CN113973646A - Culture medium and culture method for improving yield of pleurotus eryngii - Google Patents
Culture medium and culture method for improving yield of pleurotus eryngii Download PDFInfo
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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
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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
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Abstract
The invention discloses a culture medium for improving yield of pleurotus eryngii and a culture method, wherein the culture medium comprises the following raw materials in parts by mass: 3 parts of soybean meal, 0.1 part of yeast extract, 3 parts of glucose, 0.05 part of magnesium sulfate, 0.01 part of ferric sulfate, 0.05 part of monopotassium phosphate, 10.001 parts of vitamin B and 100 parts of water; subculturing and activating pleurotus eryngii hyphae in a solid flat plate, digging hypha blocks with consistent growth conditions in the solid flat plate by using an inoculation shovel, and inoculating the hypha blocks into a culture medium; shake culturing for 7 days by setting pH, temperature, liquid loading amount and rotating speed; filtering all fermentation liquor with a 40-mesh screen, filtering to remove supernatant, vacuum filtering with weighed qualitative filter paper, filtering to remove supernatant, washing mycelium with water for 3 times, filtering, collecting, drying in a forced air drying oven, weighing to constant weight, and repeating for 3 times; by adopting the fermentation condition of the invention, the strain can grow rapidly under the process, and the quality of the pleurotus eryngii can be improved.
Description
Technical Field
The invention relates to the technical field of pleurotus eryngii culture, in particular to a culture medium and a culture method for improving yield of pleurotus eryngii.
Background
Pleurotus eryngii has high nutritive value, contains protein up to 25%, various amino acids and polysaccharides, and has various biological activities, such as cancer prevention and cancer resistance and immunoregulation activity. Meanwhile, it contains a large amount of natural products with small molecules, such as sterols, and has good cytotoxicity, osteoporosis resistance, antioxidant and anti-inflammatory effects.
The pleurotus eryngii has fleshy flesh, crisp and tender texture, particularly has compact, solid and milky stem tissues, can be completely eaten, is more crisp, smooth and tasty than pileus, is called as pleurotus ostreatus king and dried oyster mushroom, has pleasant almond fragrance and taste like abalone, and is suitable for fresh keeping and processing. The edible fungus is a rare edible fungus with high popularization value because the edible fungus has the characteristics of delicious taste, rich nutrient components, low fat and low calorie, and is favored by domestic and foreign consumers.
In order to meet the requirements of more people, the conventional pleurotus eryngii production and planting technology is simple, the strain quality is poor, and the quality of pleurotus eryngii needs to be further improved by adopting a biological optimization technology.
Disclosure of Invention
The technical problem to be solved is as follows: aiming at the defects of the prior art and solving the bottleneck problem of limiting the research on the quality improvement of the pleurotus eryngii, the invention discloses a culture medium and a culture method for improving the yield of the pleurotus eryngii.
The invention provides a culture medium for improving the yield of pleurotus eryngii, which comprises the following raw materials in parts by mass: 3 parts of soybean meal, 0.1 part of yeast extract, 3 parts of glucose, 0.05 part of magnesium sulfate, 0.01 part of ferric sulfate, 0.05 part of monopotassium phosphate, 10.001 parts of vitamin B and 100 parts of water.
Preferably, the culture medium fermentation liquid for improving the yield of the pleurotus eryngii is subpackaged in 250mL conical flasks.
Preferably, the fermentation liquor in the conical flask is sterilized in a sterilizing pot at 121 ℃ for 30min, and then is cooled for use.
A culture method of a culture medium for improving yield of pleurotus eryngii comprises the following steps:
the first step is as follows: weighing 3 parts of soybean meal, 0.1 part of yeast extract, 3 parts of glucose, 0.05 part of magnesium sulfate, 0.01 part of ferric sulfate, 0.05 part of monopotassium phosphate, 10.001 parts of vitamin B and 100 parts of water according to the mass part ratio, subpackaging 80mL of fermentation liquor into 250mL of conical bottles, sterilizing in a sterilizing pot at 121 ℃ for 30min, and cooling for later use;
secondly, the operating process of shake flask fermentation is as follows: subculturing and activating Pleurotus eryngii mycelium in solid plate, and digging 0.5 × 1.5cm in solid plate with inoculating shovel2Inoculating the mycelium blocks with consistent growth conditions into the cooled standby culture medium;
the third step: setting the pH value of 5-9, the temperature of 21-29 ℃, the liquid loading amount of 80-135mL and the rotating speed of 140-160r/min, wherein other conditions are consistent, repeating 3 times for each batch, and performing shake culture for 7 days on a shaking table;
fourthly, measuring the biomass of the thalli: filtering all fermentation liquor with 40 mesh screen, filtering to remove supernatant, vacuum filtering with weighed qualitative filter paper, filtering to remove supernatant, washing mycelium with water for 3 times, filtering, collecting, drying in 80 deg.C air-blast drying oven for 2 hr, weighing to constant weight, and repeating for 3 times;
the fifth step: finally, the detection finds that the pleurotus eryngii biomass reaches the maximum value when the pH value is 5, the liquid loading amount is 110mL, the rotating speed is 150r/min and the temperature is 25 ℃.
Further, in the third step, the pH values are set to be 5, 6, 7, 8 and 9, other conditions are consistent, each batch is repeated for 3 times, and shaking culture is carried out for 7 days.
Further, in the third step, the pH is set to 6, the temperature is set to 21 ℃, 23 ℃, 25 ℃, 27 ℃ and 29 ℃, other conditions are consistent, 3 times of each batch are repeated, and shaking culture is carried out for 7 days.
Further, in the third step, the pH value is set to be 6, the temperature is set to be 27 ℃, the liquid loading amount is set to be 80mL, 90mL, 105mL, 120mL and 135mL, other conditions are consistent, 3 times of each batch are repeated, and shaking culture is carried out for 7 days.
Further, in the third step, the pH value is set to be 6, the temperature is set to be 27 ℃, the liquid loading amount is 90mL, the rotating speed is set to be 140r/min, 145r/min, 150r/min, 155r/min and 160r/min, other conditions are consistent, each batch is repeated for 3 times, and shaking culture is carried out on a shaking table for 7 days.
Has the advantages that:
compared with the prior art, the culture medium and the culture method for improving the yield of the pleurotus eryngii provided by the invention have the following advantages:
1. can be used in the field of cultivation and planting of pleurotus eryngii, in particular to a laboratory scale fermentation condition optimization method for fermentation cultivation of pleurotus eryngii;
2. the culture medium of the culture mode has simple components, controllable culture conditions and no need of special equipment, and the yield of the pleurotus eryngii fermented under the optimized culture conditions can be obviously improved;
3. the raw materials are easy to obtain, the price is low, and the popularization and the application are facilitated;
4. through optimization of fermentation conditions in a laboratory, 4 conditions of rotating speed, pH, liquid loading amount and temperature are optimized, and finally, through optimization of experimental conditions in an orthogonal test, the quality of pleurotus eryngii can be remarkably improved;
5. the preparation of the culture medium only needs to dissolve the raw materials by tap water, and the method is simple and low in difficulty and does not need special technical training and special instruments.
6. By adopting the fermentation condition of the invention, the strain can grow rapidly under the process, the quality of the pleurotus eryngii can be improved, and the final fermentation condition is determined to be that the pH is 5, the liquid loading amount is 110mL, the rotating speed is 150r/min, the temperature is 25 ℃, and the dry weight of the pleurotus eryngii reaches 1.1848 g.
Drawings
FIG. 1 is a graph showing the variation of the dry weight of liquid culture of Pleurotus eryngii according to pH in example 1.
FIG. 2 is a graph showing the trend of dry weight of liquid strain of Pleurotus eryngii according to example 2 with temperature.
FIG. 3 is a graph showing the variation of the dry weight of liquid culture of Pleurotus eryngii according to the liquid content in example 3.
FIG. 4 is a graph showing the variation of the dry weight of liquid culture of Pleurotus eryngii with the rotation speed in example 4.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
Before describing the examples, it is necessary to provide some remarks:
in small-scale experiments, in order to ensure the repeatability among parallel experiments, the corn steep liquor and the starch in the culture medium are firstly heated and dissolved so as to prevent the interference of granular nutrient substances.
The features and properties of the present invention will be described in further detail below with reference to the accompanying drawings and examples.
Example 1
The embodiment provides a culture medium and a culture method for improving yield of pleurotus eryngii, and the culture medium comprises the following specific steps:
step one, preparing a fermentation medium: accurately weighing 30g/L of soybean meal, 1g/L of yeast extract, 30g/L of glucose, 0.5g/L of magnesium sulfate, 0.1g/L of ferric sulfate, 0.5g/L of monopotassium phosphate and 10.01g/L of vitamin B, subpackaging 80mL of fermentation liquor into 250mL of conical bottles, sterilizing in a sterilization pot at 121 ℃ for 30min, and cooling for use;
in the second step, the first step is that,the operating process of the shake flask fermentation is as follows: subculturing and activating Pleurotus eryngii mycelium in solid plate, and digging 0.5 × 1.5cm in solid plate with inoculating shovel2The mycelium blocks with consistent growth conditions are inoculated into the culture medium;
the third step: setting pH values to 5, 6, 7, 8 and 9, keeping the other conditions consistent, repeating 3 times for each batch, and shake-culturing for 7 days on a shaking table;
fourthly, measuring the biomass of the thalli: the whole fermentation broth was filtered through a 40 mesh screen and the supernatant was filtered off. Vacuum filtering with weighed qualitative filter paper, filtering to remove supernatant, washing mycelium with water for 3 times, filtering, and collecting. Then putting the mixture into a forced air drying oven at the temperature of 80 ℃ for drying for 2h, weighing the mixture to constant weight, and repeating the weighing for 3 times;
the fifth step: finally, the pleurotus eryngii biomass reaches the maximum value of 9.43g/L at the pH value of 7.
Example 2
The embodiment provides a culture medium and a culture method for improving yield of pleurotus eryngii, and the culture medium comprises the following specific steps:
step one, preparing a fermentation medium: the same as example 1;
secondly, the operating process of shake flask fermentation is as follows: the same as example 1;
the third step: setting pH to 6, setting temperature to 21 deg.C, 23 deg.C, 25 deg.C, 27 deg.C, 29 deg.C, repeating for 3 batches, and shake culturing for 7 days;
fourthly, measuring the biomass of the thalli: the same as example 1;
the fifth step: finally, the detection shows that the biomass of the pleurotus eryngii reaches the maximum value of 11.86g/L at the temperature of 27 ℃.
Example 3
The embodiment provides a culture medium and a culture method for improving yield of pleurotus eryngii, and the culture medium comprises the following specific steps:
step one, preparing a fermentation medium: the same as example 1;
secondly, the operating process of shake flask fermentation is as follows: the same as example 1;
the third step: setting the pH value to be 6, setting the temperature to be 27 ℃, setting the liquid loading amounts to be 80mL, 90mL, 105mL, 120mL and 135mL, keeping the other conditions consistent, repeating 3 times for each batch, and performing shake culture for 7 days by using a shaking table;
fourthly, measuring the biomass of the thalli: the same as example 1;
the fifth step: finally, the detection shows that the pleurotus eryngii biomass reaches the maximum value of 12.26g/L when the liquid loading amount is 90 mL.
Example 4
The embodiment provides a culture medium and a culture method for improving yield of pleurotus eryngii, and the culture medium comprises the following specific steps:
step one, preparing a fermentation medium: the same as example 1;
secondly, the operating process of shake flask fermentation is as follows: the same as example 1;
the third step: setting the pH value to be 6, the temperature to be 27 ℃, the liquid loading amount to be 90mL, the rotating speed to be 140r/min, 145r/min, 150r/min, 155r/min and 160r/min, the other conditions to be consistent, repeating 3 times in each batch, and shaking and culturing for 7 days by a shaking table;
fourthly, measuring the biomass of the thalli: the same as example 1;
the fifth step: finally, the detection shows that the biomass of the pleurotus eryngii reaches the maximum value of 12.80g/L when the rotating speed is 150 r/min.
Example 5
The embodiment provides a culture medium and a culture method for improving yield of pleurotus eryngii, and the culture medium comprises the following specific steps:
step one, preparing a fermentation medium: the same as example 1;
secondly, the operating process of shake flask fermentation is as follows: the same as example 1;
the third step: setting three levels of pH, temperature, liquid loading amount and rotating speed as shown in table 1, wherein other conditions are consistent, each batch is repeated for 3 times, and shaking culture is carried out for 7 days by a shaking table;
fourthly, measuring the biomass of the thalli: the same as example 1;
the fifth step: finally, the optimal combination is A1B2C2D2, and the dry weight reaches 23.696 g/L.
TABLE 1 orthogonal test factor level design
From example 1 it can be found that: as shown in FIG. 1, the dry weight of the liquid strain of Pleurotus eryngii gradually increased with the increase of pH, and reached the maximum dry weight of the mycelium at pH 6. As can be seen from the change trend, the Pleurotus eryngii cens is suitable for growing in a weakly acidic environment, the dry weight of the pleurotus eryngii cens is gradually reduced along with the increase of the pH, the suitable pH is 6 when the dry weight of the pleurotus eryngii cens is the maximum, the pH is continuously increased, the growth pH environment and the metabolic isoelectric point are changed, and the growth of the pleurotus eryngii cens is unfavorable. Therefore, the best pH of the shake flask for Pleurotus eryngii is 6.
From example 2 it can be found that: the response of the dry weight of the pleurotus eryngii liquid strain to the change of the temperature is different from the pH, the dry weight of the hyphae is gradually increased and then decreased along with the increase of the temperature, and the dry weight of the hyphae reaches the maximum at the temperature of 27 ℃ as shown in figure 2. As can be seen from the trend, the growth of the Pleurotus eryngii pellet is too slow at room temperature of 21 ℃ and the dry weight is only 1.40g/L, while when the temperature is increased to 29 ℃, the metabolic enzyme system in the Pleurotus eryngii mycelium is affected by high temperature due to too high temperature, resulting in the dry weight being reduced to 4.00 g/L. When the temperature is 27 ℃, the dry weight of the pleurotus eryngii can reach the maximum value (the dry weight is 11.86g/L), and the dry weight is close to the suitable growth temperature (28 ℃) of the fungi. It can be seen that the temperature has a greater effect on fungal growth.
From example 3 it can be found that: the liquid loading amount influences the contact area of the fermentation liquid and air in the shaking culture process, and the larger the liquid loading amount is, the smaller the air contact area is, namely, the less oxygen is obtained by the growth of the thallus. The dry weight of the liquid strain of Pleurotus eryngii showed a tendency of increasing first and then gradually decreasing with the increase of the liquid content, and as shown in FIG. 3, the dry weight of the mycelia reached the maximum when the liquid content was 90 mL. As can be seen from the trend, when the liquid loading is 80mL, the contact area between the pleurotus eryngii hyphae and the air is the largest, but the oxygen concentration is too high to inhibit the hyphae growth, and the dry weight is only 7.90 g/L. When the liquid content increased and the liquid content was 90mL, the dry weight was the largest (12.26g/L), and when the liquid content continued to increase to 105mL, the area of hyphae contacting the air decreased and the oxygen concentration decreased to inhibit hyphae growth.
From example 4 it can be found that: as shown in FIG. 4, the rotation speed of the Pleurotus eryngii with the liquid strain dry weight increasing trend is at 140-150r/min, and the dry weight of the mycelium reaches the maximum at 150 r/min. The increase of the rotating speed increases the dissolved oxygen in the culture bottle, the oxygen has strong promotion effect on the growth of mycelium of the mycelium pellet, the rotating speed reaches 150r/min when the dry weight of the mycelium pellet is maximum, the rotating speed continues to increase, the shearing force has larger and larger influence on the mycelium pellet, and the growth of the mycelium pellet is unfavorable. Therefore, the optimal rotating speed of the shake flask of the pleurotus eryngii is 150 r/min.
From example 5 it can be found that: as shown in Table 2, the optimum combination is A1B2C2D2, the dry weight reaches 23.696g/L, namely pH is 5, the liquid loading is 110mL, the rotation speed is 150r/min, the temperature is 25 ℃, and the optimum conditions are determined. The influence of all factors on the dry weight of hyphae is as follows in sequence: temperature is higher than liquid loading quantity and rotating speed is higher than pH value.
TABLE 2 analysis of results of orthogonal experiments
It should be noted that the above-mentioned preferred embodiments are only for illustrating the present invention, and not for limiting the present invention, and those skilled in the art can make various changes or modifications without departing from the spirit and scope of the present invention.
Claims (8)
1. The culture medium for improving the yield of the pleurotus eryngii is characterized by comprising the following raw materials in parts by mass: 3 parts of soybean meal, 0.1 part of yeast extract, 3 parts of glucose, 0.05 part of magnesium sulfate, 0.01 part of ferric sulfate, 0.05 part of monopotassium phosphate, 10.001 parts of vitamin B and 100 parts of water.
2. The culture medium and the culture method for improving the yield of pleurotus eryngii according to claim 1, wherein the culture medium comprises: and the culture medium fermentation liquid for improving the yield of the pleurotus eryngii is subpackaged in 250mL conical flasks.
3. The culture medium and the culture method for improving the yield of pleurotus eryngii according to claim 2, wherein the culture medium comprises: and sterilizing the fermentation liquor in the conical flask at 121 ℃ for 30min in a sterilizing pot, and cooling for use.
4. The method for culturing the culture medium for increasing the yield of pleurotus eryngii according to claim 1, wherein the culture medium comprises the following components: the method specifically comprises the following steps:
the first step is as follows: weighing 3 parts of soybean meal, 0.1 part of yeast extract, 3 parts of glucose, 0.05 part of magnesium sulfate, 0.01 part of ferric sulfate, 0.05 part of monopotassium phosphate, 10.001 parts of vitamin B and 100 parts of water according to the mass part ratio, subpackaging 80mL of fermentation liquor into 250mL of conical bottles, sterilizing in a sterilizing pot at 121 ℃ for 30min, and cooling for later use;
secondly, the operating process of shake flask fermentation is as follows: subculturing and activating Pleurotus eryngii mycelium in solid plate, and digging 0.5 × 1.5cm in solid plate with inoculating shovel2Inoculating the mycelium blocks with consistent growth conditions into the cooled standby culture medium;
the third step: setting the pH value of 5-9, the temperature of 21-29 ℃, the liquid loading amount of 80-135mL and the rotating speed of 140-160r/min, wherein other conditions are consistent, repeating 3 times for each batch, and performing shake culture for 7 days on a shaking table;
fourthly, measuring the biomass of the thalli: filtering all fermentation liquor with 40 mesh screen, filtering to remove supernatant, vacuum filtering with weighed qualitative filter paper, filtering to remove supernatant, washing mycelium with water for 3 times, filtering, collecting, drying in 80 deg.C air-blast drying oven for 2 hr, weighing to constant weight, and repeating for 3 times;
the fifth step: finally, the detection finds that the pleurotus eryngii biomass reaches the maximum value when the pH value is 5, the liquid loading amount is 110mL, the rotating speed is 150r/min and the temperature is 25 ℃.
5. The culture medium and the culture method for improving the yield of pleurotus eryngii according to claim 4, wherein the culture medium comprises: in the third step, the pH values are set to be 5, 6, 7, 8 and 9, other conditions are consistent, each batch is repeated for 3 times, and shaking culture is carried out for 7 days.
6. The culture medium and the culture method for improving the yield of pleurotus eryngii according to claim 4, wherein the culture medium comprises: setting the pH value to be 6, setting the temperature to be 21 ℃, 23 ℃, 25 ℃, 27 ℃ and 29 ℃ in the third step, keeping the other conditions consistent, repeating the steps for 3 times in each batch, and carrying out shake culture on a shaking table for 7 days.
7. The culture medium and the culture method for improving the yield of pleurotus eryngii according to claim 4, wherein the culture medium comprises: setting the pH value to be 6 and the temperature to be 27 ℃ in the third step, setting the liquid loading amounts to be 80mL, 90mL, 105mL, 120mL and 135mL, keeping the other conditions consistent, repeating 3 times in each batch, and shaking and culturing for 7 days by a shaking table.
8. The culture medium and the culture method for improving the yield of pleurotus eryngii according to claim 4, wherein the culture medium comprises: in the third step, the pH value is set to be 6, the temperature is 27 ℃, the liquid loading amount is 90mL, the rotating speed is set to be 140r/min, 145r/min, 150r/min, 155r/min and 160r/min, other conditions are consistent, 3 times of each batch are repeated, and shaking culture is carried out on a shaking table for 7 days.
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