CN101699969A - Submerged fermentation culture method of straw mushroom liquid strain and culture medium thereof - Google Patents
Submerged fermentation culture method of straw mushroom liquid strain and culture medium thereof Download PDFInfo
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- CN101699969A CN101699969A CN200910211742A CN200910211742A CN101699969A CN 101699969 A CN101699969 A CN 101699969A CN 200910211742 A CN200910211742 A CN 200910211742A CN 200910211742 A CN200910211742 A CN 200910211742A CN 101699969 A CN101699969 A CN 101699969A
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Abstract
The invention relates to a submerged fermentation culture method of straw mushroom liquid strains and culture mediums thereof. The invention solves the technical defects of the prior art, such as long mother species propagation time, poor hypha quality, long fungus ball production time, different fungus ball sizes, higher production cost and the like. The method is used for the submerged fermentation culture of the straw mushroom liquid strains. The method comprises the following steps: 1. carrying out mother species culture in a special culture medium of straw mushroom mother species; 2. carrying out transition culture in a special culture medium of straw mushroom stocks; 3. homogenizing the stocks to obtain homogeneous succulent strains; 4. carrying out primary shaking culture and amplification in a shaking liquid culture medium; 5. carrying out secondary shaking culture and amplification; and 6. carrying out fermentation treatment in a submerged fermentation liquid culture medium to obtain the finished products of the straw mushroom liquid strains. The invention relates to the special culture medium of straw mushroom mother species, the special culture medium of straw mushroom stocks, the shaking liquid culture medium and the submerged fermentation liquid culture medium.
Description
Technical field:
The present invention relates to the submerged culturing method for making and the medium thereof of bacterial classification submerged culturing method for making and medium thereof, particularly straw mushroom liquid strain.
Background technology:
Straw mushroom uses solid spawn to produce always.In the straw mushroom industrial and annual was produced, because the straw mushroom production cycle is extremely short, so bacterial classification is in great demand, so solid spawn can not adapt to the needs that the straw mushroom industrial and annual is produced fully.Also there are a lot of problems in the straw mushroom liquid strain fermented and cultured technology of using at present, is specially: the universal medium of (1) use carries out mother and plants breeding, and therefore female kind breeding time is longer, and mycelia is of low quality; (2) be mostly that the solid spawn that will cultivate with mother culture media without the processing that homogenizes, directly carries out the one-level shake-flask culture, the time of producing the bacterium ball like this is long, the bacterium ball is not of uniform size, is unfavorable for the growth of bacterium ball, influences the quality of straw mushroom liquid strain.(3) the used medium of liquid spawn contains compositions such as a large amount of glucose, sucrose, makes culture environment and planting environment produce very big contrast, and the liquid spawn of production often produces the phenomenon of not material feeding after the cultivation inoculation, have a strong impact on finished product production.
Summary of the invention:
The present invention overcomes the deficiency of existing straw mushroom liquid strain fermentation culture method, design a kind ofly can turn out the straw mushroom liquid strain submerged culturing method for making that the even moderate bacterial classification of size can shorten incubation time again, specifically comprises:
A kind of female special-purpose mother culture media of planting breeding that is used in the straw mushroom liquid strain incubation is provided, and this mother culture media can impel that mycelial growth is vigorous, growth time is short, mycelia is strong, mycelia quality height.
A kind of straw mushroom original seed special culture media based on straw powder and cotton seed hulls powder that uses in the straw mushroom liquid strain incubation is provided, and this medium can provide the mycelia fragment vigorous, eurhythmia of growing; Can be easy in the one-level shake-flask culture, carry out homogenizing of bacterial classification again and handle back formation slurry matter bacterial classification uniformly.This medium simultaneously can be with the preservation of original seed long-term safety.
A kind of cultivation amplification culture medium that uses in the straw mushroom liquid strain incubation is provided.
A kind of submerged fermentation culture medium that uses in straw mushroom liquid strain fermented and cultured process is provided, and this medium can reduce cost, and is beneficial to produce high quality liquid straw mushroom bacterial classification.
Technical scheme of the present invention is:
(1) in the special-purpose mother culture media of straw mushroom, carries out mother and plant cultivation.
(2) will cultivate the mother who covers with test tube in mother culture media and plant the mycelia culture transferring and carry out transition in the straw mushroom original seed special culture media and cultivate, condition of culture is: 30 ± 1 ℃ of temperature, dark culturing treats that mycelia covers with.
(3) special-purpose original seed is fully smashed after immediately culture transferring handle the back and form slurry matter bacterial classification uniformly to shaking to homogenize in bottle liquid nutrient medium.
(4) in shaking bottle liquid nutrient medium, carry out the cultivation amplification that one-level is shaken bottle.
(5) one-level being shaken in the bottle 2-4 days bacterium liquid of growth moves into and to fill the fresh secondary that shakes bottle liquid nutrient medium and shake and carry out the amplification of secondary shake-flask culture in the bottle.
(6) secondary being shaken the bacterium liquid that comes out of bottle moves into and carries out fermentation process in the serum bottle that fills liquid nutrient medium and can obtain the finished product straw mushroom liquid strain.
The composition and the weight percent content scope of above-mentioned straw mushroom special mother kind culture medium raw material part are: apple juice (pulverize and squeeze the juice) 10%-30% agar 1%-3% straw mushroom bacterium chaff leachate 70%-90%; The condition of culture of regulating in pH to the 6.4-7.4. mother culture media with sodium hydroxide is: 30 ± 1 ℃ of temperature, dark culturing is treated that mycelia is covered with to get final product.
The composition and the weight percent content scope of the special-purpose pedigree seed culture medium raw material part of above-mentioned straw mushroom are: straw powder 30%-50%, cotton seed hulls powder 30%-50%; Wheat bran or rice bran 4%-16%, gypsum 1%-4%, water content 50%-70%.Condition of culture in the special-purpose pedigree seed culture medium of straw mushroom is: 30 ± 1 ℃ of temperature; Treating that mycelia is long gets final product at the bottom of the test tube time.
The method for preserving of the special-purpose original seed of above-mentioned straw mushroom is: test tube is wrapped 20 ℃ of constant temperature preservations with plastic sack, and straw mushroom special use original seed can preservation about 4 months like this.Can need be used to carry out the cultivation amplification that one-level is shaken bottle at any time by producing.
The above-mentioned bottle liquid nutrient medium that shakes is the cultivation amplification culture medium that firsts and seconds shakes bottle, and the composition and the weight percent content scope of its raw material part are: analysis for soybean powder 1%-4% corn flour 1-4% sucrose 0.5%-2% glucose 0.5%-2% yeast extract 0.15%-0.6% peptone 0.05-0.2% potassium dihydrogen phosphate 0.05%-0.2% magnesium sulfate 0.025%-0.1%Ph7.8-8.
Above-mentioned firsts and seconds shakes the cultivation amplification of bottle to carry out in triangular flask.
Above-mentioned firsts and seconds shakes bottle condition of culture of interior strain cultivation amplification: temperature: 32 ± 1 ℃; Shake frequency: 125-155 rev/min; Inoculum concentration: 10%-15%; Incubation time: firsts and seconds shakes bottle and was respectively 3-4 days and 2-3 days.
The aforesaid liquid bacterial classification carries out fermentation process in serum bottle liquid nutrient medium is a submerged fermentation culture medium, and the composition and the weight percent content scope of its raw material part are: soybean cake powder 1%-5% corn flour 1%-5% sucrose 0.5%-2% glucose 0.5%-2% peptone 0.15-0.6% leguminous plant oil 0.25%-0.5%; Potassium dihydrogen phosphate 0.05%-0.2% magnesium sulfate 0.025%-0.1%Ph7.8-8.
The aforesaid liquid bacterial classification carries out the condition of culture of fermentation process in serum bottle: temperature: 32-35 ± 1 ℃ throughput 1: 0.5 (volume ratio); Venting pressure 0.5kg/cm2; Incubation time: 50 hours.
Technique effect:
The cultural method of straw mushroom liquid strain of the present invention, in common liquid fungus seed culture technology, increased the treatment step that homogenizes, i.e. transition in straw mushroom original seed special culture media is cultivated and is shaking in bottle liquid nutrient medium original seed processings that homogenize, and starches the matter bacterial classification uniformly and form.This step has effectively solved the technical barrier that inoculation homogenizes and handles with bacterial classification in the liquid fungus seed culture technology, because straw powder and cotton seed hulls powder in the special-purpose pedigree seed culture medium of straw mushroom itself have tiny gap, the straw mushroom mycelia can fully be smashed when inoculation cultivate a period of time in this medium after, a large amount of mycelia fragments have been increased, in shaking bottle liquid nutrient medium, form uniform pulpous state bacterial classification earlier, and then turn out the evenly moderate bacterial classification of size and shorten incubation time.Formed a large amount of tiny effective bacterium balls behind the bacterial classification inoculation by the special-purpose pedigree seed culture medium cultivation of straw mushroom, diameter reaches more than 90% at the effective bacterium ball that has most vigor between the 0-2mm.And the bacterium ball of the bacterial classification inoculation of directly cultivating by mother culture media is not of uniform size, and big bacterium ball loses vigor because diameter is too big.
Straw mushroom original seed special culture media is a kind of straw powder that added, cotton seed hulls powder, wheat bran or rice bran, the medium of gypsum, straw powder, cotton seed hulls powder, the slight gap of wheat bran or rice bran helps the lace growth, and the adding of other nutrient components such as wheat bran or rice bran can make the lace growth vigorous.
Original seed in the straw mushroom original seed special culture media can preservation under 20 ℃ of constant temperature storage conditions about 4 months, need be used to carry out the cultivation amplification that one-level is shaken bottle at any time by producing after the breeding.Be particularly suitable for the production of straw mushroom industrial and annual.
Because the special-purpose mother culture media of straw mushroom is a kind of special culture media at straw mushroom liquid strain cultivation master clock, has therefore added straw mushroom bacterium chaff leachate in this medium.Because used hormone substance and contain the holoside nutrient material that is decomposed by primordial hypha but be not absorbed when in straw mushroom bacterium chaff, containing original mycelia and decomposing polysaccharide, so added the special-purpose mother culture media of straw mushroom bacterium chaff leachate, both shortened female kind of incubation time, make female mycelial growth of planting vigorous especially again, the mycelia particularly robust.
Submerged fermentation culture medium and original liquid nutrient medium difference, be to have reduced monose composition (glucose), the required sugared source of bacterium ball growth is from the corn flour and the starch of polysaccharide, the use of these two kinds of materials, both reduced the cost of medium, the C/N ratio that also makes medium is at more approaching natural environment.Improved the output of cultivation inoculation back straw mushroom.
Embodiment:
Present embodiment is a preferred embodiment, and its concrete steps are.
(1) in the special-purpose mother culture media of straw mushroom, carries out mother and plant cultivation.Get 0.5cm
2Square straw mushroom is female to be planted, and is seeded to special mother kind medium culture, and under 30 ± 1 ℃ of conditions, dark culturing is carried out actication of culture and subculture, forms the female kind of a test tube.The content and the composition of the special-purpose mother culture media preparation of this step are: apple juice (pulverize and squeeze the juice) 200ml agar 30g straw mushroom bacterium chaff leachate 800ml; Regulate pH to 7.4. with sodium hydroxide
(2) will cultivate the mother who covers with test tube in mother culture media plants the mycelia culture transferring and carries out transition in the straw mushroom original seed special culture media and cultivate: by the full bottle mycelia culture transferring of step (1) gained in the triangular flask of 150ml, can transplant six bottles, secretly cultivate then, temperature condition is 30 ± 1 ℃, treats to get final product when mycelia is expired triangular flask.The content and the composition of the preparation of straw mushroom original seed special culture media are: straw powder 45% cotton seed hulls powder, 45% wheat bran or rice bran 8%, gypsum 2% water content 50%-70%.Collocation method is: straw and cotton seed hulls pulverizing back are sifted out with 40 orders and 60 mesh sieve; The straw powder that sifts out and cotton seed hulls powder and wheat bran or rice bran and gypsum are fully mixed thoroughly; Raw material is mixed in proportion with water and get, regulating pH value is 7.4.
(3) cultured special-purpose original seed is fully smashed after immediately culture transferring form slurry matter bacterial classification uniformly to shaking the processing that homogenizes in bottle liquid nutrient medium.The one-level that above-mentioned six bottles of cultured original seeds are inoculated into six 500 milliliters is shaken bottle, will fully original seed be smashed in the time of inoculation, makes original seed form a large amount of mycelia fragments, one-level is shaken bottle shake at a high speed until forming slurry matter bacterial classification uniformly
(4) carry out the cultivation amplification that one-level is shaken bottle in shaking bottle liquid nutrient medium: the amplification cultivation condition that one-level is shaken bottle is. temperature: 32 ± 1 ℃; Shake frequency: 125-155 rev/min; Inoculum concentration: 10%, the liquid nutrient medium coefficient is 100ml-150ml/500ml (triangular flask), cultivating fate is 3 days.
(5) one-level being shaken bacterium liquid in the bottle moves into and to fill the fresh secondary that shakes bottle liquid nutrient medium and shake in the bottle and increase: the bacterium liquid that one-level is shaken in the bottle is inoculated into six 5000 milliliters shaking in the bottle, and it is bottle identical that condition of culture and one-level are shaken.Firsts and seconds shakes the used bottle content and the composition of liquid nutrient medium preparation that shake of cultivation amplification of bottle and is in above-mentioned (4) (5) step: analysis for soybean powder 2% corn flour 2% sucrose 1% glucose 1% yeast extract 0.3% peptone 0.1% potassium dihydrogen phosphate 0.1% magnesium sulfate 0.05%Ph7.8.
(6) secondary being shaken the bacterium liquid that comes out of bottle moves in the serum bottle that fills liquid nutrient medium and carries out fermentation process.The liquid nutrient medium that carries out fermentation process in serum bottle is a submerged fermentation culture medium, and the content of its preparation and composition are: soybean cake powder 2.5% corn flour 2.5% sucrose 1% glucose 1% peptone 0.3% leguminous plant oil 0.5%; Potassium dihydrogen phosphate 0.1% magnesium sulfate 0.05%Ph7.8.At first secondary is shaken the fine and soft ball that comes out of bottle and be inoculated into six 200L and contain in the serum bottle of 80% submerged fermentation culture medium, then in temperature: 32-35 ± 1 ℃ throughput 1: 0.5 (volume ratio); The condition of culture of venting pressure 0.5kg/cm2 is cultivated down can generate straw mushroom liquid strain in 50 hours.
Claims (9)
1. the submerged culturing method for making of a straw mushroom liquid strain.It is characterized in that comprising the steps: that (1) carry out mother and plant cultivation in the special-purpose mother culture media of straw mushroom.(2) the qualified mother that will cultivate in mother culture media plants the mycelia culture transferring and carries out transition in the straw mushroom original seed special culture media and cultivate, after fully smash special-purpose original seed (3) immediately culture transferring handle the back and form slurry matter bacterial classification uniformly to shaking to homogenize in bottle liquid nutrient medium.(4) in shaking bottle liquid nutrient medium, carry out the cultivation amplification that one-level is shaken bottle; (5) one-level being shaken in the bottle 2-4 days bacterium liquid of growth moves into the fresh bottle liquid nutrient medium that shakes and carry out the amplification of secondary shake-flask culture: the bacterium liquid that (6) amplify the secondary shake-flask culture moves into and carries out fermentation process in the serum bottle that fills the submerged fermentation liquid nutrient medium and can obtain the finished product straw mushroom liquid strain.
2. the submerged culturing method for making of straw mushroom liquid strain according to claim 1: the condition of culture that it is characterized in that each step of submerged culturing method for making of described straw mushroom liquid strain is: the condition of culture in the special-purpose mother culture media of straw mushroom is: 30 ± 1 ℃ of temperature, dark culturing; The condition of culture of the special-purpose pedigree seed culture medium of straw mushroom is: 30 ± 1 ℃ of temperature, dark culturing.Shake and carry out one-level in bottle liquid nutrient medium and shake the condition of culture that bottle and secondary shake-flask culture increase and be: temperature: 32 ± 1 ℃; Shake frequency: 125-155 rev/min; Inoculum concentration: 10%-15%; Incubation time: firsts and seconds shakes bottle and was respectively 3-4 days and 2-3 days; Straw mushroom liquid strain carries out fermentation process in serum bottle condition of culture is: temperature: 32-35 ± 1 ℃; Throughput 1: 0.5 (volume ratio); Venting pressure: 0.5kg/cm2; Incubation time: 50 hours.
3. the related mother culture media of the submerged culturing method for making of the described straw mushroom liquid strain of one of claim 1-2 is the special-purpose mother culture media of straw mushroom, it is characterized in that the composition and the weight percent content scope of straw mushroom special mother kind culture medium raw material part is: apple juice (pulverize and squeeze the juice) 10%-30%; Agar 1%-3%; Straw mushroom bacterium chaff leachate 70%-90%; Regulate pH to 6.4-7.4 with sodium hydroxide.
4. the related pedigree seed culture medium of the submerged culturing method for making of the described straw mushroom liquid strain of one of claim 1-2 is a straw mushroom original seed special culture media: it is characterized in that described straw mushroom original seed special culture media is the medium that contains the straw powder.
5. the related pedigree seed culture medium of the submerged culturing method for making of the described straw mushroom liquid strain of one of claim 1-2 is a straw mushroom original seed special culture media: it is characterized in that described straw mushroom original seed special culture media is the medium that contains the cotton seed hull powder.
6. the related pedigree seed culture medium of the submerged culturing method for making of the described straw mushroom liquid strain of one of claim 1-2 is a straw mushroom original seed special culture media: it is characterized in that the special-purpose pedigree seed culture medium of described straw mushroom is the medium that contains straw powder, cotton seed hull powder and wheat bran or rice bran.
7. the related pedigree seed culture medium of the submerged culturing method for making of the described straw mushroom liquid strain of claim 1-2 is a straw mushroom original seed special culture media, it is characterized in that: the composition and the weight percent content scope of this culture medium raw material part are: straw powder 30%-50%, cotton seed hulls powder 30%-50%; Wheat bran or rice bran 4%-16%, gypsum 1%-4%, water content 50%-70%.
8. the described bottle liquid nutrient medium that shakes of one of claim 1-2 is that firsts and seconds shakes the related medium of the cultivation amplification of bottle for shaking a bottle liquid nutrient medium, and it is characterized in that: the raw material composition and the weight percent content scope partly of shaking bottle liquid nutrient medium is: analysis for soybean powder 1%-4% corn flour 1-4% sucrose 0.5%-2% glucose 0.5%-2% yeast extract 0.15%-0.6% peptone 0.05-0.2% potassium dihydrogen phosphate 0.05%-0.2% magnesium sulfate 0.025%-0.1%Ph7.8-8.
9. to carry out the liquid nutrient medium of fermentation process in serum bottle be the submerged fermentation liquid nutrient medium to the described straw mushroom liquid strain of one of claim 1-2, it is characterized in that: straw mushroom liquid strain carries out the raw material part of the submerged fermentation liquid nutrient medium that submerged fermentation handles in serum bottle composition and weight percent content scope are: soybean cake powder 1%-5% corn flour 1%-5% sucrose 0.5%-2% glucose 0.5%-2% peptone 0.15-0.6% leguminous plant oil 0.25%-0.5%; Potassium dihydrogen phosphate 0.05%-0.2% magnesium sulfate 0.025%-0.1%Ph7.8-8.
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| CN102523917A (en) * | 2011-12-14 | 2012-07-04 | 广东省微生物研究所 | Method for cultivating straw mushroom |
| CN102523914A (en) * | 2010-11-30 | 2012-07-04 | 国立大学法人信州大学 | Mushroom culture medium capable of being reused in livestock feed and method for making the same |
| CN103396231A (en) * | 2013-07-21 | 2013-11-20 | 邬金飞 | Culture material composition for straw mushroom breeder seeds, and preparation method of culture material |
| CN104710206A (en) * | 2013-07-12 | 2015-06-17 | 鲁东大学 | Preparation method for volvariella volvacea liquid strain |
| CN106011044A (en) * | 2016-06-27 | 2016-10-12 | 合肥福泉现代农业科技有限公司 | Method for preparing pleurotus citrinopileatus liquid strain through pleurotus citrinopileatus culture medium based on eupatorium adenophorum |
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2009
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