CN109609420B

CN109609420B - Helicobacter pylori-resistant probiotic composition and preparation method thereof

Info

Publication number: CN109609420B
Application number: CN201910101339.3A
Authority: CN
Inventors: 张起凡; 曹崇仁
Original assignee: Shandong Huanyi Biotechnology Co ltd
Current assignee: Shandong Huanyi Biotechnology Co ltd
Priority date: 2019-01-31
Filing date: 2019-01-31
Publication date: 2021-04-06
Anticipated expiration: 2039-01-31
Also published as: CN109609420A; CN109609420A8

Abstract

The invention discloses a probiotic composition for resisting helicobacter pylori, which comprises lactobacillus plantarum powder and lactobacillus plantarum metabolite powder as active ingredients. The embodiment of the invention also provides a method for preparing the composition, which is characterized in that lactobacillus plantarum is subjected to expanded strain culture, primary seed culture, seed tank culture and fermentation culture to obtain lactobacillus plantarum zymocyte liquid. The embodiment of the invention provides a probiotic composition for resisting helicobacter pylori, which can effectively inhibit the helicobacter pylori until helicobacter pylori in a human body is eliminated, reduces the occurrence of gastric cancer and is harmless to the human body.

Description

Helicobacter pylori-resistant probiotic composition and preparation method thereof

Technical Field

The invention relates to the technical field of microbial preparations, in particular to a probiotic composition for resisting helicobacter pylori and a preparation method thereof.

Background

Helicobacter pylori (Hp) is a gram-negative spirobacterium. Helicobacter pylori has been shown to be a major causative agent of acute and chronic gastritis and gastric and duodenal ulcers, and may be associated with the development of gastric cancer and malignant lymphoma of gastric mucosa-associated lymphoid tissue (MALT). The world health organization has classified Hp as a class I carcinogen, which plays a leading role in the development of gastric cancer. The current protocol for treating Hp infection is a triple therapy with simultaneous administration of a Proton Pump Inhibitor (PPI) plus two antibiotics (two selected from clarithromycin, amoxicillin, tetracycline, metronidazole, etc.). The most major factor affecting triple therapy is thought to be the resistance of Hp to antibacterial agents; another serious problem is that proton pump inhibitors can induce dyspepsia, and large amounts of antibacterial agents cause severe destruction of the flora in the digestive tract.

In summary, there are drug resistance and other flora damage defects in the intestinal tract in drug therapy of helicobacter pylori, and a novel product for resisting helicobacter pylori is urgently needed to be developed.

Disclosure of Invention

The embodiment of the invention aims to provide a helicobacter pylori-resistant probiotic composition and a preparation method thereof, which are used for overcoming the defects that the existing helicobacter pylori-resistant product is easy to generate drug resistance and has toxic and side effects.

In order to achieve the above purpose, the embodiment of the present invention provides a probiotic composition for resisting helicobacter pylori, wherein the composition includes lactobacillus plantarum powder and the lactobacillus plantarum metabolite powder as active ingredients.

Preferably, the lactobacillus plantarum is preserved in the China center for type culture Collection with the preservation number: CCTCC NO: and M2018268.

Preferably, the composition also comprises a fungus powder carrier, wherein the fungus powder carrier comprises microcrystalline cellulose, medicinal corn starch, inulin and magnesium stearate;

the composition comprises the following raw materials in parts by weight: 9-11 parts of lactobacillus plantarum powder, 4-6 parts of lactobacillus plantarum metabolite powder, 18-22 parts of microcrystalline cellulose, 80-120 parts of medicinal corn starch, 8-12 parts of inulin and 8-12 parts of magnesium stearate.

Preferably, the composition comprises the following raw materials in parts by mass: 9.5-10.5 parts of lactobacillus plantarum powder, 4.5-5.5 parts of lactobacillus plantarum metabolite powder, 19-21 parts of microcrystalline cellulose, 90-110 parts of medicinal corn starch, 9-11 parts of inulin and 9-11 parts of magnesium stearate.

The embodiment of the invention also provides a method for preparing the composition, which comprises the steps of carrying out expanded strain culture, primary seed culture, seed tank culture and fermentation culture on lactobacillus plantarum to obtain lactobacillus plantarum zymocyte liquid;

centrifuging the lactobacillus plantarum zymocyte liquid to respectively collect lactobacillus plantarum bacterial sludge and centrifuged fermentation liquid, and freeze-drying and crushing the lactobacillus plantarum bacterial sludge and the centrifuged fermentation liquid to obtain lactobacillus plantarum bacterial powder and lactobacillus plantarum metabolite powder;

and mixing the lactobacillus plantarum powder and the lactobacillus plantarum metabolite powder with a bacterium powder carrier to obtain the composition.

Preferably, the preparation method of the lactobacillus plantarum fungus powder specifically comprises the following steps:

and mixing the lactobacillus plantarum bacterium paste, sterile water, skimmed milk powder, fructo-oligosaccharide, glycerol, maltodextrin and isomaltooligosaccharide to obtain lactobacillus plantarum suspended bacterium liquid, and freeze-drying and crushing the suspended bacterium liquid to obtain lactobacillus plantarum bacterium powder.

Preferably, the freeze-drying condition of the lactobacillus plantarum bacterial suspension is pre-freezing for 2-5 hours at-35 ℃ to-30 ℃, vacuum freeze-drying at-50 ℃ to-40 ℃, and crushing into lactobacillus plantarum bacterial powder with 100 meshes.

Preferably, the centrifuged fermentation liquid is subjected to low-temperature concentration to obtain a fermentation liquid concentrated paste with the water content of at most 20%, and the fermentation liquid concentrated paste and the medicinal corn starch are mixed in a ratio of 5: 1-4: 1, obtaining a mixture, then drying the mixture for 40 to 50 hours at the temperature of 55 to 60 ℃, and crushing the mixture to obtain the lactobacillus plantarum powder with the water content of at most 3 percent.

The application of the composition prepared by the method of the embodiment of the invention in preparing products for resisting helicobacter pylori also belongs to the protection scope of the invention.

The embodiment of the invention has the following advantages:

the embodiment of the invention provides a probiotic composition for resisting helicobacter pylori, which can effectively inhibit the helicobacter pylori until helicobacter pylori in a human body is eliminated, reduces the occurrence of gastric cancer and is harmless to the human body.

Detailed Description

The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention.

The embodiment of the invention provides a probiotic composition for resisting helicobacter pylori, which can effectively inhibit the helicobacter pylori until helicobacter pylori in a human body is eliminated, and the occurrence of gastric cancer is reduced. Firstly, a new strain is obtained, the strain is separated from fresh apple epidermis, is identified as lactobacillus plantarum through 16S DNA, is subjected to patent program biological preservation and is preserved in China center for type culture Collection with the preservation number as follows: CCTCC NO: and M2018268. The nucleotide sequence of the strain is shown as SEQ ID 1.

Example 1

One, expanding strain culture

The strain of the embodiment of the invention adopts lactobacillus plantarum, and the equipment used in the production is as follows: a strain incubator, an ultra-clean workbench, a constant-temperature incubator, various test tubes, a triangular flask, a seeding tank, a fermentation tank, a vacuum freeze dryer, a three-dimensional stirrer, a low-temperature crusher and a vacuum packaging machine.

And (3) strain culture: enlarged culture of-80 ℃ preservation strain, preparation of enlarged strain culture medium: 20 g of glucose, 10g of peptone, 8 g of beef extract powder, 3 g of sodium acetate, 801 ml of tween, 0.5 g of urea, 1000 ml of tap water and natural pH value. Adding glucose and other raw materials into 1000 ml of water in the formula, stirring for 30min to dissolve, then subpackaging in 18 x 180 test tubes, each test tube containing 10 ml, sealing with rubber plug kraft paper, sterilizing in a medical sterilizer at 0.1-0.12 MPa for 25min, and sterilizing the culture medium.

Opening a freezing tube stored at the temperature of minus 80 ℃ in a super-clean workbench under the aseptic condition, melting at room temperature, then taking 1 part of the sterilized test tube culture medium, adding 1ml of the melted-80 ℃ storage bacterium liquid into a test tube, sealing the tube opening by using a rubber plug and kraft paper, shaking up by hand, putting into a constant-temperature incubator, standing and culturing at the temperature of 37 ℃ for 15 hours, and obtaining the expanded strain culture bacterium liquid.

Second and first order seed culture

The formula of the first-level seed culture medium is as follows: 20 g of glucose, 10g of peptone, 10g of beef extract powder, 2 g of corn steep liquor, 3 g of sodium acetate, 801 ml of tween and 1000 ml of tap water, and the pH value is natural.

Adding the above materials into tap water, stirring for 20min, and dissolving to obtain primary seed culture solution. Adding the stirred and dissolved primary seed culture solution into a triangular flask according to the using amount, then binding a bottle mouth with 6 layers of gauze and a layer of kraft paper, putting the triangular flask into a medical sterilizer for sterilization, before sterilization, firstly opening an exhaust valve on the sterilizer, closing when a small amount of steam is exhausted, opening the exhaust valve on the sterilizer for exhausting for 6-8min when the pressure on the sterilizer reaches 0.05 MPa, then closing the exhaust valve, continuing to heat, starting timing when the steam pressure in the sterilizer reaches 0.1-0.12 MPa, keeping the sterilization time for 25min, leaving a heat source after the sterilization is finished, naturally cooling the heat source, opening the sterilizer when the pressure gauge on the sterilizer returns to zero, taking out the sterilized primary seed culture medium, putting the sterilized primary seed culture medium into an ultra-clean workbench for naturally cooling, transferring the cultured expanded strain culture solution into the sterilized primary seed culture solution in the triangular flask when the temperature is reduced to 37 ℃, the inoculation amount is 1 percent, namely 100 milliliters of first-stage seed culture solution is inoculated into 1 milliliter of enlarged bacterial solution, after the inoculation is finished, the bottle mouth is sealed by using original gauze and kraft paper, the bottle mouth is slightly shaken by hands to uniformly mix the inoculated strains and the culture solution, and then the mixture is cultured for 15 hours in a thermostat at 37 ℃ to obtain the first-stage seed culture solution.

Third, seeding tank culture

Sterilizing fermentation equipment, pipelines and a sterile filtration system, firstly opening valves of each inlet and outlet pipeline and a sterile air pipeline, introducing 0.12-0.14 MPa steam, communicating the steam with the pipeline valves and discharging a small amount of steam, introducing the steam for 40min, keeping the steam pressure in the pipelines at 0.12-0.14 MPa for 40min, and then closing valves of each inlet and outlet pipeline for later use.

Sterilizing empty seed tank and fermentation tank, closing valves, opening drain valve at bottom of the tank and drain valve in interlayer, opening direct steam valve, introducing 0.12-0.14 MPa steam into the tank, timing when the temperature in the tank reaches 121 deg.C, sterilizing for 40min, closing the drain valve and interlayer drain valve at bottom of the tank, and naturally cooling to 37 deg.C.

The culture medium formula of the seeding tank (weight percentage of each component of the culture medium) comprises 7 percent of glucose, 2 percent of peptone, 0.5 percent of urea, 0.5 percent of dipotassium phosphate, 1.5 percent of corn steep liquor, 0.5 percent of sodium acetate, 800.4 percent of Tween, 2 percent of polydextrose, 0.1 percent of polyether defoamer, 85.5 percent of tap water and natural pH value.

Firstly, putting tap water used in a formula into a seeding tank sterilized in an empty tank, starting a stirrer, respectively adding raw materials required in the formula, then introducing steam for heating in continuous stirring, firstly heating to 95 ℃ by using tank interlayer steam, then heating by using direct steam, and keeping for 30min when the temperature in the tank reaches 121 ℃ to achieve the sterilization effect; the steam was then turned off and the tap water cooling through the tank insert was started and was ready for use when the tank temperature reached 37 c, which was called the seeding tank medium. Then, inoculating the cultured primary seed culture solution into a seed tank under an aseptic condition, wherein the inoculation amount is 1% of the culture medium in the seed tank, namely, 1 kg of primary strain seed solution can be respectively inoculated into 100 kg of seed tank culture medium. Then, the inoculation cap on the tank is covered to start stirring culture, and the culture conditions are as follows: the temperature is 37 ℃, the pH value is natural, the stirring speed is 80 r/min, the tank pressure is 0.05 MPa, the culture time can reach the requirement after 24 hours, if the tank pressure is reduced, sterile air can be introduced to maintain the tank pressure, and the tank pressure can be adjusted by a vent valve on the tank top when the tank pressure is too high, which is called as the culture of strains in a seeding tank.

Fourthly, fermentation culture

The fermentation medium comprises the following components in percentage by weight: 12% of glucose, 1% of peptone, 3% of polydextrose, 12% of isomaltooligosaccharide, 2% of sodium acetate, 800.2% of tween, 5% of potato starch, 0.5% of conjugated linoleic acid, 0.3% of anhydrous calcium chloride, 0.5% of magnesium sulfate, 3% of corn steep liquor, 0.1% of polyether defoamer, and 60.4% of tap water, wherein the total weight is 100 kg.

Firstly, putting tap water used in a formula into a fermentation tank sterilized in an empty tank, starting a stirrer, putting raw materials used in the formula into the fermentation tank, then introducing steam for heating, firstly, heating to 95 ℃ by using interlayer steam, then, directly heating by using the steam, starting timing when the temperature in the tank reaches 121 ℃, keeping for 30min to achieve a sterilization effect, then, cooling and cooling by using the interlayer tap water, starting inoculating strains when the temperature in the tank is reduced to 37 ℃, firstly, increasing the tank pressure of a seeding tank to 0.1 MPa, keeping the tank pressure of the fermentation tank to 0.05 MPa before inoculation, then, opening an inoculation pipeline valve, conveying seeds cultured in the seeding tank into the fermentation tank in a pressure difference mode, and then, closing the inoculation pipeline valve.

The inoculation amount is 3 kg of strains cultured in each 100 kg of culture medium in the fermentation tank, the culture condition is that the fermentation temperature is 37 ℃, the stirring speed is 120 r/min, the fermentation time is 20 hours, the tank pressure is kept at 0.05 MPa, the pH value is natural, if the tank pressure is reduced, sterile air can be introduced to keep the tank pressure, and if the tank pressure is too high, the exhaust valve on the tank top can be used for adjustment, so that the zymogen liquid is obtained.

Fifthly, freeze drying of lactobacillus plantarum

And centrifuging the fermentation liquor, collecting wet bacterial sludge, pumping the fermentation liquor into a storage tank after fermentation is finished, and selecting a centrifugal rotating speed is very important in order to ensure the integrity of bacteria in the fermentation liquor. Starting a tubular centrifuge, adjusting the rotating speed to 12000 r/min, centrifuging at a feeding speed of 15 kg per minute, collecting wet bacterial sludge after centrifuging, freeze-drying the centrifugally collected wet bacterial sludge, performing low-temperature vacuum concentration on the centrifuged fermentation liquor, adding a carrier, and compounding a finished product after vacuum drying.

1. Preparing a suspension liquid: 1000 g of wet bacterial sludge is taken, and 3000 ml of sterile distilled water, 300 g of skim milk powder, 200 g of fructo-oligosaccharide, 100ml of 98% glycerol, 50 g of maltodextrin and 300 g of isomalto-oligosaccharide are added. During the specific operation, 1500 ml of water in the formula is firstly heated to 50-55 ℃, maltodextrin is added under stirring and dissolved, then the rest water in the formula is added, and then other raw materials in the formula are added. Stirring thoroughly for 30min (stirring at 25-35 ℃). The suspension liquid is put into a thermostatic chamber or a box with the temperature of 30 ℃ for 30min for pre-freezing. Pre-freezing before freeze drying, namely sub-packaging the suspension liquid with constant temperature for 30min into a freeze drying tray, then pre-freezing the suspension liquid in a pre-freezing chamber of a freeze dryer for 3 hours at the temperature of minus 30-35 ℃, and vacuumizing and drying after freezing.

And (3) vacuum drying, namely putting the pre-frozen material disc into a vacuum drying chamber to start vacuum drying, wherein the drying conditions are that the vacuum degree is 2-5 Pa, the temperature is-45 ℃, the temperature of the drying chamber is 28 ℃, and the drying time is 45 hours. And after freeze-drying, collecting freeze-dried materials, crushing the freeze-dried materials into bacterial powder with the fineness of 100 meshes by using an aseptic crusher, and sealing and packaging the bacterial powder for later use to obtain the lactobacillus plantarum bacterial powder.

2. Low-temperature concentration of centrifugal fermentation liquor: concentrating under vacuum degree of 0.06 MPa at 55-65 deg.C to obtain paste with solid content of 80% and water content of 20%, adding medicinal corn starch as carrier, mixing, and vacuum drying. Taking 1000 g of fermentation liquor concentrated paste, adding 200 g of medicinal corn starch, uniformly mixing, and drying in a vacuum drying oven, wherein the drying conditions are as follows: drying at 55-60 deg.C under 0.06 MPa for 45 hr until the water content is 3%, pulverizing to 100 mesh powder, and sealing and packaging to obtain Lactobacillus plantarum powder. Wherein the mass fraction ratio of the fermentation liquor concentrated paste to the medicinal corn starch is 5: 1-4: 1.

the finished product of the helicobacter pylori-resistant probiotic composition is compounded as follows: taking 5 g of lactobacillus plantarum powder of 100 meshes, 3 g of lactobacillus plantarum metabolite powder, 10g of microcrystalline cellulose, 20 g of medicinal corn starch, 5 g of inulin and 5 g of magnesium stearate, wherein the microcrystalline cellulose, the medicinal corn starch, the inulin and the magnesium stearate are carriers of the bacterium powder.

Mixing the above materials in solid stirring mixer, packaging under aseptic condition into capsule, and bottling or tabletting to obtain the final product.

Example 2

The finished product of the helicobacter pylori-resistant probiotic composition is compounded as follows: 9 g of lactobacillus plantarum powder of 100 meshes, 6 g of lactobacillus plantarum metabolite powder, 22 g of microcrystalline cellulose, 120 g of medicinal corn starch, 12 g of inulin and 12 g of magnesium stearate are taken.

Wherein, the lactobacillus plantarum powder and the lactobacillus plantarum metabolite powder are both the lactobacillus plantarum powder and the lactobacillus plantarum metabolite powder prepared in the method of example 1.

Example 3

The finished product of the helicobacter pylori-resistant probiotic composition is compounded as follows: 9 g of lactobacillus plantarum powder of 100 meshes, 4 g of lactobacillus plantarum metabolite powder, 22 g of microcrystalline cellulose, 120 g of medicinal corn starch, 8 g of inulin and 8 g of magnesium stearate are taken.

Example 4

The finished product of the helicobacter pylori-resistant probiotic composition is compounded as follows: taking 11 g of lactobacillus plantarum powder of 100 meshes, 6 g of lactobacillus plantarum metabolite powder, 18 g of microcrystalline cellulose, 80 g of medicinal corn starch, 8 g of inulin and 8 g of magnesium stearate.

Example 5

The finished product of the helicobacter pylori-resistant probiotic composition is compounded as follows: taking 11 g of lactobacillus plantarum powder of 100 meshes, 6 g of lactobacillus plantarum metabolite powder, 18 g of microcrystalline cellulose, 80 g of medicinal corn starch, 12 g of inulin and 12 g of magnesium stearate.

Comparative example 1

The probiotic composition for helicobacter pylori resistance of the comparative example is different from example 3 in that lactobacillus casei bacterial powder and lactobacillus casei fermentation metabolite powder are not contained.

Test examples antagonistic activity of Lactobacillus plantarum powder of the present invention against helicobacter pylori

Columbia medium (Oxiod, UK) was poured onto plates 15mL per plate, 9mm standard plates, and H.pyri 43504, H.pyri 11637, and H.pyri 26695 were spread onto plates (1X 10 concentration)⁸CFU/plate), immediately placing the mixture into 9 Oxford cups, respectively adding 100 mu L of the suspension of the plant lactobacillus powder in the embodiments 2, 3 and 4 of the invention into the Oxford cups, fixing the suspension for 1.5h under the microaerophilic condition, placing the suspension into a three-gas incubator for culturing for 48 to 72h, and measuring the size of the bacteriostatic ring. MRS and a culture medium without lactobacillus plantarum are used as negative controls, the experiment is repeated 3 times, and the test results of the lactobacillus plantarum powder for inhibiting helicobacter pylori in the embodiment of the invention are shown in Table 1.

TABLE 1

ZOI is the diameter of the zone of inhibition; SEM is standard deviation;

the test results in table 1 show that the lactobacillus plantarum of the present invention has inhibitory effect on several strains of helicobacter pylori, but the lactobacillus plantarum of the present invention has different bacteriostatic ability due to the specificity of the strains.

Test example

1 clinical trial

1.1 general data

A total of 900 cases with sleep disorders, depression, anxiety symptoms were selected from 2016-8-2016 to receive the test pharmaceutical composition, wherein treatment was performed in group 1: 200 middle-aged and elderly men, age 30-80 years, average age 56 years; treatment 2 groups: 200 middle-aged and elderly women, age 28-78 years, average age 52 years; treatment 3 groups: 150 adolescent males, aged 15-19 years, with an average age of 17 years; treatment 4 groups: 150 adolescent women, age 14-18 years, mean age 16; a control group of 200 persons, with an average age of 30 years, was used as the study subject. The difference of the basic data of each group of patients, such as sex, age, disease type and the like, has no statistical significance (P is more than 0.05) and is comparable.

1.2 evaluation criteria

And (3) curing: the clinical symptoms substantially improve or disappear entirely after treatment;

the method has the following advantages: the clinical symptoms improved after treatment;

and (4) invalidation: the symptoms are not significantly reduced or exacerbated.

5.3 methods of treatment

Treatment groups 1-4: the probiotic compositions against helicobacter pylori of examples 2 to 5 were administered to patients 1 time a day, 1 dose/10 g each time, and each gram of lactobacillus plantarum contained 50 billion/CFU of live lactobacillus plantarum.

Control group: the composition of comparative example 1 was administered to the patient 1 time a day at 1 dose/10 g.

5.4 results

After 20 days of treatment, the results of comparing the clinical treatment effects of the four groups are shown in Table 1

TABLE 1

Description of the drawings: the statistical effect started in 200 men after 3 days of administration, and 82 men with different ages after 3 days of administration were tested for a 20% reduction in H.pylori. After 9 days of administration, 172 persons had a 40% reduction in H.pylori detected. 183 people after taking the medicine for 15 days have extremely obvious helicobacter pylori elimination, 188 people after taking the medicine for 20 days have 78% of the detected helicobacter pylori elimination, 4 people have complete helicobacter pylori elimination, and 8 people after taking the medicine for lactobacillus plantarum have no effect on eliminating the helicobacter pylori.

Female: the statistical effect was initiated in 200 people who took the drug for 3 days, and in 102 people of different ages who took the drug for 3 days, the reduction in H.pylori was 20% by test. After 9 days, 169 persons had a 40% reduction in H.pylori detected. 182 people take the medicine for 15 days to eliminate the helicobacter pylori extremely obviously, 189 people take the medicine for 20 days to reduce the helicobacter pylori by more than 76 percent through detection, 2 people completely eliminate the helicobacter pylori, and 9 people take the lactobacillus plantarum to fail to eliminate the helicobacter pylori.

Teenager male: the statistical effect was initiated in 150 people who took the drug for 3 days, and in 62 people of different ages who took the drug for 3 days, the reduction in H.pylori was 20% by test. After 9 days, there were 116 subjects who had a 40% reduction in H.pylori detected. After taking the medicine for 15 days, 136 people can eliminate helicobacter pylori obviously, 140 people can reduce the helicobacter pylori by more than 80% through detection after taking the medicine for 20 days, 4 people can completely eliminate the helicobacter pylori, and 6 people can not eliminate the helicobacter pylori after taking the lactobacillus plantarum.

Teenager women: the statistical effect was initiated in 150 people taking 3 days, and 68 people of different ages taking 3 days were tested for a 20% reduction in H.pylori. After 9 days of administration, there were 108 subjects who had a 40% reduction in H.pylori detected. After taking the medicine for 15 days, 136 people can eliminate helicobacter pylori obviously, after taking the medicine for 20 days, 143 people can reduce helicobacter pylori by over 75 percent through detection, 3 people can completely eliminate the helicobacter pylori, and 4 people can not eliminate the helicobacter pylori after taking the lactobacillus plantarum.

Control group: the statistical effect was initiated in 200 persons who took the drug for 3 days, and 2 persons of different ages took the drug for 3 days were tested for a 20% reduction in H.pylori. After 9 days of administration, 2 persons had a 30% reduction in H.pylori detected. After the patient takes the medicine for 15 days, 3 people can eliminate helicobacter pylori obviously, after the patient takes the medicine for 20 days, 3 people can reduce the helicobacter pylori by over 75 percent through detection, and in total, 197 people can not eliminate the helicobacter pylori after taking the lactobacillus plantarum.

Although the invention has been described in detail above with reference to a general description and specific examples, it will be apparent to one skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.

Sequence listing

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Claims

1. A method for preparing a probiotic composition against helicobacter pylori is characterized in that,

the probiotic composition comprises lactobacillus plantarum powder and lactobacillus plantarum metabolite powder as active ingredients; the lactobacillus plantarum is preserved in China center for type culture Collection with the preservation number: CCTCC NO: m2018268;

carrying out expanded strain culture, primary seed culture, seed tank culture and fermentation culture on the lactobacillus plantarum to obtain lactobacillus plantarum zymocyte liquid;

mixing the lactobacillus plantarum powder and the lactobacillus plantarum metabolite powder with a bacterium powder carrier to obtain the probiotic composition;

the probiotic composition comprises the following raw materials in parts by weight: 9-11 parts of lactobacillus plantarum powder, 4-6 parts of lactobacillus plantarum metabolite powder, 18-22 parts of microcrystalline cellulose, 80-120 parts of medicinal corn starch, 8-12 parts of inulin and 8-12 parts of magnesium stearate;

the preparation method of the lactobacillus plantarum bacterium powder comprises the following specific steps:

mixing the lactobacillus plantarum bacterium paste, sterile water, skimmed milk powder, fructo-oligosaccharide, glycerol, maltodextrin and isomaltooligosaccharide to obtain lactobacillus plantarum suspended bacterium liquid, and freeze-drying and crushing the suspended bacterium liquid to obtain lactobacillus plantarum bacterium powder;

the freeze drying condition of the lactobacillus plantarum suspended bacterium liquid is that the lactobacillus plantarum suspended bacterium liquid is pre-frozen for 2 to 5 hours at the temperature of between 35 ℃ below zero and 30 ℃ below zero, is subjected to vacuum freeze drying at the temperature of between 50 ℃ below zero and 40 ℃ below zero, and is crushed into lactobacillus plantarum powder of 100 meshes;

and (3) concentrating the centrifuged fermentation liquor at low temperature to obtain a concentrated fermentation liquor paste with the water content of at most 20%, mixing the concentrated fermentation liquor paste with medicinal corn starch in a ratio of 5: 1-4: 1 to obtain a mixture, then drying the mixture for 40 to 50 hours at the temperature of 55 to 60 ℃, and crushing the mixture to obtain the lactobacillus plantarum metabolite powder with the water content of at most 3 percent.

2. The method for preparing the probiotic composition for resisting helicobacter pylori according to claim 1, wherein the probiotic composition comprises the following raw materials in parts by mass: 9.5-10.5 parts of lactobacillus plantarum powder, 4.5-5.5 parts of lactobacillus plantarum metabolite powder, 19-21 parts of microcrystalline cellulose, 90-110 parts of medicinal corn starch, 9-11 parts of inulin and 9-11 parts of magnesium stearate.

3. Use of a probiotic composition prepared according to the process of claim 1 or 2 for the preparation of a product against helicobacter pylori.