CN104988070A - Pleurotus eryngii liquid strain with small and many mycelium pellets and fermentation method of pleurotus eryngii liquid strain - Google Patents
Pleurotus eryngii liquid strain with small and many mycelium pellets and fermentation method of pleurotus eryngii liquid strain Download PDFInfo
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Abstract
The invention discloses a pleurotus eryngii liquid strain with small and many mycelium pellets and a fermentation method of the pleurotus eryngii liquid strain, and belongs to the technical field of pleurotus eryngii cultivation. Chinese herb extracts capable of improving cold resistance, heat irritability resistance, immunity and bacterium-resisting capacity of the strain are scientifically compounded in culture mediums for liquid seeds, shaking flask seeds, seeding tank seeds and fermentation tank liquid strains in all stages, and meanwhile wood powder which has the feedstock adaptability and has the function of an antifoaming agent is scientifically compounded; manners of supplemental inoculation and segmentation variable temperature fermentation are adopted, and therefore the difference between the strain cell age and the multiplication algebra is shortened; and packaged chitosan with a special function is properly fed, and the pleurotus eryngii liquid strain with the thick and solid fermentative hyphae, high activity, small mycelium pellet diameter, large density and uniform distribution is cultured. The density of the mycelium pellets ranges from 4.5*10<5>piece/L to 5.5*10<5>piece/L, the diameter of the mycelium pellets ranges from 0.5mm to 0.7mm, the dry weight of the mycelium pellets ranges from 90g/L to 100g/L, the germination time ranges from 4 hours to 5 hours, and the exopolysaccharide ranges from 3.0g/L to 3.5g/L.
Description
Technical field
The present invention relates to planting almond abalone mushroom technical field, be specifically related to the little and many pleurotus eryngii liquid strains of a kind of mycelium pellet and fermentation process thereof.
Background technology
Pleurotus eryngii (PleurotuseryngiiQuel.) has another name called pleurotus eryngii, there is bacterial context plumpness, the advantages such as quality is tender and crisp, particularly its stem dense structure, solid, milky white, tasty and refreshing, be called as " flat mushroom king ", " dried scallop mushroom ", there is the mouthfeel of almond flavor and abalone, be applicable to fresh-keeping, processing, firmly get liking of people, it is nutritious, rich in proteins, carbohydrate, VITAMIN and calcium, magnesium, copper, the mineral substance such as zinc, immune function of human body can be improved, have anticancer to human body, reducing blood-fat, the effects such as ease constipation stomach and beauty treatment, there is high cultivating value and commercial value.
Domestic and international planting almond abalone mushroom is based on solid spawn at present; and liquid spawn relative to solid spawn have with short production cycle, cell age is consistent, fruiting is neat, be convenient to management, bacterial classification cost is low, inoculation convenient and the advantage such as quick, is conducive to the mass-producing of Edible Fungi, batch production.In recent years due to the fast development of industrial cultivation technique, for shortening bacterial classification preparation cycle, improve strain quality and production efficiency, domestic and international manufacturer research and development liquid spawn is used for substituting during current Pleurotus eryngii is produced the solid spawn generally used, and in actual production, start application.
Pleurotus eryngii liquid strain produces great-hearted Pleurotus eryngii mycelium by liquid fermentation process.At present, domesticly not yet promulgate quality and quality that unified national standard carrys out specification pleurotus eryngii liquid strain, but people sum up effect and have worked out some region standards in actual application, as Liaoning Province's provincial standard " DB21/T 1693-2008 edible fungi liquid strain production technology regulation " etc., clear stipulaties in these standards, qualified edible fungi liquid strain should " visible distinctive hypha form, spherical and plexi mycelium distributes in a large number, mycelia is sturdy, in mycelia, protoplasma is evenly distributed " " bacterium liquid slightly thickness, there is a large amount of sheet or spherical mycelium suspended, be evenly distributed ", that is excellent liquid spawn mycelium pellet density is high, diameter is little, be evenly distributed, on the other hand, in production application, pleurotus eryngii liquid strain carries out mechanical inoculation operation by production line inoculation device, because inoculation device spout is narrow, for preventing bacterium liquid blocking pipe, also need the pretreatment technology before increasing liquid-spawn inoculation (the general method of mechanical disintegration that adopts makes the mycelium pellet diameter in bacterium liquid diminish, and bacterium liquid is become evenly).Therefore, the physical property such as size, density, uniformity coefficient of Pleurotus eryngii liquid fermenting mycelium pellet is the crucial Con trolling index of high-quality liquid spawn.
At present, about the research report of Pleurotus eryngii liquid culture and patented technology major part concentrate on shaking flask level, mass-producing fermentation research less, substantially the optimization aspect of zymotechnique is concentrated on, especially more in the improvement of fermention medium: application number be 201410568173.3 Chinese patent disclose a kind of Pleurotus eryngii liquid fermentation medium of improvement and utilize it to cultivate the method for pleurotus eryngii liquid strain, add the gelatin of 0.1-0.3g/100ml in the fermentation medium, the mixture of the one or both in agar, adopt and stir, shaking table concussion is cultivated, in above-mentioned disclosed Patent media, agar addition solidifies more than 0.4g/100ml, liquid culture cannot be carried out, wayward, there is the failed hidden danger of fermentation, and it is few that the liquid spawn amount obtained cultivated by shaking table, be not suitable for mass-producing, mechanization production, strain quality is not fine simultaneously.Application number be 201410244283.4 Chinese patent disclose a kind of production method of edible fungi liquid strain, cultivate at solid medium after activated for the edible mushroom strains preserved, then pulverize, continue sealing on solid medium and cultivate 2-3 days, contact mixing with the agar-agar soln of 0.15-0.2% and obtain liquid spawn, in fact still, there is the defect of solid fermentation in above-mentioned disclosed patent, and pulverizing is wherein culture process and the easy pollution microbes of allocating technology again, high-speed stirring making beating and the high shear force pulverized injure quite large to bacterial classification.North gardening. an edible mushrooms section discloses the preparation research [2009 (11) 230-232] of the pleurotus eryngii liquid strain that Liu Guanhui etc. delivers, pleurotus eryngii liquid strain fermention medium and fermentation condition are optimized, but still it is not comprehensive, well can not be applicable to growth, breeding, the physical property such as size, density, uniformity coefficient of the zymophyte pompon of the liquid spawn obtained is not bery desirable, and same exist stirring shearing force having a strong impact on mycelium pellet quality.
Air-blowing fermentation is fermentation mode oxygen or sterile air passed into from airlift fermentation pot bottom, possesses the effect of logical oxygen and soft stirring simultaneously, logical oxygen efficiency is high, dissolved oxygen is effective, effectively can prevent stirring-type from fermenting shearing force to the impact of pleurotus eryngii liquid strain quality, the liquid spawn mycelia obtained is sturdy, modest viscosity, mycelium pellet diameter is little, density is high, be evenly distributed.Application number be 201310479951.7 Chinese patent disclose a kind of cultural method of pleurotus eryngii liquid strain, traditional potato carbon source and wheat bran nitrogenous source are replaced with bean cake powder, pour substratum into fermentor tank, 121 DEG C of sterilizing 60-120 minute, after sterilizing, water drenches and cools and pass into sterile air; The liquid mother that inoculation culture is good plants, inoculum size is 1-2 ‰, fermentor cultivation temperature is 21-26 DEG C, venting pressure is 0.5-1.5 MPa, cultivate and obtain pleurotus eryngii liquid strain in 7-10 days, but above-mentioned disclosed Patent media not comprehensive nutrition, zymotechnique is delayed, fermentation period is long, and the liquid spawn quality obtained is still undesirable.
To sum up, according to the strain properties of Pleurotus eryngii, by Optimal Medium and culture condition, air-blowing fermentation is adopted to be still the unremitting pursue of those skilled in the art for high quality, high yield, pleurotus eryngii liquid strain that cost is low.
Summary of the invention
Technical problem solved by the invention overcomes the defect of existing air-blowing fermentation for pleurotus eryngii liquid strain, in spawn culture various stages substratum, the composite bacterial classification that improves of science resists cold, the Chinese herbal medicine extract of heat stress and immunity and adaptability raw material wood dust, segmentation is adopted to add the mode of inoculation and temperature-variable fermentation, shorten bacterial classification cell age, propagation algebraic difference distance, in good time stream add bag by after chitosan adjustment fermention medium viscosity, turn out a kind of fermented hypha sturdy, active high, mycelium pellet diameter is little, density is large, be evenly distributed, the pleurotus eryngii liquid strain that fermentation period is short.
In order to achieve the above object, the present invention is by the following technical solutions:
A fermentation process for the pleurotus eryngii liquid strain that mycelium pellet is little and many, comprises the steps:
1) go bail for the Pleurotus eryngii slant strains 0.2cm kept
2bacterial classification block 2 pieces be inoculated in plate culture medium and activate, 25 DEG C of constant temperature culture 10 days, so activation 2 times, obtains dull and stereotyped seed;
2) by dull and stereotyped seed diameter be 0.5cm punch tool intercept 0.2cm
2bacterial classification block 3 pieces access 250mL triangular flask in, liquid seed culture medium liquid amount is 100mL, in 25 DEG C of constant temperature culture 2 days, then puts into constant-temperature table, 25 DEG C, 100r/min shaking culture 3 days liquid seeds;
3) by liquid seeds with 10% inoculum size be inoculated in 500mL shaking flask, Shake flask medium liquid amount 250mL, in 25 DEG C, 150r/min vibrate constant temperature culture 4 days shake-flask seed;
4) by shake-flask seed with 10% inoculum size be inoculated in 5L seeding tank, seed tank culture base liquid amount 3L, in 25 DEG C, under air flow 4L/min condition constant temperature culture 4 days seeding tank seed;
5) first seeding tank seed is inoculated in 30L fermentor tank with 8% inoculum size, fermention medium liquid amount 15L, in 30-32 DEG C, air flow 3-5L/min ferment at constant temperature 12-24h, then 18-22 DEG C is cooled to the speed of 0.15-0.25 DEG C/min, 13-15g bag is added by chitosan and 900mL seeding tank seed in fermentor tank, ferment at constant temperature 10-16h under air flow 6-8L/min condition, last with the ramp of 0.05-0.15 DEG C/min to 24-26 DEG C, under air flow 5-7L/min condition, namely ferment at constant temperature 45-52h obtains the little and many pleurotus eryngii liquid strains of mycelium pellet;
Described Pleurotus eryngii slant strains is commercial Pleurotus eryngii purifying bacterial strain;
Described plate culture medium consists of: potato 200g/L, glucose 20g/L, peptone 5g/L, potassium primary phosphate 3g/L, magnesium sulfate 1.5g/L, agar 20g/L, VB
10.02g/L, surplus is water, pH value 6.0;
Described liquid seed culture medium consists of: glucose 30g/L, Yeast diffusion juice 5g/L, peptone 2g/L, agar 2g/L, Chinese herbal medicine extract 0.1-0.3g/L, potassium primary phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.05-0.15g/L, VB
10.01g/L, surplus is water, pH value 6.0;
Described Shake flask medium consists of: soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine extract 0.5-1.0g/L, potassium primary phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.2-0.4g/L, VB
10.01g/L, surplus is water, pH value 6.0;
Described seed tank culture base consists of: soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine extract 0.8-1.2g/L, potassium primary phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.3-0.5g/L, VB
10.01g/L, surplus is water, pH value 6.0;
Consisting of of described fermention medium: soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine extract 1.5-2g/L, wood dust 0.6-1.0g/L, potassium primary phosphate 0.5g/L, magnesium sulfate 0.5g/L, VB
10.01g/L, surplus is water, pH value 6.0;
Preferably, the preparation method of described Chinese herbal medicine extract, comprises the steps: to count by weight, take the Radix Astragali 30 parts, rhizoma zingiberis 30 parts, dried orange peel 30 parts, cow-bezoar 25 parts, the root bark of tree peony 25 parts, 20 parts, Radix Glycyrrhizae, the wind-weed 18 parts, Japanese Honeysuckle 18 parts, Herba Houttuyniae 17 parts, the radix paeoniae rubrathe 15 parts, the root of large-flowered skullcap 15 parts, Ligusticum wallichii 10 parts, Radix Angelicae Sinensis 8 parts; Put in the Ultrasonic Cleaners filling 0.2% sodium hydrogen carbonate solution and clean 12min in 200W, 30KHz, drain, said herbal medicine being crushed to particle diameter is less than 2 millimeters, puts Homogeneous phase mixing in container and adds the water of 5 times of weight, obtaining mixture, control temperature 80 DEG C keeps 3h, then being cooled to 45 DEG C, is 4 by lactic acid adjust ph, under power 200W condition, carry out microwave extraction 12min, under power 250W, frequency 35KHz condition, carry out ultrasonic-assisted extraction simultaneously; Then the mixed enzyme adding mixture gross weight 1.0% carries out enzymolysis, be 6.2 by lactic acid adjust ph, enzymolysis 3h, finally add the mixture of mixture 2 times of w ethanol and propyl alcohol, the mass ratio of ethanol and propyl alcohol mixing is 1:2, control temperature to 70 DEG C keeps 3.5h, filters, obtains the first filtrate; Add the water of filter residue 2 times of weight, control temperature 90 DEG C keeps 2h, is then cooled to 30 DEG C, filters, obtains the second filtrate; First filtrate and the second filtrate are merged according to mass ratio 3:2, filter vacuum concentrates postlyophilization, pulverizes and obtains Chinese herbal medicine extract;
Described mixed enzyme is dextranase, zytase, pentosanase, polygalacturonase 3:2:2:1 Homogeneous phase mixing in mass ratio.
Preferably, the preparation method of described wood dust comprises the steps: without mouldy, anosis worm sawmilling end or waste wood removing foreign material, be placed in the Ultrasonic Cleaners of the sodium hydrogen carbonate solution that mass percent concentration 0.5% is housed in 200W, 30KHz cleans 10min, rinse, drain, put into steam still in 0.3-0.4MPa boiling 20-30min, take out, drain, with vegetables oil 100:1 Homogeneous phase mixing in mass ratio, then mixture is put microwave dryer in 2000W, 120 DEG C of microwave exposure 5min, make it moisture and reach 8-10%, last micronizing is to particle diameter 0.3-0.5mm, pack and obtain wood dust,
More preferably, described drying process is: microwave exposure 10s, stops 10s.
Preferably, the preparation method of described fermention medium is: according to each raw material of culture medium prescription precise, first soybean cake powder quality 4-6 water is doubly got, be 5-6 by lactic acid adjust ph, add the thermostableα-amylase of 5u/g soybean cake powder, add soybean cake powder while stirring, be warming up to 90-95 DEG C, insulation, liquefaction 10-15min, then 50-60 DEG C is cooled to, insulation, add the saccharifying enzyme of 40u/g soybean cake powder and the protease hydrolysis 20-30min of 30u/g soybean cake powder while stirring, add residue water and other raw material while stirring, Homogeneous phase mixing, adjusted to ph is 6.0, namely 121-123 DEG C of sterilizing 30-40min obtain fermention medium,
Preferably, described bag is by the preparation method of chitosan, comprising the steps: to get the mass percent concentration that chitosan adds its quality 30-50% is in the cyclohexaamylose aqueous solution of 10-12%, make softwood, granulate with 10-20 mesh sieve, obtain wet granular, put in power 2000W, 60-80 DEG C of dry 4-6min in microwave dryer, namely obtain bag by chitosan with the quick whole grain of 10-20 mesh sieve.
The detection method of liquid spawn primary quality measure of the present invention:
The mensuration of mycelium pellet dry weight
Get 50ml fermented liquid 80 object screen clothes to filter, collect bacterium ball, repeatedly rinse with clear water, and to be placed in constant temp. drying box 80 DEG C and to dry after 4 hours and weigh, and weigh once every half an hour, until twice weighing difference is no more than 2mg, be constant weight.Finally calculate the mean value of dry mycelial weight.
The mensuration of mycelium pellet density
Get fermented liquid 1ml in culture dish with transfer pipet, bacterium ball quantity is counted, repeat numeration three times, finally calculate the mean value of Peloton density.
The mensuration of mycelium pellet diameter
Get fermented liquid 1ml with transfer pipet, get 10 bacterium balls with tweezers and line up a queue of, measure total length, then average, replicate measurement three times, calculate the mean value of bacterium spherical diameter.
The mensuration of the dull and stereotyped sprout time of mycelium pellet tieback
From the nutrient solution of fermention medium, get a bacterium ball be placed in PDA culture medium flat plate central authorities, each fermention medium does three repetitions, the PDA flat board being placed with bacterium ball is placed in incubator 25 DEG C cultivate, observed once every 1 hour, by the time after having observed one group of bacterium ball sprouting, change observation in every 0.5 hour into once, and record the sprout time of each experimental group, finally calculate the mean value of sprout time after bacterium ball tieback flat board in each fermention medium.
The mensuration of exocellular polysaccharide content
Get fermented liquid 50ml, suction filtration, get supernatant liquor, add the industrial spirit of two volumes, precipitate polysaccharides spends the night, then centrifugal 10 minutes of 3600r/min, collects polysaccharide, dries at being placed in 40 DEG C, weigh once every half an hour after weighing after 2 hours, until twice weighing difference is no more than 2mg, be constant weight.
Another object of the present invention is to provide the little and many pleurotus eryngii liquid strains of mycelium pellet prepared by aforesaid method.
Little and the many pleurotus eryngii liquid strain of described mycelium pellet is through ferment tank 67-92h, and mycelium pellet density is 4.5-5.5 X 10
5individual/L, mycelium pellet diameter is 0.5-0.7mm, and mycelium pellet dry weight is 90-100g/L, and sprout time is 4-5h, and exocellular polysaccharide is 3.0-3.5g/L.
A further object of the invention is to provide the application of the little and many pleurotus eryngii liquid strains of above-mentioned mycelium pellet in planting almond abalone mushroom.
Beneficial effect:
The present invention is according to the characteristic of the former bacterial strain of Pleurotus eryngii, at liquid seeds, shake-flask seed, resist cold from few composite bacterial classification that improves of science at the most in seeding tank seed and fermentor liquid bacterial classification each stage substratum, heat stress, the Chinese herbal medicine extract of immunity and germ resistance, science is composite simultaneously both had adaptability to raw material, there is again the wood dust of defoamer function, by taming step by step, rejuvenation enlarged culturing, enhance pleurotus eryngii quel strains high temperature resistance and Cold stress ability, improve the ability of producing multiple degrading enzyme, excite its multiplication capacity and propagation density, shorten sprout time, improve the output of mycelia and exocellular polysaccharide, improve the adaptability of bacterial strain to yeasting and follow-up planting environment, adopt the mode adding inoculation and segmentation temperature-variable fermentation simultaneously, shorten bacterial classification cell age and propagation algebraic difference distance, in good time stream add bag by after there is thickening property, intermiscibility, stability, the viscosity of the chitosan adjustment fermention medium of the specific function such as adsorptivity and biocidal property and stability, improve homogeneity and dissolved oxygen concentration that mycelium pellet distributes in fermented liquid, and break the preparation method of traditional zymotic substratum, to the further biological enzymolysis of soybean cake powder wherein, shorten fermentation period, turn out a kind of fermented hypha sturdy, active high, mycelium pellet diameter is little, density is large, the pleurotus eryngii liquid strain be evenly distributed, its mycelium pellet density is 4.5-5.5 X 10
5individual/L, mycelium pellet diameter is 0.5-0.7mm, and mycelium pellet dry weight is 90-100g/L, and sprout time is 4-5h, and exocellular polysaccharide is 3.0-3.5g/L.Improve the Quality and yield of pleurotus eryngii liquid strain comprehensively, reduce production cost, for the high quality of Pleurotus eryngii, high yield and large-scale planting have established solid basis, show through experiment in cultivation: pleurotus eryngii liquid strain prepared by the present invention is healthy and strong, vigor is high, fast growth, mycelial yield and quality high, anti-miscellaneous bacteria ability is strong, long quality-guarantee period, bacterium time shorten 22.22% is sent out compared with commercial liquid bacterial classification, miscellaneous bacteria infection rate reduces by 96.67%, one-level mushroom rate improves 5%, throughput is commercially available pleurotus eryngii liquid strain more than 2 times, embody higher economic worth, more be applicable to batch production, large-scale planting.
Concrete know-why is:
1. the composite bacterial classification that significantly improves of Chinese herbal medicine extract science of the present invention resists cold, heat stress, the herbal medicine of immunity and disease resistance is raw material, through ultrasonic cleaning, low temperature enzymolysis, prepared by water extraction alcohol extracting and ultrasonic assistant microwave extraction, extract active constituent content is high, cost is low, can effectively by taming step by step, rejuvenation enlarged culturing, enhance pleurotus eryngii quel strains high temperature resistance and Cold stress ability, improve cellulase-producing, proteolytic enzyme, the ability of the degrading enzymes such as amylase, excite its multiplication capacity and propagation density, shorten sprout time, effectively improve the density of pleurotus eryngii liquid strain, the output of dry mycelial weight and exocellular polysaccharide, reduce mycelium pellet diameter, enhance the adaptability of bacterial strain to yeasting and follow-up planting environment, enhance the anti-stress of the pleurotus eryngii liquid strain of improvement comprehensively, disease resistance and immunological competence, adopt ultrasonic cleaning effectively can kill worm's ovum in raw material and miscellaneous bacteria simultaneously, reduce heavy metal ion content and pesticide residue, the raw material reducing Pleurotus eryngii enriching heavy metal ion may, improve the food safety of edible mushrooms and the quality guaranteed period of liquid spawn.
2. wood dust of the present invention with the wood chip of high cellulose content and high lignin content and vegetables oil for raw material, adopt ultrasonic cleaning, steaming and decocting under high pressure and microwave drying process preparation, its quality is fine and soft soft, nutrient fibre cellulose content is high, be very beneficial for pleurotus eryngii quel strains fermentation, grid structure is enriched simultaneously, evenly, adsorptive power is strong, vegetables oil adhesion amount is large, both can be the nutrition that Pleurotus eryngii provides abundant, also can solve air-blowing under high ventilation to ferment the problem of foam easy to foaming, also for the cultivation in later stage provides environmental compatibility ability, and adopt ultrasonic cleaning effectively can kill worm's ovum in raw material and miscellaneous bacteria, reduce heavy metal ion content and pesticide residue, the raw material reducing Pleurotus eryngii enriching heavy metal ion may, improve the food safety of edible mushrooms.
3. instant invention overcomes the method that tradition prepares fermention medium, soybean cake powder is wherein liquefied and enzymolysis further, starch wherein and protein macromolecule nutritive substance are degraded to small molecules, are convenient to Pleurotus eryngii and play ferment and fermentation, shorten Pleurotus eryngii fermentation period.
4. bag of the present invention is by chitosan, there is the multi-functional of chitosan and cyclohexaamylose simultaneously, appropriateness have adjusted the viscosity of Pleurotus eryngii fermented liquid, intermiscibility, stability, adsorptivity and biocidal property, improves homogeneity and dissolved oxygen concentration that mycelium pellet distributes in fermented liquid.
5. the fermentation process of the pleurotus eryngii liquid strain that mycelium pellet of the present invention is little and many adopts air-blowing zymotechnique, technique is simple, easy to operate, fermentation period short (fermentor tank 67-92h), can mass-producing and mechanization production, spread cultivation and fermenting process substratum and zymotechnique by optimizing bacterial classification, segmentation is adopted to add the mode of inoculation and temperature-variable fermentation, shorten cell age and propagation algebraic difference distance, effectively improve biological activity and the fermenting power of pleurotus eryngii liquid strain, significantly improve the density of mycelium pellet, the output of dry mycelial weight and exocellular polysaccharide, reduce mycelium pellet diameter, improve the Quality and yield of pleurotus eryngii liquid strain comprehensively, reduce production cost.
Embodiment
The present invention is described below by specific embodiment.Unless stated otherwise, technique means used in the present invention is method known in those skilled in the art.In addition, embodiment is interpreted as illustrative, but not limits the scope of the invention, and the spirit and scope of the invention only limited by claims.To those skilled in the art, under the prerequisite not deviating from essence of the present invention and scope, the various change carry out the material component in these embodiments and consumption or change also belong to protection scope of the present invention.
Prepared by embodiment 1 raw material
1. the preparation of Chinese herbal medicine extract:
The preparation method of described Chinese herbal medicine extract, comprises the steps: to count by weight, takes the Radix Astragali 30 parts, rhizoma zingiberis 30 parts, dried orange peel 30 parts, cow-bezoar 25 parts, the root bark of tree peony 25 parts, 20 parts, Radix Glycyrrhizae, the wind-weed 18 parts, Japanese Honeysuckle 18 parts, Herba Houttuyniae 17 parts, the radix paeoniae rubrathe 15 parts, the root of large-flowered skullcap 15 parts, Ligusticum wallichii 10 parts, Radix Angelicae Sinensis 8 parts; Put in the Ultrasonic Cleaners filling 0.2% sodium hydrogen carbonate solution and clean 12min in 200W, 30KHz, drain, said herbal medicine being crushed to particle diameter is less than 2 millimeters, puts Homogeneous phase mixing in container and adds the water of 5 times of weight, obtaining mixture, control temperature 80 DEG C keeps 3h, then being cooled to 45 DEG C, is 4 by lactic acid adjust ph, under power 200W condition, carry out microwave extraction 12min, under power 250W, frequency 35KHz condition, carry out ultrasonic-assisted extraction simultaneously; Then the mixed enzyme adding mixture gross weight 1.0% carries out enzymolysis, be 6.2 by lactic acid adjust ph, enzymolysis 3h, finally add the mixture of mixture 2 times of w ethanol and propyl alcohol, the mass ratio of ethanol and propyl alcohol mixing is 1:2, control temperature to 70 DEG C keeps 3.5h, filters, obtains the first filtrate; Add the water of filter residue 2 times of weight, control temperature 90 DEG C keeps 2h, is then cooled to 30 DEG C, filters, obtains the second filtrate; First filtrate and the second filtrate are merged according to mass ratio 3:2, filter vacuum concentrates postlyophilization, pulverizes and obtains Chinese herbal medicine extract;
Described mixed enzyme is dextranase, zytase, pentosanase, polygalacturonase 3:2:2:1 Homogeneous phase mixing in mass ratio.
2. the preparation of wood dust:
The preparation method of described wood dust, comprising the steps: will without mouldy, anosis worm sawmilling end or waste wood removing foreign material, be placed in the Ultrasonic Cleaners of the sodium hydrogen carbonate solution that mass percent concentration 0.5% is housed in 200W, 30KHz cleans 10min, rinse, drain, put into steam still in 0.35MPa boiling 25min, take out, drain, with vegetables oil 100:1 Homogeneous phase mixing in mass ratio, then mixture is put microwave dryer in 2000W, 120 DEG C of microwave exposure 5min, wherein microwave exposure 10s, stop 10s, last micronizing is to particle diameter 0.4mm, pack and obtain wood dust.
3. bag is by the preparation of chitosan:
Described bag is by the preparation method of chitosan, comprising the steps: to get the mass percent concentration that chitosan adds its quality 40% is in the cyclohexaamylose aqueous solution of 11%, make softwood, granulate with 20 mesh sieves, obtain wet granular, put in power 2000W, 70 DEG C of dry 5min in microwave dryer, namely obtain bag by chitosan with the quick whole grain of 20 mesh sieve.
The Chinese herbal medicine extract used in following embodiment 2-7, wood dust and bag are embodiment 1 by chitosan to be prepared, and " No. 3, Longhai City " that pleurotus eryngii quel strains provides for edible mushrooms institute of Longhai City of Zhangzhou City of Fujian Province, other raw material is commercial.
Embodiment 2
A fermentation process for the pleurotus eryngii liquid strain that mycelium pellet is little and many, comprises the steps:
1) go bail for the Pleurotus eryngii slant strains 0.2cm kept
2bacterial classification block 2 pieces be inoculated in plate culture medium and activate, 25 DEG C of constant temperature culture 10 days, so activation 2 times, obtains dull and stereotyped seed;
2) by dull and stereotyped seed diameter be 0.5cm punch tool intercept 0.2cm
2bacterial classification block 3 pieces access 250mL triangular flask in, liquid seed culture medium liquid amount is 100mL, in 25 DEG C of constant temperature culture 2 days, then puts into constant-temperature table, 25 DEG C, 100r/min shaking culture 3 days liquid seeds;
3) by liquid seeds with 10% inoculum size be inoculated in 500mL shaking flask, Shake flask medium liquid amount 250mL, in 25 DEG C, 150r/min vibrate constant temperature culture 4 days shake-flask seed;
4) by shake-flask seed with 10% inoculum size be inoculated in 5L seeding tank, seed tank culture base liquid amount 3L, in 25 DEG C, under air flow 4L/min condition constant temperature culture 4 days seeding tank seed;
5) first seeding tank seed is inoculated in 30L fermentor tank with 8% inoculum size, fermention medium liquid amount 15L, in 31 DEG C, air flow 4L/min ferment at constant temperature 18h, then 20 DEG C are cooled to 0.2 DEG C/min speed, 14g bag is added by chitosan and 900mL seeding tank seed in fermentor tank, ferment at constant temperature 13h under air flow 7L/min condition, last with 0.1 DEG C/min ramp to 25 DEG C, under air flow 6L/min condition, namely ferment at constant temperature 48h obtains the little and many pleurotus eryngii liquid strains of mycelium pellet;
Described plate culture medium consists of: potato 200g/L, glucose 20g/L, peptone 5g/L, potassium primary phosphate 3g/L, magnesium sulfate 1.5g/L, agar 20g/L, VB
10.02g/L, surplus is water, pH value 6.0;
Described liquid seed culture medium consists of: glucose 30g/L, Yeast diffusion juice 5g/L, peptone 2g/L, agar 2g/L, Chinese herbal medicine extract 0.2g/L, potassium primary phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.1g/L, VB
10.01g/L, surplus is water, pH value 6.0;
Described Shake flask medium consists of: soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine extract 0.8g/L, potassium primary phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.3g/L, VB
10.01g/L, surplus is water, pH value 6.0;
Described seed tank culture base consists of: soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine extract 1.0g/L, potassium primary phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.4g/L, VB
10.01g/L, surplus is water, pH value 6.0;
Consisting of of described fermention medium: soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine extract 1.8g/L, wood dust 0.8g/L, potassium primary phosphate 0.5g/L, magnesium sulfate 0.5g/L, VB
10.01g/L, surplus is water, pH value 6.0;
The preparation method of described fermention medium is: according to each raw material of culture medium prescription precise, first the water of soybean cake powder quality 5 times is got, be 5.5 by lactic acid adjust ph, add the thermostableα-amylase of 5u/g soybean cake powder, add soybean cake powder while stirring, be warming up to 93 DEG C, insulation, liquefaction 12min, then 55 DEG C are cooled to, insulation, add the saccharifying enzyme of 40u/g soybean cake powder and the protease hydrolysis 25min of 30u/g soybean cake powder while stirring, add residue water and other raw material while stirring, Homogeneous phase mixing, adjusted to ph is 6.0, namely 121 DEG C of sterilizing 40min obtain fermention medium.
The pleurotus eryngii liquid strain mycelium pellet density that the mycelium pellet prepared through aforesaid method is little and many is 5.5 X 10
5individual/L, mycelium pellet diameter is 0.5mm, and mycelium pellet dry weight is 100g/L, and exocellular polysaccharide is 3.5g/L.
Embodiment 3
A fermentation process for the pleurotus eryngii liquid strain that mycelium pellet is little and many, comprises the steps:
Step 1) to 4) with embodiment 2;
5) first seeding tank seed is inoculated in 30L fermentor tank with 8% inoculum size, fermention medium liquid amount 15L, in 30 DEG C, air flow 3L/min ferment at constant temperature 12h, then 18 DEG C are cooled to the speed of 0.15 DEG C/min, 13g bag is added by chitosan and 900mL seeding tank seed in fermentor tank, ferment at constant temperature 10h under air flow 6L/min condition, last with the ramp to 24 DEG C of 0.05 DEG C/min, under air flow 5L/min condition, namely ferment at constant temperature 45h obtains the little and many pleurotus eryngii liquid strains of mycelium pellet;
Described plate culture medium composition is with embodiment 2;
Described liquid seed culture medium consists of: glucose 30g/L, Yeast diffusion juice 5g/L, peptone 2g/L, agar 2g/L, Chinese herbal medicine extract 0.1g/L, potassium primary phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.05g/L, VB
10.01g/L, surplus is water, pH value 6.0;
Described Shake flask medium consists of: soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine extract 0.5g/L, potassium primary phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.2g/L, VB
10.01g/L, surplus is water, pH value 6.0;
Described seed tank culture base consists of: soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine extract 0.8g/L, potassium primary phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.3g/L, VB
10.01g/L, surplus is water, pH value 6.0;
Consisting of of described fermention medium: soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine extract 1.5g/L, wood dust 0.6g/L, potassium primary phosphate 0.5g/L, magnesium sulfate 0.5g/L, VB
10.01g/L, surplus is water, pH value 6.0;
The preparation method of described fermention medium is: according to each raw material of culture medium prescription precise, first the water of soybean cake powder quality 4 times is got, be 5 by lactic acid adjust ph, add the thermostableα-amylase of 5u/g soybean cake powder, add soybean cake powder while stirring, be warming up to 90 DEG C, insulation, liquefaction 10min, then 50 DEG C are cooled to, insulation, add the saccharifying enzyme of 40u/g soybean cake powder and the protease hydrolysis 20min of 30u/g soybean cake powder while stirring, add residue water and other raw material while stirring, Homogeneous phase mixing, adjusted to ph is 6.0, namely 123 DEG C of sterilizing 30min obtain fermention medium.
The pleurotus eryngii liquid strain mycelium pellet density that the mycelium pellet prepared through aforesaid method is little and many is 5.0 X 10
5individual/L, mycelium pellet diameter is 0.6mm, and mycelium pellet dry weight is 96g/L, and exocellular polysaccharide is 3.2g/L.
Embodiment 4
A fermentation process for the pleurotus eryngii liquid strain that mycelium pellet is little and many, comprises the steps:
Step 1) to 4) with embodiment 2;
5) first seeding tank seed is inoculated in 30L fermentor tank with 8% inoculum size, fermention medium liquid amount 15L, in 32 DEG C, air flow 5L/min ferment at constant temperature 24h, then 22 DEG C are cooled to the speed of 0.25 DEG C/min, 15g bag is added by chitosan and 900mL seeding tank seed in fermentor tank, ferment at constant temperature 16h under air flow 8L/min condition, last with the ramp to 26 DEG C of 0.15 DEG C/min, under air flow 7L/min condition, namely ferment at constant temperature 52h obtains the little and many pleurotus eryngii liquid strains of mycelium pellet;
Described plate culture medium composition is with embodiment 2;
Described liquid seed culture medium consists of: glucose 30g/L, Yeast diffusion juice 5g/L, peptone 2g/L, agar 2g/L, Chinese herbal medicine extract 0.3g/L, potassium primary phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.15g/L, VB
10.01g/L, surplus is water, pH value 6.0;
Described Shake flask medium consists of: soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine extract 1.0g/L, potassium primary phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.4g/L, VB
10.01g/L, surplus is water, pH value 6.0;
Described seed tank culture base consists of: soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine extract 1.2g/L, potassium primary phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.5g/L, VB
10.01g/L, surplus is water, pH value 6.0;
Consisting of of described fermention medium: soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine extract 2g/L, wood dust 1.0g/L, potassium primary phosphate 0.5g/L, magnesium sulfate 0.5g/L, VB
10.01g/L, surplus is water, pH value 6.0;
The preparation method of described fermention medium is: according to each raw material of culture medium prescription precise, first the water of soybean cake powder quality 6 times is got, be 6 by lactic acid adjust ph, add the thermostableα-amylase of 5u/g soybean cake powder, add soybean cake powder while stirring, be warming up to 95 DEG C, insulation, liquefaction 15min, then 60 DEG C are cooled to, insulation, add the saccharifying enzyme of 40u/g soybean cake powder and the protease hydrolysis 30min of 30u/g soybean cake powder while stirring, add residue water and other raw material while stirring, Homogeneous phase mixing, adjusted to ph is 6.0, namely 121 DEG C of sterilizing 30min obtain fermention medium.
The pleurotus eryngii liquid strain mycelium pellet density that the mycelium pellet prepared through aforesaid method is little and many is 4.8 X 10
5individual/L, mycelium pellet diameter is 0.5mm, and mycelium pellet dry weight is 92g/L, and exocellular polysaccharide is 3.1g/L.
Embodiment 5
A fermentation process for the pleurotus eryngii liquid strain that mycelium pellet is little and many, comprises the steps:
Step 1) to 4) with embodiment 2;
5) first seeding tank seed is inoculated in 30L fermentor tank with 8% inoculum size, fermention medium liquid amount 15L, in 32 DEG C, air flow 3L/min ferment at constant temperature 24h, then 22 DEG C are cooled to the speed of 0.15 DEG C/min, 13g bag is added by chitosan and 900mL seeding tank seed in fermentor tank, ferment at constant temperature 10h under air flow 8L/min condition, last with the ramp to 24 DEG C of 0.15 DEG C/min, under air flow 7L/min condition, namely ferment at constant temperature 45h obtains the little and many pleurotus eryngii liquid strains of mycelium pellet;
Described plate culture medium composition is with embodiment 2;
Described liquid seed culture medium consists of: glucose 30g/L, Yeast diffusion juice 5g/L, peptone 2g/L, agar 2g/L, Chinese herbal medicine extract 0.1g/L, potassium primary phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.15g/L, VB
10.01g/L, surplus is water, pH value 6.0;
Described Shake flask medium consists of: soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine extract 0.5g/L, potassium primary phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.4g/L, VB
10.01g/L, surplus is water, pH value 6.0;
Described seed tank culture base consists of: soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine extract 0.8g/L, potassium primary phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.5g/L, VB
10.01g/L, surplus is water, pH value 6.0;
Consisting of of described fermention medium: soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine extract 1.5g/L, wood dust 1.0g/L, potassium primary phosphate 0.5g/L, magnesium sulfate 0.5g/L, VB
10.01g/L, surplus is water, pH value 6.0;
The preparation method of described fermention medium is: according to each raw material of culture medium prescription precise, first the water of soybean cake powder quality 4 times is got, be 6 by lactic acid adjust ph, add the thermostableα-amylase of 5u/g soybean cake powder, add soybean cake powder while stirring, be warming up to 90 DEG C, insulation, liquefaction 15min, then 50 DEG C are cooled to, insulation, add the saccharifying enzyme of 40u/g soybean cake powder and the protease hydrolysis 30min of 30u/g soybean cake powder while stirring, add residue water and other raw material while stirring, Homogeneous phase mixing, adjusted to ph is 6.0, namely 121 DEG C of sterilizing 40min obtain fermention medium.
The pleurotus eryngii liquid strain mycelium pellet density that the mycelium pellet prepared through aforesaid method is little and many is 4.6 X 10
5individual/L, mycelium pellet diameter is 0.7mm, and mycelium pellet dry weight is 90g/L, and exocellular polysaccharide is 3.1g/L.
Embodiment 6
A fermentation process for the pleurotus eryngii liquid strain that mycelium pellet is little and many, comprises the steps:
Step 1) to 4) with embodiment 2;
5) first seeding tank seed is inoculated in 30L fermentor tank with 8% inoculum size, fermention medium liquid amount 15L, in 30 DEG C, air flow 5L/min ferment at constant temperature 12h, then 18 DEG C are cooled to the speed of 0.25 DEG C/min, 15g bag is added by chitosan and 900mL seeding tank seed in fermentor tank, ferment at constant temperature 16h under air flow 6L/min condition, last with the ramp to 26 DEG C of 0.05 DEG C/min, under air flow 5L/min condition, namely ferment at constant temperature 52h obtains the little and many pleurotus eryngii liquid strains of mycelium pellet;
Described plate culture medium composition is with embodiment 2;
Described liquid seed culture medium consists of: glucose 30g/L, Yeast diffusion juice 5g/L, peptone 2g/L, agar 2g/L, Chinese herbal medicine extract 0.3g/L, potassium primary phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.05g/L, VB
10.01g/L, surplus is water, pH value 6.0;
Described Shake flask medium consists of: soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine extract 1.0g/L, potassium primary phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.2g/L, VB
10.01g/L, surplus is water, pH value 6.0;
Described seed tank culture base consists of: soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine extract 1.2g/L, potassium primary phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.3g/L, VB
10.01g/L, surplus is water, pH value 6.0;
Consisting of of described fermention medium: soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine extract 2g/L, wood dust 0.6g/L, potassium primary phosphate 0.5g/L, magnesium sulfate 0.5g/L, VB
10.01g/L, surplus is water, pH value 6.0;
The preparation method of described fermention medium is: according to each raw material of culture medium prescription precise, first the water of soybean cake powder quality 6 times is got, be 5 by lactic acid adjust ph, add the thermostableα-amylase of 5u/g soybean cake powder, add soybean cake powder while stirring, be warming up to 95 DEG C, insulation, liquefaction 10min, then 60 DEG C are cooled to, insulation, add the saccharifying enzyme of 40u/g soybean cake powder and the protease hydrolysis 20min of 30u/g soybean cake powder while stirring, add residue water and other raw material while stirring, Homogeneous phase mixing, adjusted to ph is 6.0, namely 123 DEG C of sterilizing 30min obtain fermention medium.
The pleurotus eryngii liquid strain mycelium pellet density that the mycelium pellet prepared through aforesaid method is little and many is 5.2 X 10
5individual/L, mycelium pellet diameter is 0.6mm, and mycelium pellet dry weight is 98g/L, and exocellular polysaccharide is 3.4g/L.
Embodiment 7
A fermentation process for the pleurotus eryngii liquid strain that mycelium pellet is little and many, comprises the steps:
Step 1) to 4) with embodiment 2;
5) first seeding tank seed is inoculated in 30L fermentor tank with 8% inoculum size, fermention medium liquid amount 15L, in 31 DEG C, air flow 4L/min ferment at constant temperature 18h, then 20 DEG C are cooled to 0.2 DEG C/min speed, 14g bag is added by chitosan and 900mL seeding tank seed in fermentor tank, ferment at constant temperature 13h under air flow 7L/min condition, last with 0.1 DEG C/min ramp to 25 DEG C, under air flow 6L/min condition, namely ferment at constant temperature 48h obtains the little and many pleurotus eryngii liquid strains of mycelium pellet;
Described plate culture medium consists of: potato 200g/L, glucose 20g/L, peptone 5g/L, potassium primary phosphate 3g/L, magnesium sulfate 1.5g/L, agar 20g/L, VB
10.02g/L, surplus is water, pH value 6.0;
Described liquid seed culture medium consists of: glucose 30g/L, Yeast diffusion juice 5g/L, peptone 2g/L, agar 2g/L, Chinese herbal medicine extract 0.2g/L, potassium primary phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.1g/L, VB
10.01g/L, surplus is water, pH value 6.0;
Described Shake flask medium consists of: soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine extract 0.8g/L, potassium primary phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.3g/L, VB
10.01g/L, surplus is water, pH value 6.0;
Described seed tank culture base consists of: soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine extract 1.0g/L, potassium primary phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.4g/L, VB
10.01g/L, surplus is water, pH value 6.0;
Consisting of of described fermention medium: soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine extract 1.8g/L, wood dust 0.8g/L, potassium primary phosphate 0.5g/L, magnesium sulfate 0.5g/L, VB
10.01g/L, surplus is water, pH value 6.0.
The pleurotus eryngii liquid strain mycelium pellet density that the mycelium pellet prepared through aforesaid method is little and many is 4.5 X 10
5individual/L, mycelium pellet diameter is 0.7mm, and mycelium pellet dry weight is 90g/L, and exocellular polysaccharide is 3.0g/L.
The experiment in cultivation of the pleurotus eryngii liquid strain that embodiment 8 mycelium pellet of the present invention is little and many
1. test strain: the liquid spawn of preparing with the embodiment of the present invention 2 and commercially available " No. 3, Longhai City " liquid spawn, for test strain, are divided into test group and control group;
2. inoculation method: test group adopts ordinary method inoculation with control group with identical inoculum size and inoculum density, inoculates 2000 bottles respectively;
3. production management: test group and control group adopt same management process
3.1 hair's tube reasons
After inoculation, bacterium bottle is placed on constant temperature culture indoor, lucifuge is cultivated.Temperature controls at 23 DEG C, relative humidity 70% ~ 75%.The nutrient that this stage is used in planting material decomposes completely and accumulates.The key of bacteria developing period management is insulation, moisturizing, if early stage, temperature was too low, the time length is longer, can reduce spawn activity, and mycelia production rate slows down.The dense Bai Houzai of all bacterium bottle mycelia continues to cultivate 15d, and nutrient is thoroughly decomposed, and mycelia reaches physiological maturity.
3.2 uncorks, mycelium stimulation
After bacterium bottle latter stage of ripening, carry out uncork, mycelium stimulation process according to a conventional method.In mycelium stimulation process, during liquid spawn mycelium stimulation, remove surperficial thin layer.After mycelium stimulation completes, bacterium bottle is moved into mushroom room, bacterium bottle is inverted, and makes mycelia restoration ecosystem.After mycelia recovers, temperature controls at 14 DEG C ~ 16 DEG C, and relative humidity is 90 ~ 95%, illumination 500 ~ 800Lx, CO
2concentration 600 ~ 1500ppm, fruiting to be buddingged.
3.3 sporophore growth management
Temperature controls at 15 ~ 18 DEG C, and atmospheric moisture remains on 85% ~ 90%, CO
2concentration is between 1500 ~ 1800ppm, and sporophore growth is rapid.When mushroom lid launches substantially, a damp mushroom of gathering when spore not yet launches.
4. quality and production index measure
The mycelial concentration of observed and recorded test group and control group culture bottle, mycelia color, a bacterium time, miscellaneous bacteria infection state, observation and comparison sporophore proterties, measure sporophore stem length, cap size and stem diameter, single bottle of output, and statistical computation miscellaneous bacteria infection rate, one-level mushroom rate and biological transformation ratio, result is as table 1-3:
Biological conversion rate (%)=sporophore fresh goods output (g)/culture material dry weight (g) × 100%
Table 1
| Project | Send out the bacterium time | Mycelia color | Mycelial concentration | Miscellaneous bacteria infection rate |
| Test group | 14 days | White | Dense | 0.5％ |
| Control group | 18 days | White partially yellow | Lighter | 15％ |
Above result shows: pleurotus eryngii liquid strain of the present invention is healthy and strong, vigor is high, fast growth, mycelial yield and quality high, anti-miscellaneous bacteria ability is strong, sends out bacterium time shorten 22.22% compared with control group, and miscellaneous bacteria infection rate reduces by 96.67%.
Table 2
| Project | Stem mean length (cm) | Cap mean diameter (cm) | Stem mean diameter (cm) | One-level mushroom rate |
| Test group | 15.17 | 7.34 | 6.28 | 97％ |
| Control group | 10.36 | 7.79 | 3.42 | 92％ |
Above result shows: it is suitable that the sporophore cap after pleurotus eryngii liquid strain cultivation of the present invention and stem grow ratio, attractive appearance, and quality grade is high, and one-level mushroom rate improves 5% compared with control group.
Table 3
| Project | Single bottle of weight in average (g) | Planting material dry weight (g) | Biological transformation ratio (%) |
| Test group | 123.41 | 180 | 68.56 |
| Control group | 91.73 | 180 | 50.96 |
Above result shows: the sporophore after pleurotus eryngii liquid strain cultivation of the present invention is high only according to output, and biological transformation ratio is high, has good economic benefit, is more applicable to batch production, large-scale planting.Compared with control group, biological transformation ratio improves 34.79%.
Meanwhile, pleurotus eryngii liquid strain mycelium pellet density of the present invention is high is 5.5 X 10
5individual/L, commercially available pleurotus eryngii liquid strain mycelium pellet density is up to 2.7 X 10
5individual/L, compared with control group, its throughput is commercially available pleurotus eryngii liquid strain more than 2 times, embodies higher economic worth.
It should be noted that: abalone mushroom liquid culture prepared by embodiment of the present invention 3-7 has above-mentioned experiment effect equally, between each embodiment and little with above-mentioned experiment effect otherness.
Claims (10)
1. the fermentation process of the pleurotus eryngii liquid strain that a mycelium pellet is little and many, it is characterized in that, comprise the steps: by preserve Pleurotus eryngii slant strains through plate culture medium activation after successively through liquid seed culture medium, shake-flask seed substratum and seed tank culture base step by step enlarged culturing obtain seeding tank seed, first seeding tank seed is inoculated in 30L fermentor tank with 8% inoculum size, fermention medium liquid amount 15L, in 30-32 DEG C, air flow 3-5L/min ferment at constant temperature 12-24h, then 18-22 DEG C is cooled to the speed of 0.15-0.25 DEG C/min, 13-15g bag is added by chitosan and 900mL seeding tank seed in fermentor tank, ferment at constant temperature 10-16h under air flow 6-8L/min condition, last with the ramp of 0.05-0.15 DEG C/min to 24-26 DEG C, under air flow 5-7L/min condition, namely ferment at constant temperature 45-52h obtains the little and many pleurotus eryngii liquid strains of mycelium pellet,
It is in the cyclohexaamylose aqueous solution of 10-12% that described bag is comprised the steps: to get by the preparation method of chitosan the mass percent concentration that chitosan adds its quality 30-50%, make softwood, granulate with 10-20 mesh sieve, obtain wet granular, put in power 2000W, 60-80 DEG C of dry 4-6min in microwave dryer, namely obtain bag by chitosan with the quick whole grain of 10-20 mesh sieve.
2. the fermentation process of the pleurotus eryngii liquid strain that a kind of mycelium pellet as claimed in claim 1 is little and many, it is characterized in that, described liquid seed culture medium consists of: glucose 30g/L, Yeast diffusion juice 5g/L, peptone 2g/L, agar 2g/L, Chinese herbal medicine extract 0.1-0.3g/L, potassium primary phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.05-0.15g/L, VB
10.01g/L, surplus is water, pH value 6.0.
3. the fermentation process of the pleurotus eryngii liquid strain that a kind of mycelium pellet as claimed in claim 1 is little and many, it is characterized in that, described Shake flask medium consists of: soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine extract 0.5-1.0g/L, potassium primary phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.2-0.4g/L, VB
10.01g/L, surplus is water, pH value 6.0.
4. the fermentation process of the pleurotus eryngii liquid strain that a kind of mycelium pellet as claimed in claim 1 is little and many, it is characterized in that, described seed tank culture base consists of: soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine extract 0.8-1.2g/L, potassium primary phosphate 0.5g/L, magnesium sulfate 0.5g/L, wood dust 0.3-0.5g/L, VB
10.01g/L, surplus is water, pH value 6.0.
5. the fermentation process of the pleurotus eryngii liquid strain that a kind of mycelium pellet as claimed in claim 1 is little and many, it is characterized in that, consisting of of described fermention medium: soybean cake powder 60g/L, glucose 10g/L, peptone 5g/L, Chinese herbal medicine extract 1.5-2g/L, wood dust 0.6-1.0g/L, potassium primary phosphate 0.5g/L, magnesium sulfate 0.5g/L, VB
10.01g/L, surplus is water, pH value 6.0.
6. the fermentation process of the pleurotus eryngii liquid strain that a kind of mycelium pellet as described in as arbitrary in claim 2-5 is little and many, is characterized in that, the preparation method of described Chinese herbal medicine extract, comprise the steps: to count by weight, take the Radix Astragali 30 parts, rhizoma zingiberis 30 parts, dried orange peel 30 parts, cow-bezoar 25 parts, the root bark of tree peony 25 parts, 20 parts, Radix Glycyrrhizae, the wind-weed 18 parts, Japanese Honeysuckle 18 parts, Herba Houttuyniae 17 parts, the radix paeoniae rubrathe 15 parts, the root of large-flowered skullcap 15 parts, Ligusticum wallichii 10 parts, Radix Angelicae Sinensis 8 parts; Put in the Ultrasonic Cleaners filling 0.2% sodium hydrogen carbonate solution and clean 12min in 200W, 30KHz, drain, said herbal medicine being crushed to particle diameter is less than 2 millimeters, puts Homogeneous phase mixing in container and adds the water of 5 times of weight, obtaining mixture, control temperature 80 DEG C keeps 3h, then being cooled to 45 DEG C, is 4 by lactic acid adjust ph, under power 200W condition, carry out microwave extraction 12min, under power 250W, frequency 35KHz condition, carry out ultrasonic-assisted extraction simultaneously; Then the mixed enzyme adding mixture gross weight 1.0% carries out enzymolysis, be 6.2 by lactic acid adjust ph, enzymolysis 3h, finally add the mixture of mixture 2 times of w ethanol and propyl alcohol, the mass ratio of ethanol and propyl alcohol mixing is 1:2, control temperature to 70 DEG C keeps 3.5h, filters, obtains the first filtrate; Add the water of filter residue 2 times of weight, control temperature 90 DEG C keeps 2h, is then cooled to 30 DEG C, filters, obtains the second filtrate; First filtrate and the second filtrate are merged according to mass ratio 3:2, filter vacuum concentrates postlyophilization, pulverizes and obtains Chinese herbal medicine extract;
Described mixed enzyme is dextranase, zytase, pentosanase, polygalacturonase 3:2:2:1 Homogeneous phase mixing in mass ratio.
7. the fermentation process of the pleurotus eryngii liquid strain that a kind of mycelium pellet as described in as arbitrary in claim 2-5 is little and many, it is characterized in that, the preparation method of described wood dust comprises the steps: without mouldy, anosis worm sawmilling end or waste wood removing foreign material, be placed in the Ultrasonic Cleaners of the sodium hydrogen carbonate solution that mass percent concentration 0.5% is housed in 200W, 30KHz cleans 10min, rinse, drain, put into steam still in 0.3-0.4MPa boiling 20-30min, take out, drain, with vegetables oil 100:1 Homogeneous phase mixing in mass ratio, then mixture is put microwave dryer in 2000W, 120 DEG C of microwave exposure 5min, last micronizing is to particle diameter 0.3-0.5mm, pack and obtain wood dust.
8. the fermentation process of the pleurotus eryngii liquid strain that a kind of mycelium pellet as claimed in claim 7 is little and many, is characterized in that, microwave exposure 10s during microwave drying, stops 10s.
9. the pleurotus eryngii liquid strain that the mycelium pellet that the fermentation process as described in as arbitrary in claim 1-8 obtains is little and many.
10. the application of pleurotus eryngii liquid strain in planting almond abalone mushroom that mycelium pellet as claimed in claim 9 is little and many.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
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| CN105647725A (en) * | 2016-03-28 | 2016-06-08 | 天津中天精科科技有限公司 | Dry red wine with good aging effects and method for manufacturing dry red wine |
| CN105647724A (en) * | 2016-03-28 | 2016-06-08 | 天津中天精科科技有限公司 | Dry white wine and production method thereof |
| CN105647723A (en) * | 2016-03-28 | 2016-06-08 | 天津中天精科科技有限公司 | Dry red wine and production method thereof |
| CN105647722A (en) * | 2016-03-28 | 2016-06-08 | 天津中天精科科技有限公司 | Flavored dry red wine and preparation method thereof |
| CN105647721A (en) * | 2016-03-28 | 2016-06-08 | 天津中天精科科技有限公司 | Probiotic dry red wine and preparation method thereof |
| CN105861616A (en) * | 2016-03-29 | 2016-08-17 | 广东省农业科学院蔬菜研究所 | Agrocybe Cylindracea liquid strain quality detection method |
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