CN107916231A - A kind of fermentation process of the pleurotus eryngii liquid strain of mycelia stalwartness - Google Patents
A kind of fermentation process of the pleurotus eryngii liquid strain of mycelia stalwartness Download PDFInfo
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- CN107916231A CN107916231A CN201711350950.7A CN201711350950A CN107916231A CN 107916231 A CN107916231 A CN 107916231A CN 201711350950 A CN201711350950 A CN 201711350950A CN 107916231 A CN107916231 A CN 107916231A
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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
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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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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
Abstract
The invention discloses a kind of fermentation process of the pleurotus eryngii liquid strain of mycelia stalwartness, belong to planting almond abalone mushroom technical field, in liquid seeds, shake-flask seed, in seeding tank seed and each stage culture medium of fermentor liquid strain science compounding can improve strain resist it is cold, heat stress, the Chinese herbal medicine extract of immunity and anti-miscellaneous bacteria ability, science, which has compounded, at the same time both has adaptability to raw material, there is the wood dust of defoamer again, and by the way of additional inoculation and segmentation temperature-variable fermentation, shorten strain cell age and propagation algebraic step away from, stream adds the chitosan with specific function after coating in due course, it is sturdy to turn out fermented hypha, it is active high, mycelia bulb diameter is small, density is big, the pleurotus eryngii liquid strain being evenly distributed, its mycelium pellet density is 4.5 5.5 X 105A/L, a diameter of 0.5 0.7mm of mycelium pellet, mycelium pellet dry weight are 90 100g/L, and sprout time is 4 5h, and exocellular polysaccharide is 3.0 3.5g/L.
Description
The application is the divisional application of following application：The applying date is on June 11st, 2015, Application No.
CN201510317162.2, the entitled a kind of pleurotus eryngii liquid strain and its fermentation process of mycelia stalwartness.
Technical field
The present invention relates to planting almond abalone mushroom technical field, and in particular to a kind of pleurotus eryngii liquid strain of mycelia stalwartness and its
Fermentation process.
Background technology
Pleurotus eryngii (PleurotuseryngiiQuel.) also known as pleurotus eryngii, it is excellent to have that bacterial context is plump, quality is tender and crisp etc.
Point, particularly its stem dense structure, solid, milky white, tasty and refreshing, are referred to as " oyster mushroom king ", " dried scallop mushroom ", have almond flavor and
The mouthfeel of abalone, is adapted to fresh-keeping, processing, firmly gets liking for people, its is full of nutrition, rich in protein, carbohydrate, dimension life
Element and the mineral matter such as calcium, magnesium, copper, zinc, can improve immune function of human body, have to human body anticancer, reducing blood lipid, ease constipation stomach and
Beauty etc. acts on, and has high cultivating value and commercial value.
Domestic and international planting almond abalone mushroom is based on solid spawn at present, and liquid spawn has production week relative to solid spawn
Phase is short, cell age is consistent, fruiting is neat, easy to manage, the advantages that strain cost is low, inoculation is convenient and quick, be conducive to edible mushroom life
Scale, the batch production of production.In recent years due to the fast development of industrial cultivation technique, to shorten strain manufacturing cycle, improve
Strain quality and production efficiency, domestic and international manufacturer's research and development liquid spawn are used to substitute universal in current Pleurotus eryngii production
The solid spawn used, and start to apply in actual production.
Pleurotus eryngii liquid strain is to produce great-hearted Pleurotus eryngii mycelium by liquid fermentation process.At present, it is domestic still
Do not promulgate that unified national standard comes the quality and quality of specification pleurotus eryngii liquid strain, but the people in actual application
Summarize application effect and worked out some region standards, such as Liaoning Province's provincial standard《DB21/T 1693-2008 edible fungus liquids
Strain production technology regulation》Deng clearly stipulate that qualified edible fungi liquid strain should be " visible distinctive in these standards
Hypha form, spherical and plexi mycelium are largely distributed, and mycelia is sturdy, and plasm is evenly distributed in mycelia " " bacterium solution is slightly sticky thick,
Have a large amount of sheets or it is spherical it is mycelium suspended, be evenly distributed ", that is to say, that excellent liquid spawn mycelium pellet density is high, diameter
It is small, it is evenly distributed；On the other hand, pleurotus eryngii liquid strain is to carry out machinery by production line inoculation device in production application
Change inoculation operation, since inoculation device spout is narrow, to prevent bacterium solution from blocking pipeline, it is also necessary to increase pre- before liquid spawn inoculation
Treatment process (general to make the mycelia bulb diameter in bacterium solution diminish using the method for mechanical crushing, to make bacterium solution become uniform).Therefore, apricot
The physical characteristics such as the size of abalone mushroom liquid fermentation mycelium pellet, density, the uniformity are the crucial Con trolling index of high-quality liquid spawn.
At present, research report and patented technology in relation to Pleurotus eryngii Liquid Culture are largely focused on shaking flask level, scale
Change the less of fermentation research, it is especially more in terms of the improvement of fermentation medium in terms of the optimization for concentrating on zymotechnique substantially：
The Chinese patent of Application No. 201410568173.3 discloses Pleurotus eryngii liquid fermentation medium and the utilization of a kind of improvement
It cultivates the method for pleurotus eryngii liquid strain, in the fermentation medium one in the gelatin of addition 0.1-0.3g/100ml, agar
Kind or the mixture of the two, using stirring, shaking table shake culture, agar addition exceedes in the above-mentioned published patent culture medium
0.4g/100ml solidifies, and can not carry out Liquid Culture, is difficult to control, and there is fermentation failure hidden danger, and shaking table culture obtains
Liquid spawn amount is few, is not suitable for scale, mechanization production, while strain quality is not fine.Application No.
201410244283.4 Chinese patent discloses a kind of production method of edible fungi liquid strain, is by the edible fungi of preservation
In solid medium culture after strain is activated, then crush, continue sealing culture 2-3 days on solid medium, with 0.15-
0.2% agar solution contact is uniformly mixed so as to obtain liquid spawn, the above-mentioned published patent substantially lacking there are still solid fermentation
Fall into, and crushing therein is further cultured for technique and allocating technology miscellaneous bacteria easy to pollute, high-speed stirred mashing and the high shear force crushed
Strain is injured quite big.A northern gardening edible mushrooms piece discloses the preparation research for the pleurotus eryngii liquid strain that Liu Guanhui etc. is delivered
【2009(11)230-232】, pleurotus eryngii liquid strain fermentation medium and fermentation condition are optimized, but still not comprehensively,
It cannot be adapted to growth, the thing such as breeding, the size of the zymophyte pompon of obtained liquid spawn, density, the uniformity well
It is not highly desirable to manage characteristic, equally exists stirring shearing force and mycelium pellet quality is seriously affected.
Air-blowing fermentation is the fermentation method for being passed through oxygen or filtrated air from airlift fermentation pot bottom, is provided simultaneously with leading to
Oxygen and the effect of soft stirring, logical oxygen efficiency is high, dissolved oxygen effect is good, can effectively prevent stirring-type fermentation shearing force to Pleurotus eryngii liquid
The influence of body strain quality, obtained liquid spawn mycelia is sturdy, modest viscosity, mycelia bulb diameter is small, density is high, distribution is equal
It is even.The Chinese patent of Application No. 201310479951.7 discloses a kind of cultural method of pleurotus eryngii liquid strain, by tradition
Potato carbon source and wheat bran nitrogen source replace with bean cake powder, culture medium is poured into fermentation tank, 121 DEG C sterilize 60-120 minutes, go out
Water leaching cools down and is passed through filtrated air after bacterium；The good liquid parent species of inoculated and cultured, inoculum concentration are 1-2 ‰, fermentation tank culture temperature
For 21-26 DEG C, venting pressure is 0.5-1.5 megapascal, and culture obtains pleurotus eryngii liquid strain, but the above-mentioned published patent in 7-10 days
Culture medium not comprehensive nutrition, zymotechnique hysteresis, fermentation period length, obtained liquid spawn quality are still undesirable.
To sum up, the strain properties according to Pleurotus eryngii, by Optimal Medium and condition of culture, being fermented using air-blowing, it is high to prepare
The low pleurotus eryngii liquid strain of quality, high yield, cost is still the unremitting pursuit of those skilled in the art.
The content of the invention
Technical problem solved by the invention is the defects of overcoming existing air-blowing fermentation to prepare pleurotus eryngii liquid strain, in bacterium
Science compounding can improve the Chinese herbal medicine extract that strain resists hot and cold irritability and immunity in kind each stage culture medium of incubation
And adaptability raw material wood dust, by the way of the additional inoculation of segmentation and temperature-variable fermentation, shorten strain cell age, propagation algebraic step
Away from, the viscosity of the chitosan adjustment fermentation medium after stream plus coating in due course, turn out a kind of fermented hypha it is sturdy, it is active it is high,
Mycelia bulb diameter is small, density is big, is evenly distributed, the pleurotus eryngii liquid strain that fermentation period is short.
In order to achieve the above object, the present invention uses following technical scheme：
A kind of fermentation process of the pleurotus eryngii liquid strain of mycelia stalwartness, includes the following steps：
1) go bail for the Pleurotus eryngii slant strains 0.2cm kept22 pieces of strain block be inoculated in plating medium and activated,
25 DEG C of constant temperature incubations 10 days, so activation 2 times, obtain tablet seed；
2) the tablet seed card punch of a diameter of 0.5cm is intercepted into 0.2cm23 pieces of strain block access 250mL triangular flasks
In, liquid seed culture medium liquid amount is 100mL, in 25 DEG C of constant temperature incubations 2 days, is then placed in constant-temperature table, 25 DEG C,
100r/min shaken cultivations obtain liquid seeds in 3 days；
3) liquid seeds are inoculated in 500mL shaking flasks with 10% inoculum concentration, Shake flask medium liquid amount 250mL, in
25 DEG C, 150r/min vibration constant temperature incubations 4 days shake-flask seed；
4) shake-flask seed is inoculated in 5L seeding tanks with 10% inoculum concentration, seed tank culture base liquid amount 3L, in 25
DEG C, under the conditions of throughput 4L/min constant temperature incubation 4 days seeding tank seed；
5) seeding tank seed is inoculated in 30L fermentation tanks with 8% inoculum concentration first, fermentation medium liquid amount 15L, in
30-32 DEG C, throughput 6-8L/min ferment at constant temperature 12-24h, are then cooled to 18-22 DEG C with the speed of 0.3-0.5 DEG C/min,
7-9g coating chitosans and 900mL seeding tank seeds, the ferment at constant temperature under the conditions of throughput 6-8L/min are added into fermentation tank
10-16h, is finally warming up to 24-26 DEG C with the speed of 0.2-0.4 DEG C/min, the ferment at constant temperature under the conditions of throughput 5-7L/min
45-52h up to mycelia stalwartness pleurotus eryngii liquid strain；
The Pleurotus eryngii slant strains purify bacterial strain for Pleurotus eryngii purchased in market；
The plating medium forms：Potato 200g/L, glucose 20g/L, peptone 5g/L, potassium dihydrogen phosphate
3g/L, magnesium sulfate 1.5g/L, agar 20g/L, VB10.02g/L, surplus are water, pH value 6.0；
The liquid seed culture medium forms：Glucose 30g/L, Yeast diffusion juice 5g/L, peptone 2g/L, agar
2g/L, Chinese herbal medicine extract 0.3-0.5g/L, potassium dihydrogen phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.1-0.3g/L,
VB10.01g/L, surplus are water, pH value 6.0；
The Shake flask medium forms：Soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine extraction
Thing 0.5-1.0g/L, potassium dihydrogen phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.2-0.4g/L, VB10.01g/L, surplus
For water, pH value 6.0；
The seed tank culture base forms：Soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine carry
Take thing 0.9-1.1g/L, potassium dihydrogen phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.3-0.5g/L, VB10.01g/L, it is remaining
Measure as water, pH value 6.0；
The composition of the fermentation medium is：Soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine carry
Take thing 1.6-1.8g/L, wood dust 0.3-0.5g/L, potassium dihydrogen phosphate 0.5g/L, magnesium sulfate 0.5g/L, VB10.01g/L, it is remaining
Measure as water, pH value 6.0；
Preferably, the preparation method of the Chinese herbal medicine extract, includes the following steps：Count in parts by weight, weigh radix astragali
30 parts, 30 parts of rhizoma zingiberis, 30 parts of dried orange peel, 25 parts of cow-bezoar, 25 parts of the root bark of tree peony, 20 parts of radix glycyrrhizae, 18 parts of rhizoma anemarrhenae, 18 parts of honeysuckle, cordate houttuynia
17 parts, 15 parts of the radix paeoniae rubrathe, 15 parts of radix scutellariae, 10 parts of Ligusticum wallichii, 8 parts of Angelica sinensis；Put the supersonic wave cleaning machine for filling 0.2% sodium bicarbonate solution
In in 200W, 30KHz clean 12min, drain, said herbal medicine be crushed to particle diameter as less than 2 millimeters, puts in container and uniformly mixes
Merge the water of 5 times of weight of addition, obtain mixed material, 80 DEG C of holding 3h of control temperature, are then cooled to 45 DEG C, with newborn acid for adjusting pH
It is worth for 4, Microwave Extraction 12min is carried out under the conditions of power 200W, while surpassed under the conditions of power 250W, frequency 35KHz
Sound wave assisted extraction；Then the mixed enzyme for adding mixed material gross weight 1.0% is digested, and is 6.2 with newborn acid for adjusting pH value,
3h is digested, finally adds the mixture of 2 times of w ethanols of mixed material and propyl alcohol, the mass ratio that ethanol and propyl alcohol mix is 1:2,
Controlling temperature, filtering, obtains the first filtrate to 70 DEG C of holding 3.5h；Add the water of 2 times of weight of filter residue, the 90 DEG C of holdings of control temperature
2h, is then cooled to 30 DEG C, filtering, obtains the second filtrate；By the first filtrate and the second filtrate according to mass ratio 3:2 merge, filtrate
It is freeze-dried, is crushed up to Chinese herbal medicine extract after vacuum concentration；
The mixed enzyme is dextranase, zytase, pentosanase, pectase in mass ratio 3:2:2:1 uniformly mixing.
Preferably, the preparation method of the wood dust includes the following steps：Will without it is mouldy, without disease pest sawmilling end or waste wood
Remove debris, be placed in the supersonic wave cleaning machine of the sodium bicarbonate solution equipped with mass percent concentration 0.5% in 200W,
30KHz cleans 10min, rinses, drains, and is put into steam kettle in 0.3-0.4MPa boiling 20-30min, takes out, drain, with plant
Thing oil in mass ratio 100:1 uniformly mixing, then puts microwave dryer in 2000W, 120 DEG C of microwave irradiation 5min by mixture,
It is allowed to moisture and reaches 8-10%, last ultramicro grinding to particle diameter 0.3-0.5mm, packs up to wood dust；
It is highly preferred that the drying process is：Microwave irradiation 10s, stops 10s.
Preferably, the preparation method of the fermentation medium is：According to each raw material of culture medium prescription precise, take first
The water of 4-6 times of soybean cake powder quality, is 5-6 with newborn acid for adjusting pH value, adds the thermostableα-amylase of 5u/g soybean cake powders, side
Stir side and add soybean cake powder, be warming up to 90-95 DEG C, insulation, liquefy 10-15min, is then cooled to 50-60 DEG C, keeps the temperature, side
Stir side and add the carbohydrase of 40u/g soybean cake powders and the protease hydrolytic 20-30min of 30u/g soybean cake powders, add while stirring
Enter remaining water and other raw materials, uniformly mixing, adjustment pH value is 6.0,121-123 DEG C of sterilizing 30-40min up to fermented and cultured
Base；
Preferably, the preparation method of the coating chitosan, includes the following steps：Chitosan is taken to add its quality 30-
50% mass percent concentration is in the cyclohexaamylose aqueous solution of 10-12%, and softwood is made, and is pelletized, obtained with 10-20 mesh sieves
Wet granular, puts in 2000W, 60-80 DEG C of dry 4-6min of power in microwave dryer, with the quick whole grain of 10-20 mesh sieves up to bag
By chitosan.
The detection method of fluid present invention strain primary quality measure：
The measure of mycelium pellet dry weight
Take 50ml zymotic fluids to be filtered with the sieve of 80 mesh, collect bacterium ball, rinsed repeatedly with clear water, and be placed on
Weigh after when 80 DEG C of drying 4 are small in constant temperature drying box, and weighed once every half an hour, until weighing difference twice is no more than 2mg,
As constant weight.Finally calculate the average value of dry mycelial weight.
The measure of mycelium pellet density
Zymotic fluid 1ml is taken in culture dish with pipette, bacterium ball quantity is counted, and is repeated numeration three times, is finally counted
Calculate the average value of Peloton density.
The measure of mycelia bulb diameter
Zymotic fluid 1ml is taken with pipette, takes 10 bacterium balls to line up with tweezers a queue of, total length is measured, is then averaging
Value, duplicate measurements three times, calculate the average value of bacterium bulb diameter.
The measure of mycelium pellet tieback tablet sprout time
A bacterium ball is taken to be placed in PDA culture medium tablet center, each fermented and cultured from the nutrient solution of fermentation medium
Base does three repetitions, and the PDA plate for being placed with bacterium ball is placed in incubator and is cultivated for 25 DEG C, is observed once when 1 is small, etc.
To after having observed that one group of bacterium ball is sprouted, be changed to every 0.5 it is small when observation once, and record the sprout time of each experimental group, finally
Calculate the average value of sprout time after bacterium ball tieback tablet in each fermentation medium.
The measure of exocellular polysaccharide content
Zymotic fluid 50ml is taken, is filtered, is taken supernatant, add the industrial alcohol of two volumes, precipitate polysaccharides are overnight, then
3600r/min centrifuge 10 minutes, collect polysaccharide, be placed at 40 DEG C and dry, 2 it is small when after weigh after every half an hour weigh one
It is secondary, until weighing difference twice is no more than 2mg, it is constant weight.
Another object of the present invention is to provide the pleurotus eryngii liquid strain of the mycelia stalwartness of above method preparation.
The fermented tank fermentation 67-92h of pleurotus eryngii liquid strain of the mycelia stalwartness, mycelium pellet density is 4.5-5.5 X
105A/L, a diameter of 0.5-0.7mm of mycelium pellet, mycelium pellet dry weight are 90-100g/L, sprout time 4-5h, and exocellular polysaccharide is
3.0-3.5g/L。
Pleurotus eryngii liquid strain it is a still further object of the present invention to provide above-mentioned mycelia stalwartness is in planting almond abalone mushroom
Using.
Beneficial effect：
Characteristic of the invention according to Pleurotus eryngii opportunistic pathogen strain, in liquid seeds, shake-flask seed, seeding tank seed and fermentation tank liquid
Strain can be improved from as little as more science compoundings resist hot and cold irritability, immunity and antibiotic property in each stage culture medium of body strain
Chinese herbal medicine extract, while science has compounded not only has adaptability to raw material, but also there is the wood dust of defoamer, by step by step
Domestication, rejuvenation expand culture, enhance pleurotus eryngii quel strains high temperature resistance and Cold stress ability, improve the energy for producing a variety of degrading enzymes
Power, excites its multiplication capacity and propagation density, shortens sprout time, improve the yield of mycelia and exocellular polysaccharide, improve
Adaptability of the bacterial strain to yeasting and follow-up planting environment, while by the way of additional inoculation and segmentation temperature-variable fermentation,
Strain cell age and propagation algebraic step are shortened away from stream, which adds after being coated with, in due course has thickening property, intermiscibility, stability, adsorptivity
With the viscosity and stability of the chitosan adjustment fermentation medium of the specific function such as biocidal property, mycelium pellet is improved in zymotic fluid
The homogeneity and dissolved oxygen concentration of distribution, and broken the preparation method of traditional zymotic culture medium, to soybean cake powder therein into
One step biological enzymolysis, shortens fermentation period, turn out a kind of fermented hypha it is sturdy, it is active it is high, mycelia bulb diameter is small, density
Greatly, the pleurotus eryngii liquid strain being evenly distributed, its mycelium pellet density are 4.5-5.5 X 105A/L, a diameter of 0.5- of mycelium pellet
0.7mm, mycelium pellet dry weight are 90-100g/L, sprout time 4-5h, exocellular polysaccharide 3.0-3.5g/L.Improve apricot comprehensively
The quality and yield of abalone mushroom liquid culture, reduce production cost, are established for high quality, high yield and the large-scale planting of Pleurotus eryngii
Solid foundation is determined, has shown through experiment in cultivation：Pleurotus eryngii liquid strain prepared by the present invention is healthy and strong, vigor is high, the speed of growth
It hurry up, mycelial yield and quality are high, and anti-miscellaneous bacteria ability is strong, long shelf-life, the bacterium germination time shortening compared with commercial liquid strain
22.22%, miscellaneous bacteria infection rate reduces by 96.67%, and level-one mushroom rate improves 5%, and production capacity is commercially available 2 times of pleurotus eryngii liquid strain
More than, higher economic value is embodied, is more suitable for batch production, large-scale planting.
Particular technique principle is：
1. the Chinese herbal medicine extract science compounding of the present invention significantly improves strain and resists hot and cold irritability, immunity and disease-resistant
Property Chinese herbal medicine be raw material, through be cleaned by ultrasonic, low temperature enzymolysis, water extracting alcohol carry with ultrasonic wave assisted microwave synthesis extract and prepare, carry
Take thing active constituent content high, cost is low, can effectively by taming step by step, rejuvenation expand culture, enhance pleurotus eryngii quel strains and resist
High temperature and Cold stress ability, improve the ability of the degrading enzymes such as cellulase-producing, protease, amylase, excite its propagation
Ability and propagation density, shorten sprout time, effectively increase the density of pleurotus eryngii liquid strain, dry mycelial weight and extracellular more
The yield of sugar, reduces mycelia bulb diameter, enhances adaptability of the bacterial strain to yeasting and follow-up planting environment, comprehensively enhancing
The resisting stress of pleurotus eryngii liquid strain, disease resistance and immunocompetence, while can effectively be killed using ultrasonic cleaning in raw material
Worm's ovum and miscellaneous bacteria, reduce heavy metal ion content and persticide residue, reduce the raw material of Pleurotus eryngii enriching heavy metal ion
May, improve the foodsafety of edible mushroom and the shelf-life of liquid spawn.
2. wood dust of the present invention is using the sawdust of high cellulose content and high lignin content and vegetable oil as raw material, using super
Prepared by sound cleaning, autoclaving and microwave drying process, its quality is fine and soft soft, and trophic fiber cellulose content is high, is very beneficial for apricot
Abalone mushroom strain fermentation, while network is enriched, uniformly, strong adsorption force, vegetable oil adhesion amount is big, may be either that Pleurotus eryngii offer is rich
Rich nutrition, can also solve the problems, such as air-blowing fermentation foam easy to foaming under high ventilation quantity, also provide ring for the cultivation in later stage
Border adaptive capacity, and worm's ovum and the miscellaneous bacteria that can be effectively killed in raw material using being cleaned by ultrasonic, reduce heavy metal ion content
And persticide residue, reduce Pleurotus eryngii enriching heavy metal ion raw material may, improve the foodsafety of edible mushroom.
3. the present invention overcomes the method that tradition prepares fermentation medium, to the further liquefaction of soybean cake powder therein and enzyme
Solution, small molecule is degraded to by starch therein and protein macromolecule nutriment, is played ferment and fermentation easy to Pleurotus eryngii, is shortened
Pleurotus eryngii fermentation period.
4. the coating chitosan of the present invention, while there is the multi-functional of chitosan and cyclohexaamylose, moderately it have adjusted
Viscosity, intermiscibility, stability, adsorptivity and the biocidal property of Pleurotus eryngii zymotic fluid, improve what mycelium pellet was distributed in zymotic fluid
Homogeneity and dissolved oxygen concentration.
5. the fermentation process of the pleurotus eryngii liquid strain of mycelia stalwartness of the present invention uses air-blowing zymotechnique, technique is simple,
It is easy to operate, fermentation period is short (fermentation tank 67-92h), can scale and mechanization production, spread cultivation and ferment by optimizing strain
Process culture medium and zymotechnique, by the way of the additional inoculation of segmentation and temperature-variable fermentation, shorten cell age and propagation algebraic step
Away from effectively increasing the bioactivity and fermenting power of pleurotus eryngii liquid strain, significantly improve density, the dry mycelial weight of mycelium pellet
With the yield of exocellular polysaccharide, mycelia bulb diameter is reduced, the quality and yield of pleurotus eryngii liquid strain is improved comprehensively, reduces
Production cost.
Embodiment
The present invention is described below by specific embodiment.Unless stated otherwise, technological means used in the present invention
It is method known in those skilled in the art.In addition, embodiment is interpreted as illustrative, it is not intended to limit the present invention
Scope, the spirit and scope of the invention are limited only by the claims that follow.To those skilled in the art, without departing substantially from this
The various changes carried out on the premise of invention spirit and scope to the material component in these embodiments and dosage or change
Belong to protection scope of the present invention.
It is prepared by 1 raw material of embodiment
1. the preparation of Chinese herbal medicine extract：
The preparation method of the Chinese herbal medicine extract, includes the following steps：Count in parts by weight, weigh 30 parts of radix astragali, do
30 parts of ginger, 30 parts of dried orange peel, 25 parts of cow-bezoar, 25 parts of the root bark of tree peony, 20 parts of radix glycyrrhizae, 18 parts of rhizoma anemarrhenae, 18 parts of honeysuckle, 17 parts of cordate houttuynia are red
15 parts of Chinese herbaceous peony, 15 parts of radix scutellariae, 10 parts of Ligusticum wallichii, 8 parts of Angelica sinensis；Put in the supersonic wave cleaning machine for filling 0.2% sodium bicarbonate solution in
200W, 30KHz clean 12min, drain, and said herbal medicine is crushed to particle diameter for less than 2 millimeters, puts in container and uniformly mixes simultaneously
The water of 5 times of weight is added, obtains mixed material, 80 DEG C of holding 3h of control temperature, are then cooled to 45 DEG C, are with newborn acid for adjusting pH value
4, Microwave Extraction 12min is carried out under the conditions of power 200W, while ultrasonic wave is carried out under the conditions of power 250W, frequency 35KHz
Assisted extraction；Then the mixed enzyme for adding mixed material gross weight 1.0% is digested, and is 6.2 with newborn acid for adjusting pH value, enzymolysis
3h, finally adds the mixture of 2 times of w ethanols of mixed material and propyl alcohol, and the mass ratio that ethanol and propyl alcohol mix is 1:2, control
For temperature to 70 DEG C of holding 3.5h, filtering, obtains the first filtrate；The water of 2 times of weight of filter residue is added, 90 DEG C of control temperature keeps 2h, so
After be cooled to 30 DEG C, filtering, obtains the second filtrate；By the first filtrate and the second filtrate according to mass ratio 3:2 merge, and filter vacuum is dense
It is freeze-dried, is crushed up to Chinese herbal medicine extract after contracting；
The mixed enzyme is dextranase, zytase, pentosanase, pectase in mass ratio 3:2:2:1 uniformly mixing.
2. the preparation of wood dust：
The preparation method of the wood dust, includes the following steps：Will without it is mouldy, without disease pest sawmilling end or waste wood remove impurity elimination
Thing, is placed in the supersonic wave cleaning machine of the sodium bicarbonate solution equipped with mass percent concentration 0.5% and is cleaned in 200W, 30KHz
10min, rinses, drains, and is put into steam kettle in 0.35MPa boiling 25min, takes out, drain, with vegetable oil in mass ratio 100:
1 uniformly mixing, then puts microwave dryer in 2000W, wherein 120 DEG C of microwave irradiation 5min, microwave irradiation 10s by mixture,
10s is stopped, last ultramicro grinding to particle diameter 0.4mm, packs up to wood dust.
3. it is coated with the preparation of chitosan：
The preparation method of the coating chitosan, includes the following steps：Chitosan is taken to add the quality hundred of its quality 40%
Divide in the cyclohexaamylose aqueous solution that specific concentration is 11%, softwood is made, pelletized with 20 mesh sieves, obtain wet granular, put microwave drying
In power 2000W, 70 DEG C of dry 5min in machine, chitosan is coated with to obtain the final product with the quick whole grain of 20 mesh sieves.
Chinese herbal medicine extract, wood dust and coating chitosan used in following embodiment 2-7 are that embodiment 1 is made
Standby, pleurotus eryngii quel strains are " Longhai City 3 " that edible mushroom research institute of Longhai City of Zhangzhou City of Fujian Province provides, and other raw materials are purchased in market.
Embodiment 2
A kind of fermentation process of the pleurotus eryngii liquid strain of mycelia stalwartness, includes the following steps：
1) go bail for the Pleurotus eryngii slant strains 0.2cm kept22 pieces of strain block be inoculated in plating medium and activated,
25 DEG C of constant temperature incubations 10 days, so activation 2 times, obtain tablet seed；
2) the tablet seed card punch of a diameter of 0.5cm is intercepted into 0.2cm23 pieces of strain block access 250mL triangular flasks
In, liquid seed culture medium liquid amount is 100mL, in 25 DEG C of constant temperature incubations 2 days, is then placed in constant-temperature table, 25 DEG C,
100r/min shaken cultivations obtain liquid seeds in 3 days；
3) liquid seeds are inoculated in 500mL shaking flasks with 10% inoculum concentration, Shake flask medium liquid amount 250mL, in
25 DEG C, 150r/min vibration constant temperature incubations 4 days shake-flask seed；
4) shake-flask seed is inoculated in 5L seeding tanks with 10% inoculum concentration, seed tank culture base liquid amount 3L, in 25
DEG C, under the conditions of throughput 4L/min constant temperature incubation 4 days seeding tank seed；
5) seeding tank seed is inoculated in 30L fermentation tanks, fermentation medium liquid amount 15L, in 31 with 8% inoculum concentration first
DEG C, throughput 7L/min ferment at constant temperature 18h, be then cooled to 20 DEG C with 0.4 DEG C/min speed, into fermentation tank add 8g coating
Chitosan and 900mL seeding tank seeds, the ferment at constant temperature 13h under the conditions of throughput 7L/min, finally with 0.3 DEG C/min speed liters
Temperature to 25 DEG C, under the conditions of throughput 6L/min ferment at constant temperature 48h up to mycelia stalwartness pleurotus eryngii liquid strain；
The plating medium forms：Potato 200g/L, glucose 20g/L, peptone 5g/L, potassium dihydrogen phosphate
3g/L, magnesium sulfate 1.5g/L, agar 20g/L, VB10.02g/L, surplus are water, pH value 6.0；
The liquid seed culture medium forms：Glucose 30g/L, Yeast diffusion juice 5g/L, peptone 2g/L, agar
2g/L, Chinese herbal medicine extract 0.4g/L, potassium dihydrogen phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.2g/L, VB1 0.01g/
L, surplus are water, pH value 6.0；
The Shake flask medium forms：Soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine extraction
Thing 0.8g/L, potassium dihydrogen phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.3g/L, VB10.01g/L, surplus are water, pH value
6.0；
The seed tank culture base forms：Soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine carry
Take thing 1.0g/L, potassium dihydrogen phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.4g/L, VB10.01g/L, surplus are water, pH
Value 6.0；
The composition of the fermentation medium is：Soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine carry
Take thing 1.7g/L, wood dust 0.4g/L, potassium dihydrogen phosphate 0.5g/L, magnesium sulfate 0.5g/L, VB10.01g/L, surplus are water, pH
Value 6.0；
The preparation method of the fermentation medium is：According to each raw material of culture medium prescription precise, soyabean cake is taken first
The water that 5 times of silty amount, is 5.5 with newborn acid for adjusting pH value, adds the thermostableα-amylase of 5u/g soybean cake powders, adds while stirring
Enter soybean cake powder, be warming up to 93 DEG C, insulation, liquefy 12min, is then cooled to 55 DEG C, insulation, it is yellow to add 40u/g while stirring
The carbohydrase of beancake powder and the protease hydrolytic 25min of 30u/g soybean cake powders, add remaining water and other raw materials while stirring,
Uniformly mixing, adjustment pH value are 6.0,121 DEG C of sterilizing 40min up to fermentation medium.
The pleurotus eryngii liquid strain mycelium pellet density of the mycelia stalwartness prepared through the above method is 5.5 X 105A/L, mycelia
Bulb diameter is 0.5mm, and mycelium pellet dry weight is 100g/L, exocellular polysaccharide 3.5g/L.
Embodiment 3
A kind of fermentation process of the pleurotus eryngii liquid strain of mycelia stalwartness, includes the following steps：
Step 1) is to 4) the same as embodiment 2；
5) seeding tank seed is inoculated in 30L fermentation tanks, fermentation medium liquid amount 15L, in 30 with 8% inoculum concentration first
DEG C, throughput 6L/min ferment at constant temperature 12h, be then cooled to 18 DEG C with the speed of 0.3 DEG C/min, into fermentation tank add 7g bags
By chitosan and 900mL seeding tank seeds, the ferment at constant temperature 10h under the conditions of throughput 6L/min, finally with the speed of 0.2 DEG C/min
Rate is warming up to 24 DEG C, under the conditions of throughput 5L/min ferment at constant temperature 45h up to mycelia stalwartness pleurotus eryngii liquid strain；
The plating medium composition is the same as embodiment 2；
The liquid seed culture medium forms：Glucose 30g/L, Yeast diffusion juice 5g/L, peptone 2g/L, agar
2g/L, Chinese herbal medicine extract 0.3g/L, potassium dihydrogen phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.1g/L, VB1 0.01g/
L, surplus are water, pH value 6.0；
The Shake flask medium forms：Soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine extraction
Thing 0.5g/L, potassium dihydrogen phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.2g/L, VB10.01g/L, surplus are water, pH value
6.0；
The seed tank culture base forms：Soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine carry
Take thing 0.9g/L, potassium dihydrogen phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.3g/L, VB10.01g/L, surplus are water, pH
Value 6.0；
The composition of the fermentation medium is：Soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine carry
Take thing 1.6g/L, wood dust 0.3g/L, potassium dihydrogen phosphate 0.5g/L, magnesium sulfate 0.5g/L, VB10.01g/L, surplus are water, pH
Value 6.0；
The preparation method of the fermentation medium is：According to each raw material of culture medium prescription precise, soyabean cake is taken first
The water that 4 times of silty amount, is 5 with newborn acid for adjusting pH value, adds the thermostableα-amylase of 5u/g soybean cake powders, add while stirring
Soybean cake powder, is warming up to 90 DEG C, and insulation, liquefy 10min, is then cooled to 50 DEG C, insulation, adds 40u/g soya beans while stirring
The carbohydrase of cake powder and the protease hydrolytic 20min of 30u/g soybean cake powders, add remaining water and other raw materials, while stirring
Even mixing, adjustment pH value are 6.0,123 DEG C of sterilizing 30min up to fermentation medium.
The pleurotus eryngii liquid strain mycelium pellet density of the mycelia stalwartness prepared through the above method is 5.0 X 105A/L, mycelia
Bulb diameter is 0.6mm, and mycelium pellet dry weight is 96g/L, exocellular polysaccharide 3.2g/L.
Embodiment 4
A kind of fermentation process of the pleurotus eryngii liquid strain of mycelia stalwartness, includes the following steps：
Step 1) is to 4) the same as embodiment 2；
5) seeding tank seed is inoculated in 30L fermentation tanks, fermentation medium liquid amount 15L, in 32 with 8% inoculum concentration first
DEG C, throughput 8L/min ferment at constant temperature 24h, be then cooled to 22 DEG C with the speed of 0.5 DEG C/min, into fermentation tank add 9g bags
By chitosan and 900mL seeding tank seeds, the ferment at constant temperature 16h under the conditions of throughput 8L/min, finally with the speed of 0.4 DEG C/min
Rate is warming up to 26 DEG C, under the conditions of throughput 7L/min ferment at constant temperature 52h up to mycelia stalwartness pleurotus eryngii liquid strain；
The plating medium composition is the same as embodiment 2；
The liquid seed culture medium forms：Glucose 30g/L, Yeast diffusion juice 5g/L, peptone 2g/L, agar
2g/L, Chinese herbal medicine extract 0.5g/L, potassium dihydrogen phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.3g/L, VB1 0.01g/
L, surplus are water, pH value 6.0；
The Shake flask medium forms：Soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine extraction
Thing 1.0g/L, potassium dihydrogen phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.4g/L, VB10.01g/L, surplus are water, pH value
6.0；
The seed tank culture base forms：Soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine carry
Take thing 1.1g/L, potassium dihydrogen phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.5g/L, VB10.01g/L, surplus are water, pH
Value 6.0；
The composition of the fermentation medium is：Soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine carry
Take thing 1.8g/L, wood dust 0.5g/L, potassium dihydrogen phosphate 0.5g/L, magnesium sulfate 0.5g/L, VB10.01g/L, surplus are water, pH
Value 6.0；
The preparation method of the fermentation medium is：According to each raw material of culture medium prescription precise, soyabean cake is taken first
The water that 6 times of silty amount, is 6 with newborn acid for adjusting pH value, adds the thermostableα-amylase of 5u/g soybean cake powders, add while stirring
Soybean cake powder, is warming up to 95 DEG C, and insulation, liquefy 15min, is then cooled to 60 DEG C, insulation, adds 40u/g soya beans while stirring
The carbohydrase of cake powder and the protease hydrolytic 30min of 30u/g soybean cake powders, add remaining water and other raw materials, while stirring
Even mixing, adjustment pH value are 6.0,121 DEG C of sterilizing 30min up to fermentation medium.
The pleurotus eryngii liquid strain mycelium pellet density of the mycelia stalwartness prepared through the above method is 4.8 X 105A/L, mycelia
Bulb diameter is 0.5mm, and mycelium pellet dry weight is 92g/L, exocellular polysaccharide 3.1g/L.
Embodiment 5
A kind of fermentation process of the pleurotus eryngii liquid strain of mycelia stalwartness, includes the following steps：
Step 1) is to 4) the same as embodiment 2；
5) seeding tank seed is inoculated in 30L fermentation tanks, fermentation medium liquid amount 15L, in 32 with 8% inoculum concentration first
DEG C, throughput 6L/min ferment at constant temperature 24h, be then cooled to 22 DEG C with the speed of 0.3 DEG C/min, into fermentation tank add 7g bags
By chitosan and 900mL seeding tank seeds, the ferment at constant temperature 10h under the conditions of throughput 8L/min, finally with the speed of 0.4 DEG C/min
Rate is warming up to 24 DEG C, under the conditions of throughput 7L/min ferment at constant temperature 45h up to mycelia stalwartness pleurotus eryngii liquid strain；
The plating medium composition is the same as embodiment 2；
The liquid seed culture medium forms：Glucose 30g/L, Yeast diffusion juice 5g/L, peptone 2g/L, agar
2g/L, Chinese herbal medicine extract 0.3g/L, potassium dihydrogen phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.3g/L, VB1 0.01g/
L, surplus are water, pH value 6.0；
The Shake flask medium forms：Soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine extraction
Thing 0.5g/L, potassium dihydrogen phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.4g/L, VB10.01g/L, surplus are water, pH value
6.0；
The seed tank culture base forms：Soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine carry
Take thing 0.9g/L, potassium dihydrogen phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.5g/L, VB10.01g/L, surplus are water, pH
Value 6.0；
The composition of the fermentation medium is：Soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine carry
Take thing 1.6g/L, wood dust 0.5g/L, potassium dihydrogen phosphate 0.5g/L, magnesium sulfate 0.5g/L, VB10.01g/L, surplus are water, pH
Value 6.0；
The preparation method of the fermentation medium is：According to each raw material of culture medium prescription precise, soyabean cake is taken first
The water that 4 times of silty amount, is 6 with newborn acid for adjusting pH value, adds the thermostableα-amylase of 5u/g soybean cake powders, add while stirring
Soybean cake powder, is warming up to 90 DEG C, and insulation, liquefy 15min, is then cooled to 50 DEG C, insulation, adds 40u/g soya beans while stirring
The carbohydrase of cake powder and the protease hydrolytic 30min of 30u/g soybean cake powders, add remaining water and other raw materials, while stirring
Even mixing, adjustment pH value are 6.0,121 DEG C of sterilizing 40min up to fermentation medium.
The pleurotus eryngii liquid strain mycelium pellet density of the mycelia stalwartness prepared through the above method is 4.6 X 105A/L, mycelia
Bulb diameter is 0.7mm, and mycelium pellet dry weight is 90g/L, exocellular polysaccharide 3.1g/L.
Embodiment 6
A kind of fermentation process of the pleurotus eryngii liquid strain of mycelia stalwartness, includes the following steps：
Step 1) is to 4) the same as embodiment 2；
5) seeding tank seed is inoculated in 30L fermentation tanks, fermentation medium liquid amount 15L, in 30 with 8% inoculum concentration first
DEG C, throughput 8L/min ferment at constant temperature 12h, be then cooled to 18 DEG C with the speed of 0.5 DEG C/min, into fermentation tank add 9g bags
By chitosan and 900mL seeding tank seeds, the ferment at constant temperature 16h under the conditions of throughput 6L/min, finally with the speed of 0.2 DEG C/min
Rate is warming up to 26 DEG C, under the conditions of throughput 5L/min ferment at constant temperature 52h up to mycelia stalwartness pleurotus eryngii liquid strain；
The plating medium composition is the same as embodiment 2；
The liquid seed culture medium forms：Glucose 30g/L, Yeast diffusion juice 5g/L, peptone 2g/L, agar
2g/L, Chinese herbal medicine extract 0.5g/L, potassium dihydrogen phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.1g/L, VB1 0.01g/
L, surplus are water, pH value 6.0；
The Shake flask medium forms：Soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine extraction
Thing 1.0g/L, potassium dihydrogen phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.2g/L, VB10.01g/L, surplus are water, pH value
6.0；
The seed tank culture base forms：Soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine carry
Take thing 1.1g/L, potassium dihydrogen phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.3g/L, VB10.01g/L, surplus are water, pH
Value 6.0；
The composition of the fermentation medium is：Soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine carry
Take thing 1.8g/L, wood dust 0.3g/L, potassium dihydrogen phosphate 0.5g/L, magnesium sulfate 0.5g/L, VB10.01g/L, surplus are water, pH
Value 6.0；
The preparation method of the fermentation medium is：According to each raw material of culture medium prescription precise, soyabean cake is taken first
The water that 6 times of silty amount, is 5 with newborn acid for adjusting pH value, adds the thermostableα-amylase of 5u/g soybean cake powders, add while stirring
Soybean cake powder, is warming up to 95 DEG C, and insulation, liquefy 10min, is then cooled to 60 DEG C, insulation, adds 40u/g soya beans while stirring
The carbohydrase of cake powder and the protease hydrolytic 20min of 30u/g soybean cake powders, add remaining water and other raw materials, while stirring
Even mixing, adjustment pH value are 6.0,123 DEG C of sterilizing 30min up to fermentation medium.
The pleurotus eryngii liquid strain mycelium pellet density of the mycelia stalwartness prepared through the above method is 5.2 X 105A/L, mycelia
Bulb diameter is 0.6mm, and mycelium pellet dry weight is 98g/L, exocellular polysaccharide 3.4g/L.
Embodiment 7
A kind of fermentation process of the pleurotus eryngii liquid strain of mycelia stalwartness, includes the following steps：
Step 1) is to 4) the same as embodiment 2；
5) seeding tank seed is inoculated in 30L fermentation tanks, fermentation medium liquid amount 15L, in 31 with 8% inoculum concentration first
DEG C, throughput 7L/min ferment at constant temperature 18h, be then cooled to 20 DEG C with 0.4 DEG C/min speed, into fermentation tank add 8g coating
Chitosan and 900mL seeding tank seeds, the ferment at constant temperature 13h under the conditions of throughput 7L/min, finally with 0.3 DEG C/min speed liters
Temperature to 25 DEG C, under the conditions of throughput 6L/min ferment at constant temperature 48h up to mycelia stalwartness pleurotus eryngii liquid strain；
The plating medium forms：Potato 200g/L, glucose 20g/L, peptone 5g/L, potassium dihydrogen phosphate
3g/L, magnesium sulfate 1.5g/L, agar 20g/L, VB10.02g/L, surplus are water, pH value 6.0；
The liquid seed culture medium forms：Glucose 30g/L, Yeast diffusion juice 5g/L, peptone 2g/L, agar
2g/L, Chinese herbal medicine extract 0.4g/L, potassium dihydrogen phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.2g/L, VB1 0.01g/
L, surplus are water, pH value 6.0；
The Shake flask medium forms：Soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine extraction
Thing 0.8g/L, potassium dihydrogen phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.3g/L, VB10.01g/L, surplus are water, pH value
6.0；
The seed tank culture base forms：Soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine carry
Take thing 1.0g/L, potassium dihydrogen phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.4g/L, VB10.01g/L, surplus are water, pH
Value 6.0；
The composition of the fermentation medium is：Soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine carry
Take thing 1.7g/L, wood dust 0.4g/L, potassium dihydrogen phosphate 0.5g/L, magnesium sulfate 0.5g/L, VB10.01g/L, surplus are water, pH
Value 6.0.
The pleurotus eryngii liquid strain mycelium pellet density of the mycelia stalwartness prepared through the above method is 4.5 X 105A/L, mycelia
Bulb diameter is 0.7mm, and mycelium pellet dry weight is 90g/L, exocellular polysaccharide 3.0g/L.
The experiment in cultivation of the pleurotus eryngii liquid strain of 8 mycelia of the present invention stalwartness of embodiment
1. test strain：The liquid spawn and commercially available " Longhai City 3 " liquid spawn prepared using the embodiment of the present invention 2 is examination
Strain is tested, is divided into test group and control group；
2. inoculation method：Test group and control group are inoculated with identical inoculum concentration and inoculum density using conventional method, point
Jie Zhong not be 2000 bottles；
3. production management：Test group and control group use same management method
3.1 bacterium germination management
Bacterium bottle is placed in constant temperature incubation room after inoculation, lucifuge culture.Temperature control at 23 DEG C, relative humidity 70%~
75%.The nutrient that this stage is used in culture material decomposes completely and accumulation.The key of bacterium germination period management is insulation, moisturizing, early period
If temperature is too low, the duration is longer, can reduce spawn activity, and mycelia speed of production slows down.All bacterium bottle mycelia it is dense it is white after
15d is persistently cultivated again, nutrient is thoroughly decomposed, and mycelia reaches physiological maturity.
3.2 corkages, mycelium stimulation
After bacterium bottle latter stage of ripening, opened according to a conventional method, mycelium stimulation processing.During mycelium stimulation, during liquid spawn mycelium stimulation
Remove surface thin layer.Bacterium bottle is moved into mushroom room after the completion of mycelium stimulation, bacterium bottle is inverted, and makes mycelia restoration ecosystem.Treat bacterium
After silk recovers, temperature control is at 14 DEG C~16 DEG C, and relative humidity is 90~95%, illumination 500~800Lx, CO2Concentration 600~
1500ppm, fruiting to be buddingged.
3.3 sporophore growth management
For temperature control at 15~18 DEG C, air humidity is maintained at 85%~90%, CO2Concentration 1500~1800ppm it
Between, sporophore growth is rapid.When mushroom lid is unfolded substantially, a damp mushroom is harvested when spore not yet launches.
4. mass and yield index measure
Observe and record mycelial concentration, mycelia color, bacterium germination time, the miscellaneous bacteria infection shape of test group and control group culture bottle
Condition, fructification character, measurement fructification stem length, cap size and stem diameter, single bottle yield are compared in observation, and count meter
Miscellaneous bacteria infection rate, level-one mushroom rate and biological transformation ratio are calculated, as a result such as table 1-3：
Biological conversion rate (%)=fructification fresh goods yield (g)/compost dry weight (g) × 100%
Table 1
| Project | The bacterium germination time | Mycelia color | Mycelial concentration | Miscellaneous bacteria infection rate |
| Test group | 14 days | White | It is dense | 0.5% |
| Control group | 18 days | It is partially yellow in vain | It is thin | 15% |
Result above shows：Pleurotus eryngii liquid strain of the present invention is healthy and strong, vigor is high, and the speed of growth is fast, mycelial yield and matter
Amount is high, and anti-miscellaneous bacteria ability is strong, and the bacterium germination time shortening 22.22% compared with control group, miscellaneous bacteria infection rate reduces by 96.67%.
Table 2
| Project | Stem average length (cm) | Cap average diameter (cm) | Stem average diameter (cm) | Level-one mushroom rate |
| Test group | 15.17 | 7.34 | 6.28 | 97% |
| Control group | 10.36 | 7.79 | 3.42 | 92% |
Result above shows：Fructification cap and stem growth ratio after pleurotus eryngii liquid strain cultivation of the present invention close
Suitable, attractive appearance, credit rating is high, and level-one mushroom rate improves 5% compared with control group.
Table 3
| Project | Single bottle average weight (g) | Culture material dry weight (g) | Biological transformation ratio (%) |
| Test group | 123.41 | 180 | 68.56 |
| Control group | 91.73 | 180 | 50.96 |
Result above shows：Fructification after pleurotus eryngii liquid strain cultivation of the present invention is high only according to yield, biological transformation ratio
Height, has preferable economic benefit, is more suitable for batch production, large-scale planting.Compared with control group, biological transformation ratio improves
34.79%.
Meanwhile a height of 5.5 X 10 of pleurotus eryngii liquid strain mycelium pellet density of the present invention5A/L, commercially available Pleurotus eryngii liquid bacteria
Kind mycelium pellet density is up to 2.7 X 105A/L, compared with control group, its production capacity is commercially available 2 times of pleurotus eryngii liquid strain
More than, embody higher economic value.
It should be noted that：Abalone mushroom liquid culture prepared by 3-7 of the embodiment of the present invention equally has above-mentioned experiment effect, respectively
It is between embodiment and little with above-mentioned experiment effect otherness.
Claims (5)
1. a kind of fermentation process of the pleurotus eryngii liquid strain of mycelia stalwartness, it is characterised in that include the following steps：By preservation
Pleurotus eryngii slant strains are trained through liquid seed culture medium, shake-flask seed culture medium and seeding tank successively after plating medium activates
Foster base expands culture and obtains seeding tank seed step by step, and seeding tank seed is inoculated in 30L fermentation tanks with 8% inoculum concentration first, hair
Ferment culture medium liquid amount 15L, in 31 DEG C, throughput 7L/min ferment at constant temperature 18h, is then cooled to 20 with 0.4 DEG C/min speed
DEG C, 8g coating chitosans and 900mL seeding tank seeds, the ferment at constant temperature under the conditions of throughput 7L/min are added into fermentation tank
13h, is finally warming up to 25 DEG C with 0.3 DEG C/min speed, and ferment at constant temperature 48h is healthy and strong up to mycelia under the conditions of throughput 6L/min
Pleurotus eryngii liquid strain；
The composition of the fermentation medium is：Soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine extract
1.7g/L, wood dust 0.4g/L, potassium dihydrogen phosphate 0.5g/L, magnesium sulfate 0.5g/L, VB10.01g/L, surplus are water, pH value
6.0；
The preparation method of the Chinese herbal medicine extract, includes the following steps：Count in parts by weight, weigh 30 parts of radix astragali, rhizoma zingiberis 30
Part, 30 parts of dried orange peel, 25 parts of cow-bezoar, 25 parts of the root bark of tree peony, 20 parts of radix glycyrrhizae, 18 parts of rhizoma anemarrhenae, 18 parts of honeysuckle, 17 parts of cordate houttuynia, the radix paeoniae rubrathe 15
Part, 15 parts of radix scutellariae, 10 parts of Ligusticum wallichii, 8 parts of Angelica sinensis；Put in the supersonic wave cleaning machine for filling 0.2% sodium bicarbonate solution in 200W,
30KHz cleans 12min, drains, and said herbal medicine is crushed to particle diameter for less than 2 millimeters, puts and 5 is uniformly mixed and added in container
The water of times weight, obtains mixed material, and 80 DEG C of holding 3h of control temperature, are then cooled to 45 DEG C, are 4 with newborn acid for adjusting pH value,
Microwave Extraction 12min is carried out under the conditions of power 200W, while ultrasonic wave auxiliary is carried out under the conditions of power 250W, frequency 35KHz
Extraction；Then the mixed enzyme for adding mixed material gross weight 1.0% is digested, and is 6.2 with newborn acid for adjusting pH value, is digested 3h,
The mixture of 2 times of w ethanols of mixed material and propyl alcohol is finally added, the mass ratio that ethanol and propyl alcohol mix is 1:2, control temperature
For degree to 70 DEG C of holding 3.5h, filtering, obtains the first filtrate；The water of 2 times of weight of filter residue is added, 90 DEG C of control temperature keeps 2h, then
30 DEG C are cooled to, filtering, obtains the second filtrate；By the first filtrate and the second filtrate according to mass ratio 3:2 merge, filter vacuum concentration
After be freeze-dried, crush up to Chinese herbal medicine extract；
The mixed enzyme is dextranase, zytase, pentosanase, pectase in mass ratio 3:2:2:1 uniformly mixing；
The preparation method of the wood dust includes the following steps：Will without it is mouldy, without disease pest sawmilling end or waste wood remove debris, put
10min is cleaned in 200W, 30KHz in the supersonic wave cleaning machine of the sodium bicarbonate solution equipped with mass percent concentration 0.5%,
Rinse, drain, be put into steam kettle in 0.3-0.4MPa boiling 20-30min, take out, drain, with vegetable oil in mass ratio 100:
1 uniformly mixing, then puts microwave dryer in 2000W, 120 DEG C of microwave irradiation 5min, last ultramicro grinding to grain by mixture
Footpath 0.3-0.5mm, packs up to wood dust；
The preparation method of the fermentation medium is：According to each raw material of culture medium prescription precise, soyabean cake silty is taken first
The water of 5 times of amount, is 5.5 with newborn acid for adjusting pH value, adds the thermostableα-amylase of 5u/g soybean cake powders, is added while stirring yellow
Beancake powder, is warming up to 93 DEG C, and insulation, liquefy 12min, is then cooled to 55 DEG C, insulation, adds 40u/g soyabean cakes while stirring
The carbohydrase of powder and the protease hydrolytic 25min of 30u/g soybean cake powders, add remaining water and other raw materials, uniformly while stirring
Mixing, adjustment pH value are 6.0,121 DEG C of sterilizing 40min up to fermentation medium；
The preparation method of the coating chitosan includes the following steps：Chitosan is taken to add the quality percentage of its quality 30-50%
Specific concentration is in the cyclohexaamylose aqueous solution of 10-12%, and softwood is made, and is pelletized with 10-20 mesh sieves, obtains wet granular, put microwave
In 2000W, 60-80 DEG C of dry 4-6min of power in drying machine, chitosan is coated with to obtain the final product with the quick whole grain of 10-20 mesh sieves；
The shake-flask seed culture medium forms：Soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine extraction
Thing 0.8g/L, potassium dihydrogen phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.3g/L, VB10.01g/L, surplus are water, pH value
6.0。
2. a kind of fermentation process of the pleurotus eryngii liquid strain of mycelia stalwartness as claimed in claim 1, it is characterised in that described
Liquid seed culture medium forms：Glucose 30g/L, Yeast diffusion juice 5g/L, peptone 2g/L, agar 2g/L, Chinese herbal medicine carry
Take thing 0.3-0.5g/L, potassium dihydrogen phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.1-0.3g/L, VB10.01g/L, it is remaining
Measure as water, pH value 6.0.
3. a kind of fermentation process of the pleurotus eryngii liquid strain of mycelia stalwartness as claimed in claim 1, it is characterised in that described
Shake-flask seed culture medium forms：Soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine extract 0.5-
1.0g/L, potassium dihydrogen phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.2-0.4g/L, VB10.01g/L, surplus are water, pH
Value 6.0.
4. a kind of fermentation process of the pleurotus eryngii liquid strain of mycelia stalwartness as claimed in claim 1, it is characterised in that described
Seed tank culture base forms：Soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine extract 0.9-1.1g/
L, potassium dihydrogen phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.3-0.5g/L, VB10.01g/L, surplus are water, pH value
6.0。
A kind of 5. fermentation process of the pleurotus eryngii liquid strain of mycelia stalwartness as claimed in claim 1, it is characterised in that microwave
Microwave irradiation 10s when dry, stops 10s.
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| CN109089728A (en) * | 2018-08-09 | 2018-12-28 | 合肥福泉现代农业科技有限公司 | A kind of method of soybean highland barley enzymatic hydrolysis and fermentation Efficient Cultivation edible mushroom |
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| JP2011078394A (en) * | 2009-10-06 | 2011-04-21 | Kimio Munemura | Fermentation medium for edible mushroom and production method therefor |
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| CN109089728A (en) * | 2018-08-09 | 2018-12-28 | 合肥福泉现代农业科技有限公司 | A kind of method of soybean highland barley enzymatic hydrolysis and fermentation Efficient Cultivation edible mushroom |
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| CN108251312A (en) | 2018-07-06 |
| CN107815420A (en) | 2018-03-20 |
| CN104938212B (en) | 2018-05-11 |
| CN108094049A (en) | 2018-06-01 |
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Application publication date: 20180417 |