CN104478515A - Fermentation liquid for culturing pleurotus eryngii matrix, pleurotus eryngii culture medium and preparation method thereof - Google Patents

Abstract

The invention relates to a fermentation liquid for cultivating a pleurotus eryngii substrate, a pleurotus eryngii culture medium and a preparation method thereof, belonging to the technical field of edible fungus cultivation. The culture substrate uses cottonseed hulls, sawdust, corncobs, bran, bagasse, bean stalks, rice bran, calcium carbonate and quicklime as raw materials, mixed with the microbial fermentation liquid of Bacillus subtilis and Bacillus circulans to build a heap for fermentation, and use inoculation Starter composting can accelerate the decomposition and transformation of organic matter such as lignin and cellulose, and directly supply nutrients such as sugars and carbon nitrogen compounds required for the growth of Pleurotus eryngii mycelia. Not only the cultivation medium of Pleurotus eryngii has been fully utilized, but also the fruiting time has been shortened, the biological transformation rate has been improved, and the investment cost has been saved.

Description

Fermentation liquid for cultivating pleurotus eryngii substrate, pleurotus eryngii culture medium and preparation method thereof

technical field

The invention relates to the technical field of edible fungus cultivation, in particular to a fermentation liquid for cultivating a pleurotus eryngii substrate, a pleurotus eryngii culture medium and a preparation method thereof.

Background technique

Pleurotus eryngii (Pleurotu seryngii), also known as Pleurotus eryngii, belongs to the fungus phylum, Eubasidiomycetes Agaricaceae, Pleurotus genus, and is a precious fungus suitable for both medicine and food. Pleurotus eryngii has thick meat, crisp and tender texture, especially the white stipe, thick and long, extremely dense, firm, and strong crispness. It has a unique taste and is one of the best-tasting mushrooms. It is known as "Oyster Mushroom King", "Scallop Mushroom". Pleurotus eryngii is rich in nutrition, contains a variety of amino acids and trace elements, and has anti-oxidation, anti-virus and other active effects. It is a precious edible fungus with high nutritional value and health care value. One of the 16 rare edible fungi recommended by the Food and Drug Administration (FAO).

The cultivation of Pleurotus eryngii generally uses miscellaneous wood chips, bagasse, cottonseed hulls, corn cobs, bran, bean stems, etc. as the culture substrate, and the raw materials are rich in nutrients such as lignin, cellulose, and hemicellulose. At present, the domestic cultivation of Pleurotus eryngii mostly uses raw materials to be sterilized and then inoculated directly, so that a large amount of macromolecular organic matter is not completely decomposed and rationally utilized, which not only causes a waste of resources, but also prolongs the time for fruiting. A small number of enterprises use the cultivation substrate of Pleurotus eryngii to ferment and then inoculate, so that the organic matter of the raw materials can be decomposed and transformed under the action of microorganisms so that Pleurotus eryngii can be better utilized. However, natural stacking fermentation is generally used now, which takes a long time and the product quality is not controlled.

Contents of the invention

The purpose of the invention of the present invention is to provide a kind of fermented liquid for cultivating Pleurotus eryngii substrate, Pleurotus eryngii culture medium and preparation method thereof, utilize inoculation starter compost to be able to accelerate the decomposition conversion of organic matter such as lignin, cellulose, directly supply Nutrients such as sugars, carbon and nitrogen compounds required for the growth of Pleurotus eryngii mycelium. Not only the cultivation medium of Pleurotus eryngii has been fully utilized, but also the fruiting time has been shortened, the biological transformation rate has been improved, and the investment cost has been saved.

The invention provides a fermentation liquid for cultivating Pleurotus eryngii substrate, which is obtained by culturing Bacillus subtilis CMCC 63501 and Bacillus circulans ATCC 4513 in LB liquid at a ratio of 1-1.5:1. Wherein, the pH of the culture solution is 6.8-7.2, the culture temperature is 35°C-37°C, and the culture is 22h-24h.

The present invention also provides a kind of Pleurotus eryngii culture medium, it is made up of culture material and the fermented liquid as claimed in claim 1 or 2 inoculated on the culture material;

Wherein the water content of the compost is 64wt%-66wt%; the proportion of solids to remove water in the compost is: 10-40 parts by weight of cottonseed hulls, 10-30 parts by weight of sawdust, 10-30 parts by weight of corn cobs, 10-30 parts by weight of bran, 5-15 parts by weight of bagasse, 5-15 parts by weight of bean stems, 5-15 parts by weight of rice bran, 1-5 parts by weight of calcium carbonate, and 1-5 parts by weight of lime.

The present invention also provides a preparation method of Pleurotus eryngii culture medium, is characterized in that, comprises the steps:

Crush the compost into soybean grain-sized particles, then mix and mix well;

Mix the fermented liquid and the compost evenly with an inoculum amount of 10%-15% according to the volume/mass ratio, select a sunny and sheltered place to build a heap, and cover it with a plastic film for fermentation; when the compost is 200kg-500kg Build a steamed bun-shaped pile, and build a trapezoidal pile when the compost is 500kg-1000kg. Preferably, the pile has a height of 1.0m-1.5m and a width of 1.5m-2m.

During the fermentation period, the piles are turned every day. As a preference, when the temperature of the material pile is 18cm-22cm from the ground and the temperature reaches 58°C-62°C, the first turnover is carried out. Among them, the steps of turning the pile are: make some holes on the top of the pile, then make holes on the surface at an oblique angle to the ground at an interval of 25cm-40cm to the center of the pile, and then make some holes parallel to the ground at the bottom of the pile at a distance of 8cm-12cm from the ground . Preferably, the angle of the bevel is 45 degrees.

After turning over, cover with plastic film to continue heat preservation and fermentation, and stop fermentation after 3d-5d; after fermentation, the pH of the cultivation substrate is 7.0-7.5, the mass percentage of water is 62%-66%, the color is brown, soft and elastic, The smell is normal, no sour, no smelly, no sticky, no mildew, moderate humidity, balls in the hand, no prickly feeling, and it will fall apart when you let go of your hand.

Pleurotus eryngii can be inoculated for cultivation after the fermented cultivation medium is bagged, sterilized and cooled to 30°C-37°C.

Among them, as a preference, after the cultivation medium is packed into bags, it is sterilized in an autoclave at 0.13MP-0.15MPa for 2.5h-3h.

The present invention has the following advantages:

(1) After adopting this method to inoculate the Bacillus subtilis CMCC 63501 bacterial strain and the Bacillus circulans ATCC 4513 bacterial strain to carry out compost fermentation to the compost, the degradation rate of lignin and cellulose in the compost reaches 90%-98%, compared with The compost of non-inoculated bacterial strains has improved by 20%-30%; (2) the pleurotus eryngii cultivation substrate that adopts this method to make is fast, and mycelia grows vigorously, stoutly, and fruiting time is short, output height, good quality, Economic benefits have been significantly improved.

Detailed ways

The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Apparently, the described embodiments are only some of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without making creative efforts belong to the protection scope of the present invention.

Example 1

Preparation and Application of Cultivation Substrate Ⅰ

(1) Preparation of fermentation broth

Bacillus subtilis CMCC 63501 and Bacillus circulans ATCC 4513 were mixed with peptone 1% (W/V), NaCl 1% (W/V) and yeast powder 0.5% (W/V) in a 1:1 ratio. The LB liquid medium was cultured for 24 hours under the culture conditions of pH 7.0 and culture temperature of 37° C. to obtain a fermentation broth.

(2) Raw material ratio and mixing

The ratio of raw materials is: 20 parts by weight of cottonseed hulls, 10 parts by weight of wood chips, 12 parts by weight of corncobs, 28 parts by weight of bran, 15 parts by weight of bagasse, 13 parts by weight of bean stems, 10 parts by weight of rice bran, and 1 part by weight of calcium carbonate , 1 part by weight of lime.

The above raw materials are crushed into small granules the size of soybean grains, then mixed and evenly mixed, water is added to the material, the water content of the culture material is adjusted to 64% by weight, and the mixture is fully stirred evenly.

(3) Construction of compost

Add 11 parts by weight of fermented liquid to the above-mentioned compost and mix evenly, select a sunny and sheltered place to build a heap, and build a steamed bun-shaped heap. The pile is 1m high and 1.5m wide. After the stockpile is built, it is covered with plastic film for fermentation. When the temperature of the material pile 20cm from the ground reaches 60°C, turn the pile for the first time, gently flatten the surface with a shovel, and pierce 2 holes downward with a wooden stick with a diameter of 8cm on the top of the pile, and then use 25cm on the surface. A wooden stick with a diameter of about 2 cm and the ground is at an angle of 45 degrees to the center of the pile, and then two holes are made parallel to the ground at the bottom of the pile at a distance of 10 cm from the ground, and then covered with a plastic film for heat preservation and fermentation.

(4) Fermentation of compost

Turn over once a day, after 4 days of fermentation, when the compost is brown, soft and elastic, with normal smell, no sour, no stink, no sticky, no mildew, moderate humidity, balls in hand, no thorns Feeling, loose when you let go, pH 7.2, stop fermentation when the mass percentage of water is 62%, and then get Pleurotus eryngii cultivation substrate I.

(5) Application of Cultivation Substrate I in Cultivation of Pleurotus eryngii

The cultivation medium I prepared in (4) was bagged, sterilized in an autoclave at 0.14 MPa for 3 hours, and cooled to 37° C. to cultivate Pleurotus eryngii. The management of inoculation, germination and fruiting is carried out according to the routine management of Pleurotus eryngii.

100 bags of Pleurotus eryngii are cultivated in this cultivation medium Ⅰ, the total dry weight of each bag of compost is 70kg, the average fruiting time is 48d, the fresh weight of the total fruiting bodies is 85kg, and the biotransformation rate is 121%.

Example 2

Preparation and Application of Cultivation Substrate Ⅱ

(1) Preparation of fermentation broth

Put Bacillus subtilis and Bacillus circulans in the ratio of 3:2 in the LB liquid of peptone 1% (W/V)+NaCl 1% (W/V)+yeast powder 0.5% (W/V), pH 7.2, cultivated for 24 hours under the culture condition of 37° C. to obtain a fermentation broth.

(2) Raw material ratio and mixing

The ratio of raw materials is: 40 parts by weight of cottonseed hulls, 20 parts by weight of wood chips, 20 parts by weight of corncobs, 15 parts by weight of bran, 10 parts by weight of bagasse, 8 parts by weight of bean stems, 5 parts by weight of rice bran, and 2 parts by weight of calcium carbonate , 2 parts by weight of lime.

The above-mentioned raw materials are crushed into small particles the size of soybean grains, then mixed and evenly mixed, water is added to the material, the water content of the culture material is adjusted to 66% by weight, and the mixture is fully stirred evenly.

(3) Construction of compost

Add 18 parts by weight of fermented liquid to the above-mentioned compost and mix evenly, select a sunny and sheltered place to build a heap, and build a trapezoidal heap. The height of the stack is 1.2m, and the width is 1.8m. After the stockpile is built, it is covered with plastic film for fermentation. When the temperature at 22cm from the ground reaches 58°C, turn the pile for the first time, gently flatten the surface with a shovel, pierce 3 holes on the top of the pile with a wooden stick with a diameter of 8cm, and then use 33cm on the surface. A wooden stick with a diameter of about 2 cm and the ground is at an angle of 45 degrees to the center of the pile, and then 3 holes are made parallel to the ground at the bottom of the pile at a distance of 12 cm from the ground, and then covered with a plastic film for heat preservation and fermentation.

(4) Fermentation of compost

Turn over once a day, after 5 days of fermentation, when the compost is brown, soft and elastic, with normal smell, no sour, no stink, no sticky, no mildew, moderate humidity, balls in hand, no thorns Feeling, it looses up when you let go of your hand, the pH is 7.0, and the fermentation is stopped when the mass percentage of water is 66%, and the Pleurotus eryngii cultivation substrate II is obtained.

(5) Application of Cultivation Substrate II in Cultivation of Pleurotus eryngii

Pack the cultivation medium II prepared in (4), sterilize in an autoclave at 0.15MPa for 2.5h, cool to 30°C, and cultivate Pleurotus eryngii. The management of inoculation, germination and fruiting is carried out according to the routine management of Pleurotus eryngii.

This cultivation medium II cultivates 100 bags of Pleurotus eryngii, the total dry weight of each bag of compost is 70kg, the average fruiting time is 45d, the total fresh weight of fruit bodies is 89kg, and the biotransformation rate is 127%.

Example 3

Preparation and Application of Cultivation Substrate Ⅲ

(1) Preparation of fermentation broth

Bacillus subtilis and Bacillus circulans are in the LB liquid of peptone 1% (W/V)+NaCl 1% (W/V)+yeast powder 0.5% (W/V) in the ratio of 6:5, The pH is 6.8 and the culture temperature is 36° C. for 23 hours to obtain a fermentation broth.

(2) Raw material ratio and mixing

The ratio of raw materials is: 10 parts by weight of cottonseed hulls, 30 parts by weight of wood chips, 28 parts by weight of corn cobs, 10 parts by weight of bran, 5 parts by weight of bagasse, 5 parts by weight of bean stems, 15 parts by weight of rice bran, and 5 parts by weight of calcium carbonate , 4 parts by weight of lime.

The above-mentioned raw materials are crushed into small particles the size of soybean grains, then mixed and evenly mixed, water is added to the material, the water content of the culture material is adjusted to 65% by weight, and the mixture is fully stirred evenly.

(3) Construction of compost

Add 14 parts by weight of fermented liquid to the above-mentioned compost and mix evenly, select a sunny and sheltered place to build a heap, and build a trapezoidal heap. The pile height is 1.5m and the width is 2m. After the stockpile is built, it is covered with plastic film for fermentation. When the temperature of the material pile at 18cm from the ground reaches 62°C, turn the pile for the first time, gently flatten the surface with a shovel, pierce 4 holes downward with a wooden stick with a diameter of 8cm on the top of the pile, and then use 40cm on the surface to use A wooden stick with a diameter of about 2 cm and the ground is at an angle of 60 degrees to the center of the pile, and then 4 holes are made parallel to the ground at the bottom of the pile at a distance of 8 cm from the ground, and then covered with a plastic film for heat preservation and fermentation.

(4) Fermentation of compost

Turn the heap twice a day, after 3 days of fermentation, when the compost is brown, soft and elastic, with a normal smell, no sour, no smell, no sticky, no mildew, moderate humidity, balls in the hand, no thorns Feeling, loose when you let go, pH 7.5, stop fermentation when the mass percentage of water is 64%, and obtain the Pleurotus eryngii cultivation substrate III.

(5) Application of Cultivation Substrate III in Cultivation of Pleurotus eryngii

The cultivation medium III prepared in (4) was packed into bags, sterilized at 0.13 MPa for 2.8 hours in an autoclave, cooled to 35° C., and cultivated Pleurotus eryngii. The management of inoculation, germination and fruiting is carried out according to the routine management of Pleurotus eryngii.

This cultivation medium III cultivates 100 bags of Pleurotus eryngii, the total dry weight of each bag of compost is 70kg, the average fruiting time is 47d, the total fresh weight of fruit bodies is 90kg, and the biotransformation rate is 129%.

comparative example

Preparation and Application of Cultivation Substrate Ⅳ

(1) Raw material ratio and mixing

The ratio of raw materials is: 20 parts by weight of cottonseed hulls, 10 parts by weight of wood chips, 12 parts by weight of corncobs, 28 parts by weight of bran, 15 parts by weight of bagasse, 13 parts by weight of bean stems, 10 parts by weight of rice bran, and 1 part by weight of calcium carbonate , 1 part by weight of lime.

The above raw materials are crushed into small granules the size of soybean grains, then mixed and evenly mixed, water is added to the material, the water content of the culture material is adjusted to 64% by weight, and the mixture is fully stirred evenly.

(2) Building up compost

The above-mentioned compost is selected to build a heap in a place facing the sun and sheltered from the wind, and builds a steamed bun-shaped heap. The pile is 1m high and 1.5m wide. After the stockpile is built, it is covered with plastic film for fermentation. When the temperature of the material pile at 20cm from the ground reaches 62°C, turn the pile for the first time, gently flatten the surface with a shovel, and pierce 2 holes downward with a wooden stick with a diameter of 8cm on the top of the pile, and then use a wooden stick with a diameter of 8cm on the top of the pile. A wooden stick with a diameter of about 2 cm and the ground is at an angle of 30 degrees to the center of the pile, and then two holes are made parallel to the ground at the bottom of the pile at a distance of 10 cm from the ground, and then covered with a plastic film for heat preservation and fermentation.

(3) Fermentation of compost

Turn the pile once a day, stop fermentation after 4 days, and obtain Pleurotus eryngii cultivation substrate IV.

(4) Application of Cultivation Substrate IV in Cultivation of Pleurotus eryngii

The cultivation medium IV prepared in (3) is packed into bags, sterilized at 0.14 MPa for 3 hours in an autoclave, and cultivated Pleurotus eryngii after cooling. The management of inoculation, germination and fruiting is carried out according to the routine management of Pleurotus eryngii.

100 bags of Pleurotus eryngii are cultivated in this cultivation medium Ⅰ, the total dry weight of each bag of compost is 70kg, the average fruiting time is 60d, the total fruit body fresh weight is 56kg, and the biotransformation rate is 80%.

The above embodiments are intended to illustrate that the present invention can be implemented or used by those skilled in the art. It will be obvious to those skilled in the art to modify the above embodiments, so the present invention includes but is not limited to the above embodiments. Any method, process, or product that conforms to the claims or the description of the specification, and conforms to the principles, novelty, and creative features disclosed herein falls within the protection scope of the present invention.

Claims (10)

1. a fermented liquid for cultivating Pleurotus eryngii substrate is characterized in that it is that bacillus subtilis CMCC 63501 and bacillus circulans ATCC 4513 are cultivated in LB liquid in proportion to 1～1.5:1. 2. The fermentation liquid for cultivating Pleurotus eryngii substrate according to claim 1, characterized in that the pH of the culture liquid is 6.8 to 7.2, the culture temperature is 35°C to 37°C, and the culture time is 22h to 24h. 3. a pleurotus eryngii culture medium is characterized in that it is made up of culture material and the fermented liquid as claimed in claim 1 or 2 that is inoculated on the culture material; Wherein the water content of the compost is 64wt%～66wt%; the proportion of solid material for removing water in the compost is: 10～40 parts by weight of cottonseed hulls, 10～30 parts by weight of sawdust, 10～30 parts by weight of corn cobs, 10-30 parts by weight of bran, 5-15 parts by weight of bagasse, 5-15 parts by weight of bean stems, 5-15 parts by weight of rice bran, 1-5 parts by weight of calcium carbonate, and 1-5 parts by weight of lime. 4. Pleurotus eryngii culture medium according to claim 3, is characterized in that described fermented liquid and compost volume/mass ratio are 10%～15%. 5. a preparation method of Pleurotus eryngii medium, is characterized in that, comprises the steps: (1) compost as claimed in claim 3 is pulverized to the particle size of soybean grain, then mix and mix thoroughly; (2) mixing the fermented liquid as claimed in claim 1 or 2 with the inoculum amount of 10% to 15% by volume/mass ratio, selecting a sunny place to build a heap, and covering it with a plastic film to ferment ; (3) Turn over the pile every day during the fermentation period, cover the pile with a plastic film to continue the heat preservation and fermentation, and stop the fermentation after 3d to 5d; (4) Pleurotus eryngii can be inoculated with Pleurotus eryngii for cultivation after the fermented cultivation medium is bagged, sterilized and cooled to 30°C to 37°C. 6. the preparation method of Pleurotus eryngii cultivation substrate according to claim 5 is characterized in that, when compost is 200kg～500kg, builds steamed bun shape heap, when compost is 500kg～1000kg, builds trapezoidal heap. 7. The preparation method of Pleurotus eryngii cultivation substrate according to claim 5 or 6, characterized in that, the pile height of the pile is 1.0m～1.5m, and the width is 1.5m～2m. 8. according to the microbial pretreatment method of Pleurotus eryngii cultivation substrate described in claim 5 or 6, it is characterized in that, when stockpile is apart from ground 18cm～22cm place temperature reaches 58 ℃～62 ℃, carry out the 1st turning over . 9. the preparation method of Pleurotus eryngii cultivation substrate according to claim 5 is characterized in that, described turning over is to pierce some holes at the top of the pile, and then form an oblique angle to the pile center with the ground every 25cm～40cm on the surface Prick holes, and then pierce several holes parallel to the ground at the bottom of the stockpile at a distance of 8cm to 12cm from the ground. 10. The preparation method of Pleurotus eryngii cultivation substrate according to claim 5, characterized in that, after the cultivation substrate is bagged, it is sterilized at 0.13MPa～0.15MPa for 2.5h～3h in an autoclave.