CN112375698A - Chinese herbal medicine bacterial flora culture solution - Google Patents
Chinese herbal medicine bacterial flora culture solution Download PDFInfo
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- CN112375698A CN112375698A CN202011219518.6A CN202011219518A CN112375698A CN 112375698 A CN112375698 A CN 112375698A CN 202011219518 A CN202011219518 A CN 202011219518A CN 112375698 A CN112375698 A CN 112375698A
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
Abstract
The invention discloses a Chinese herbal medicine bacterial flora culture solution, and particularly relates to the field of preparation of bacterial flora culture solutions, wherein the preparation steps are as follows: the method comprises the following steps of preparing raw materials, wherein the raw materials comprise 10 g of wormwood, 8 g of dandelion, 5 g of houttuynia cordata, 4 g of folium isatidis, 15 g of folium artemisiae argyi, 12 g of honeysuckle, 2 g of fructus forsythiae, 8 g of dandelion, 4 g of folium isatidis, 5 g of bezoar, 5 g of houttuynia cordata, 4 g of blackberry lily, 5 g of Chinese pulsatilla root, 5 g of radix isatidis, 5 g of indigo naturalis, 5 g of rhizoma smilacis glabrae, 5 g of subprostrate sophora, 2 g of oldenlandia diffusa, 8 g of philippine violet herb, 10 g of common andrographis herb, 11 g. The Chinese herbal medicine bacterial flora culture solution prepared by the invention can provide various nutrient substances for more than 100 microbial floras, is beneficial to the co-existence of various floras, achieves the optimal living environment of Chinese herbal medicine bacteria, and is more beneficial to the propagation of Chinese herbal medicine bacterial flora.
Description
Technical Field
The invention belongs to the field of preparation of a bacterial flora culture solution, and particularly relates to a Chinese herbal medicine bacterial flora culture solution.
Background
The deodorizing method for refuse landfill is to use automatic spraying system and spraying truck to eliminate the bad smell from refuse. Spraying two deodorizers, namely plant liquid and biological bacteria, by covering. At present, most of the plant liquid is used. When landfill summer, produce the filtration liquid easily and leak, filtration liquid is harmful again to the environment, so generally all can carry out recovery processing, but filtration liquid can distribute very smelly gas. Conventional deodorants do not solve these problems.
The condition that leachate leaks can be solved fast in microbial deodorization, and because the product compounds many bacterial species, so to this high compound foul smell material gas of leachate, have apparent dispelling effect, but current technique is difficult to cultivate multiple bacterial flora in same culture solution, all contain carbohydrate in a culture solution generally, nitrogenous material, inorganic salt (including microelement) and vitamin and water, because multiple bacterial flora is different to the demand of nutrient substance, can cause some flora poor nutrition, so put forward more rigorous requirement to the preparation of culture solution, will satisfy multiple bacterial flora and live together and breed.
Disclosure of Invention
The invention provides a Chinese herbal medicine bacterial flora culture solution, and aims to solve the problem that multiple bacterial flora are difficult to culture in the same culture solution in the prior art.
The invention is realized in such a way, and provides the following technical scheme: a Chinese herbal medicine bacterial flora culture solution is prepared by the following steps:
step one, preparing raw materials, namely 10 g of wormwood, 8 g of dandelion, 5 g of houttuynia cordata, 4 g of folium isatidis, 15 g of folium artemisiae argyi, 12 g of honeysuckle, 2 g of fructus forsythiae, 8 g of dandelion, 4 g of folium isatidis, 5 g of bezoar, 5 g of houttuynia cordata, 4 g of blackberry lily, 5 g of Chinese pulsatilla root, 5 g of radix isatidis, 5 g of indigo naturalis, 5 g of rhizoma smilacis glabrae, 5 g of subprostrate sophora, 2 g of oldenlandia diffusa, 8 g of Chinese violet, 10 g of andrographis paniculata, 11 g of purslane and 13 g of cortex dictamn;
step two, crushing the raw materials, namely mixing all the raw materials in the step one, putting the raw materials into a crusher for crushing after mixing, controlling the rotating speed of the crusher to be 1000-1200 rpm, controlling the crushing time to be 10-14min, controlling the particle sizes of all the crushed raw materials to be 100-180 um, and then fully mixing and shaking the raw materials uniformly to obtain powder of the mixed raw materials;
step three, heating, namely putting the powder of the mixed raw material obtained in the step two into steam for heating, wherein the temperature of the steam is controlled to be 60-100 ℃, and the heating time is controlled to be 10-14 minutes;
step four, high-temperature high-pressure heating, namely placing the powder of the mixed raw material heated in the step three in a ventilated place for natural cooling, then placing the powder in a closed container for high-temperature high-pressure heating, wherein the heating temperature is controlled to be 300-340 ℃, and the pressure is controlled to be 200-240 KPa, so that the cell walls of the Chinese herbal medicines are broken, the single-layer membrane of vacuole is broken, and the cell sap is vaporized;
step five, cooling and depressurizing, wherein the closed container in the step four is cooled and depressurized, the temperature is controlled to be 20-60 ℃, the pressure is controlled to be 20-60 KPa, so that vaporized cell sap is liquefied, the cell sap is continuously liquefied to form liquid drops, and the liquid drops are collected by the container;
step six, filtering, namely filtering the liquid drops collected in the step five, and separating carbon-containing impurities, organelles and cell walls in the liquid drops;
step seven, centrifugal purification, pouring the filtered liquid drops in the step six into a centrifugal tube, then placing the centrifugal tube into a centrifugal machine for separation and purification, and removing redundant impurities to obtain purified cell sap;
step eight, weighing and mixing, namely weighing 20 g of the purified cell sap obtained in the step seven, then adding 5 g of camphor wood powder, 8 g of bamboo charcoal powder, 100 g of distilled water, 2 g of sodium chloride, 5 g of peptone, 6 g of glucose, 5 g of inorganic salt, 5 g of nucleic acid, 3 g of enzyme and 5 g of amino acid, adding, and fully mixing to finally obtain a Chinese herbal medicine bacterial flora culture solution;
and step nine, sterilizing, namely sterilizing the Chinese herbal medicine bacterial flora culture solution obtained in the step eight to obtain a sterile Chinese herbal medicine bacterial flora culture solution.
Step ten, storage, namely freezing the Chinese herbal medicine bacterial flora culture solution in the step nine by using a freezer, and taking out the culture solution in advance when in use, and matching the culture solution with microorganisms after treatment.
In a preferred embodiment, the mesh diameter of the filter screen used in the sixth step is controlled to be between 2um and 6 um.
In a preferred embodiment, the rotating speed of the centrifuge is controlled to be 8000-.
In a preferred embodiment, the inorganic salt in step eight is a mixture of calcium, phosphorus, potassium, sulfur, sodium, chlorine, magnesium, iron, zinc, selenium, molybdenum and iodine elements.
In a preferred embodiment, the amino acid in step eight consists of glycine, alanine, valine, leucine, isoleucine, methionine, proline, tryptophan, serine, tyrosine, cysteine, phenylalanine, asparagine, glutamine, threonine, aspartic acid, glutamic acid, lysine, arginine and histidine.
In a preferred embodiment, the sterilization environment in the ninth step is 0.05MPa (110 ℃), and the sterilization time is controlled to be 20-30 min.
In a preferred embodiment, the freezing temperature in the step ten is controlled within the range of-80 to-50 ℃ and the freezing time is 72 hours.
Compared with the prior art, the invention has the beneficial effects that:
1. the cell sap of the purified Chinese herbal medicine cells is obtained by extracting the Chinese herbal medicine cell sap with high purity as basic nutrient substances and removing cell walls, organelles and carbon-containing impurities of the Chinese herbal medicine cells, so that a culture solution is cleaner, the impurities are prevented from influencing the survival of microorganisms, dozens of Chinese herbal medicine raw materials are prepared, the nutrient substances contained in the purified cell sap are more complete, various nutrient substances can be provided for more than 100 microbial floras, the situation that some microbial floras cannot survive due to lack of the nutrient substances is avoided, 20 amino acids necessary for the survival of the organisms are added, the direct absorption of various microbial floras is facilitated, various trace elements are added, and the requirement of inorganic salts of various microorganisms is met;
2. the obtained Chinese herbal medicine bacterial flora culture solution is sterilized, so that other mixed bacteria and microorganisms can be removed, the influence of other mixed bacteria on the survival of the Chinese herbal medicine bacteria is prevented, the Chinese herbal medicine bacterial flora culture solution prepared by the preparation method is more favorable for the co-survival of various floras, the nutrition is more comprehensive, various impurities influencing the survival of the Chinese herbal medicine bacterial flora are removed, the optimal living environment of the Chinese herbal medicine bacteria is achieved, and the propagation of the Chinese herbal medicine bacterial flora is more favorable.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
Example 1:
the invention provides a Chinese herbal medicine bacterial flora culture solution which comprises the following components in parts by weight: the preparation steps are as follows:
step one, preparing raw materials, namely 10 g of wormwood, 8 g of dandelion, 5 g of houttuynia cordata, 4 g of folium isatidis, 15 g of folium artemisiae argyi, 12 g of honeysuckle, 2 g of fructus forsythiae, 8 g of dandelion, 4 g of folium isatidis, 5 g of bezoar, 5 g of houttuynia cordata, 4 g of blackberry lily, 5 g of Chinese pulsatilla root, 5 g of radix isatidis, 5 g of indigo naturalis, 5 g of rhizoma smilacis glabrae, 5 g of subprostrate sophora, 2 g of oldenlandia diffusa, 8 g of Chinese violet, 10 g of andrographis paniculata, 11 g of purslane and 13 g of cortex dictamn;
step two, crushing the raw materials, namely mixing all the raw materials in the step one, putting the raw materials into a crusher for crushing after mixing, controlling the rotating speed of the crusher to be 1000-1200 rpm, controlling the crushing time to be 10-14min, controlling the particle sizes of all the crushed raw materials to be 100um, and then fully mixing and shaking the raw materials uniformly to obtain the powder of the mixed raw materials;
step three, heating, namely putting the powder of the mixed raw material obtained in the step two into steam for heating, wherein the temperature of the steam is controlled to be 60-100 ℃, and the heating time is controlled to be 10-14 minutes;
step four, high-temperature high-pressure heating, namely placing the powder of the mixed raw material heated in the step three in a ventilated place for natural cooling, then placing the powder in a closed container for high-temperature high-pressure heating, wherein the heating temperature is controlled at 300 ℃, and the pressure is controlled at 200KPa-240KPa, so that the cell walls of the Chinese herbal medicines are broken, and the single-layer membrane of vacuole is broken, thereby vaporizing the cell sap;
step five, cooling and depressurizing, namely cooling and depressurizing the closed container in the step four, controlling the temperature to be 20-60 ℃ and the pressure to be 20KPa, so that the vaporized cell sap is liquefied, the cell sap is continuously liquefied to form liquid drops, and collecting the liquid drops by using the container;
step six, filtering, namely filtering the liquid drops collected in the step five, separating carbon-containing impurities, organelles and cell walls in the liquid drops, and controlling the mesh diameter of a filter screen to be between 2 and 6 um;
step seven, centrifugal purification, pouring the filtered liquid drops in the step six into a centrifugal tube, then placing the centrifugal tube into a centrifugal machine for separation and purification, removing redundant impurities to obtain purified cell sap, controlling the rotating speed of the centrifugal machine to 8000-12000r/min, and controlling the time to 20-24 min;
step eight, weighing and mixing, namely weighing 20 g of purified cell sap in the step seven, then adding 5 g of camphor wood powder, 8 g of bamboo charcoal powder, 100 g of distilled water, 2 g of sodium chloride, 5 g of peptone, 6 g of glucose, 5 g of inorganic salt, 5 g of nucleic acid, 3 g of enzyme and 5 g of amino acid, and fully mixing after adding to obtain a Chinese herbal medicine bacterial flora culture solution, wherein the inorganic salt is a mixture of calcium, phosphorus, potassium, sulfur, sodium, chlorine, magnesium, iron, zinc, selenium, molybdenum and iodine elements, and the amino acid consists of glycine, alanine, valine, leucine, isoleucine, methionine, proline, tryptophan, serine, tyrosine, cysteine, phenylalanine, asparagine, glutamine, threonine, aspartic acid, glutamic acid, lysine, arginine and histidine;
step nine, sterilizing, namely sterilizing the Chinese herbal medicine bacterial flora culture solution obtained in the step eight to obtain a sterile Chinese herbal medicine bacterial flora culture solution, wherein the sterilization environment is 0.05MPa (110 ℃), and the sterilization time is controlled to be 20-30 min;
step ten, storing, namely freezing the Chinese herbal medicine bacterial flora culture solution in the step nine by using a freezer, taking out the culture solution in advance when in use, matching the culture solution with microorganisms after treatment, controlling the freezing temperature within the range of-80 to-50 ℃, and freezing for 72 hours.
Example 2:
the invention provides a Chinese herbal medicine bacterial flora culture solution which comprises the following components in parts by weight:
step one, preparing raw materials, namely 10 g of wormwood, 8 g of dandelion, 5 g of houttuynia cordata, 4 g of folium isatidis, 15 g of folium artemisiae argyi, 12 g of honeysuckle, 2 g of fructus forsythiae, 8 g of dandelion, 4 g of folium isatidis, 5 g of bezoar, 5 g of houttuynia cordata, 4 g of blackberry lily, 5 g of Chinese pulsatilla root, 5 g of radix isatidis, 5 g of indigo naturalis, 5 g of rhizoma smilacis glabrae, 5 g of subprostrate sophora, 2 g of oldenlandia diffusa, 8 g of Chinese violet, 10 g of andrographis paniculata, 11 g of purslane and 13 g of cortex dictamn;
step two, crushing the raw materials, namely mixing all the raw materials in the step one, putting the raw materials into a crusher for crushing after mixing, controlling the rotating speed of the crusher to be 1000-1200 rpm, controlling the crushing time to be 11min, controlling the particle sizes of all the crushed raw materials to be 100-180 um, and then fully mixing and shaking the raw materials to obtain the powder of the mixed raw materials;
step three, heating, namely putting the powder of the mixed raw material obtained in the step two into steam for heating, wherein the temperature of the steam is controlled to be 60-100 ℃, and the heating time is controlled to be 10-14 minutes;
step four, high-temperature high-pressure heating, namely placing the powder of the mixed raw material heated in the step three in a ventilated place for natural cooling, then placing the powder in a closed container for high-temperature high-pressure heating, wherein the heating temperature is controlled at 310 ℃, and the pressure is controlled at 200KPa-240KPa, so that the cell walls of the Chinese herbal medicines are broken, and the single-layer membrane of vacuole is broken, thereby vaporizing the cell sap;
step five, cooling and depressurizing, namely cooling and depressurizing the closed container in the step four, controlling the temperature to be 20-60 ℃ and the pressure to be 30KPa, so that the vaporized cell sap is liquefied, the cell sap is continuously liquefied to form liquid drops, and collecting the liquid drops by using the container;
step six, filtering, namely filtering the liquid drops collected in the step five, separating carbon-containing impurities, organelles and cell walls in the liquid drops, and controlling the mesh diameter of a filter screen to be between 2 and 6 um;
step seven, centrifugal purification, pouring the filtered liquid drops in the step six into a centrifugal tube, then placing the centrifugal tube into a centrifugal machine for separation and purification, removing redundant impurities to obtain purified cell sap, controlling the rotating speed of the centrifugal machine to 8000-12000r/min, and controlling the time to 20-24 min;
step eight, weighing and mixing, namely weighing 20 g of purified cell sap in the step seven, then adding 5 g of camphor wood powder, 8 g of bamboo charcoal powder, 100 g of distilled water, 2 g of sodium chloride, 5 g of peptone, 6 g of glucose, 5 g of inorganic salt, 5 g of nucleic acid, 3 g of enzyme and 5 g of amino acid, and fully mixing after adding to obtain a Chinese herbal medicine bacterial flora culture solution, wherein the inorganic salt is a mixture of calcium, phosphorus, potassium, sulfur, sodium, chlorine, magnesium, iron, zinc, selenium, molybdenum and iodine elements, and the amino acid consists of glycine, alanine, valine, leucine, isoleucine, methionine, proline, tryptophan, serine, tyrosine, cysteine, phenylalanine, asparagine, glutamine, threonine, aspartic acid, glutamic acid, lysine, arginine and histidine;
step nine, sterilizing, namely sterilizing the Chinese herbal medicine bacterial flora culture solution obtained in the step eight to obtain a sterile Chinese herbal medicine bacterial flora culture solution, wherein the sterilization environment is 0.05MPa (110 ℃), and the sterilization time is controlled to be 20-30 min;
step ten, storing, namely freezing the Chinese herbal medicine bacterial flora culture solution in the step nine by using a freezer, taking out the culture solution in advance when in use, matching the culture solution with microorganisms after treatment, controlling the freezing temperature within the range of-80 to-50 ℃, and freezing for 72 hours.
Example 3:
the invention provides a Chinese herbal medicine bacterial flora culture solution which comprises the following components in parts by weight:
step one, preparing raw materials, namely 10 g of wormwood, 8 g of dandelion, 5 g of houttuynia cordata, 4 g of folium isatidis, 15 g of folium artemisiae argyi, 12 g of honeysuckle, 2 g of fructus forsythiae, 8 g of dandelion, 4 g of folium isatidis, 5 g of bezoar, 5 g of houttuynia cordata, 4 g of blackberry lily, 5 g of Chinese pulsatilla root, 5 g of radix isatidis, 5 g of indigo naturalis, 5 g of rhizoma smilacis glabrae, 5 g of subprostrate sophora, 2 g of oldenlandia diffusa, 8 g of Chinese violet, 10 g of andrographis paniculata, 11 g of purslane and 13 g of cortex dictamn;
step two, crushing the raw materials, namely mixing all the raw materials in the step one, putting the raw materials into a crusher for crushing after mixing, controlling the rotating speed of the crusher to be 1000-1200 rpm, controlling the crushing time to be 12min, controlling the particle sizes of all the crushed raw materials to be 100-180 um, and then fully mixing and shaking the raw materials to obtain the powder of the mixed raw materials;
step three, heating, namely putting the powder of the mixed raw material obtained in the step two into steam for heating, wherein the temperature of the steam is controlled to be 60-100 ℃, and the heating time is controlled to be 10-14 minutes;
step four, high-temperature high-pressure heating, namely placing the powder of the mixed raw material heated in the step three in a ventilated place for natural cooling, then placing the powder in a closed container for high-temperature high-pressure heating, wherein the heating temperature is controlled at 320 ℃, and the pressure is controlled at 200KPa-240KPa, so that the cell walls of the Chinese herbal medicines are broken, and the single-layer membrane of vacuole is broken, thereby vaporizing the cell sap;
step five, cooling and depressurizing, namely cooling and depressurizing the closed container in the step four, controlling the temperature to be 20-60 ℃ and the pressure to be 40KPa, so that the vaporized cell sap is liquefied, the cell sap is continuously liquefied to form liquid drops, and collecting the liquid drops by using the container;
step six, filtering, namely filtering the liquid drops collected in the step five, separating carbon-containing impurities, organelles and cell walls in the liquid drops, and controlling the mesh diameter of a filter screen to be between 2 and 6 um;
step seven, centrifugal purification, pouring the filtered liquid drops in the step six into a centrifugal tube, then placing the centrifugal tube into a centrifugal machine for separation and purification, removing redundant impurities to obtain purified cell sap, controlling the rotating speed of the centrifugal machine to 8000-12000r/min, and controlling the time to 20-24 min;
step eight, weighing and mixing, namely weighing 20 g of purified cell sap in the step seven, then adding 5 g of camphor wood powder, 8 g of bamboo charcoal powder, 100 g of distilled water, 2 g of sodium chloride, 5 g of peptone, 6 g of glucose, 5 g of inorganic salt, 5 g of nucleic acid, 3 g of enzyme and 5 g of amino acid, and fully mixing after adding to obtain a Chinese herbal medicine bacterial flora culture solution, wherein the inorganic salt is a mixture of calcium, phosphorus, potassium, sulfur, sodium, chlorine, magnesium, iron, zinc, selenium, molybdenum and iodine elements, and the amino acid consists of glycine, alanine, valine, leucine, isoleucine, methionine, proline, tryptophan, serine, tyrosine, cysteine, phenylalanine, asparagine, glutamine, threonine, aspartic acid, glutamic acid, lysine, arginine and histidine;
step nine, sterilizing, namely sterilizing the Chinese herbal medicine bacterial flora culture solution obtained in the step eight to obtain a sterile Chinese herbal medicine bacterial flora culture solution, wherein the sterilization environment is 0.05MPa (110 ℃), and the sterilization time is controlled to be 20-30 min;
step ten, storing, namely freezing the Chinese herbal medicine bacterial flora culture solution in the step nine by using a freezer, taking out the culture solution in advance when in use, matching the culture solution with microorganisms after treatment, controlling the freezing temperature within the range of-80 to-50 ℃, and freezing for 72 hours.
Example 4:
the invention provides a Chinese herbal medicine bacterial flora culture solution which comprises the following components in parts by weight:
step one, preparing raw materials, namely 10 g of wormwood, 8 g of dandelion, 5 g of houttuynia cordata, 4 g of folium isatidis, 15 g of folium artemisiae argyi, 12 g of honeysuckle, 2 g of fructus forsythiae, 8 g of dandelion, 4 g of folium isatidis, 5 g of bezoar, 5 g of houttuynia cordata, 4 g of blackberry lily, 5 g of Chinese pulsatilla root, 5 g of radix isatidis, 5 g of indigo naturalis, 5 g of rhizoma smilacis glabrae, 5 g of subprostrate sophora, 2 g of oldenlandia diffusa, 8 g of Chinese violet, 10 g of andrographis paniculata, 11 g of purslane and 13 g of cortex dictamn;
step two, crushing the raw materials, namely mixing all the raw materials in the step one, putting the raw materials into a crusher for crushing after mixing, controlling the rotating speed of the crusher to be 1000-1200 rpm, controlling the crushing time to be 13min, controlling the particle sizes of all the crushed raw materials to be 100-180 um, and then fully mixing and shaking the raw materials to obtain the powder of the mixed raw materials;
step three, heating, namely putting the powder of the mixed raw material obtained in the step two into steam for heating, wherein the temperature of the steam is controlled to be 60-100 ℃, and the heating time is controlled to be 10-14 minutes;
step four, high-temperature high-pressure heating, namely placing the powder of the mixed raw material heated in the step three in a ventilated place for natural cooling, then placing the powder in a closed container for high-temperature high-pressure heating, wherein the heating temperature is controlled to be 330 ℃, and the pressure is controlled to be 200KPa-240KPa, so that the cell walls of the Chinese herbal medicines are broken, and the single-layer membrane of vacuole is broken, thereby vaporizing the cell sap;
step five, cooling and depressurizing, namely cooling and depressurizing the closed container in the step four, controlling the temperature to be 20-60 ℃ and the pressure to be 50KPa, so that the vaporized cell sap is liquefied, the cell sap is continuously liquefied to form liquid drops, and collecting the liquid drops by using the container;
step six, filtering, namely filtering the liquid drops collected in the step five, separating carbon-containing impurities, organelles and cell walls in the liquid drops, and controlling the mesh diameter of a filter screen to be between 2 and 6 um;
step seven, centrifugal purification, pouring the filtered liquid drops in the step six into a centrifugal tube, then placing the centrifugal tube into a centrifugal machine for separation and purification, removing redundant impurities to obtain purified cell sap, controlling the rotating speed of the centrifugal machine to 8000-12000r/min, and controlling the time to 20-24 min;
step eight, weighing and mixing, namely weighing 20 g of purified cell sap in the step seven, then adding 5 g of camphor wood powder, 8 g of bamboo charcoal powder, 100 g of distilled water, 2 g of sodium chloride, 5 g of peptone, 6 g of glucose, 5 g of inorganic salt, 5 g of nucleic acid, 3 g of enzyme and 5 g of amino acid, and fully mixing after adding to obtain a Chinese herbal medicine bacterial flora culture solution, wherein the inorganic salt is a mixture of calcium, phosphorus, potassium, sulfur, sodium, chlorine, magnesium, iron, zinc, selenium, molybdenum and iodine elements, and the amino acid consists of glycine, alanine, valine, leucine, isoleucine, methionine, proline, tryptophan, serine, tyrosine, cysteine, phenylalanine, asparagine, glutamine, threonine, aspartic acid, glutamic acid, lysine, arginine and histidine;
step nine, sterilizing, namely sterilizing the Chinese herbal medicine bacterial flora culture solution obtained in the step eight to obtain a sterile Chinese herbal medicine bacterial flora culture solution, wherein the sterilization environment is 0.05MPa (110 ℃), and the sterilization time is controlled to be 20-30 min;
step ten, storing, namely freezing the Chinese herbal medicine bacterial flora culture solution in the step nine by using a freezer, taking out the culture solution in advance when in use, matching the culture solution with microorganisms after treatment, controlling the freezing temperature within the range of-80 to-50 ℃, and freezing for 72 hours.
Example 5:
the invention provides a Chinese herbal medicine bacterial flora culture solution which comprises the following components in parts by weight:
step one, preparing raw materials, namely 10 g of wormwood, 8 g of dandelion, 5 g of houttuynia cordata, 4 g of folium isatidis, 15 g of folium artemisiae argyi, 12 g of honeysuckle, 2 g of fructus forsythiae, 8 g of dandelion, 4 g of folium isatidis, 5 g of bezoar, 5 g of houttuynia cordata, 4 g of blackberry lily, 5 g of Chinese pulsatilla root, 5 g of radix isatidis, 5 g of indigo naturalis, 5 g of rhizoma smilacis glabrae, 5 g of subprostrate sophora, 2 g of oldenlandia diffusa, 8 g of Chinese violet, 10 g of andrographis paniculata, 11 g of purslane and 13 g of cortex dictamn;
step two, crushing the raw materials, namely mixing all the raw materials in the step one, putting the raw materials into a crusher for crushing after mixing, controlling the rotating speed of the crusher to be 1000-1200 rpm, controlling the crushing time to be 14min, controlling the particle sizes of all the crushed raw materials to be 100-180 um, and then fully mixing and shaking the raw materials to obtain the powder of the mixed raw materials;
step three, heating, namely putting the powder of the mixed raw material obtained in the step two into steam for heating, wherein the temperature of the steam is controlled to be 60-100 ℃, and the heating time is controlled to be 10-14 minutes;
step four, high-temperature high-pressure heating, namely placing the powder of the mixed raw material heated in the step three in a ventilated place for natural cooling, then placing the powder in a closed container for high-temperature high-pressure heating, wherein the heating temperature is controlled to be 340 ℃, and the pressure is controlled to be 200KPa-240KPa, so that the cell walls of the Chinese herbal medicines are broken, and the single-layer membrane of vacuole is broken, thereby vaporizing the cell sap;
step five, cooling and depressurizing, namely cooling and depressurizing the closed container in the step four, controlling the temperature to be 20-60 ℃ and the pressure to be 60KPa, so that the vaporized cell sap is liquefied, the cell sap is continuously liquefied to form liquid drops, and collecting the liquid drops by using the container;
step six, filtering, namely filtering the liquid drops collected in the step five, separating carbon-containing impurities, organelles and cell walls in the liquid drops, and controlling the mesh diameter of a filter screen to be between 2 and 6 um;
step seven, centrifugal purification, pouring the filtered liquid drops in the step six into a centrifugal tube, then placing the centrifugal tube into a centrifugal machine for separation and purification, removing redundant impurities to obtain purified cell sap, controlling the rotating speed of the centrifugal machine to 8000-12000r/min, and controlling the time to 20-24 min;
step eight, weighing and mixing, namely weighing 20 g of purified cell sap in the step seven, then adding 5 g of camphor wood powder, 8 g of bamboo charcoal powder, 100 g of distilled water, 2 g of sodium chloride, 5 g of peptone, 6 g of glucose, 5 g of inorganic salt, 5 g of nucleic acid, 3 g of enzyme and 5 g of amino acid, and fully mixing after adding to obtain a Chinese herbal medicine bacterial flora culture solution, wherein the inorganic salt is a mixture of calcium, phosphorus, potassium, sulfur, sodium, chlorine, magnesium, iron, zinc, selenium, molybdenum and iodine elements, and the amino acid consists of glycine, alanine, valine, leucine, isoleucine, methionine, proline, tryptophan, serine, tyrosine, cysteine, phenylalanine, asparagine, glutamine, threonine, aspartic acid, glutamic acid, lysine, arginine and histidine;
step nine, sterilizing, namely sterilizing the Chinese herbal medicine bacterial flora culture solution obtained in the step eight to obtain a sterile Chinese herbal medicine bacterial flora culture solution, wherein the sterilization environment is 0.05MPa (110 ℃), and the sterilization time is controlled to be 20-30 min;
step ten, storing, namely freezing the Chinese herbal medicine bacterial flora culture solution in the step nine by using a freezer, taking out the culture solution in advance when in use, matching the culture solution with microorganisms after treatment, controlling the freezing temperature within the range of-80 to-50 ℃, and freezing for 72 hours.
Taking the Chinese herbal medicine bacterial flora culture solution prepared in the above examples 1-5, respectively, changing the crushing time of a crusher, changing the temperature during high-temperature and high-pressure heating, and changing the pressure during cooling and depressurization, then proportioning the Chinese herbal medicine bacterial flora culture solution prepared in the examples 1-5 and microorganisms to culture Chinese herbal medicine bacterial flora, after culturing for a period of time, testing the total number increase rate of the Chinese herbal medicine bacterial flora, the species increase rate of the Chinese herbal medicine bacterial flora, the survival ability increase rate of the Chinese herbal medicine bacterial flora and the reproduction increase rate of the Chinese herbal medicine bacterial flora, and then carrying out statistics and calculation to obtain the following data:
as can be seen from the above table, in example 3, the time for pulverizing by the pulverizer, the temperature during high-temperature and high-pressure heating, and the pressure during temperature and pressure reduction are all moderate, and then the total number increase rate of the Chinese herbal medicine bacterial flora, the species increase rate of the Chinese herbal medicine bacterial flora, the survival capacity increase rate of the Chinese herbal medicine bacterial flora, and the propagation increase rate of the Chinese herbal medicine bacterial flora in example 3 are tested, so that the total number increase rate of the Chinese herbal medicine bacterial flora, the species increase rate of the Chinese herbal medicine bacterial flora, the survival capacity increase rate of the Chinese herbal medicine bacterial flora, and the propagation increase rate of the Chinese herbal medicine bacterial flora in example 3 are all the highest.
The present invention is not limited to the above preferred embodiments, and any modifications, equivalent substitutions and improvements made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (7)
1. A Chinese herbal medicine bacterial flora culture solution is characterized in that: the preparation steps are as follows:
step one, preparing raw materials, namely 10 g of wormwood, 8 g of dandelion, 5 g of houttuynia cordata, 4 g of folium isatidis, 15 g of folium artemisiae argyi, 12 g of honeysuckle, 2 g of fructus forsythiae, 8 g of dandelion, 4 g of folium isatidis, 5 g of bezoar, 5 g of houttuynia cordata, 4 g of blackberry lily, 5 g of Chinese pulsatilla root, 5 g of radix isatidis, 5 g of indigo naturalis, 5 g of rhizoma smilacis glabrae, 5 g of subprostrate sophora, 2 g of oldenlandia diffusa, 8 g of Chinese violet, 10 g of andrographis paniculata, 11 g of purslane and 13 g of cortex dictamn;
step two, crushing the raw materials, namely mixing all the raw materials in the step one, putting the raw materials into a crusher for crushing after mixing, controlling the rotating speed of the crusher to be 1000-1200 rpm, controlling the crushing time to be 10-14min, controlling the particle sizes of all the crushed raw materials to be 100-180 um, and then fully mixing and shaking the raw materials uniformly to obtain powder of the mixed raw materials;
step three, heating, namely putting the powder of the mixed raw material obtained in the step two into steam for heating, wherein the temperature of the steam is controlled to be 60-100 ℃, and the heating time is controlled to be 10-14 minutes;
step four, high-temperature high-pressure heating, namely placing the powder of the mixed raw material heated in the step three in a ventilated place for natural cooling, then placing the powder in a closed container for high-temperature high-pressure heating, wherein the heating temperature is controlled to be 300-340 ℃, and the pressure is controlled to be 200-240 KPa, so that the cell walls of the Chinese herbal medicines are broken, the single-layer membrane of vacuole is broken, and the cell sap is vaporized;
step five, cooling and depressurizing, wherein the closed container in the step four is cooled and depressurized, the temperature is controlled to be 20-60 ℃, the pressure is controlled to be 20-60 KPa, so that vaporized cell sap is liquefied, the cell sap is continuously liquefied to form liquid drops, and the liquid drops are collected by the container;
step six, filtering, namely filtering the liquid drops collected in the step five, and separating carbon-containing impurities, organelles and cell walls in the liquid drops;
step seven, centrifugal purification, pouring the filtered liquid drops in the step six into a centrifugal tube, then placing the centrifugal tube into a centrifugal machine for separation and purification, and removing redundant impurities to obtain purified cell sap;
step eight, weighing and mixing, namely weighing 20 g of the purified cell sap obtained in the step seven, then adding 5 g of camphor wood powder, 8 g of bamboo charcoal powder, 100 g of distilled water, 2 g of sodium chloride, 5 g of peptone, 6 g of glucose, 5 g of inorganic salt, 5 g of nucleic acid, 3 g of enzyme and 5 g of amino acid, adding, and fully mixing to finally obtain a Chinese herbal medicine bacterial flora culture solution;
step nine, sterilization, namely sterilizing the Chinese herbal medicine bacterial flora culture solution obtained in the step eight to obtain a sterile Chinese herbal medicine bacterial flora culture solution;
step ten, storage, namely freezing the Chinese herbal medicine bacterial flora culture solution in the step nine by using a freezer, and taking out the culture solution in advance when in use, and matching the culture solution with microorganisms after treatment.
2. The Chinese herbal medicine bacterial flora culture solution of claim 1, wherein: and in the sixth step, the diameter of the filter screen is controlled between 2um and 6 um.
3. The Chinese herbal medicine bacterial flora culture solution of claim 1, wherein: in the seventh step, the rotating speed of the centrifuge is controlled to be 8000-12000r/min, and the time is controlled to be 20-24 min.
4. The Chinese herbal medicine bacterial flora culture solution of claim 1, wherein: and in the step eight, the inorganic salt is a mixture of calcium, phosphorus, potassium, sulfur, sodium, chlorine, magnesium, iron, zinc, selenium, molybdenum and iodine elements.
5. The Chinese herbal medicine bacterial flora culture solution of claim 1, wherein: the amino acid in step eight consists of glycine, alanine, valine, leucine, isoleucine, methionine, proline, tryptophan, serine, tyrosine, cysteine, phenylalanine, asparagine, glutamine, threonine, aspartic acid, glutamic acid, lysine, arginine and histidine.
6. The Chinese herbal medicine bacterial flora culture solution of claim 1, wherein: in the ninth step, the sterilization environment is 0.05MPa (110 ℃), and the sterilization time is controlled to be 20-30 min.
7. The Chinese herbal medicine bacterial flora culture solution of claim 1, wherein: the freezing temperature in the step ten is controlled within the range of-80 to-50 ℃, and the freezing time is 72 hours.
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| US20090208470A1 (en) * | 2005-10-22 | 2009-08-20 | Se-Joon Park | Microorganisms Having Bad Smell Removal Activity of Organic Waste and Use Thereof |
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| CN111876362A (en) * | 2020-08-14 | 2020-11-03 | 浙江九田环保科技有限公司 | Preparation process of ecological microbial agent |
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