CN106148201A - A kind of preparation method of Agaricus Bisporus liquid spawn - Google Patents
A kind of preparation method of Agaricus Bisporus liquid spawn Download PDFInfo
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- CN106148201A CN106148201A CN201610698556.1A CN201610698556A CN106148201A CN 106148201 A CN106148201 A CN 106148201A CN 201610698556 A CN201610698556 A CN 201610698556A CN 106148201 A CN106148201 A CN 106148201A
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- green tea
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
Abstract
The invention belongs to the preparing technical field of edible fungus species, relate to the preparation method of a kind of Agaricus Bisporus liquid spawn, this preparation method is to be cultivated by slant medium, one-level secondary medium and fermentation medium expanding propagation to form, the formula of the various culture medium that the present invention provides can provide nutritional labeling required in Agaricus Bisporus strain growth course, shorten the growth cycle of Agaricus Bisporus liquid spawn, strengthen spawn activity；The present invention is by adding a small amount of green tea extractive liquor, it is possible to the generation of mycete in effectively preventing Agaricus Bisporus growth course, miscellaneous bacteria etc., and promotes the growth of Agaricus Bisporus strain, and mycelial growth rate is fast, quality better, and purity is high.
Description
Technical field
The invention belongs to the preparing technical field of edible fungus species, relate to the preparation method of a kind of Agaricus Bisporus liquid spawn.
Background technology
Agaricus Bisporus, also known as white mushroom or mushroom, is that cultivated area is maximum, yield is most in the world, consumes most common one
Edible fungi.Agaricus Bisporus color and luster is pure white, quality is tender, nutritious, delicious flavour, containing rich in protein, several amino acids
With vitamin and mineral, it is not only one grade of high-grade mushroom vegetable, and there is heat-clearing and toxic substances removing, antiinflammatory lung moistening, brain-strengthening, reduction
The plurality of health care functions such as cholesterol, are preferable health foods.
Along with the rise of the prices such as wood flour, the price cultivating solid spawn goes up the most therewith, and solid spawn incubation time
Long, send out bacterium speed slow.Owing to mycelial growth is vigorous, cell age is short, fruiting gas, change of tide is fast, and quality and yield are apparently higher than traditional
The liquid spawn of the mode of production has slowly won market and benefit.At present, there is cultivation during Agaricus Bisporus strain cultivation
The each component ratio of base is unreasonable, easily grow the problems such as miscellaneous bacteria, therefore, research and develop a kind of can high yield and high quality produce Agaricus Bisporus
Liquid spawn has big market value.
Summary of the invention
The problem existed for prior art, the invention provides the preparation method of a kind of Agaricus Bisporus liquid spawn.
The technical scheme that the present invention is used to achieve these goals is:
The invention provides the preparation method of a kind of Agaricus Bisporus liquid spawn, comprise the following steps:
(1) Agaricus Bisporus mother kind is inoculated in primary inclined plane culture medium, cultivates 5-6d at 24-26 DEG C, obtain Agaricus Bisporus slant strains,
It is divided into slant strains block；
(2) being inoculated in first cell culture medium by 5-6 block one-level Pleurotus eryngii slant strains block, at temperature 20-22 DEG C, shaking table is cultivated
1-2d, then quiescent culture 1-2d at 20-22 DEG C, obtains one-level culture fluid；
(3) one-level culture fluid is inoculated in secondary medium according to the inoculum concentration of volumn concentration 10%, at 20-22 DEG C
2-3d cultivated by shaking table, obtains two grades of culture fluid；
(4) two grades of culture fluid are inoculated in fermentation medium cultivation according to volumn concentration 15%, at 25 DEG C, cultivate 4-
5d, ventilation 2.5m3/ h, i.e. obtains Agaricus Bisporus liquid spawn after fermentation ends；
Further, described slant medium is addition 8-10mL green tea extractive liquor, 20-28g Semen Maydis in every 1000mL pure water
Starch, 1-2g agar, 0.2-0.5g casein hydrolysate peptides, 0.8-1.2g peptone, 0.2-0.5g potassium dihydrogen phosphate, stirring is all
Even, cool down after sterilization processing and get final product.
Further, described first cell culture medium every 1000mL pure water adds 15-20mL green tea extractive liquor, peptone 7-
10g, soluble starch 50-55g, glycerol 5-8g, magnesium sulfate 0.2-0.8g, pulullan polysaccharide 0.2-0.3g, sucrose 8-12g, salt
Allithiamine element 0.06-0.08g.
Further, described secondary medium every 1000mL pure water adds 20-25mL green tea extractive liquor, peptone
30-40g, wheat bran 45-60g, sucrose 18-20g, potassium dihydrogen phosphate 1.5-2.0g, carbomer 0.06-0.08g.
Further, described fermentation medium every 1000mL pure water adds yeast extract 12-15g, 15-20g sucrose, 8-
10g treaster, 10-15mL green tea extractive liquor, 40-50g peptone, zinc citrate 0.02-0.05g, potassium dihydrogen phosphate 1g, sulphuric acid
Magnesium 0.8-1.2g.
Green tea extractive liquor used in the present invention uses following methods to be prepared from: the green tea of fresh harvesting is added food
Grind after salt, then encase the green tea after grinding with gauze, be subsequently adding the water accounting for green tea weight 8 times, add medical stone powder,
Polyethylene Glycol, is heated to 90-100 DEG C, and insulated and stirred 2h filters to get filtrate, and concentrates the filtrate to the half of original volume.
Further, the addition of described Sal accounts for the 1% of green tea weight；The addition of described medical stone powder accounts for addition
The 5% of the weight of water.
Further, the addition of Polyethylene Glycol accounts for the 1.5% of green tea weight.
The length and width of slant strains block of the present invention are 2cm.
Treaster used in the present invention is the skin slag garbage after the processing such as grape juice, mainly Pericarpium Vitis viniferae, seed
Composition.
The green tea extractive liquor contained in culture medium of the present invention, in preparation process, is firstly added Sal and is ground, then
Extract, it is possible to accelerate the dissolution of active substance in green tea, containing substantial amounts of trace element in medical stone powder, and antibacterial is had
There is strong adsorption, by adding green tea extractive liquor in the medium, it is possible to prevent the generation of miscellaneous bacteria, unleavened green tea
Containing nutritional labelings such as tea polyphenols, catechin, caffeine, aminoacid, accelerate the fruiting speed of Agaricus Bisporus.
Advantages of the present invention is:
(1) formula of the various culture medium that the present invention provides can provide nutrition required in Agaricus Bisporus strain growth course to become
Point, shorten the growth cycle of Agaricus Bisporus liquid spawn, strengthen spawn activity；
(2) present invention is by adding a small amount of green tea extractive liquor, it is possible to mycete in effectively preventing Agaricus Bisporus growth course, miscellaneous
The generation of bacterium etc., and promote the growth of Agaricus Bisporus strain, mycelial growth rate is fast, quality better, and purity is high；
(3) the method low-carbon (LC) of present invention offer, environmental protection, yield is high.
Detailed description of the invention
Below by embodiment, the present invention will be further elaborated, it should be appreciated that, the description below merely to
Explain the present invention, its content is not defined.
Green tea extractive liquor used herein uses following methods to be prepared from: account for green by the green tea addition of fresh harvesting
Grind (being ground to without obvious lamellar) after the Sal of Folium Camelliae sinensis weight 1%, then encase the green tea after grinding, then with gauze
Add the water accounting for green tea weight 8 times, add the medical stone powder of the weight 5% accounting for water, account for the poly-second two of green tea weight 1.5%
Alcohol, is heated to 90-100 DEG C, and insulated and stirred 2h filters to get filtrate, and concentrates the filtrate to the half of original volume.
Embodiment 1
The preparation method of a kind of Agaricus Bisporus liquid spawn, comprises the following steps:
(1) Agaricus Bisporus mother kind is inoculated in primary inclined plane culture medium, cultivates 5d at 24-26 DEG C, obtain Agaricus Bisporus slant strains, draw
It is divided into length and width and is the slant strains block of 2cm；
Described slant medium is addition 8mL green tea extractive liquor in every 1000mL pure water, 25g corn starch, 2g agar,
0.5g casein hydrolysate peptides, 0.8g peptone, 0.35g potassium dihydrogen phosphate, stir, cool down after sterilization processing and get final product；
(2) being inoculated in first cell culture medium by 5-6 block one-level Pleurotus eryngii slant strains block, at temperature 20-22 DEG C, shaking table is cultivated
2d, then quiescent culture 1d at 20-22 DEG C, obtains one-level culture fluid；
Addition 20mL green tea extractive liquor in described first cell culture medium every 1000mL pure water, peptone 10g, soluble starch 50g,
Glycerol 7g, magnesium sulfate 0.6g, pulullan polysaccharide 0.2g, sucrose 12g, thiamine hydrochloride 0.06g；
(3) one-level culture fluid is inoculated in secondary medium according to the inoculum concentration of volumn concentration 10%, at 20-22 DEG C
2d cultivated by shaking table, obtains two grades of culture fluid；
Described secondary medium every 1000mL pure water adds 23mL green tea extractive liquor, peptone 30g, wheat bran 56g, sucrose
18g, potassium dihydrogen phosphate 1.5g, carbomer 0.08g；
(4) two grades of culture fluid are inoculated in fermentation medium cultivation according to volumn concentration 15%, at 25 DEG C, cultivate 5d,
Ventilation 2.5m3/ h, i.e. obtains Agaricus Bisporus liquid spawn after fermentation ends；
Adding yeast extract 12g in described fermentation medium every 1000mL pure water, 15g sucrose, 10g treaster, 12mL green tea carries
Take liquid, 50g peptone, zinc citrate 0.03g, potassium dihydrogen phosphate 1g, magnesium sulfate 1.2g.
Embodiment 2
The preparation method of a kind of Agaricus Bisporus liquid spawn, comprises the following steps:
(1) Agaricus Bisporus mother kind is inoculated in primary inclined plane culture medium, cultivates 6d at 24-26 DEG C, obtain Agaricus Bisporus slant strains, draw
It is divided into length and width and is the slant strains block of 2cm；
Described slant medium is addition 9mL green tea extractive liquor, 20g corn starch, 1g agar, 0.3g in every 1000mL pure water
Casein hydrolysate peptides, 1.0g peptone, 0.5g potassium dihydrogen phosphate, stir, cool down after sterilization processing and get final product；
(2) being inoculated in first cell culture medium by 5-6 block one-level Pleurotus eryngii slant strains block, at temperature 20-22 DEG C, shaking table is cultivated
1d, then quiescent culture 2d at 20-22 DEG C, obtains one-level culture fluid；
Addition 15mL green tea extractive liquor in described first cell culture medium every 1000mL pure water, peptone 9g, soluble starch 53g,
Glycerol 5g, magnesium sulfate 0.7g, pulullan polysaccharide 0.3g, sucrose 8g, thiamine hydrochloride 0.07g；
(3) one-level culture fluid is inoculated in secondary medium according to the inoculum concentration of volumn concentration 10%, at 20-22 DEG C
2d cultivated by shaking table, obtains two grades of culture fluid；
Described secondary medium every 1000mL pure water adds 20mL green tea extractive liquor, peptone 35g, wheat bran 60g, sucrose
19g, potassium dihydrogen phosphate 1.8g, carbomer 0.07g；
(4) two grades of culture fluid are inoculated in fermentation medium cultivation according to volumn concentration 15%, at 25 DEG C, cultivate 5d,
Ventilation 2.5m3/ h, i.e. obtains Agaricus Bisporus liquid spawn after fermentation ends；
Adding yeast extract 13g, 18g sucrose, 9g treaster in described fermentation medium every 1000mL pure water, 15mL green tea extracts
Liquid, 46g peptone, zinc citrate 0.05g, potassium dihydrogen phosphate 1g, magnesium sulfate 0.8g.
Embodiment 3
The preparation method of a kind of Agaricus Bisporus liquid spawn, comprises the following steps:
(1) Agaricus Bisporus mother kind is inoculated in primary inclined plane culture medium, cultivates 5d at 24-26 DEG C, obtain Agaricus Bisporus slant strains, draw
It is divided into length and width and is the slant strains block of 2cm；
Described slant medium is addition 10mL green tea extractive liquor in every 1000mL pure water, 28g corn starch, 2g agar,
0.2g casein hydrolysate peptides, 1.2g peptone, 0.2g potassium dihydrogen phosphate, stir, cool down after sterilization processing and get final product；
(2) being inoculated in first cell culture medium by 5-6 block one-level Pleurotus eryngii slant strains block, at temperature 20-22 DEG C, shaking table is cultivated
2d, then quiescent culture 1d at 20-22 DEG C, obtains one-level culture fluid；
Addition 18mL green tea extractive liquor in described first cell culture medium every 1000mL pure water, peptone 7g, soluble starch 55g,
Glycerol 8g, magnesium sulfate 0.8g, pulullan polysaccharide 0.3g, sucrose 10g, thiamine hydrochloride 0.08g；
(3) one-level culture fluid is inoculated in secondary medium according to the inoculum concentration of volumn concentration 10%, at 20-22 DEG C
3d cultivated by shaking table, obtains two grades of culture fluid；
Described secondary medium every 1000mL pure water adds 25mL green tea extractive liquor, peptone 40g, wheat bran 45g, sucrose
20g, potassium dihydrogen phosphate 2.0g, carbomer 0.06g；
(4) two grades of culture fluid are inoculated in fermentation medium cultivation according to volumn concentration 15%, at 25 DEG C, cultivate 4d,
Ventilation 2.5m3/ h, i.e. obtains Agaricus Bisporus liquid spawn after fermentation ends；
Adding yeast extract 15g, 20g sucrose, 8g treaster in described fermentation medium every 1000mL pure water, 10mL green tea extracts
Liquid, 40g peptone, zinc citrate 0.02g, potassium dihydrogen phosphate 1g, magnesium sulfate 1.0g.
Comparative example 1
The preparation method of a kind of Agaricus Bisporus liquid spawn, comprises the following steps:
(1) Agaricus Bisporus mother kind is inoculated in primary inclined plane culture medium, cultivates 4-5d at 24-26 DEG C, obtain Agaricus Bisporus slant strains,
It is divided into length and width and is the slant strains block of 2cm；
Described slant medium is addition 20g corn starch in every 1000mL pure water, 1g agar, 0.3g casein hydrolysate peptides,
1.0g peptone, 0.5g potassium dihydrogen phosphate, stir, cool down after sterilization processing and get final product；
(2) being inoculated in first cell culture medium by 5-6 block one-level Pleurotus eryngii slant strains block, at temperature 20-22 DEG C, shaking table is cultivated
3d, obtains one-level culture fluid；
Described first cell culture medium every 1000mL pure water adds peptone 9g, soluble starch 53g, glycerol 5g, magnesium sulfate
0.7g, pulullan polysaccharide 0.3g, sucrose 8g, thiamine hydrochloride 0.07g；
(3) one-level culture fluid is inoculated in secondary medium according to the inoculum concentration of volumn concentration 10%, at 20-22 DEG C
3d cultivated by shaking table, obtains two grades of culture fluid；
Described secondary medium every 1000mL pure water adds peptone 35g, wheat bran 60g, sucrose 19g, potassium dihydrogen phosphate
1.8g, carbomer 0.07g；
(4) two grades of culture fluid are inoculated in fermentation medium cultivation according to volumn concentration 15%, at 25 DEG C, cultivate 7d,
Ventilation 2.5m3/ h, i.e. obtains Agaricus Bisporus liquid spawn after fermentation ends；
Addition yeast extract 13g in described fermentation medium every 1000mL pure water, 18g sucrose, 9g treaster, 46g peptone,
Zinc citrate 0.05g, potassium dihydrogen phosphate 1g, magnesium sulfate 0.8g.
Comparative example 2
The preparation method of a kind of Agaricus Bisporus liquid spawn, comprises the following steps:
(1) Agaricus Bisporus mother kind is inoculated in primary inclined plane culture medium, cultivates 6d at 24-26 DEG C, obtain Agaricus Bisporus slant strains, draw
It is divided into length and width and is the slant strains block of 2cm；
Described slant medium is addition 20g corn starch, 1g agar, 1.0g peptone, 0.5g phosphorus in every 1000mL pure water
Acid dihydride potassium, stirs, and cools down and get final product after sterilization processing；
(2) being inoculated in first cell culture medium by 5-6 block one-level Pleurotus eryngii slant strains block, at temperature 20-22 DEG C, shaking table is cultivated
4d, obtains one-level culture fluid；
Described first cell culture medium every 1000mL pure water adds peptone 9g, soluble starch 53g, glycerol 5g, magnesium sulfate
0.7g, sucrose 8g, thiamine hydrochloride 0.07g；
(3) one-level culture fluid is inoculated in secondary medium according to the inoculum concentration of volumn concentration 10%, at 20-22 DEG C
3d cultivated by shaking table, obtains two grades of culture fluid；
Described secondary medium every 1000mL pure water adds peptone 35g, wheat bran 60g, sucrose 19g, potassium dihydrogen phosphate
1.8g；
(4) two grades of culture fluid are inoculated in fermentation medium cultivation according to volumn concentration 15%, at 25 DEG C, cultivate 6d,
Ventilation 2.5m3/ h, i.e. obtains Agaricus Bisporus liquid spawn after fermentation ends；
Described fermentation medium every 1000mL pure water adds yeast extract 13g, 18g sucrose, 46g peptone, zinc citrate
0.05g, potassium dihydrogen phosphate 1g, magnesium sulfate 0.8g.
Effect example:
1, embodiment 1-3 and the female of comparative example 1-2 are planted the growing state of mycelia, concrete outcome on different slant mediums
It is shown in Table 1.
Table 1
In upper table, ++++represent that mycelia is dense white, sturdy；+++ represent that mycelia is dense in vain, fine and closely woven；++ represent that the dense leucorrhea of mycelia is yellowish, carefully
Close have shoestring；+ representing that mycelia is greyish white, shoestring is many, and mycelial growth potential enters weak.
2, Agaricus Bisporus liquid-spawn inoculation embodiment 1-3 and comparative example 1 prepared is on bacterium bag, arranges matched group, right
Agaricus Bisporus liquid spawn according to group inoculation is commercially available, and the specification of bacterium bag is the most identical, and inoculum concentration is identical, by Agaricus Bisporus total output and residual
Mushroom rate is added up, and concrete outcome is shown in Table 2.
Table 2
It addition, find through substantial amounts of experiment, when the content of each component of culture medium is beyond scope provided by the present invention, effect
Being more or less the same, cross the waste causing resource at most, during less than scope provided by the present invention, possible nutrition is insufficient, and effect is relatively
Difference, yield reduces.
Specific embodiment described herein is only to illustrate spirit of the present invention.The technology people of art
Member it is understood that above-mentioned specific embodiment is not limited to the present invention, all within the spirit and principles in the present invention, done
Any equivalent, improvement etc., should be included within the scope of the present invention.
Claims (9)
1. the preparation method of an Agaricus Bisporus liquid spawn, it is characterised in that comprise the following steps:
(1) Agaricus Bisporus mother kind is inoculated in primary inclined plane culture medium, cultivates 5-6d at 24-26 DEG C, obtain Agaricus Bisporus slant strains,
It is divided into slant strains block；
(2) being inoculated in first cell culture medium by 5-6 block one-level Pleurotus eryngii slant strains block, at temperature 20-22 DEG C, shaking table is cultivated
1-2d, then quiescent culture 1-2d at 20-22 DEG C, obtains one-level culture fluid；
(3) one-level culture fluid is inoculated in secondary medium according to the inoculum concentration of volumn concentration 10%, at 20-22 DEG C
2-3d cultivated by shaking table, obtains two grades of culture fluid；
(4) two grades of culture fluid are inoculated in fermentation medium cultivation according to volumn concentration 15%, at 25 DEG C, cultivate 4-
5d, ventilation 2.5m3/ h, i.e. obtains Agaricus Bisporus liquid spawn after fermentation ends.
Preparation method the most according to claim 1, it is characterised in that described slant medium is in every 1000mL pure water
Add 8-10mL green tea extractive liquor, 20-28g corn starch, 1-2g agar, 0.2-0.5g casein hydrolysate peptides, 0.8-1.2g egg
White peptone, 0.2-0.5g potassium dihydrogen phosphate, stir, cool down after sterilization processing and get final product.
Preparation method the most according to claim 1, it is characterised in that add in described first cell culture medium every 1000mL pure water
Enter 15-20mL green tea extractive liquor, peptone 7-10g, soluble starch 50-55g, glycerol 5-8g, magnesium sulfate 0.2-0.8g, Pu Lu
Blue polysaccharide 0.2-0.3g, sucrose 8-12g, thiamine hydrochloride 0.06-0.08g.
Preparation method the most according to claim 1, it is characterised in that add in described secondary medium every 1000mL pure water
Enter 20-25mL green tea extractive liquor, peptone 30-40g, wheat bran 45-60g, sucrose 18-20g, potassium dihydrogen phosphate 1.5-2.0g, card
Ripple nurse 0.06-0.08g.
Preparation method the most according to claim 1, it is characterised in that add in described fermentation medium every 1000mL pure water
Enter yeast extract 12-15g, 15-20g sucrose, 8-10g treaster, 10-15mL green tea extractive liquor, 40-50g peptone, zinc citrate
0.02-0.05g, potassium dihydrogen phosphate 1g, magnesium sulfate 0.8-1.2g.
6. according to the preparation method described in any one of claim 2-5, it is characterised in that described green tea extractive liquor uses with lower section
Method is prepared from: grinds after the green tea of fresh harvesting adds Sal, then encases the green tea after grinding with gauze, then add
Enter to account for the water of green tea weight 8 times, add medical stone powder, Polyethylene Glycol, be heated to 90-100 DEG C, insulated and stirred 2h, filters
Filtrate, concentrates the filtrate to the half of original volume.
Preparation method the most according to claim 6, it is characterised in that the addition of described Sal accounts for green tea weight
1%；The addition of described medical stone powder accounts for the 5% of the weight adding water.
Preparation method the most according to claim 7, it is characterised in that the addition of described Polyethylene Glycol accounts for green tea weight
1.5%.
Method the most according to claim 1, it is characterised in that the length and width of described slant strains block are 2cm.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108865899A (en) * | 2018-07-06 | 2018-11-23 | 上海市农业科学院 | A kind of agaricus bisporus bacterial strain and its selection |
| CN114503873A (en) * | 2020-11-16 | 2022-05-17 | 上海国森生物科技有限公司 | Agaricus bisporus liquid strain reduction strain and cultivation method thereof |
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| CN102550293A (en) * | 2012-02-03 | 2012-07-11 | 连云港市农业科学院 | Method for liquid fermentation cultivation of Agaricus bisporus strain |
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2016
- 2016-08-22 CN CN201610698556.1A patent/CN106148201A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102550293A (en) * | 2012-02-03 | 2012-07-11 | 连云港市农业科学院 | Method for liquid fermentation cultivation of Agaricus bisporus strain |
Non-Patent Citations (2)
| Title |
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| FRANCISCO J. GEA ＥＴ ＡＬ．: "Effect of spent mushroom compost tea on mycelial growth and yield of button mushroom (Agaricus bisporus) ", 《WORLD J MICROBIOL BIOTECHNOL》 * |
| 杨宁等: "双孢菇常用保存方法", 《储藏加工》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108865899A (en) * | 2018-07-06 | 2018-11-23 | 上海市农业科学院 | A kind of agaricus bisporus bacterial strain and its selection |
| CN114503873A (en) * | 2020-11-16 | 2022-05-17 | 上海国森生物科技有限公司 | Agaricus bisporus liquid strain reduction strain and cultivation method thereof |
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