CN111789252A - Method for improving kidney-tonifying and yang-strengthening effects of maca by enzymolysis fermentation method - Google Patents
Method for improving kidney-tonifying and yang-strengthening effects of maca by enzymolysis fermentation method Download PDFInfo
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- CN111789252A CN111789252A CN202010405741.3A CN202010405741A CN111789252A CN 111789252 A CN111789252 A CN 111789252A CN 202010405741 A CN202010405741 A CN 202010405741A CN 111789252 A CN111789252 A CN 111789252A
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- maca
- fermentation
- yang
- tonifying
- kidney
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Classifications
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Abstract
The invention discloses a method for improving kidney-tonifying and yang-strengthening effects of maca by an enzymolysis fermentation method, and belongs to the technical field of biology. The maca biological product with the high in-vitro kidney tonifying and yang strengthening effects is developed by taking maca, moringa seeds and rhizoma curculiginis as raw materials and integrating a modern bioseparation technology, a biological enzymolysis technology, a mixed bacteria microbial fermentation technology and a Chinese herbal medicine compatibility application technology, wherein mixed bacteria aerobic fermentation strains mainly comprise aspergillus niger, candida utilis and bacillus subtilis; the method can fully utilize flavone and polyphenol components in the medicinal materials such as maca and the like, enhance the medicinal effect, reduce the waste of the medicinal materials and improve the treatment effect of the medicinal materials; the kidney-tonifying and yang-strengthening effects of the maca biological product are remarkably improved through the synergistic effect of the Chinese herbal medicines and the mixed bacteria microorganisms, and the application requirements of the health care, medical treatment, food and other fields on maca are met; the process is simple and efficient, the obtained product has strong kidney-tonifying and yang-strengthening effects and free radical scavenging capacity, the production cost is low, and the industrial mass production is easy to realize.
Description
Technical Field
The invention relates to a method for improving kidney-tonifying and yang-strengthening effects of maca by an enzymolysis fermentation method, in particular to a maca biological product with high kidney-tonifying and yang-strengthening effects prepared by synergistic fermentation of microorganisms, and belongs to the technical field of biology.
Background
Maca is a cruciferous plant native to south america. Oval leaves and roots like radish, is edible, is a pure natural food with rich nutrient components and reputed by 'south American ginseng'. The original high-altitude mountain area of maca is suitable for growing in high-altitude, low-latitude, high day and night temperature difference, slightly acidic sandy soil and sunny land; the planting method is distributed in south America Andes mountains, is artificially planted in the middle and south of Peru, Yunnan and Xinjiang in China and other areas, and has a large area of suitable planting land. The maca is rich in high-unit nutrients and has the functions of nourishing and strengthening the body. Research shows that the maca mainly comprises the following nutritional ingredients: carbohydrate with protein content of more than 10% and 59%; 8.5% of fiber contains abundant mineral substances such as zinc, calcium, ferrum, titanium, rubidium, potassium, sodium, copper, manganese, magnesium, strontium, phosphorus, iodine, etc., and vitamin C, B1、B2、B6、A、E、B12、B5The fat content is not high but is mostly unsaturated fatty acid, the content of linoleic acid and linolenic acid is more than 53 percent, and the natural active ingredients comprise alkaloid, glucosinolate and decomposition products thereof, namely benzyl isothiocyanate, sterol, polyphenol substances and the like. In 1999, american scientists discovered that maca contains two new plant active ingredients, macamide and maca jelly, and confirmed that these two substances have a significant effect on balancing the secretion of hormones in the human body, so maca is also called a natural hormone engine. In the targeted research of maca products, the maca products are foundWith the research of several specific maca alkaloids, more active novel substances in maca can be separated and identified.
At present, researches on maca mainly focus on the development and utilization of maca, an extraction method mainly comprises a chemical method, an enzymolysis method and a microbial fermentation method, researches such as enzymolysis and microbial fermentation are lacked, particularly, related researches on the improvement of the kidney tonifying and yang strengthening effects of maca due to the lack of compatibility of Chinese herbal medicines restrict the application of maca in the fields of food, health care products, medicines and the like. The application discusses a method for improving the efficacy of maca in tonifying kidney and strengthening yang in vitro by matching maca and other Chinese herbal medicines, so as to solve the technical defects of low extraction rate of nutrient substances, resource utilization and the like in high-value utilization of maca in the prior art.
Disclosure of Invention
In order to overcome the defects of the prior art and realize the high-efficiency extraction of functional active substances of maca and the remarkable improvement of the kidney-tonifying and yang-strengthening effects, the application provides a method for improving the kidney-tonifying and yang-strengthening effects of maca by an enzymolysis fermentation method, integrates a modern biological separation technology, a biological enzymolysis technology, a mixed bacteria microbial fermentation technology and a Chinese herbal medicine compatibility application technology, develops and develops maca biological products with high in-vitro kidney-tonifying and yang-strengthening effects, meets the application requirements of the fields of health care, medical treatment, food and the like on maca, and has the advantages of simple and high-efficiency fermentation process, high extraction rate of active substances such as flavone and the like of the obtained products, strong kidney-tonifying and yang-strengthening.
The invention realizes the technical effects through the following technical scheme:
a method for improving kidney-tonifying and yang-strengthening efficacy of maca by an enzymolysis fermentation method is characterized in that moringa seeds and curculigo orchioides are added in the maca extraction process, and the compatibility proportion is as follows: 100 parts of maca, 2-5 parts of moringa seeds and 1-2 parts of curculigo orchioides.
The Chinese herbal medicine raw materials are subjected to the following pre-extraction treatment: removing impurities from raw materials of maca, moringa seeds and curculigo orchioides, crushing, sieving with a 80-150-mesh sieve, adding purified water with the mass ratio of 10 times of that of the raw materials of the Chinese herbal medicines, and soaking for 30min at 40-60 ℃; adding cellulase accounting for 2% of the mass of the Chinese herbal medicine raw materials, extracting for 2min under the assistance of ultrasonic waves, wherein the power of the ultrasonic waves is 250-400W, and then standing for more than 30 min.
The application provides a method for improving kidney-tonifying and yang-strengthening effects of maca by an enzymolysis fermentation method, which is characterized in that pre-extracted Chinese herbal medicines are subjected to mixed-bacterium aerobic fermentation, wherein aerobic fermentation strains mainly comprise aspergillus niger, candida utilis and bacillus subtilis.
The mixed bacteria aerobic fermentation process specifically comprises the following steps:
(1) fermentation culture preparation: adding purified water which is 10 times of the mass ratio of the Chinese herbal medicine raw materials into the pre-extracted Chinese herbal medicine, then adding glucose, ammonium sulfate, potassium dihydrogen phosphate and magnesium sulfate, wherein the mass ratio of the fermentation liquor is 3-5%, 0.3-0.5%, 0.1-0.2% and 0.01%, and uniformly stirring;
(2) aerobic fermentation: inoculating aspergillus niger, candida utilis and bacillus subtilis into the fermentation liquor, wherein the inoculation amounts of the aspergillus niger, the candida utilis and the bacillus subtilis are respectively 2-3%, 3-5% and 5-8% of the volume ratio of the fermentation liquor; controlling the temperature to be 28-35 ℃ for aerobic fermentation, wherein the rotating speed of a shaking table is 100-150 r/min, and the fermentation time is 12-24 h;
(3) solid-liquid separation: and (4) after the fermentation is finished, carrying out solid-liquid separation to obtain the maca biological product.
Furthermore, the ventilation rate is 0.1-0.2 m when the aerobic fermentation process is carried out in a fermentation tank3/m3.min。
Preferably, the residue after the solid-liquid separation is recycled and added to the next fermentation medium.
Preferably, 1-2% of xylanase and 0.5-1% of amylase can be added in the process of configuring the fermentation medium.
The preparation method of the aspergillus niger seed liquid comprises the following steps: culturing for 24h at 35-40 deg.C in potato culture medium under aseptic condition to obtain Aspergillus niger seed liquid with effective viable count not less than 109cfu/mL。
The preparation method of the yeast seed liquid comprises the following steps: under the aseptic operation condition, two rings of strains are picked from the inclined plane of the yeast YDP by using an inoculating ring, inoculated into a 500mL triangular flask containing 300mLPDA liquid culture medium, vibrated at the temperature of 28-32 ℃ and at the speed of 140r/minCulturing for 24 hr, inoculating seed tank, and enlarging culture to obtain yeast seed liquid with effective viable count not less than 109cfu/mL。
The preparation method of the bacillus subtilis seed solution comprises the following steps: under the aseptic condition, LB culture medium is adopted to culture for 24 hours at the temperature of 28-32 ℃ to prepare liquid bacillus seed liquid, and the effective viable count of the seed liquid is required to be more than or equal to 109cfu/mL。
The invention provides a method for improving the kidney-tonifying and yang-strengthening effects of maca by an enzymolysis fermentation method, the obtained maca has the advantages that the extraction rate and the in-vitro kidney-tonifying and yang-strengthening effects are remarkably improved, the extracted solution can be subjected to concentration, adsorption drying, alcohol precipitation or resin purification and other treatments according to application ways, and the efficient utilization in the fields of health care, food, medicine, fresh keeping, feed, fertilizer and the like is met.
The invention provides a method for improving kidney-tonifying and yang-strengthening efficacy of maca by an enzymolysis fermentation method, which has the following remarkable advantages compared with the prior art:
(1) the maca, the moringa seeds and the curculigo orchioides are scientifically matched, so that the growth promoting effect of the Chinese herbal medicines such as the moringa seeds and the curculigo orchioides on microorganisms can be fully utilized; the kidney-tonifying and yang-strengthening effects of the maca biological product are remarkably improved through the synergistic effect of the Chinese herbal medicines, and the technical effect of '1 +1> 2' is achieved;
(2) the method utilizes aerobic bacteria (aspergillus niger, saccharomycetes and bacillus subtilis) to jointly ferment the Chinese herbal medicine raw materials to extract the maca, makes full use of the synergistic effect among microorganisms, and improves the extraction rate of the maca and the effects of tonifying kidney and strengthening yang in vitro, wherein: aspergillus niger can decompose cellulose at high temperature, and break the constraint of cellulose on the escape of active ingredients (flavone, polyphenol, etc.) of maca function; the yeast can generate various enzymes for degrading cell walls in the fermentation process, so that more active substances such as flavone and the like are released from the cell walls, and the yeast can generate the enzymes capable of releasing combined state phenolic acid, so that the in-vitro kidney tonifying and yang strengthening effects of the maca can be improved; the bacillus subtilis can kill pathogenic bacteria in fermentation liquor in the high-temperature fermentation process, has strong capabilities of producing amylase, protease and the like, and can degrade macromolecular starch and protein substances into microorganisms for utilization;
(3) the liquid fermentation is adopted, so that the growth rate of the thalli is improved, the fermentation time is shortened, the production efficiency is improved, the fermentation process conditions are mild, the production cost is low, the large-scale production is easy, and the method plays an important role in improving the industrial economic benefit and large-scale popularization and application.
Detailed Description
Example 1
A method for improving kidney-tonifying and yang-strengthening effects of maca by an enzymolysis fermentation method comprises the following specific process steps:
(1) the compatibility of the Chinese herbal medicines is that the raw materials are accurately mixed according to the proportion of 100 parts of maca, 3 parts of moringa seeds and 1 part of curculigo orchioides;
(2) chinese herbal medicine pretreatment: removing impurities from raw materials of maca, moringa seeds and curculigo orchioides, crushing, sieving with a 80-150-mesh sieve, adding purified water with the mass ratio of 10 times of that of the raw materials of the Chinese herbal medicines, and soaking for 30min at 40-60 ℃; adding cellulase accounting for 2% of the mass ratio of the Chinese herbal medicine raw materials, extracting for 2min under the assistance of ultrasonic waves, wherein the ultrasonic wave power is 250-400W, and then standing for more than 30 min;
(3) fermentation culture preparation: adding purified water which is 10 times of the mass ratio of the Chinese herbal medicine raw materials into the pre-extracted Chinese herbal medicine, then adding glucose which is 4% of the mass ratio of the fermentation liquor, 0.45% of ammonium sulfate, 0.12% of monopotassium phosphate and 0.01% of magnesium sulfate, and uniformly stirring; adding 1% of xylanase and 0.5% of amylase into the fermentation liquor;
(4) preparing a fermentation seed solution:
the preparation method of the aspergillus niger seed liquid comprises the following steps: culturing for 24h at 35-40 deg.C in potato culture medium under aseptic condition to obtain Aspergillus niger seed liquid with effective viable count not less than 109cfu/mL。
The preparation method of the yeast seed liquid comprises the following steps: under the aseptic operation condition, two rings of strains are picked from the inclined plane of the yeast YDP by using an inoculating ring, inoculated into a 500mL triangular flask containing 300mLPDA liquid culture medium, subjected to shaking culture at the temperature of between 28 and 32 ℃ and at the speed of 140r/min for 24 hours, and then inoculated into a seed tank for expanded culture according to the process to prepare the yeast seed liquid, wherein the effective viable count of the seed liquid is required to be more than or equal to109cfu/mL。
The preparation method of the bacillus subtilis seed solution comprises the following steps: under the aseptic condition, LB culture medium is adopted to culture for 24 hours at the temperature of 28-32 ℃ to prepare liquid bacillus seed liquid, and the effective viable count of the seed liquid is required to be more than or equal to 109cfu/mL。
(5) Aerobic fermentation: inoculating aspergillus niger, candida utilis and bacillus subtilis into the fermentation liquor, wherein the inoculation amounts of the aspergillus niger, the candida utilis and the bacillus subtilis are respectively 3%, 5% and 8% of the volume ratio of the fermentation liquor; controlling the temperature to be 28-35 ℃ for aerobic fermentation, wherein the rotating speed of a shaking table is 100-150 r/min, and the fermentation time is 16 h;
(6) solid-liquid separation: and (4) after the fermentation is finished, carrying out solid-liquid separation to obtain the maca biological product.
Example 2
A method for improving kidney-tonifying and yang-strengthening effects of maca by an enzymolysis fermentation method comprises the following specific process steps:
(1) the compatibility of the Chinese herbal medicines is that the raw materials are accurately mixed according to the proportion of 100 parts of maca, 5 parts of moringa seeds and 2 parts of curculigo orchioides;
(2) chinese herbal medicine pretreatment: removing impurities from raw materials of maca, moringa seeds and curculigo orchioides, crushing, sieving with a 80-150-mesh sieve, adding purified water with the mass ratio of 10 times of that of the raw materials of the Chinese herbal medicines, and soaking for 30min at 40-60 ℃; adding cellulase accounting for 2% of the mass ratio of the Chinese herbal medicine raw materials, extracting for 2min under the assistance of ultrasonic waves, wherein the ultrasonic wave power is 250-400W, and then standing for more than 30 min;
(3) fermentation culture preparation: adding purified water which is 10 times of the mass ratio of the Chinese herbal medicine raw materials into the pre-extracted Chinese herbal medicine, then adding glucose which is 4% of the mass ratio of the fermentation liquor, 0.45% of ammonium sulfate, 0.12% of monopotassium phosphate and 0.01% of magnesium sulfate, and uniformly stirring; adding 1% of xylanase and 0.5% of amylase into the fermentation liquor;
(4) preparing a fermentation seed solution:
the preparation method of the aspergillus niger seed liquid comprises the following steps: culturing for 24h at 35-40 deg.C in potato culture medium under aseptic condition to obtain Aspergillus niger seed liquid with effective viable count not less than 109cfu/mL。
The preparation method of the yeast seed liquid comprises the following steps: in a sterile operating stripUnder the condition, two rings of strains are picked from the inclined plane of the yeast YDP by using an inoculating ring, inoculated into a 500mL triangular flask filled with 300mLPDA liquid culture medium, subjected to shaking culture at the temperature of between 28 and 32 ℃ for 24 hours at 140r/min, and then inoculated into a seed tank for expanded culture according to the process to prepare the yeast seed liquid, wherein the effective viable count of the seed liquid is required to be more than or equal to 109cfu/mL。
The preparation method of the bacillus subtilis seed solution comprises the following steps: under the aseptic condition, LB culture medium is adopted to culture for 24 hours at the temperature of 28-32 ℃ to prepare liquid bacillus seed liquid, and the effective viable count of the seed liquid is required to be more than or equal to 109cfu/mL。
(5) Aerobic fermentation: inoculating aspergillus niger, candida utilis and bacillus subtilis into the fermentation liquor, wherein the inoculation amounts of the aspergillus niger, the candida utilis and the bacillus subtilis are respectively 3%, 5% and 8% of the volume ratio of the fermentation liquor; controlling the temperature to be 28-35 ℃ for aerobic fermentation, wherein the rotating speed of a shaking table is 100-150 r/min, and the fermentation time is 16 h;
(6) solid-liquid separation: and (4) after the fermentation is finished, carrying out solid-liquid separation to obtain the maca biological product.
Example 3
A method for improving kidney-tonifying and yang-strengthening effects of maca by an enzymolysis fermentation method comprises the following specific process steps:
(1) the compatibility of the Chinese herbal medicines is that the raw materials are accurately mixed according to the proportion of 100 parts of maca, 3 parts of moringa seeds and 1 part of curculigo orchioides;
(2) chinese herbal medicine pretreatment: removing impurities from raw materials of maca, moringa seeds and curculigo orchioides, crushing, sieving with a 80-150-mesh sieve, adding purified water with the mass ratio of 10 times of that of the raw materials of the Chinese herbal medicines, and soaking for 30min at 40-60 ℃; adding cellulase accounting for 2% of the mass ratio of the Chinese herbal medicine raw materials, extracting for 2min under the assistance of ultrasonic waves, wherein the ultrasonic wave power is 250-400W, and then standing for more than 30 min;
(3) fermentation culture preparation: adding purified water which is 10 times of the mass ratio of the Chinese herbal medicine raw materials into the pre-extracted Chinese herbal medicine, then adding glucose which is 4% of the mass ratio of the fermentation liquor, 0.45% of ammonium sulfate, 0.12% of monopotassium phosphate and 0.01% of magnesium sulfate, and uniformly stirring;
(4) preparing a fermentation seed solution:
the preparation method of the aspergillus niger seed liquid comprises the following steps: under the aseptic condition, adopting potatoCulturing the culture medium at 35-40 ℃ for 24h to prepare the Aspergillus niger seed liquid, wherein the effective viable count of the seed liquid is required to be more than or equal to 109cfu/mL。
The preparation method of the yeast seed liquid comprises the following steps: under the aseptic operation condition, two rings of strains are picked from the inclined plane of the yeast YDP by using an inoculating ring, inoculated into a 500mL triangular flask containing 300mLPDA liquid culture medium, subjected to shaking culture at the temperature of between 28 and 32 ℃ and at the speed of 140r/min for 24 hours, and then inoculated into a seed tank for expanded culture according to the process to prepare the yeast seed liquid, wherein the effective viable count of the seed liquid is required to be more than or equal to 109cfu/mL。
The preparation method of the bacillus subtilis seed solution comprises the following steps: under the aseptic condition, LB culture medium is adopted to culture for 24 hours at the temperature of 28-32 ℃ to prepare liquid bacillus seed liquid, and the effective viable count of the seed liquid is required to be more than or equal to 109cfu/mL。
(5) Aerobic fermentation: inoculating aspergillus niger, candida utilis and bacillus subtilis into the fermentation liquor, wherein the inoculation amounts of the aspergillus niger, the candida utilis and the bacillus subtilis are respectively 3%, 5% and 8% of the volume ratio of the fermentation liquor; controlling the temperature to be 28-35 ℃ for aerobic fermentation, wherein the rotating speed of a shaking table is 100-150 r/min, and the fermentation time is 16 h;
(6) solid-liquid separation: and (4) after the fermentation is finished, carrying out solid-liquid separation to obtain the maca biological product.
Example 4 Effect of different Process conditions on in vitro Kidney-invigorating and Yang-strengthening efficacy of maca biological products 1, index determination method
(1) The effect of maca on scavenging DPPH free radicals: DPPH9.7mg is accurately weighed, a small amount of absolute ethyl alcohol is added for dissolving, and then the volume is determined to be 250mL by 95% ethyl alcohol, thus preparing DPPH solution with the concentration of 0.2 mmol/L. Respectively adding 1mL of maca solutions with different concentrations into 2mL of prepared DPPH solution, supplementing to 4mL with 95% ethanol, mixing uniformly, standing for 30min, measuring the light absorption value at 517nm with 95% ethanol as a blank control, and calculating the inhibition rate;
(2) maca pair Fe3+Reduction of (2): the reduction force detection of maca refers to the Oyaizu method. Sequentially adding 0.75mL of 0.2mol/L phosphate buffer solution (pH6.6), 0.75mL of 1% potassium ferrocyanide and 0.75mL of 1mg/mL maca solution, uniformly mixing, and carrying out water bath at 50 ℃ for 20 min; after rapid cooling in ice bath, 0.75mL 10% trichloroacetic acid was added in 3000 preparedg. Centrifuging at 4 deg.C for 10 min; taking 1.5mL of the supernatant, adding 0.1% FeCl30.5mL and ddH2Mixing with O10 mL, standing at room temperature for 10min, and measuring absorbance A at 700nm1At 1mg/mLVCControl A, and zero tubes A0Calculating the relative reducing power of the maca;
2. test grouping
Examples 1-3 test groups; comparative examples 1-3 groups, the raw materials contained only maca, moringa seeds and curculigo orchioides, respectively, and the preparation process was the same as in example 1;
3. test results
TABLE 2 influence of different process conditions on the in vitro kidney-invigorating and yang-strengthening efficacy of maca biological products
| Test group | DPPH radical inhibition (%) | Fe3+Reducing power (%) |
| Example 1 test group | 82.54 | 333.89 |
| Example 2 test group | 84.18 | 341.27 |
| Example 3 test group | 79.37 | 315.82 |
| Comparative example 1 test group | 81.65 | 332.62 |
| Comparative example 2 test group | 79.74 | 300.66 |
| Comparative example 3 test group | 82.05 | 318.25 |
Example 5 Effect of different Process conditions on in vitro Kidney-invigorating and Yang-strengthening efficacy of maca biological products
Test animals: male ICR mice, body mass (20 ± 2) g;
grouping and administration: 110 ICR male mice, after 5 days of acclimation under experimental conditions, were divided into 7 experimental groups of 10 mice each. Wherein the test groups of examples 1-3 were administered the biologicals of the present invention; comparative examples 1-3 test groups were given samples of comparative examples; the gavage dosage of the same volume of physiological saline for the blank control group ig mice is 2g-1. Measuring various indexes after 30 days;
and (3) carrying out a load swimming experiment: the load swimming test and the normal pressure hypoxia tolerance test are carried out on male mice according to the requirements of 'health food inspection and evaluation technical specification'. After the mice are subjected to last gastric lavage administration for 30min, a 5% lead skin of the weight is loaded at 1/3-2/3 parts of the tail root of the mouse, the mouse swims in water with the water depth of 30cm and the water temperature of (30 +/-2) DEG C, a plurality of 60cm multiplied by 40cm multiplied by 50cm water tanks are prepared at the same time, the time from swimming to death of the mouse is recorded by a stopwatch, and the time for timing is not started when the head of the mouse sinks into the water for 7 s;
and (3) normal-pressure hypoxia tolerance experiment, namely performing the normal-pressure hypoxia tolerance experiment on male mice according to the requirements of 'health food inspection and evaluation technical specifications'. After the mice are subjected to last intragastric administration for 1 hour, 5g of soda lime is wrapped by gauze and placed into a 250mL wide-mouth bottle, the mice are placed into the bottle (1 per bottle), vaseline is immediately smeared on the bottle mouth to be tightly covered, the time from the time when the mice are placed into the bottle to the time when the mice have general spasm symptoms is recorded by a stopwatch, and the mice are quickly taken out;
the serum NO and testosterone (T) value is measured by resting the mouse for 60min after a normal pressure hypoxia-resistant experiment, taking eyeball and taking blood to store in a 1.5mL centrifuge tube, centrifuging for 5min at 4000r/min, taking upper serum, processing the sample according to the NO and testosterone kit instruction and measuring the content by using an enzyme-linked immunosorbent assay;
the test results are shown in the following table:
test results show that the maca biological product has remarkable effects of tonifying kidney and strengthening yang, and the embodiment 1 is the best embodiment of the invention.
The above embodiments are only used for illustrating the technical solution of the present invention, and not for limiting the same; although the invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that various changes may be made and equivalents may be substituted for elements thereof; such modifications and substitutions do not depart from the spirit and scope of the present invention as set forth in the appended claims.
Claims (10)
1. A method for improving kidney-tonifying and yang-strengthening effects of maca by an enzymolysis fermentation method is characterized in that moringa seeds and curculigo orchioides are added in the maca extraction process, and the compatibility proportion is as follows: 100 parts of maca, 2-5 parts of moringa seeds and 1-2 parts of curculigo orchioides; the Chinese herbal medicine raw materials are subjected to the following pre-extraction treatment: removing impurities from raw materials of maca, moringa seeds and curculigo orchioides, crushing, sieving with a 80-150-mesh sieve, adding purified water with the mass ratio of 10 times of that of the raw materials of the Chinese herbal medicines, and soaking for 30min at 40-60 ℃; adding cellulase accounting for 2% of the mass of the Chinese herbal medicine raw materials, extracting for 2min under the assistance of ultrasonic waves, wherein the power of the ultrasonic waves is 250-400W, and then standing for more than 30 min.
2. The method for improving kidney-tonifying and yang-strengthening effects of maca by using the enzymolysis fermentation method according to claim 1, wherein the pre-extracted Chinese herbal medicines are subjected to mixed-bacteria aerobic fermentation, wherein aerobic fermentation strains mainly comprise aspergillus niger, candida utilis and bacillus subtilis; the method specifically comprises the following steps:
(1) fermentation culture preparation: adding purified water which is 10 times of the mass ratio of the Chinese herbal medicine raw materials into the pre-extracted Chinese herbal medicine, then adding glucose, ammonium sulfate, potassium dihydrogen phosphate and magnesium sulfate, wherein the mass ratio of the fermentation liquor is 3-5%, 0.3-0.5%, 0.1-0.2% and 0.01%, and uniformly stirring;
(2) aerobic fermentation: inoculating aspergillus niger, candida utilis and bacillus subtilis into the fermentation liquor, wherein the inoculation amounts of the aspergillus niger, the candida utilis and the bacillus subtilis are respectively 2-3%, 3-5% and 5-8% of the volume ratio of the fermentation liquor; controlling the temperature to be 28-35 ℃ for aerobic fermentation, wherein the rotating speed of a shaking table is 100-150 r/min, and the fermentation time is 12-24 h;
(3) solid-liquid separation: and (4) after the fermentation is finished, carrying out solid-liquid separation to obtain the maca biological product.
3. The method for improving kidney-tonifying and yang-strengthening effects of maca by using the enzymolysis fermentation method according to claim 2, wherein the ventilation rate is 0.1-0.2 m when the aerobic fermentation process is carried out in a fermentation tank3/m3.min。
4. The method for improving the kidney-tonifying and yang-strengthening effects of maca according to claim 2, wherein the residues after solid-liquid separation can be recycled and added to a next fermentation medium.
5. The method for improving the kidney-tonifying and yang-strengthening effects of maca by the enzymolysis fermentation method according to claim 2, wherein 1-2% of xylanase and 0.5-1% of amylase can be added in the preparation process of the fermentation medium.
6. The method for improving the kidney-tonifying and yang-strengthening efficacy of maca according to claim 2, wherein the preparation method of the aspergillus niger seed solution comprises the following steps: culturing for 24h at 35-40 deg.C in potato culture medium under aseptic condition to obtain Aspergillus niger seed liquid with effective viable count not less than 109cfu/mL。
7. The method for improving the kidney-tonifying and yang-strengthening efficacy of maca according to claim 2, wherein the yeast seed solution is prepared by the following steps: under the aseptic operation condition, two-ring strains are picked from the inclined plane of the yeast YDP by using an inoculating ring, inoculated into a 500mL triangular flask containing 300mLPDA liquid culture medium, subjected to shaking culture at the temperature of between 28 and 32 ℃ at 140r/min for 24 hours, and then inoculated with seeds according to the processPreparing yeast seed liquid by tank enlargement culture, wherein the effective viable count of the seed liquid is required to be more than or equal to 109cfu/mL。
8. The method for improving the kidney-tonifying and yang-strengthening effects of maca by the enzymolysis fermentation method according to claim 2, wherein the preparation method of the bacillus subtilis seed solution comprises the following steps: under the aseptic condition, LB culture medium is adopted to culture for 24 hours at the temperature of 28-32 ℃ to prepare liquid bacillus seed liquid, and the effective viable count of the seed liquid is required to be more than or equal to 109cfu/mL。
9. Maca biologics prepared by the process of claims 1-8.
10. The maca biological product of claim 9, wherein the maca biological product is used in the fields of food, health products, medicines and feeds.
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| CN112251469A (en) * | 2020-10-28 | 2021-01-22 | 付鹏磊 | Method for improving oxidation resistance of maca by enzymolysis fermentation method |
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