one or more of sugar resistance, moisture retention or whitening.
The invention also provides a medicament, which comprises any of the following items and acceptable auxiliary materials:
(I) The Lactobacillus plantarum (Lactplantibibacillus plantarum) LP-K0901; and/or
(II) the above-mentioned products; and/or
(III) the above products.
The invention also provides a medicine combination which comprises the medicine and other any effective components.
The bacterial strain and the application of the invention have the following effects:
the results of the biological activity experiments show that the OD values of the lactobacillus plantarum LP-K0901 cultured for 16h and 24h are higher than those of other strains, and the biological activity of the lactobacillus plantarum LP-K0901 is higher than that of the other strains. The acid resistance experiment result shows that the survival rate of 5 lactobacillus plantarum strains is reduced to different degrees along with the time prolongation under the condition of pH 2.0, wherein the survival rate of lactobacillus plantarum LP-K0901 is the highest, which indicates that the acid resistance of the lactobacillus plantarum strain is the strongest under the condition of pH 2.0. Wherein the survival rate of the lactobacillus plantarum LP-K0901 is inversely increased along with the prolonging of time under the condition of pH 3.0, and is more than 100% after 4h, which indicates that the lactobacillus plantarum LP-K0901 has the strongest acid resistance under the condition of pH 3.0. The survival rates of the other 4 strains showed a downward trend with time. The results of the bile salt resistance experiment show that the survival rate of 5 lactobacillus plantarum strains is reduced under the condition of 0.3% of bile salt, wherein the survival rate of the lactobacillus plantarum LP-K0901 is the highest, which shows that the lactobacillus plantarum has the strongest capacity of resisting 0.3% of bile salt and can be quickly adapted to high bile salt concentration. The experimental result of removing DPPH free radicals shows that the lactobacillus plantarum LP-K0901 fermentation liquor has the highest DPPH free radical removal rate, and the bacteria has the strongest DPPH free radical removal capacity. The bacteriostatic ability experiment result shows that the bacteriostatic circle diameter of 5 strains of lactobacillus plantarum on staphylococcus aureus shows that the lactobacillus plantarum LP-K0901 has the strongest ability of inhibiting staphylococcus aureus. The strongest inhibition ability of the lactobacillus plantarum LP-K0901 to escherichia coli is shown by comparing the inhibition zone diameter of 5 strains of lactobacillus plantarum to escherichia coli. The experimental result of the fermented traditional Chinese medicine shows that the poria cocos is fermented for 24h to 48h by using the lactobacillus plantarum LP-K0901, the OD value is highest, the pH value is lowest, and the fact that the bacteria grow best in the poria cocos fermentation liquor and produce the most acid is shown; fermenting Ganoderma with Lactobacillus plantarum LP-K0901 for 24h to 48h, wherein OD value is highest and pH value is lowest, which indicates that the strain grows best in Ganoderma fermentation liquid and produces most acid; pueraria lobata is fermented for 24h to 48h by using lactobacillus plantarum LP-K0901, the OD value is highest, and the pH value is lowest, which shows that the bacterium grows best in lucid ganoderma fermentation liquor and produces most acid. The experimental result of immunity enhancement shows that the phagocytic index of the fermentation liquor is higher than that of the fermentation liquor of other strains after the lactobacillus plantarum LP-K0901 ferments the traditional Chinese medicines of poria cocos, lucid ganoderma and radix puerariae for 24 hours, which shows that the immunity of the fermentation liquor is improved more than that of other strains after the fermentation of the strains.
Description of biological preservation
Biological material: lactplantibacillus plantarum LP-K0901, under taxonomic nomenclature: lactplantibacillus plantarum, deposited at 28/11/2022 in the Guangdong province collection of microorganisms at the following locations: the microbiological research institute of the department of sciences, guangzhou, no. 59 building, no. 5 building, zhonglu, xieli, dazhou, guangzhou, mingzhou, ministry of sciences; the collection number is GDMCC No. 63007.
The LP-K0901 is the strain with the preservation number of GDMCC No. 63007.
Detailed Description
The invention discloses traditional Chinese medicine fermentation and application by utilizing a novel lactobacillus plantarum strain, and a person skilled in the art can realize the fermentation by properly improving process parameters by referring to the content. It is specifically noted that all such substitutions and modifications will be apparent to those skilled in the art and are intended to be included herein. While the methods and applications of this invention have been described in terms of preferred embodiments, it will be apparent to those of ordinary skill in the art that variations and modifications in the methods and applications described herein, as well as other suitable variations and combinations, may be made to implement and use the techniques of this invention without departing from the spirit and scope of the invention.
Aiming at the problems of unsatisfactory functions of the prior probiotic strains such as biological activity, acid resistance, cholate resistance, oxidation resistance, plant fermentation, flora regulation and the like, the invention provides a strain, namely lactobacillus plantarum LP-K0901, of which the comprehensive performance is superior to that of the lactobacillus plantarum in the market.
Based on the defects and the requirements of the prior art in the field, the invention aims to provide a novel lactobacillus plantarum LP-K0901, which is preserved in Guangdong province microorganism strain preservation center with the preservation number: GDMCC No. 63007. The lactobacillus plantarum strain is superior to lactobacillus plantarum strains on the market and other lactobacillus plantarum strains screened by the inventor in the aspects of bioactivity, gastric acid and bile salt resistance, harmful bacterium inhibition, beneficial bacterium promotion, intestinal flora balance regulation, oxidation resistance, plant fermentation capacity, fermented product function, disease prevention, health care and the like.
The flora balance is regulated, and the flora balance comprises human flora such as intestinal flora, vaginal flora, skin flora, oral flora, stomach flora, urinary system flora, etc.
The inhibition has bacteriostasis, and comprises staphylococcus aureus, escherichia coli and the like.
The product can promote the growth of beneficial bacteria, including Bifidobacterium, lactobacillus, etc.
The antioxidant, including scavenging free radicals, further includes scavenging DPPH free radicals.
The plant fermentation capacity comprises traditional Chinese medicine fermentation, fruit and vegetable fermentation, new food raw material fermentation and the like, and the traditional Chinese medicine fermentation further comprises medicine-food homologous traditional Chinese medicine fermentation.
The function of the plant fermentation product is improved by fermenting the plant, and further comprises the functions of improving safety and enhancing immunity.
The application of the product in disease prevention and treatment and health care.
The health product can be used for preventing and treating diseases, including digestive system diseases, motor system diseases, respiratory system diseases, urinary system diseases, reproductive system diseases, circulatory system diseases, endocrine system diseases, nervous system diseases, immune system diseases, nourishing, protecting health, caring skin, etc.
The digestive system diseases comprise spleen deficiency, stomach nourishing, diarrhea, irritable bowel syndrome, ulcerative colitis, crohn's disease, antibiotic-associated diarrhea, pseudomembranous enteritis, infantile pneumonia secondary diarrhea, intestinal cancer, tumor radiotherapy and chemotherapy adjuvant treatment, abdominal distension and dyspepsia, constipation, excrement odor, liver and gall diseases, viral diarrhea, feeding intolerance, necrotizing enterocolitis, jaundice, oral ulcer, thrush, infantile intestinal tract development maturation promotion, digestive absorption, nutrition health care or growth and development, and nutritional absorption disorder diseases.
The liver and gall diseases comprise liver diseases or hangover and liver protection, further comprise fatty liver, hepatitis, cirrhosis or liver cancer, further comprise alcoholic liver diseases or non-alcoholic liver diseases, and further comprise alcoholic fatty liver, alcoholic hepatitis, non-alcoholic fatty liver or non-alcoholic steatohepatitis and the like.
The diseases of the locomotor system comprise arthritis, osteoporosis, rickets, degenerative arthritis, rheumatoid arthritis, ankylosing spondylitis, etc.
The respiratory diseases comprise nourishing lung, moistening lung for arresting cough, pulmonary nodule, pneumonia, new crown pneumonia, recurrent respiratory tract infection, antivirus infection, etc.
The urinary system diseases comprise kidney deficiency, further comprise kidney yin deficiency, kidney yang deficiency and the like.
The reproductive system diseases include vaginitis, hysteromyoma, dysmenorrhea, breast nodule, breast carcinoma, climacteric syndrome, etc
The circulatory system diseases comprise cardiovascular and cerebrovascular diseases, hyperlipidemia, hypertension, etc.
The endocrine system diseases comprise hyperglycemia, blood sugar abnormality, diabetes, hyperuricemia, gout, thyroid nodule and the like.
The nervous system diseases comprise insomnia, mood relief, abnormal sleep, headache, depression, anxiety and the like.
The immune system diseases comprise allergic diseases, allergy, skin diseases, immunity enhancement, allergic rhinitis and the like.
The health care includes dampness elimination, qi tonifying, yang tonifying, blood enriching, yin tonifying, oxidation resistance and the like.
The beautifying comprises sugar resistance, moisture retention, whitening and the like.
A micro-ecological, probiotic, synbiotic or post-biotic product comprising an active ingredient; the active ingredients comprise lactobacillus plantarum LP-K0901, with a deposit number: GDMCC No. 63007. The micro-ecological, probiotic or post-biotic product further comprises: an auxiliary material; the auxiliary materials are selected from medicinal auxiliary materials or edible auxiliary materials; the product comprises medicines, health-care foods, foods for special medical use, foods, cosmetics, feeds, feed additives, pet foods and the like; the dosage form of the product comprises powder, granules, capsules, tablets, pills, extractum, liquid preparations, sugar and jelly, and the sugar further comprises soft sweets and tabletting candies.
The fermented plant product has fermented strain at least comprising Lactobacillus plantarum LP-K0901 with the preservation number: GDMCC No. 63007. The plant fermentation product also comprises: an auxiliary material; the auxiliary materials are selected from medicinal auxiliary materials or edible auxiliary materials; the product comprises medicines, health-care foods, foods for special medical use, foods, cosmetics, feeds, feed additives, pet foods and the like; the dosage form of the product comprises powder, granules, capsules, tablets, pills, extractum, liquid preparations, sugar and jelly, and the sugar further comprises soft sweets and tabletting candies.
The lactobacillus plantarum LP-K0901 comprises live thalli, inactivated thalli or metabolites and the like.
The invention screens lactobacillus plantarum from natural conditions.
The examples illustrate that: the invention is described with the representative of Lactobacillus plantarum LP-K0901, but the invention is not limited thereto.
The lactobacillus plantarum provided by the invention is Lactplantibibacillus plantarum.
Unless otherwise specified, the raw materials, reagents, consumables and instruments according to the present invention are all commercially available products, and are all commercially available.
The invention is further illustrated by the following examples:
example 1
Description of the preparation examples: the powder of Lactobacillus plantarum LP-K0901 is specifically described as an example, the preparation method of other preparations of Lactobacillus plantarum LP-K0901 can be easily grasped by those skilled in the art through the example, and the preparation method of other preparations can be easily grasped by those skilled in the art through the implementation, and the description is omitted here. The preparation method is not limited to the examples of the invention, and any known method capable of achieving the preparation purpose can be used, and the preparation description of the examples is only illustrative of the invention and is not limiting to the protection scope of the invention.
1 preparation of fungal powder
Inoculating Lactobacillus plantarum LP-K0901 into MRS liquid culture medium, culturing at 37 deg.C for 24 hr, centrifuging the obtained culture solution to separate out thallus, and freeze drying the thallus under vacuum to obtain dry powder.
2 making into powder
Preparing the plant lactobacillus LP-K0901 into bacterial powder, adding related adjuvants as required, and making into various dosage forms, preferably powder with viable count not less than 1 × 10 7 CFU/g, and then bagging.
Example 2: biological Activity assay
The experimental method comprises the following steps: the strain LP-K0901 is activated in MRS liquid culture medium for 7h, diluted with sterile water to have OD value of 1%, inoculated into 100mL of MRS liquid culture medium according to 1% inoculation amount, subjected to static culture at 37 ℃ for 16-24 hours, and sampled at 16h and 24h to detect the OD value. The results are shown in Table 1.
Table 1: OD value of biological activity of strain
And (4) conclusion: the lactobacillus plantarum LP-K0901 group was statistically significantly different from the other groups (P < 0.05). The OD values of the lactobacillus plantarum LP-K0901 after 16 hours and 24 hours of culture are higher than those of other strains, which shows that the biological activity of the lactobacillus plantarum LP-K0901 is higher than that of the other strains.
Example 3: acid resistance test
The experimental method comprises the following steps: adjusting the pH value of the MRS liquid culture medium to 2.0 and 3.0 by using 1mol/L HCl respectively; sterilizing at 121 ℃ for 15min, activating an LP-K0901 strain in an MRS liquid culture medium for 16h, diluting with sterile water to obtain an OD value of 5.0, wherein the inoculation amount is 2 percent, keeping the initial viable count in the culture medium the same, culturing at the constant temperature of 37 ℃, and sampling for 0h and 4h to measure the OD value, wherein the strain survival rate (%) = Nt/N0 multiplied by 100 percent, wherein N0 represents the OD value of the strain 0h, and Nt represents the OD value of the viable count of the strain 4h. The results are shown in tables 2 and 3.
Table 2: data table of strains at pH 2.0
Table 3: data table of strains at pH 3.0
And (4) conclusion: the lactobacillus plantarum LP-K0901 group was significantly different from the other groups (P < 0.01) by statistical analysis.
1. The survival rate of 5 strains of the lactobacillus plantarum is reduced to different degrees along with the prolonging of time under the condition of pH 2.0, wherein the survival rate of the lactobacillus plantarum LP-K0901 is the highest, which indicates that the acid resistance of the lactobacillus plantarum is the strongest under the condition of pH 2.0.
2. The survival rate of the lactobacillus plantarum LP-K0901 is increased inversely with the time when the pH value is 3.0, and is more than 100% after 4h, which indicates that the lactobacillus plantarum LP-K0901 has the strongest acid resistance under the pH value of 3.0. The survival rates of the other 4 strains showed a downward trend with time.
Example 4: bile salt resistance test
The experimental method comprises the following steps: the pig bile salt is weighed, dissolved by stirring with pure water, and added into MRS broth to prepare a bile salt liquid culture medium with a concentration of 0.3%. LP-K0901 strains were activated in MRS liquid medium for 16h. Inoculating 2% seed solution with the same OD value into a liquid culture medium containing 0.3% bile salt, and culturing at 37 deg.C for 4h. And sampling at 0h and 4h respectively to measure OD, and counting the survival rate. Wherein, the strain survival rate (%) = Nt/N0 × 100%, wherein N0 represents the OD value of the strain 0h, and Nt represents the OD value of the viable bacteria number of the strain 4h. The results are shown in Table 4.
Table 4: survival rate of strain under 0.3% bile salt condition
And (4) conclusion: the Lactobacillus plantarum LP-K0901 group was significantly different from the other groups (P < 0.01) by statistical analysis. Under the condition of 0.3% of bile salt, the survival rate of 5 strains of lactobacillus plantarum is reduced, wherein the survival rate of lactobacillus plantarum LP-K0901 is the highest, which indicates that the lactobacillus plantarum has the strongest capacity of tolerating 0.3% of bile salt and can be quickly adapted to high bile salt concentration.
Example 5: experiment for eliminating DPPH free radical
The experimental method comprises the following steps: 2mL of LP-K0901 fermentation broth fermented for 24h is added into 2mL of 0.1mmol/L freshly prepared DPPH-absolute ethanol solution, the mixture is uniformly mixed, the mixture is reacted for 30min at room temperature in dark in a dark place, the mixture is centrifuged for 2min at 10000r/min, and the supernatant is taken to measure the light absorption value at the wavelength of 517nm and is recorded As As. Replacing a DPPH solution with equal volume of absolute ethyl alcohol in the blank group to measure a light absorption value, and marking as Ab; the absorbance of the control was measured by replacing the sample solution with ultrapure water and was designated as Ac.
DPPH radical clearance (%) = [1- (As-Ab)/Ac ] × 100%, where As is the sample set absorbance; ac is the absorbance of the control group (distilled water instead of the sample); ab is blank absorbance (absolute ethanol instead of DPPH). The results are shown in Table 5.
Table 5: DPPH radical clearance rate
And (4) conclusion: the lactobacillus plantarum LP-K0901 group was statistically significantly different from the other groups (P < 0.05). Experimental data show that the lactobacillus plantarum LP-K0901 fermentation broth has the highest scavenging rate on DPPH free radicals, and the bacteria strain has the strongest scavenging capacity on DPPH free radicals.
Example 6: experiment of bacteriostatic ability
The experimental method comprises the following steps:LP-K0901 strains were activated in MRS broth for 16h at 37 ℃. Pouring 20mL of MRS agar culture medium on each plate, taking 100 mu L of test bacterial liquid (escherichia coli and staphylococcus aureus) for gradient dilution, wherein the concentration of the diluted bacterial liquid is 10 5 ～10 6 About cfu, 100 μ L of diluted bacterial liquid is respectively dropped on a solid plate, and is coated by a coating rod and is left for standby. And lightly placing the sterilized Oxford cup on the surface of a plate culture medium (3 per plate), standing for 5min, sucking 100 mu L of activated bacteria liquid, adding the activated bacteria liquid into the Oxford cup, transferring the plate to a 37 ℃ incubator for culturing for 24-72 h after the bacteria liquid is fully diffused for 2h, observing the phenomenon at regular time, measuring the diameter of a bacteriostatic zone by using a ruler, and taking a picture for recording. The results are shown in Table 6.
Table 6: diameter data of bacteria inhibition zone of strain on staphylococcus aureus and escherichia coli
And (4) conclusion: the lactobacillus plantarum LP-K0901 group was statistically significantly different from the other groups (P < 0.05).
1. The comparison of the diameter of the inhibition zone of 5 strains of the lactobacillus plantarum on staphylococcus aureus shows that the lactobacillus plantarum LP-K0901 has the strongest capacity of inhibiting staphylococcus aureus.
2. The strongest inhibition ability of the lactobacillus plantarum LP-K0901 to escherichia coli is shown by comparing the inhibition zone diameter of 5 strains of lactobacillus plantarum to escherichia coli.
Example 7: experiment of fermented Chinese medicine
The experimental method comprises the following steps: respectively adding water into poria cocos, lucid ganoderma and kudzuvine root, boiling and extracting for 1-2 h, filtering and collecting supernatant, adding 4% of crystalline fructose, subpackaging, and sterilizing at 121 ℃ for 30min; LP-K0901 strains were activated in MRS broth for 16h at 37 ℃. Inoculating according to the inoculation amount of 2%, culturing at 37 ℃ for 48h, and sampling at 24h and 48h to detect the OD value and the pH value. The results are shown in tables 7 to 9.
Table 7: experimental data of fermented poria cocos
And (4) conclusion: the Lactobacillus plantarum LP-K0901 group was significantly different from the other groups (P < 0.01) by statistical analysis. The experimental result shows that the tuckahoe is fermented for 24 to 48h by the lactobacillus plantarum LP-K0901, the OD value is highest, the pH value is lowest, and the bacterium grows best in tuckahoe fermentation liquor and produces most acid.
Table 8: experimental data of fermented ganoderma
And (4) conclusion: the lactobacillus plantarum LP-K0901 group was significantly different from the other groups (P < 0.01) by statistical analysis. Experimental results show that the lactobacillus plantarum LP-K0901 is used for fermenting the lucid ganoderma for 24h to 48h, the OD value is highest, and the pH value is lowest, which shows that the strain grows best in lucid ganoderma fermentation liquor and produces most acid.
Table 9: experimental data of fermented kudzu vine root
And (4) conclusion: the lactobacillus plantarum LP-K0901 group was significantly different from the other groups (P < 0.01) by statistical analysis. Experimental results show that the kudzu root is fermented for 24h to 48h by using the lactobacillus plantarum LP-K0901, the OD value is highest, and the pH value is lowest, which shows that the bacterium grows best in the lucid ganoderma fermentation liquid, and produces most acid.
Example 8: experiment for enhancing immunity
The experimental method comprises the following steps: RAW264.7 cells were added to a 96-well plate at 200. Mu.L/well, 37 ℃ with 5% CO 2 Culturing for 16H with 0.8-1 mmol/L H 2 O 2 Adding the solution into 96-well plate to damage cells, adding culture solution into control group, and culturing at 37 deg.C with 5% CO 2 And culturing for 24 hours under the condition. Diluting the LP-K0901 fermentation broth and adding the diluted broth into a 96-well plate, H 2 O 2 Adding culture solution to group and control group, and adding 5% CO at 37 deg.C 2 And (5) carrying out conditioned culture for 24 hours. After sucking out the liquid, add per holeAnd (5) continuously culturing for 4 hours by using a 10% neutral red dye solution. Sucking out the neutral red dye solution, adding 150 μ L/hole of neutral red lysis solution, mixing for 10min, and detecting with enzyme labeling instrument at 490nm wavelength. Phagocytosis index = experiment OD value/control OD value, increase rate = (experiment phagocytosis index-H) 2 O 2 Phagocytosis index)/H 2 O 2 Group phagocytosis index 100%. The results are shown in tables 10 to 12.
Table 10: experimental data of poria cocos fermentation liquor for enhancing immunity
Table 11: data of experiment for enhancing immunity of poria cocos and ganoderma lucidum liquid
Table 12: experimental data for enhancing immunity of radix puerariae fermentation liquid
And (4) conclusion: the lactobacillus plantarum LP-K0901 group was statistically significantly different from the other groups (P < 0.05). Cell phagocytosis index experiments prove that after the lactobacillus plantarum LP-K0901 ferments the traditional Chinese medicines of poria cocos, lucid ganoderma and radix puerariae for 24 hours, the phagocytosis index of the fermentation liquor is higher than that of the fermentation liquor of other strains, which shows that the immunity of the fermentation liquor is improved more than that of the other strains after the fermentation of the strain.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (10)
1. Lactobacillus plantarum (Lactplantibibacillus plantarum) LP-K0901, with accession number GDMCC No:63007.
2. Lactobacillus plantarum LP-K0901, according to claim 1, comprising one or more of viable bacteria, inactivated bacteria, fermentation broth, exosomes or metabolites.
3. Lactobacillus plantarum LP-K0901, according to claim 1 or 2, including its higher bioactivity, greater gastric acid tolerance and greater bile salt tolerance.
4. Use of Lactobacillus plantarum (Lactplantibibacillus plantarum) LP-K0901 according to any one of claims 1 to 3 for the preparation of any one of:
(I) A probiotic preparation, a synbiotic and/or a metabiotic preparation; and/or
(II) products for plant fermentation;
the micro-ecological product, probiotic product, synbiotic product, post-biotic product and/or the product for plant fermentation include, but are not limited to, one or more of a drug, a nutraceutical, a food for special medical use, a functional food, a cosmetic, a feed additive or a pet food; and/or
The dosage form of the microecological product, the probiotic product, the synbiotic product, the metazoan product and/or the product for plant fermentation comprises one or more of but not limited to powder, granule, capsule, tablet, pill, extract, liquid preparation, sugar or jelly; and/or
Including but not limited to fondants and/or tabletted confections; and/or
The plant fermentation includes but is not limited to one or more of traditional Chinese medicine fermentation, fruit and vegetable fermentation or new food raw material fermentation;
the traditional Chinese medicine fermentation includes but is not limited to the fermentation of medicine-food homologous traditional Chinese medicines;
the traditional Chinese medicine comprises but is not limited to one or more of poria cocos, radix puerariae and lucid ganoderma.
5. Article of manufacture comprising Lactobacillus plantarum (Lactplantibibacillus plantarum) LP-K0901 according to any one of claims 1 to 3;
the article comprises any of:
(I) A probiotic preparation, a synbiotic preparation, and/or a metabiotic preparation; and/or
(II) products for plant fermentation;
the micro-ecological product, probiotic product, synbiotic product, post-biotic product and/or the product for plant fermentation include, but are not limited to, one or more of a drug, a nutraceutical, a food for special medical use, a functional food, a cosmetic, a feed additive or a pet food; and/or
The dosage form of the microecological product, the probiotic product, the synbiotic product, the metazoan product and/or the product for plant fermentation comprises one or more of but not limited to powder, granule, capsule, tablet, pill, extract, liquid preparation, sugar or jelly; and/or
Including but not limited to fondants and/or tabletted confections; and/or
The plant fermentation includes but is not limited to one or more of traditional Chinese medicine fermentation, fruit and vegetable fermentation or new food raw material fermentation;
the traditional Chinese medicine fermentation includes but is not limited to the fermentation of medicine-food homologous traditional Chinese medicines;
the traditional Chinese medicine comprises but is not limited to one or more of poria cocos, radix puerariae and lucid ganoderma.
6. Product, produced by fermentation of the Lactobacillus plantarum (Lactplantibibacillus plantarum) LP-K0901 according to any one of claims 1 to 3.
7. The product of claim 6, wherein the product includes, but is not limited to, a plant fermented product; and/or
The product includes, but is not limited to, one or more of a pharmaceutical product, a nutraceutical product, a food for special medical use, a functional food, a food product, a cosmetic product, a feed additive or a pet food; and/or
The dosage form of the product comprises but is not limited to one or more of powder, granules, capsules, tablets, pills, extractum, liquid preparation, sugar or jelly; and/or
Including but not limited to fondants and/or tabletted confections; and/or
The plant fermentation includes but is not limited to one or more of traditional Chinese medicine fermentation, fruit and vegetable fermentation or new food raw material fermentation;
the traditional Chinese medicine fermentation includes but is not limited to the fermentation of medicine-food homologous traditional Chinese medicines;
the traditional Chinese medicine comprises but is not limited to one or more of poria cocos, radix puerariae and lucid ganoderma.
8. Lactobacillus plantarum (Lactplantibibacillus plantarum) LP-K0901 according to any one of claims 1 to 3; and/or
The article of claim 5; and/or
Use of the product of claim 6 or 7 in any of:
(I) Preparing a product for inhibiting the growth of harmful bacteria; and/or
(II) preparing a product for promoting the growth of beneficial bacteria; and/or
(III) preparing a product for regulating the flora balance; and/or
(IV) preparing an antioxidant product; and/or
(V) improving the product performance of plant fermentation; and/or
(VI) preparing a medicament or formulation for the prevention and/or treatment of disease; and/or
(VII) preparing a health-care product.
9. The use of claim 8, wherein the harmful bacteria include, but are not limited to, staphylococcus aureus and/or escherichia coli; and/or
The beneficial bacteria include but are not limited to bifidobacteria and/or lactobacilli; and/or
The flora balance includes, but is not limited to, one or more of intestinal flora, vaginal flora, skin flora, oral flora, gastric flora, or urinary system flora; and/or
The antioxidant includes, but is not limited to scavenging free radicals; preferably, the scavenging of free radicals includes, but is not limited to, scavenging DPPH free radicals; and/or
Improving the product performance of plant fermentation includes but is not limited to improving safety and/or enhancing immunity function; and/or
The prevention and/or treatment of diseases include, but are not limited to, one or more of digestive system diseases, motor system diseases, respiratory system diseases, urinary system diseases, reproductive system diseases, circulatory system diseases, endocrine system diseases, nervous system diseases, or immune system diseases; and/or
The health care includes, but is not limited to, one or more of nutritional health care, maintaining physical health, or beautifying;
the digestive system diseases include but are not limited to one or more of spleen deficiency, stomach nourishing, diarrhea, irritable bowel syndrome, ulcerative colitis, crohn's disease, antibiotic-associated diarrhea, pseudomembranous enteritis, infantile secondary diarrhea of pneumonia, intestinal cancer, adjuvant treatment of tumor radiotherapy and chemotherapy, abdominal distension and dyspepsia, constipation, stool odor, diseases of liver and gall, viral diarrhea, feeding intolerance, necrotizing enterocolitis, jaundice, oral ulcer, thrush, promotion of infantile intestinal development maturation, digestive absorption, nutrition health care or growth development or nutrition absorption disorder diseases;
the liver and gall diseases comprise but are not limited to liver diseases and/or alcohol dispelling and liver protecting;
preferably, the hepatobiliary disease includes, but is not limited to, one or more of fatty liver, hepatitis, cirrhosis or liver cancer;
preferably, the hepatobiliary disease includes, but is not limited to, alcoholic and/or non-alcoholic liver disease;
preferably, the hepatobiliary disease includes, but is not limited to, one or more of alcoholic fatty liver, alcoholic hepatitis, non-alcoholic fatty liver, or non-alcoholic steatohepatitis; and/or
The motor system disease includes, but is not limited to, one or more of arthritis, osteoporosis, or rickets;
preferably, the motor system disease includes, but is not limited to, one or more of degenerative arthritis, rheumatoid arthritis, or ankylosing spondylitis; and/or
The respiratory disease includes but is not limited to one or more of nourishing lung, moistening lung to arrest cough, pulmonary nodule, pneumonia, novel coronavirus infection, recurrent respiratory tract infection or antiviral infection; and/or
The urinary system diseases include but are not limited to kidney deficiency;
preferably, the urinary system disorder includes, but is not limited to, kidney yin deficiency and/or kidney yang deficiency; and/or
The reproductive system disease includes but is not limited to one or more of vaginitis, uterine fibroids, dysmenorrhea, breast nodules, breast cancer or climacteric syndrome; and/or
The circulatory system diseases include but are not limited to one or more of cardiovascular and cerebrovascular diseases, hyperlipidemia or hypertension; and/or
The endocrine system disease includes but is not limited to one or more of hyperglycemia, dyslipidemia, diabetes, hyperuricemia, gout or thyroid nodule; and/or
The neurological disorder includes, but is not limited to, one or more of insomnia, mood relief, sleep disorder, headache, depression, or anxiety; and/or
The immune system diseases include but are not limited to one or more of allergic diseases, allergy, skin diseases, immunity enhancement, allergic rhinitis; and/or
The maintaining of body health includes, but is not limited to, one or more of eliminating dampness, invigorating qi, tonifying yang, replenishing blood, tonifying yin, or resisting oxidation; and/or
The beautifying includes but is not limited to one or more of sugar resistance, moisture retention or whitening.
10. The medicine or medicine combination is characterized by comprising any of the following active ingredients and acceptable auxiliary materials:
(I) Lactobacillus plantarum (Lactplantibibacillus plantarum) LP-K0901 according to any one of claims 1 to 3; and/or
(II) the article of claim 5; and/or
(III) a product as claimed in claim 6 or 7.
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| CN117683698A (en) * | 2024-02-04 | 2024-03-12 | 山东中科嘉亿生物工程有限公司 | Lactobacillus plantarum JYLP-376 and metabacteria agent, preparation method and application |
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