CN114437997A - Lactobacillus plantarum SF-L38 and application thereof in preparation of blood sugar control product - Google Patents
Lactobacillus plantarum SF-L38 and application thereof in preparation of blood sugar control product Download PDFInfo
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- CN114437997A CN114437997A CN202210361077.6A CN202210361077A CN114437997A CN 114437997 A CN114437997 A CN 114437997A CN 202210361077 A CN202210361077 A CN 202210361077A CN 114437997 A CN114437997 A CN 114437997A
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- lactobacillus plantarum
- juice
- ginseng
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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Abstract
The invention relates to the technical field of microorganisms, in particular to lactobacillus plantarum SF-L38 and application thereof in preparing a blood sugar control product, wherein lactobacillus plantarum SF-L38 is preserved in China general microbiological culture Collection center (CGMCC) at 23 months 9 and 2021, the preservation address is No. 3 of Beijing Shang Yang West Lu No.1 of the Korean district, the preservation number is CGMCC No.23475, and the lactobacillus plantarum SF-L38 is named as lactobacillus plantarum in classificationLactobacillus plantarum. The invention provides a plantThe lactobacillus plantarum SF-L38 has high adhesion and strong gastrointestinal adaptability, and the lactobacillus plantarum SF-L38 is used as a strain for fermentation to prepare the fermented beverage, so that the conversion of polysaccharide into monosaccharide can be inhibited, the blood sugar index can be improved, and the lactobacillus plantarum SF-L38 has important functions of preventing and treating diabetes.
Description
Technical Field
The invention relates to the technical field of microorganisms, in particular to lactobacillus plantarum SF-L38 and application thereof in preparing a blood sugar control product.
Background
Diabetes mellitus is a common, frequent and life-long-lasting disease caused by an inappropriate elevation of blood glucose (a monosaccharide) levels due to the body's inability to normally release or utilize insulin. In recent years, the prevalence rate of diabetes in China is gradually rising. According to survey data, the following steps are shown: "in the population over 20 years old in our country, the prevalence rate of diabetes of men and women respectively reaches 10.6% and 8.8%, and the prevalence rate of diabetes is 9.7%, so that it is calculated that the total prevalence rate of diabetes in the whole country is about 9200 ten thousand people. "the number of diabetes patients in 2019 in China is about 1.16 hundred million, and China has become the country with the most diabetes patients worldwide; at the same time, the number of diabetic patients is still continuously increasing rapidly. The number of people suffering from diabetes in 2040 years in China is predicted to reach 1.51 hundred million people. The growth rate of the insulin market in China also exceeds the average growth rate in the world.
The drug treatment of diabetes is divided into insulin treatment, oral hypoglycemic western medicine treatment and Chinese medicine treatment, and the biotechnology treatment is developed at present. Patent CN201610629475.6 discloses that the diabetes treatment product contains nutrient components in the terminal ileum and colon-specific release oral specific release preparation. CN201711432775.6 discloses a synthetic method of a drug for treating diabetes, namely 2- (phenylamino) -4H-benzo [ e ] [1,3] thiazin-4-one. CN201110375880.7 discloses an application of dioscin in preparing a medicament for preventing and treating diabetes. CN201110073133.8 discloses a Xinjiang achillea preparation applied to a medicine for treating diabetes and a preparation method thereof. CN200910114494.5 discloses a method for preparing alpha-glucosidase inhibitor by microbial fermentation for diabetes treatment. In conclusion, the western medicines for treating diabetes are high in price and large in toxic and side effects, and can treat symptoms and not root causes; the Chinese patent medicine has slow effect and long time for reducing blood sugar.
Disclosure of Invention
Aiming at the problems of great side effect of western medicines, slow effect of traditional Chinese medicines and the like in the prior art, the invention provides the lactobacillus plantarum SF-L38 and the application thereof in preparing blood sugar control products, which can effectively inhibit polysaccharide in human bodies from being converted into monosaccharide, improve blood sugar indexes and have the effects of preventing and treating diabetes.
In a first aspect, the present invention provides a Lactobacillus plantarum SF-L38, a Lactobacillus plantarum bacterium（Lactobacillus plantarum) The biological culture medium is preserved in the China general microbiological culture Collection center with the preservation number of CGMCC No.23475, the preservation date of 2021, 9 months and 23 days, and the preservation organization address: xilu No.1 Hospital No. 3, Beijing, Chaoyang, North.
In a second aspect, the invention provides a culture method of lactobacillus plantarum SF-L38, which comprises the steps of inoculating lactobacillus plantarum SF-L38 into a sterilized MRS liquid culture medium, wherein the inoculation amount is 1%, and culturing at 37 ℃ for 18h to obtain a lactobacillus plantarum SF-L38 culture solution.
Further, the MRS liquid culture medium comprises the following components: 10.0g of peptone, 5.0g of beef powder, 4.0g of yeast powder and K2HPO4·7H2O2.0 g, triammonium citrate 2.0g, glucose 20.0g, CH3COONa·3H2O 5.0g，MgSO4·7H2O 0.2g，MnSO4·4H2O0.05 g, Tween 801.0 mL, agar 15.0g, and distilled water 1000 mL.
In a third aspect, the invention provides an application of lactobacillus plantarum SF-L38, namely, an application of lactobacillus plantarum SF-L38 in preparing a blood sugar control product.
Furthermore, the blood sugar control product is a plant fermented drink.
In a fourth aspect, the present invention provides a method for preparing a plant fermented beverage using lactobacillus plantarum SF-L38, comprising the steps of:
(1) respectively crushing ginseng, medlar, tuckahoe, kudzu root, astragalus root, mulberry, balsam pear and fragrant solomonseal rhizome, adding water, decocting to obtain extracting solutions, concentrating the extracting solutions, and then carrying out spray drying to respectively obtain ginseng extract, medlar extract, tuckahoe extract, kudzu root extract, astragalus root extract, mulberry extract, balsam pear extract and fragrant solomonseal rhizome extract;
(2) mixing Ginseng radix extract, fructus Lycii extract, Poria extract, radix Puerariae extract, radix astragali extract, Mori fructus extract, fructus Momordicae Charantiae extract, rhizoma Polygonati Odorati extract with rhizoma Dioscoreae powder, Sucus Dauci Sativae and purified water to obtain mixed slurry, autoclaving at 120 deg.C for 30min, and cooling to 40 deg.C to obtain sterilized solution;
(3) inoculating lactobacillus plantarum SF-L38 into a sterilized MRS liquid culture medium, wherein the inoculation amount is 1%, and culturing at 37 ℃ for 18h to obtain a lactobacillus plantarum SF-L38 culture solution;
(4) inoculating lactobacillus plantarum SF-L38 culture solution into a sterilization solution, wherein the inoculation amount is 1%, and culturing at 37 ℃ for 20h to obtain fermentation liquor;
(5) and (3) after the fermentation liquor is subjected to wall breaking, adding honey peach concentrated juice and pineapple concentrated juice, mixing, and performing pasteurization under the wall breaking pressure of 20Mpa to obtain the plant fermented beverage.
Further, the plant fermented beverage comprises the following raw materials in parts by weight: 50-70 parts of purified water, 1-2 parts of a ginseng extract, 2-5 parts of a wolfberry extract, 2-5 parts of a poria cocos extract, 1-3 parts of a pueraria extract, 3-5 parts of a astragalus extract, 1-2 parts of a mulberry extract, 2-6 parts of a bitter gourd extract, 2-3 parts of a polygonatum extract, 2-4 parts of rhizoma dioscoreae powder, 2-7 parts of carrot juice, 4-6 parts of honey peach concentrated juice and 3-5 parts of pineapple concentrated juice.
Further, the plant fermented beverage comprises the following raw materials in parts by weight: 60 parts of purified water, 1 part of ginseng extract, 3 parts of medlar extract, 2 parts of tuckahoe extract, 2 parts of kudzu root extract, 3 parts of astragalus extract, 2 parts of mulberry extract, 4 parts of bitter gourd extract, 2 parts of polygonatum extract, 3 parts of Chinese yam powder, 4 parts of carrot juice, 5 parts of honey peach concentrated juice and 4 parts of pineapple concentrated juice.
The invention adopts the following medicinal action mechanisms of various auxiliary materials:
ginseng: has effects of invigorating primordial qi, invigorating spleen, benefiting lung, promoting fluid production, and tranquilizing. The nature and taste are sweet and slightly bitter.
Common yam rhizome: tonify spleen and stomach, promote the production of body fluid and benefit lung, tonify kidney and astringe essence, clear heat and remove toxicity.
Medlar: nourish liver, nourish kidney, moisten lung.
Poria cocos: sweet and bland in flavor and mild in nature. It enters heart, lung, spleen and kidney meridians. Induce diuresis and drain dampness, invigorate spleen and calm heart.
Kudzu root: sweet, pungent and cool. Has the functions of expelling pathogenic factors from muscles, allaying fever, promoting eruption, promoting the production of body fluid to quench thirst, invigorating yang and stopping diarrhea.
Astragalus root: tonify qi, strengthen superficies, expel toxin, expel pus, induce diuresis, and promote granulation.
And (3) mulberry fruit: sweet in taste and cold in nature, entering heart, liver and kidney meridians, it has the action of nourishing yin and tonifying blood, and can be used for treating yin deficiency, fluid deficiency, insomnia, etc.
Bitter gourd: has the effects of removing summer heat, clearing away heat, improving eyesight and detoxifying.
Fragrant solomonseal rhizome: to nourish yin, moisten dryness, promote the production of body fluid and quench thirst.
The lactobacillus plantarum SF-L38 has the advantages of high adhesion and strong gastrointestinal adaptability, and meanwhile, the lactobacillus plantarum SF-L38 is used as a strain for fermentation to prepare the fermented beverage, so that the lactobacillus plantarum SF-L38 can inhibit polysaccharide from being converted into monosaccharide, improves blood sugar indexes, and has important functions of preventing and treating diabetes.
Detailed Description
In order to make those skilled in the art better understand the technical solutions in the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all 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 MRS liquid medium used in the following examples included the following components:
10g of peptone, 5g of beef powder, 4g of yeast powder and K2HPO4·7H2O2g, triammonium citrate 2g, glucose 20g, CH3COONa·3H2O 5g、MgSO4·7H2O 0.2g、MnSO4·4H2O0.05 g, Tween 801 mL, and distilled water 1000 mL.
The MRS solid plate medium used in the following examples included the following components:
10g of peptone, 5g of beef powder, 4g of yeast powder and K2HPO4·7H2O2g, triammonium citrate 2g, glucose 20g, CH3COONa·3H2O5g、MgSO4·7H2O0.2g、MnSO4·4H20.05g of O, 801 mL of Tween, 15.0g of agar and 1000mL of distilled water;
the preparation method of each substance extract of the following examples is as follows: cleaning Ginseng radix, fructus Lycii, Poria, radix Puerariae, radix astragali, Mori fructus, fructus Momordicae Charantiae, and rhizoma Polygonati Odorati, crushing into coarse particles, decocting with 10 times of water for 2 times respectively for 3 hr and 1 hr, mixing the filtrates, standing for 2 hr, filtering to obtain extractive solution, concentrating the extractive solution under vacuum degree of-0.08 Mpa to 1.10 (80 deg.C), spray drying, and pulverizing into 80 mesh fine powder to obtain extract.
The preparation method of the common yam rhizome powder in the following embodiment comprises the following steps: cleaning rhizoma Dioscoreae slice with clear water, blanching in boiling water for 8min, taking out rhizoma Dioscoreae slice, rinsing with clear water to remove mucus, oven drying rhizoma Dioscoreae slice, and grinding into powder by electric grinder to obtain rhizoma Dioscoreae powder.
The preparation method of the carrot juice and the juicy peach condensed juice in the following embodiments comprises the following steps: precooking peeled radix Dauci Sativae dices and juicy peach dices for 5min, pulping, extracting juice from the pulp with horizontal screw machine, concentrating to 40% with dual-effect vacuum evaporator, sterilizing at high temperature, and cooling to obtain radix Dauci Sativae juice and juicy peach concentrated juice.
The following examples are presented to prepare pineapple juice concentrates: squeezing peeled pineapple, filtering, removing crude fiber, impurities, other suspended substances and colloidal particles which are easy to precipitate, degassing pineapple juice, sterilizing, cooling to 50 deg.C, adding sodium benzoate (0.5 g/kg of pineapple juice), and vacuum concentrating to total sugar (calculated as invert sugar) 60% to obtain concentrated pineapple juice.
Example 1
This example is the isolation and characterization of Lactobacillus plantarum SF-L38
1. A bacterium source: the collection time of the yoghourt is 2021 year and 1 month, and the collection place is Xining City in Qinghai province.
2. Isolation of the Strain
Ten times of gradient dilution is carried out on the yoghourt sample, 100 mu L of each gradient is uniformly coated on an MRS solid plate culture medium, and then the yoghourt sample is taken out and placed in an anaerobic tank to be subjected to anaerobic culture at 37 ℃ overnight;
the next day, colonies with different shapes and sizes grow on the plate, a plurality of characteristic colonies of lactic acid bacteria are selected for the second-generation streak purification treatment, the plate is subjected to anaerobic activation for 1 day to obtain a culture, and then the strain identification is carried out.
The isolated lactobacillus plantarum SF-L38 of the present invention has the following microbiological characteristics: the bacillus circulans is usually 0.9-1.2 vtm x 3.0-8.0 μm, single, paired or short chain. Usually lacking flagella, but able to move. Gram positive, no sporulation. Facultative anaerobe, surface colony diameter about 3mm, convex, round, smooth surface, fine, white, occasionally light yellow or dark yellow. Can ferment pentose or gluconate, and the lactic acid accounts for more than 85% of the final product. Can produce DL-lactic acid, grow in gluconate, and produce CO2. Fermentation of 1 molecule of ribose or other pentose sugar produces 1 molecule of lactic acid and 1 molecule of acetic acid. Can grow at 15 ℃, and the optimal growth temperature is 30-35 ℃ generally.
3. Identification of Lactobacillus plantarum SF-L38
(1) Identification unit: biometrics (Shanghai) Ltd.
(2) Primer sequences
27F：5'- AGAGTTTGATCMTGGCTCAG -3'；
1492R：5'- GGTTACCTTGTTACGACTT -3'。
(3) Identified sequences
AAGGGTTACCTTGTTACGACTTCACCCTAATCATCTGTCCCACCTTAGGCGGCTGGTTCCTAAAAGGTTACCCCACCGACTTTGGGTGTTACAAACTCTCATGGTGTGACGGGCGGTGTGTACAAGGCCCGGGAACGTATTCACCGCGGCATGCTGATCCGCGATTACTAGCGATTCCGACTTCATGTAGGCGAGTTGCAGCCTACAATCCGAACTGAGAATGGCTTTAAGAGATTAGCTTACTCTCGCGAGTTCGCAACTCGTTGTACCATCCATTGTAGCACGTGTGTAGCCCAGGTCATAAGGGGCATGATGATTTGACGTCATCCCCACCTTCCTCCGGTTTGTCACCGGCAGTCTCACCAGAGTGCCCAACTTAATGCTGGCAACTGATAATAAGGGTTGCGCTCGTTGCGGGACTTAACCCAACATCTCACGACACGAGCTGACGACACCATGCACCACCTGTATCCATGTCCCCGAAGGGAACGTCTAATCTCTTAGATTTGCATAGTATGTCAAGACCTGGTAAGGTTCTTCGCGTAGCTTCGAATTAAACCACATGCTCCACCGCTTGTGCGGGCCCCCGTCAATTCCTTTGAGTTTCAGCCTTGCGGCCGTACTCCCCAGGCGGAATGCTTAATGCGTTAGCTGCAGCACTGAAGGGCGGAAACCCTCCAACACTTAGCATTCATCGTTTACGGTATGGACTACCAGGGTATCTAATCCTGTTTGCTACCCATACTTTCGAGCCTCAGCGTCAGTTACAGACCAGACAGCCGCCTTCGCCACTGGTGTTCTTCCATATATCTACGCATTTCACCGCTACACATGGAGTTCCACTGTCCTCTTCTGCACTCAAGTTTCCCAGTTTCCGATGCACTTCTTCGGTTGAGCCGAAGGCTTTCACATCAGACTTAAAAAACCGCCTGCGCTCGCTTTACGCCCAATAAATCCGGACAACGCTTGCCACCTACGTATTACCGCGGCTGCTGGCACGTAGTTAGCCGTGGCTTTCTGGTTAAATACCGTCAATACCTGAACAGTTACTCTCAGATATGTTCTTCTTTAACAACAGAGTTTTACGAGCCGAAACCCTTCTTCACTCACGCGGCGTTGCTCCATCAGACTTTCGTCCATTGTGGAAGATTCCCTACTGCTGCCTCCCGTAGAGTTTGGGCCGTGTCTCAGTCCCAATGTGGCCGATTACCCTCTCAGTCGGCTACGTATCATTGCCATGGTGAGCCGTTACCCCACCATCTAGCTATACGCCGCGGGACCATCCAAAGTGATAGCCGAAGCCATCTTTCAAGCTCGGACATGCGGTTCCAGTGTATGCGTATAGCATCTGTTCAGTGTATCCCCGCTCTGGCCAGTTTCCCACGTGTTTACTTCAC。
(4) Identification results
This strain was identified as Lactobacillus (Lactobacillus), Lactobacillus plantarum (Lactobacillus plantarum).
Example 2
The preparation method of the lactobacillus plantarum SF-L38 fermented beverage comprises the following steps:
(1) firstly, 60 parts of purified water, 1 part of ginseng extract, 3 parts of Chinese yam powder, 3 parts of medlar extract, 2 parts of tuckahoe extract, 2 parts of kudzu root extract, 3 parts of astragalus extract, 2 parts of mulberry extract, 4 parts of bitter gourd extract, 2 parts of polygonatum extract and 4 parts of carrot juice are mixed according to a formula ratio to obtain mixed slurry.
(2) Conveying the mixed slurry prepared in the step (1) into a sterilization container, performing high-pressure sterilization at 120 ℃ for 30min, and then cooling to 40 ℃ to obtain a sterilization solution;
(3) inoculating lactobacillus plantarum SF-L38 into an MRS liquid culture medium, wherein the inoculation amount is 1%, and culturing at 37 ℃ for 18h to prepare an inoculant;
(4) inoculating the lactobacillus plantarum SF-L38 inoculant prepared in the step (3) into the sterilized solution prepared in the step (2), wherein the inoculation amount is 1%, and culturing at 37 ℃ for 20 hours to prepare a fermentation liquid;
(5) after the fermentation in the step (4) is finished, conveying the prepared fermentation liquor to a high-pressure spraying wall breaking machine, and setting the pressure to be 20MPa for wall breaking treatment;
(6) and (4) mixing and seasoning the fermentation liquor subjected to the wall breaking in the step (5) with 5 parts of juicy peach concentrated juice and 4 parts of pineapple concentrated juice, and performing pasteurization to obtain the fermented beverage.
Example 3
The preparation method of the lactobacillus plantarum SF-L38 fermented beverage comprises the following steps:
(1) firstly, mixing 50 parts of purified water, 2 parts of ginseng extract, 2 parts of Chinese yam powder, 2 parts of medlar extract, 3 parts of poria cocos extract, 1 part of kudzu root extract, 5 parts of astragalus extract, 2 parts of mulberry extract, 2 parts of bitter gourd extract, 3 parts of polygonatum extract and 7 parts of carrot juice according to a formula ratio to obtain mixed slurry;
(2) conveying the mixed slurry prepared in the step (1) into a sterilization container, performing high-pressure sterilization at 120 ℃ for 30min, and then cooling to 40 ℃ to obtain a sterilization solution;
(3) inoculating lactobacillus plantarum SF-L38 into an MRS liquid culture medium, wherein the inoculation amount is 1%, and culturing at 37 ℃ for 18h to prepare an inoculant;
(4) inoculating the lactobacillus plantarum SF-L38 inoculant prepared in the step (3) into the sterilized solution prepared in the step (2), wherein the inoculation amount is 1%, and culturing at 37 ℃ for 20h to prepare a fermentation solution;
(5) after the fermentation in the step (4) is finished, conveying the prepared fermentation liquor to a high-pressure spraying wall breaking machine, and setting the pressure to be 20MPa for wall breaking treatment;
(6) and (3) mixing and seasoning the wall-broken fermentation liquor obtained in the step (5) with 4 parts of juicy peach concentrated juice and 3 parts of pineapple concentrated juice, and performing pasteurization to obtain the fermented beverage.
Example 4
The preparation method of the lactobacillus plantarum SF-L38 fermented beverage comprises the following steps:
(1) firstly, mixing 70 parts of purified water, 1 part of ginseng extract, 4 parts of Chinese yam powder, 5 parts of medlar extract, 5 parts of poria cocos extract, 3 parts of kudzu root extract, 4 parts of astragalus extract, 1 part of mulberry extract, 6 parts of bitter gourd extract, 2 parts of polygonatum extract and 2 parts of carrot juice according to a formula ratio to obtain mixed slurry;
(2) conveying the mixed slurry prepared in the step (1) into a sterilization container, performing high-pressure sterilization at 120 ℃ for 30min, and then cooling to 40 ℃ to obtain a sterilization solution;
(3) inoculating lactobacillus plantarum SF-L38 into an MRS liquid culture medium, wherein the inoculation amount is 1%, and culturing at 37 ℃ for 18h to prepare an inoculant;
(4) inoculating the lactobacillus plantarum SF-L38 inoculant prepared in the step (3) into the sterilized solution prepared in the step (2), wherein the inoculation amount is 1%, and culturing at 37 ℃ for 20h to prepare a fermentation solution;
(5) after the fermentation in the step (4) is finished, conveying the prepared fermentation liquor to a high-pressure spraying wall breaking machine, and setting the pressure to be 20MPa for wall breaking treatment;
(6) and (3) mixing and seasoning the wall-broken fermentation liquor obtained in the step (5) with 6 parts of juicy peach concentrated juice and 5 parts of pineapple concentrated juice, and performing pasteurization to obtain the fermented beverage.
Comparative example 1
Comparative example 1 differs from the preparation method of example 2 in that lactobacillus plantarum JYLP-326 was used for fermentation, and lactobacillus plantarum JYLP-325 was purchased from China general microbiological culture Collection center (CGMCC NO. 18038), and the fermented product was prepared.
Comparative example 2
Comparative example 2 the preparation method of example 2 is different, adopt plant lactobacillus JYLP-375 to ferment, plant lactobacillus JYLP-375 is bought from China general microbiological culture Collection center of China Committee for culture Collection of microorganisms, the serial number CGMCC NO.18039, make fermented products.
Example 5 Lactobacillus plantarum SF-L38 adhesion Property testing
This experiment was conducted by testing the ability of Lactobacillus plantarum SF-L38 obtained in example 1 to adhere to extracellular mechanisms.
Inoculating Lactobacillus plantarum SF-L38 in MRS liquid culture medium according to 1% of inoculation amount, culturing at 37 ℃ for 12-14 h, centrifuging zymocyte liquid 8000r/min for 15 min, removing supernatant, collecting precipitated thallus, mixing thallus with RPMI-1640 cell culture medium in a certain weight ratio to reach 10% concentration8CFU/mL. The prepared bacterial suspension was added to a 24-well plate at 1ml per well, and the 24-well plate was incubated in 5% CO295% CO in air2Culturing at 37 deg.C for 3 hr in incubator, absorbing non-adhesive bacterial suspension, washing with phosphate buffer salt for 5 times, adding phosphate buffer solution containing polyethylene glycol octyl phenyl ether to dissolve cells, and adding sterile water to dissolve. And finally counting, wherein the culture condition is constant-temperature anaerobic culture at 37 ℃, and counting the number of viable bacteria after 2-3 days (GB4789.35 method detection).
The comparative strains were: JYLP-375, JYLP-326 and BB12, wherein BB12 is a standard product of a high-adhesion Bifidobacterium strain recognized in the industry, and the strains are all derived from an open strain depository and can be purchased from the market. As can be seen from Table 1, 0.2% is a relatively ideal index, and the results show that the adhesion of Lactobacillus plantarum SF-L38 of the present invention has a significant advantage.
TABLE 1 Lactobacillus plantarum SF-L38 adhesion characteristics test results
Example 6 Lactobacillus plantarum SF-L38 gastrointestinal tract suitability test
The experiment was carried out using Lactobacillus plantarum SF-L38 obtained in example 1.
(1) Configuration of simulated gastric fluid: pepsin was dissolved in sterile physiological saline (0.9% w/v) and the pH was adjusted to 3.0 to a final concentration of 3 g/L. Filtering with 0.22 μm sterile filter membrane, and mixing;
(2) preparation of simulated intestinal fluid: trypsin was dissolved in sterilized normal saline (0.9% w/v, pH adjusted to 8.0) to a final concentration of 1g/L, and bile salt was added to a final concentration of 0.3%. Filtering with 0.22 μm sterile filter membrane, and mixing;
(3) the experiments were carried out after three successive generations (18 h each) of different L.plantarum (JYLP-375, JYLP-326 and SF-L38) were activated. Measuring the OD value of the continuous activated third-generation strain under A600, and calculating the required amount of the OD 5;
(4) centrifuging the OD 5 bacterial liquid at 8000G for 10min, discarding the supernatant, suspending in 1mL simulated gastric juice, culturing at 37 deg.C for 3h, and counting viable bacteria on the plate;
(5) taking 8000G bacteria liquid treated by simulated gastric juice, centrifuging for 10min, discarding supernatant, suspending in simulated intestinal fluid with the same volume, culturing at 37 ℃ for 4h, and counting viable bacteria on a flat plate.
The survival rate (%) after the gastrointestinal fluid tolerance (viable cell count in bacterial suspension after the intestinal fluid simulation/original viable cell count in bacterial suspension) x 100%.
TABLE 2 Lactobacillus plantarum SF-L38 gastrointestinal tract suitability test
The result shows that the lactobacillus plantarum SF-L38 has stronger gastrointestinal adaptability than other lactobacillus plantarum.
Example 7 inhibitory Activity on alpha-glucosidase
The activity of the functional product for preventing diabetes on inhibiting alpha-glucosidase is determined: to 150. mu.L of 20mM 4-nitrophenol-. alpha. -D-glucopyranoside (PNPG) solution and 25. mu.L of the sample to be tested (fermented products prepared in example 2, example 3, example 4, comparative example 1 and comparative example 2, respectively) were added 150. mu.L of LPBS, and the mixture was water-bathed at 37 ℃ for 10 min. 50 μ L of alpha-glucosidase solution (0.17U/mL) extracted from rat intestine was added and the reaction was continued for 10 min. 1ml of 0.1MNa was added2CO3As a reaction termination solution. And measuring the light absorption value of the reaction solution at 405nm, wherein the light absorption value is in direct proportion to the free amount of 4-nitrophenol (PNP). The reaction system adopts 0.1MPBS with pH6.8As a blank control of the alpha-glucosidase solution and the sample to be tested, the inhibitory activity of the sample is calculated by the following formula:
enzyme inhibition (%) = [1- (C-D)/(A-B) ]. times.100%,
wherein A is the measured absorbance of a solution containing alpha-glucosidase but not containing a sample;
b is a measured light absorption value of a solution without alpha-glucosidase and a sample to be measured;
c is a determination light absorption value of the solution containing the alpha-glucosidase and the sample to be detected;
d is a determination light absorption value which does not contain an alpha-glucosidase solution but contains a sample to be detected;
as can be seen from Table 3, the inhibition ratios of the alpha-glucosidase activities of the lactobacillus plantarum SF-L38 fermented products prepared in examples 2-4 are obviously higher than those of the comparative examples, and the lactobacillus plantarum SF-L38 fermented product prepared in example 2 has the most obvious effect of inhibiting the alpha-glucosidase activities, and the inhibition ratio reaches 15.6%, which shows that the lactobacillus plantarum SF-L38 fermented product prepared in the invention can improve the blood glucose index.
TABLE 3 influence of fermentation products prepared in examples and comparative examples on the activity of alpha-glucosidase
Example 8
This example demonstrates the effect of the lactobacillus plantarum SF-L38 fermented product on the prevention and treatment of diabetes.
32 outpatients were treated, 17 males and 15 females, aged 35-60 years. This example uses the Lactobacillus plantarum SF-L38 fermented product of example 2, in an amount of 20ml three times a day. 15 days is a course of treatment, and six courses of treatment are taken.
Evaluation criteria:
the effect is shown: fasting plasma glucose and 2 hours post-prandial plasma glucose fall to normal ranges; or the blood glucose level of the fasting blood glucose and the blood glucose level of the 2 hours after meal are reduced by more than 40 percent before treatment; the glycated hemoglobin value drops to normal, or more than 30% before treatment.
Improvement: the blood sugar of the fasting blood sugar and the blood sugar of the 2 hours after meal are reduced by more than 20 percent before treatment, but the blood sugar does not reach the significant effect standard; the value of the glycosylated hemoglobin is reduced by more than 10 percent before treatment, but the value of the glycosylated hemoglobin does not reach the significance standard.
And (4) invalidation: the fasting blood sugar and the blood sugar after 2 hours have no decline, or the decline does not reach the improvement standard; the value of the glycosylated hemoglobin is not reduced or the value of the glycosylated hemoglobin is not reduced to reach the improvement standard.
TABLE 4 Lactobacillus plantarum SF-L38 fermented product clinical treatment efficacy
As can be seen from Table 4, the Lactobacillus plantarum SF-L38 fermented product prepared by the method has a good treatment effect on diabetes.
Although the present invention has been described in detail by way of preferred embodiments, the present invention is not limited thereto. Various equivalent modifications or substitutions can be made on the embodiments of the present invention by those skilled in the art without departing from the spirit and scope of the present invention, and these modifications or substitutions are within the scope of the present invention/any person skilled in the art can easily conceive of the changes or substitutions within the technical scope of the present invention.
Sequence listing
<110> Shandong sunflower bioengineering Co., Ltd
<120> Lactobacillus plantarum SF-L38 and application thereof in preparation of blood sugar control products
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Claims (8)
1. Lactobacillus plantarum SF-L38, characterized in that it is（Lactobacillus plantarum) SF-L38 is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No.23475, the preservation date of 2021, 9 and 23 days, and the preservation organization address: xilu No.1 Hospital No. 3, Beijing, Chaoyang, North.
2. The method for culturing Lactobacillus plantarum SF-L38, according to claim 1, comprising inoculating Lactobacillus plantarum SF-L38 in sterilized MRS broth, at an inoculum size of 1%, and culturing at 37 ℃ for 18h to obtain Lactobacillus plantarum SF-L38 broth.
3. The culture method according to claim 2, wherein the MRS liquid medium comprises the following components: 10.0g of peptone, 5.0g of beef powder, 4.0g of yeast powder and K2HPO4·7H2O2.0 g, triammonium citrate 2.0g, glucose 20.0g, CH3COONa·3H2O 5.0g，MgSO4·7H2O 0.2g，MnSO4·4H2O0.05 g, Tween 801.0 mL, and distilled water 1000 mL.
4. Use of lactobacillus plantarum SF-L38, according to claim 1, in the preparation of a glycemic control product SF-L38.
5. The use of claim 4, wherein the glycemic control product is a plant fermented drink.
6. A method for preparing a fermented plant beverage using Lactobacillus plantarum SF-L38 according to claim 1, comprising the steps of:
(1) respectively crushing ginseng, medlar, tuckahoe, kudzu root, astragalus root, mulberry, balsam pear and fragrant solomonseal rhizome, adding water, decocting to obtain extracting solutions, concentrating the extracting solutions, and then carrying out spray drying to respectively obtain ginseng extract, medlar extract, tuckahoe extract, kudzu root extract, astragalus root extract, mulberry extract, balsam pear extract and fragrant solomonseal rhizome extract;
(2) mixing Ginseng radix extract, fructus Lycii extract, Poria extract, radix Puerariae extract, radix astragali extract, Mori fructus extract, fructus Momordicae Charantiae extract, rhizoma Polygonati Odorati extract with rhizoma Dioscoreae powder, Sucus Dauci Sativae and purified water to obtain mixed slurry, autoclaving at 120 deg.C for 30min, and cooling to 40 deg.C to obtain sterilized solution;
(3) inoculating lactobacillus plantarum SF-L38 into a sterilized MRS liquid culture medium, wherein the inoculation amount is 1%, and culturing at 37 ℃ for 18h to obtain a lactobacillus plantarum SF-L38 culture solution;
(4) inoculating lactobacillus plantarum SF-L38 culture solution into a sterilization solution, wherein the inoculation amount is 1%, and culturing at 37 ℃ for 20h to obtain fermentation liquor;
(5) and after the fermentation liquor is subjected to wall breaking, adding the honey peach concentrated juice and the pineapple concentrated juice, mixing, and performing pasteurization to obtain the plant fermented beverage, wherein the wall breaking pressure is 20 Mpa.
7. The method of claim 6, wherein the plant fermented beverage comprises the following raw materials in parts by weight: 50-70 parts of purified water, 1-2 parts of a ginseng extract, 2-5 parts of a wolfberry extract, 2-5 parts of a poria cocos extract, 1-3 parts of a pueraria extract, 3-5 parts of a astragalus extract, 1-2 parts of a mulberry extract, 2-6 parts of a bitter gourd extract, 2-3 parts of a polygonatum extract, 2-4 parts of rhizoma dioscoreae powder, 2-7 parts of carrot juice, 4-6 parts of honey peach concentrated juice and 3-5 parts of pineapple concentrated juice.
8. The method of claim 6, wherein the plant fermented beverage comprises the following raw materials in parts by weight: 60 parts of purified water, 1 part of ginseng extract, 3 parts of medlar extract, 2 parts of tuckahoe extract, 2 parts of kudzu root extract, 3 parts of astragalus extract, 2 parts of mulberry extract, 4 parts of bitter gourd extract, 2 parts of polygonatum extract, 3 parts of Chinese yam powder, 4 parts of carrot juice, 5 parts of honey peach concentrated juice and 4 parts of pineapple concentrated juice.
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