CN113079948A - Culture medium and preparation method and application thereof - Google Patents
Culture medium and preparation method and application thereof Download PDFInfo
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- CN113079948A CN113079948A CN202110379235.6A CN202110379235A CN113079948A CN 113079948 A CN113079948 A CN 113079948A CN 202110379235 A CN202110379235 A CN 202110379235A CN 113079948 A CN113079948 A CN 113079948A
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- apple pomace
- culture medium
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- rhizopus
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
- A01G18/20—Culture media, e.g. compost
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Abstract
The invention discloses a culture medium and a preparation method and application thereof, wherein the main raw material components comprise apple pomace and rhizopus spore liquid, and each kilogram of apple pomace is matched with 200-400ml of rhizopus spore liquid for use. The apple pomace and the rhizopus spore liquid are mixed and fermented, corncobs and bean cake powder are added to further absorb the moisture content, and the prepared culture medium can accelerate the growth speed of mushrooms, improve the taste of the mushrooms and reduce the production cost through a mushroom inoculation test.
Description
Technical Field
The invention belongs to the technical field of agricultural waste recycling, relates to recycling of apple pomace, and particularly relates to a culture medium and a preparation method and application thereof. The culture medium is prepared from fructus Mali Pumilae residue, and can be used for culturing Agaricus campestris.
Background
The apple pomace is the remainder of fresh apples after crushing, squeezing and juice extraction, mainly comprises peel, kernel and residual pulp, and is rich in soluble sugar, vitamins, mineral substances, cellulose and other nutrient substances. Through determination, in the apple pomace, the peel pulp accounts for 96.2%, the fruit seeds account for 3.1%, the fruit stalks account for 0.7%, and the nitrogen-free extract of the apple pomace accounts for 61.5%, wherein the total sugar accounts for 15.1%, the crude fat accounts for 6.8%, and the crude protein content is 6.2%. In addition to a small amount of fruit shells and fruit stalks in the crude fibers being lignin, most of the fruit pulps and fruit peels are hemicellulose and cellulose. The fresh apple pomace has high acidity and is an excellent culture medium for acid-resistant straw rotting mushrooms (pleurotus eryngii, flammulina velutipes, oyster mushrooms, hypsizygus marmoreus and pleurotus citrinopileatus) and a small amount of wood rotting mushrooms (shiitake mushrooms). The fresh apple pomace is processed into apple pomace dry powder which can be used for preparing complete feed or granular feed and used as feed for domestic livestock and poultry such as pigs, cattle, sheep and the like.
Patent CN103044118A discloses a mushroom culture medium, which comprises the following raw materials in parts by weight: 50-70 parts of apple pomace, 30-50 parts of peanut shells and 2 parts of gypsum. The preparation method comprises the following steps: dissolving Gypsum Fibrosum in water, adding fructus Mali Pumilae residue and peanut shell according to the formula, adding water in an amount of 60-70%, stirring, placing the uniformly mixed culture medium into polypropylene cylindrical bag, sterilizing at 120 deg.C for 100min, naturally cooling to room temperature, inoculating, and placing into culture room. The apple pomace contains a large amount of reducing sugar, is easy to rot and mildew and loses the function of the apple pomace as feed, and the viscosity of fresh apple pomace is high, so that the apple pomace as feed has the defects of low protein content and low biological conversion rate. The fresh pomace is directly used for mushroom cultivation, so that the nutrient absorption of hyphae is not facilitated, the inclusion between pectin and cellulose prevents the combination of cellulase and cellulose and hemicellulose, hyphae grow slowly and cannot grow into a substrate, and the utilization of cellulose polysaccharide in the pomace is limited. The extraction agent dosage required for extracting the soluble fiber of the apple pomace is large, the energy consumption is high, and the utilization of the apple pomace is limited.
Therefore, it is known from the above analysis that the mushroom cultivation medium described in patent CN103044118A has limited utilization of cellulose-based polysaccharide in apple pomace, the comprehensive utilization efficiency of apple pomace is low, and the problem of damage to mushroom growth caused by methanol generated when apple pomace is used is not solved.
A large amount of cellulose and hemicellulose in the apple pomace are high-quality growth nutrition of mushrooms (large filamentous fungi), and although researchers do relevant research, the technical problem of using the pomace for mushroom cultivation cannot be solved well, and the comprehensive utilization efficiency of the apple pomace is low. By providing the culture medium and the preparation method and the application thereof, the utilization rate of the apple pomace can be effectively improved, the growth and the development of mushrooms are promoted, the quality is improved, and the cost is reduced.
Disclosure of Invention
The invention aims to recycle apple pomace and prepare mushroom culture medium.
Based on the aim, the invention provides a culture substrate and a preparation method and application thereof, and particularly, apple pomace and rhizopus spore liquid are mixed and then subjected to aerobic fermentation, wherein 200-400ml of rhizopus spore liquid is inoculated to every kilogram of apple pomace, and each ml of rhizopus spore liquid contains 100000 rhizopus spores of 80000-one.
The invention also provides a preparation method of the mushroom culture medium, which comprises the following steps:
1) the fresh fruit residues are further crushed without sieving and can be normally divided into fragments with the diameter of 0.3-1 cm. Adding lime water to adjust pH to 7.5-9.0, adjusting for many times, storing for 4-8h, and keeping the pH stable for later use.
2) The rhizopus spore liquid is used, the specific dosage is 100000 pieces/ml per 80000-. The rhizopus spore liquid has the advantage of counting, and can accurately control the amount of added microorganisms.
3) Mixing apple pomace and rhizopus spore liquid, inserting a tip wooden stick with the diameter of 2-8cm into the bottom of the apple pomace pile, punching an exhaust hole, carrying out aerobic fermentation at the constant temperature of 32-35 ℃ for 2-5 days, and turning over once every 7-8h to prevent the anaerobic fermentation of rhizopus to convert sugar into alcohol.
4) When more liquid is generated after the fermentation of the apple pomace begins, the pomace is not adhered and is full of water, and when the content of reducing sugar in the pomace is increased by 25-40g/kg, 40-mesh corncobs are added, the corncobs can absorb water to increase gaps of the apple pomace, oxygen is sufficient, and after the corncobs absorb water, the apple pomace is integrally loosened to facilitate the growth of later-stage hyphae. Continuing fermentation, reducing the temperature of the fermentation pile to 24-25 ℃, stabilizing the water content, and avoiding water seepage in the apple pomace fermentation pile.
5) Adjusting the pH value of the product obtained in the step 4) to 5-7 by using ammonia water or lime water, increasing the water absorption of the bean cake powder, and blending to 1-4:15-20 of the weight ratio of the bean cake to the apple pomace to ensure that the water absorption content of the total mixture of the fermented apple pomace, the bean cake powder and the corncob powder is 85% -90%. The bean cake powder is an important nitrogen source component in the ingredients, the apple pomace is lack of nitrogen source nutrition for microbial growth, raw materials containing nitrogen nutrition are artificially increased, and the aim of cultivation can be fulfilled only by adjusting the matrix component.
6) Bagging using an automatic filling machine, or bottle filling to fill naturally. After bagging, sterilizing at normal pressure and high temperature for 18-24h at 110-. Cooling to normal temperature, standing at room temperature, culturing for 85-100h without breeding of foreign bacteria, and collecting the culture medium.
Compared with the prior art, the invention has the following beneficial effects or advantages:
the invention provides a culture medium and a preparation method and application thereof, wherein the main raw materials comprise apple pomace and rhizopus spore liquid, the apple pomace and the rhizopus spore liquid are subjected to mixed aerobic fermentation, corncobs and bean cake powder are added to further reduce the moisture content, the prepared culture medium is beneficial to the rapid growth of mushrooms, the taste of the mushrooms is improved, and the toxic action on mushroom hyphae is avoided.
Detailed Description
The following examples are given to illustrate the technical aspects of the present invention, but the present invention is not limited to the following examples.
Example 1
The embodiment provides a culture substrate and a preparation method and application thereof, and raw material components of the culture substrate comprise 100g of apple pomace, 20ml of 80000 and 100000 rhizopus spore solutions, 30g of 40-mesh corn cobs and 5g of bean cake powder.
The preparation method comprises the following specific steps:
step one, further crushing 100g of apple pomace to form fragments with the diameter of 0.3-1 cm. Adding lime water to adjust pH value to 7.5-9.0, storing for 4-8h, and keeping pH value stable for use.
And step two, mixing the treated apple pomace with rhizopus spore liquid, punching an exhaust hole, carrying out aerobic fermentation at the constant temperature of 32-35 ℃ for 5 days, and turning over once every 7-8 h.
And step three, after the apple pomace begins to ferment to generate more mixed reducing sugar liquid, the pomace is not adhered and is full of water, the content of the reducing sugar in the pomace is measured to be increased by 25-40g/kg, 40-mesh corncobs are added, the water is absorbed, the fermentation is continued, the temperature of a fermentation pile is reduced to 24-25 ℃, the water is stable, and no water seeps. Measuring pH, adding lime water to adjust pH to 5-7, and adding 5g bean cake powder to absorb water.
And step four, bottle cultivation filling to fill the bottles naturally.
Step five, sterilizing the filled culture substrate for 18-24h at 110-120 ℃ under normal pressure.
And sixthly, after cooling to the normal temperature, carrying out vacant constant-temperature culture for 85-100h, observing no sundry bacteria breeding, and preparing the culture medium.
Example 2
This example provides a test of a growth substrate for promoting mushroom growth. The main raw materials of the culture medium comprise apple pomace and rhizopus spore liquid.
The tested mushroom variety is agaricus bisporus 2796, the test site is carried out in laboratories of life science college of Shanxi university of science and technology, and all groups of culture mediums are uniformly managed during the test.
The apple pomace and the rhizopus spore liquid are prepared into the culture substrate according to the steps of example 1. The invention provides a solid culture medium, and therefore, a common substitute culture medium for mushroom culture is selected as a contrast.
The preparation method of the common substitute culture medium comprises the following specific steps:
collecting 100g potato, 5g glucose, 5g maltose, and 1g KH2PO4、0.5g MgSO4And 10g of agar, 200ml of distilled water was added, and after sufficiently dissolving the mixture by heating, the volume was adjusted to 500ml by using distilled water.
Placing the fungus blocks with diameter of 0.5cm in the center of the culture medium of each treatment group, repeating the test three times for each treatment group, culturing at 24 deg.C in dark, counting the time from the completion of inoculation, counting the length of mycelium and mushroom growth condition of each treatment group, and detecting whether mycotoxin is produced, wherein the counting result is shown in Table 1.
TABLE 1 test of growth promotion of mushrooms by cultivation substrate prepared by fermenting apple pomace
As can be seen from the data in Table 1, the culture medium prepared by fermenting the apple pomace can effectively promote the growth of mushrooms by using the raw materials of the apple pomace and the rhizopus spore liquid within a certain proportion range, the half bag growth time of hypha is 20 days, the full bag growth time is 45 days, the hypha after-ripening time is 14 days, and the primordium emergence time is 78 days, which are all lower than the culture time of a common substitute culture medium.
Example 3
This example provides a test of a growth substrate for promoting the growth of mushroom hyphae. The main raw materials of the culture medium comprise apple pomace and rhizopus spore liquid.
The tested mushroom variety is agaricus bisporus 2796, the test site is carried out in laboratories of life science college of Shanxi university of science and technology, and all groups of culture mediums are uniformly managed during the test.
The apple pomace and the rhizopus spore liquid are prepared into the culture substrate according to the steps of example 1. The test set up test and control. In the test group, apple pomace and rhizopus spore liquid are mixed according to the proportion shown in the table 2 to prepare the culture medium. The invention provides a solid culture medium, which selects a common substitute culture medium for mushroom culture as a control group.
The preparation method of the common substitute culture medium comprises the following specific steps:
collecting 100g potato, 5g glucose, 5g maltose, and 1g KH2PO4、0.5g MgSO4And 10g of agar, 200ml of distilled water was added, and after sufficiently dissolving the mixture by heating, the volume was adjusted to 500ml by using distilled water.
And (3) placing a fungus block with the diameter of 0.5cm in the center of the culture medium of each treatment group, repeating the test for three times for each treatment group, culturing at 24 ℃ in the dark, timing from the completion of inoculation, and counting the time required by the fungus filaments of each treatment group to be full of bags, wherein the counting result is shown in table 2.
The promotion rate (%) was calculated as follows:
TABLE 2 test of growth promotion of mushroom hyphae by cultivation substrate prepared by fermenting apple pomace
As can be seen from the data in Table 2, the cultivation substrate prepared by fermenting the apple pomace can effectively promote the growth of mushroom hyphae by using the raw materials of the apple pomace and the rhizopus spore liquid within a certain proportion range, the minimum bag filling time of the hyphae is 1046.30h, namely 43.6 days, and the highest promotion rate reaches 13.08%.
As described above, the present invention can be preferably implemented, and the above-mentioned embodiments only describe the preferred embodiments of the present invention, and do not limit the scope of the present invention, and various changes and modifications of the technical solution of the present invention made by those skilled in the art without departing from the design spirit of the present invention shall fall within the protection scope defined by the present invention.
Claims (10)
1. The preparation method of the culture medium is characterized by comprising the steps of mixing the apple pomace and the rhizopus spore liquid and then carrying out aerobic fermentation, wherein 200-400ml of rhizopus spore liquid is inoculated to each kilogram of apple pomace, and each ml of rhizopus spore liquid contains 80000-100000 rhizopus spores.
2. The method of preparing a culture medium according to claim 1, wherein the pH of the apple pomace is adjusted to be stabilized in a range of 7.5 to 9.0 before the apple pomace is mixed with the rhizopus spore solution.
3. The method for preparing a culture medium according to claim 1, wherein the temperature condition of the aerobic fermentation is 32-35 ℃, and apple pomace is required to be turned over during the aerobic fermentation.
4. The method for preparing a culture medium according to claim 1, wherein the aerobic fermentation is carried out to produce liquid water, and then the liquid water is adsorbed by the crushed corncobs, and the fermentation is continued; when the produced liquid water is stable, adjusting pH to 5.0-7.0, adding bean cake powder to absorb water to make the water content of the fermentation product 85-90%.
5. The preparation method of the culture medium according to claim 4, wherein the mass ratio of the bean cake powder to the apple pomace is 1: 20.
6. The method for preparing a culture medium according to claim 4, wherein the pH is adjusted by using ammonia water or lime water.
7. The method for preparing a culture medium according to claim 2, wherein lime water is selected to adjust the pH of the apple pomace.
8. A method for preparing culture medium comprises adjusting pH of fructus Mali Pumilae residue to make pH of fructus Mali Pumilae residue stable in 7.5-9.0 range; mixing the apple pomace and the rhizopus spore liquid, and then carrying out aerobic fermentation, wherein 200-400ml of rhizopus spore liquid is inoculated to each kilogram of apple pomace, each ml of rhizopus spore liquid contains 80000-100000 rhizopus spores, the temperature condition of the aerobic fermentation is 32-35 ℃, and the apple pomace needs to be turned over during the aerobic fermentation; after liquid water is generated, the liquid water is absorbed by the corncobs, and the fermentation is continued; and when the generated liquid water is stable, adjusting the pH value to be 5.0-7.0, and adding bean cake powder to absorb water, so that the total mixture of the fermented apple pomace, the bean cake powder and the corncobs has the water absorption content of 85-90%.
9. A culture substrate prepared by the method of any one of claims 1-8.
10. Use of a culture medium according to claim 9 for mushroom cultivation.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104945129A (en) * | 2015-06-18 | 2015-09-30 | 曾少兰 | Mushroom culture medium |
| CN106306361A (en) * | 2016-08-25 | 2017-01-11 | 宁夏泰瑞制药股份有限公司 | Method for preparing biological fermentation feed |
| CN110144317A (en) * | 2019-06-05 | 2019-08-20 | 甘肃省畜牧兽医研究所 | A kind of composite bacteria agent, freeze-drying microbial inoculum and apple pomace protein feed for fermentation apple pomace |
| AU2020103860A4 (en) * | 2020-12-03 | 2021-02-11 | Gansu Animal Husbandry and Veterinary Medicine Institute | Complex microbial agent and freeze-dried microbial agent for fermenting apple pomace and apple pomace protein feed |
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- 2021-04-08 CN CN202110379235.6A patent/CN113079948A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104945129A (en) * | 2015-06-18 | 2015-09-30 | 曾少兰 | Mushroom culture medium |
| CN106306361A (en) * | 2016-08-25 | 2017-01-11 | 宁夏泰瑞制药股份有限公司 | Method for preparing biological fermentation feed |
| CN110144317A (en) * | 2019-06-05 | 2019-08-20 | 甘肃省畜牧兽医研究所 | A kind of composite bacteria agent, freeze-drying microbial inoculum and apple pomace protein feed for fermentation apple pomace |
| AU2020103860A4 (en) * | 2020-12-03 | 2021-02-11 | Gansu Animal Husbandry and Veterinary Medicine Institute | Complex microbial agent and freeze-dried microbial agent for fermenting apple pomace and apple pomace protein feed |
Non-Patent Citations (1)
| Title |
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| 李巨秀: "混菌种发酵果渣生产蛋白饲料的研究", 《中国优秀硕士学位论文全文数据库农业科技辑》 * |
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Application publication date: 20210709 |