CN105670942A - Method for producing ophiocordyceps sinensis powder through rapid and deep liquid state fermentation - Google Patents
Method for producing ophiocordyceps sinensis powder through rapid and deep liquid state fermentation Download PDFInfo
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Abstract
The invention discloses a method for producing ophiocordyceps sinensis powder through rapid and deep liquid state fermentation. The method comprises the steps that fermentation liquor rich in mycelia is rapidly obtained from ophiocordyceps sinensis strains through a semi-continuous fermentation technology, and then the fermentation liquor is subjected to micro-filtration-membrane treatment and separation, so that ophiocordyceps sinensis mycelia are obtained. By means of the method, the traditional intermittent batch fermentation process for ophiocordyceps sinensis is changed, and in addition, the centrifugal separation mode is replaced with efficient ceramic membrane microfiltration for separation of the mycelia. By means of the semi-continuous fermentation process, the defects that the traditional batch fermentation process is large in number of process links, long in intermediate gap time, low in equipment efficiency and the like are overcome; the fermentation liquor is filtered through an efficient ceramic membrane, the situation that filtrate enters the mycelia can be reduced, the yield of the mycelia can be raised, COD of the filtrate is lowered, sewage treatment amount is decreased, cells are not destroyed, and it is guaranteed that all effective constituents in the cells are recycled. Compared with the traditional process, the method has remarkable economic benefits, social benefits and environment protection benefits.
Description
Technical field
The invention belongs to microbial technology field, it is specifically related to a kind of method utilizing deep layer liquid state fermentation technology to produce radicula byssoidea fast, a kind of method of rapid deep zone liquid state fermentation production hirsutella sinensis fungal powder more precisely.
Background technology
Fermentable can be divided in batches, fed-batch, the various modes such as semicontinuous, continuous. The human and material resources consumption of batch fermentation is relatively big, and every batch fermentation all needs to carry out to feed, sterilizing, inoculate, put material, cleaning etc. and operate, and operation is loaded down with trivial details, and fermentation period is longer, and production efficiency is lower; Though fed-batch fermentation supplements the deficiency of nutrient or precursor by feed supplement, but the continuous accumulation due to unwanted metabolic products, and Product formation is finally checked unavoidably; Relatively batch fermentation and the fed-batch fermentation production intensity of continuously fermenting improves greatly, but easily suffers the pollution of miscellaneous bacteria, and bacterial classification is easily degenerated, and facility investment is relatively big, and Fermentation Substance Concentration is lower; In semicontinuous fermentation process, fill into fresh culture again by putting part fermented liquid, it is possible not only to supplementary nutrient and precursor, and metabolism nuisance is diluted, thus the continuation being conducive to product is synthesized.
The application of semicontinuous fermentation process can play the effect alleviated Product inhibiton and avoid metabolic by-prods to accumulate, improve the culture environment of microorganism, contribute to keeping the stable of thalline vigor, and for some secondary metabolite, its highest throughput rate only just can reach when some transient state. Adopting semicontinuous fermentation process to be possible not only to this kind of transient state condition is occurred repeatedly, but also can improve the utilization ratio of equipment, therefore semicontinuous fermentation process has important application in the production of secondary metabolite.
Namely hirsutella sinensis fungal powder ferments Cordyceps fungus powder, be separated from the fresh Cordyceps sinensis in Qinghai the section ergot fungus cordyceps sinensis obtained asexual generation--China pilose spore bacterial classification is through the dried powder of liquid fermentation and culture gained mycelium.Being mainly used in tonifying lung kidney, cough that benefit vital essence and deficiency of both the lung and kidney cause, asthma, spitting of blood, aching pain in waist and back etc. be sick and the assisting therapy of chronic bronchitis.
Tradition China pilose spore liquid-state fermentation technology mostly is the batch fermentation technique of intermittence, and the human and material resources consumption of batch fermentation technique is relatively big, every batch fermentation all needs to carry out to feed, sterilizing, inoculate, put material, cleaning etc. and operate, and operation is loaded down with trivial details, the non-fermented cycle is longer, and production efficiency is lower; And the follow-up separating technology multiplex whizzer of tradition is separated, which is because being limited by the impact of whizzer separation factor, in sepn process, mycelium suffers that shearing force is destroyed, part effective constituent is lost in filtrate to cause quality product and the decline of receipts rate, separating obtained filtrate is more muddy, the inside is containing more broken mycelia, cause outer row liquor C OD higher, if this filtrate reaches sewage drainage standard, need through complicated sewage disposal, Sewage treatment systems workload is too big, brings serious economical load to enterprise.
Summary of the invention
It is an object of the invention to the deficiency produced to solve above-mentioned hirsutella sinensis fungal powder, thus devise the green novel method of a kind of rapid deep zone liquid state fermentation production hirsutella sinensis fungal powder, the method adopts semicontinuous fermentation process can decrease process procedure, shorten the non-fermented time, improve plant factor, reduce production cost; Adopt again efficient ceramic membrane filter, it is to increase the receipts rate of mycelium simultaneously, reduce the mycelium entering filtrate, reduce the COD of filtrate, reduce sewage load; Do not destroy again cell itself, ensure whole recovery of effective constituent in born of the same parents. This invention not only can obtain the mycelium of the China pilose spore of high-quality fast, turn improves the receipts rate of mycelium in sepn process, adds economic benefit; Again reduce COD in waste water, alleviate Sewage treatment systems pressure, add social benefit.
The object of the invention is mainly achieved in the following ways:
A method for rapid deep zone liquid state fermentation production hirsutella sinensis fungal powder, the method carries out according to following step:
1) China pilose spore semicontinuous fermentation liquid preparation: the fermention medium after China pilose spore seed liquor is inoculated into sterilizing carries out deep layer liquid state fermentation and obtains China pilose spore fermentation culture, described deep layer liquid state fermentation inoculum size is the 10～20% of volume content, ventilating ratio is 1:0.5-1.5vvm, tank pressure 0.02～0.05Mpa, at 15～20 DEG C after fermentation culture 10-30 days, release the fermented liquid of the 10-50% of incubator volume, fill into the fermention medium after the sterilizing of equal volume, continue fermentation 1-10 days; So constantly putting material, feed supplement, each feed supplement continues fermentation 1-10 days, and repeated multiple times, the operational cycle can be adjusted to 12-70 days according to number of times, obtains China pilose spore fermented liquid; Just hirsutella sinensis fungal seed liquor can be prepared after activating in conventional manner by China pilose spore bacteria strain before fermention medium after hirsutella sinensis fungal seed liquor is inoculated into sterilizing; Preferred fermentation condition is ventilating ratio is 1:0.5vvm, tank pressure 0.04Mpa, fermentation culture 25 days at 20 DEG C;
2) China pilose spore fermented liquid ceramic membrane filter: by step 1) in the China pilose spore fermented liquid of gained obtained micro-filter dialyzate and the mycelium of fermented liquid by microfiltration membrane, collect and obtain mycelium;
3) cryogenic vacuum is dried: by step 2) in the mycelium of gained be-0.065～-0.095Mpa in vacuum tightness, be dried under 60～75 DEG C of conditions.
Described China pilose spore seed liquor prepares by the following method: China pilose spore seed culture medium consists of glucose 1-3%, peptone 0.5-2%, milk powder 0.1-1%, wheat bran 1-3%, potassium primary phosphate 0.1-0.5%, magnesium sulfate heptahydrate 0.1-0.5%, defoamer 0.01-0.05%; Adjust ph 5.5-6.5; Culture condition is: ventilating ratio is 1:0.5-1.5vvm, tank pressure 0.02～0.05Mpa, culture temperature 15～20 DEG C, culture cycle 5-15 days.
Preferred China pilose spore seed liquor prepares by the following method: activated shaking flask hirsutella sinensis fungal bacterial classification is accessed seed culture medium, and China pilose spore seed culture medium consists of glucose 2%, peptone 0.8%, milk powder 0.5%, wheat bran 1.5%, potassium primary phosphate 0.3%, magnesium sulfate heptahydrate 0.3%, defoamer 0.02-0.03%; Adjust ph 5.5-6.5; Culture condition is: ventilating ratio is 1:0.5-1vvm, tank pressure 0.03～0.04Mpa, culture temperature 18～20 DEG C, culture cycle 5-10 days.
China pilose spore fermention medium consists of glucose 1-3%, Semen Maydis powder 1-3%, peptone 0.5-2%, yeast powder 0.1-1%, wheat bran 1-3%, potassium primary phosphate 0.1-0.5%, magnesium sulfate heptahydrate 0.1-0.5%, defoamer 0.01-0.05%; Adjust ph 5.5-6.5.
The method step 2) described in microfiltration membrane be ceramic membrane or organic membrane, its aperture is at 0.05～0.5 μm, and service temperature is 50～60 DEG C, operating pressure: entering pressure is 2～4bar, and going out pressure is 0～5bar.
Preferred the method step 2) described in microfiltration membrane be ceramic membrane or organic membrane, its aperture is at 50nm, and service temperature is 50～60 DEG C, operating pressure: enter pressure for 3bar, goes out pressure for 1bar.
The present invention is applicable to any China pilose spore bacterial classification (Ophiocordycepssinensis), and as being preserved in by Chinese industrial Microbiological Culture Collection administrative center, preserving number CICC14088,14099,14114 etc. all can use.
China pilose spore bacterial strain of the present invention is not limited to certain concrete manufacturer, commercially available all can use.
The present invention is that rapid deep zone liquid state fermentation production hirsutella sinensis fungal powder provides a kind of new method. The method has following outstanding advantage:
1, fermentation period is short: traditional batch fermentation technique, every batch fermentation all needs to carry out to feed, sterilizing, inoculate, put the operations such as material, cleaning, operation is loaded down with trivial details, total fermentation period is long, and adopt semicontinuous fermentation process to decrease process procedure, shorten non-fermented time and fermentation time, it is to increase plant factor and ferment strength, reduce production cost.
2, receipts rate is improved: adopt 0.05～10 μm, microfiltration membrane aperture, significantly improve mycelia yield.
3, improve the quality of products: in tradition centrifuge separation process, Hirsutella sinensis mycelium suffers that shearing force is destroyed, part effective constituent is lost in filtrate to cause mycelium effective constituent to reduce, this technique adopts micro-filtrate membrane filtration, operating process can not produce shearing force, effectively ensure mycelium effective constituent.
3, filtrate COD is reduced, alleviate environmental protection pressure: the sleeping spiral shell of tradition is separated owing to mycelium rejection is low, and part mycelium enters filtrate, causes COD content in filtrate to increase, present method separation factor height, mycelium rejection greatly being increased, filtrate is clarified, and in filtrate, mycelium is few, the dialyzate COD obtained reduces greatly, its COD index can reach direct emission standard, and only sewage disposal greatly reduces sewage disposal expense just for enterprise reduces by 30 tons for the treatment of capacities every day.
4, cleaning production continuously, energy consumption is low: the inventive method can be produced continuously, film and support equipment long service life thereof, convenience for device maintenance, less investment, and operating process noise-less pollution is easy to clean.
Accompanying drawing explanation
Fig. 1 is traditional technology route map described in embodiment.
Embodiment
Below by way of specific embodiment, the present invention is described further. But the detail of embodiment is only for explaining the present invention, it should not be interpreted as limited overall technical solution.
Embodiment 1
1) China pilose spore bacterial strain (Ophiocordycepssinensis) is provided by Chinese industrial Microbiological Culture Collection administrative center and identifies, deposit number CICC14088. By the China pilose spore bacterial strain seed liquor kind cultivated to seed culture medium, inoculum size is 10%, seed culture medium consists of glucose 2%, peptone 0.8%, milk powder 0.5%, wheat bran 1.5%, potassium primary phosphate 0.3%, magnesium sulfate heptahydrate 0.3%, defoamer 0.02%, adjust ph 6.0; Culture condition is: ventilating ratio is 1:0.5vvm, tank pressure 0.03Mpa, culture temperature 18～20 DEG C, cultivates and can obtain seed liquor in about 7 days. It is inoculated into 50L fermention medium after sterilizing by having cultivated seed liquor, inoculum size 15%, fermention medium consists of glucose 2%, Semen Maydis powder 1.5%, peptone 1%, yeast powder 0.5%, wheat bran 2%, potassium primary phosphate 0.3%, magnesium sulfate heptahydrate 0.3%, defoamer 0.03%; Adjust ph 6.0; Ventilating ratio is 1:0.5vvm, tank pressure 0.04Mpa, and at 20 DEG C, fermentation culture is after 25 days, releases the fermented liquid of 20L, fills into the fermention medium after 20L sterilizing, continues fermentation 10 days; So constantly putting material, feed supplement, 3 times repeatedly, fermentation ends can obtain fermented liquid 115L, and fermentation period is 55 days.
2) micro-filtration filter: China pilose spore fermented liquid 20L, 20L, 20L and the 55L after fermenting four times respectively is by micro-filtration filter, obtain China pilose spore wet mycelium 2.2kg, 2.15kg, 2.10kg, 5.5kg (water content about 75%) respectively, micro-filter dialyzate 17.8Kg, 17.85Kg, 17.9Kg and 49.5Kg, the transmittance of dialyzate is 82-85%, COD is 40-60mg/L. Micro-filtration filter mould material used is aperture is the ceramic membrane of 50nm, and concrete processing condition are: service temperature is 50～60 DEG C, enters pressure for 3.0bar, goes out pressure for 1.0bar, and pressure difference is 2.0bar.
3) cryodrying: 2.2kg, 2.15kg, 2.10kg, 5.5kg China pilose spore wet mycelium step 2 obtained enters vacuum drying oven dries, obtain the Hirsutella sinensis mycelium of 0.567kg, 0.554kg, 0.541kg and 1.418kg moisture content about 3%, this mycelium adenosine is 0.12-0.16%, mannitol content 10-13%, aminoacids content is at 35-40%. According to WS3-181 (Z-60)-2006 (Z)-2010 standard method is checked. Concrete drying process condition is: drying temperature 65 DEG C, and vacuum tightness is-0.085Mpa.
Traditional processing technology and the embodiment of the present invention 1 handicraft product Yield and quality contrast table
Can find out from above table and adopt traditional technology fermentation in 64 days 2 batches, mycelium 2.836kg can be obtained; And adopt the total fermentation period of technique of the present invention to be 55 days, and the mycelium 3.08kg of 4 batches can be obtained, fermentation period is the 86% of traditional technology, and to obtain mycelium production be traditional 111.5%, and mycelium quality product is higher than traditional technology produces. Therefore adopt technique of the present invention both can shorten fermentation period, it is to increase seed output and quality, reduce production cost.
Embodiment 2
1) China pilose spore bacterial strain (Ophiocordycepssinensis) is provided by Chinese industrial Microbiological Culture Collection administrative center and identifies, deposit number CICC14114.By the China pilose spore bacterial strain seed liquor kind cultivated to seed culture medium, inoculum size is 10%, seed culture medium consists of glucose 2%, peptone 0.8%, milk powder 0.5%, wheat bran 1.5%, potassium primary phosphate 0.3%, magnesium sulfate heptahydrate 0.3%, defoamer 0.02%, adjust ph 6.0; Culture condition is: ventilating ratio is 1:0.5vvm, tank pressure 0.03Mpa, culture temperature 18～20 DEG C, cultivates and can obtain seed liquor in about 7 days. It is inoculated into 100L fermention medium after sterilizing by having cultivated seed liquor, inoculum size 15%, fermention medium consists of glucose 2.5%, Semen Maydis powder 2%, peptone 1%, yeast powder 0.7%, wheat bran 2.5%, potassium primary phosphate 0.3%, magnesium sulfate heptahydrate 0.3%, defoamer 0.03%; Adjust ph 6.5; Ventilating ratio is 1:0.5vvm, tank pressure 0.04Mpa, and at 20 DEG C, fermentation culture is after 25 days, releases the fermented liquid of 40L, fills into the fermention medium after 40L sterilizing, continues fermentation 10 days; So constantly putting material, feed supplement, 3 times repeatedly, fermentation ends can obtain fermented liquid 230L, and fermentation period is 55 days.
2) micro-filtration filter: China pilose spore fermented liquid 40L, 40L, 40L and the 110L after fermenting four times respectively is by micro-filtration filter, obtain China pilose spore wet mycelium 4.4kg, 4.3kg, 4.2kg, 11kg (water content about 75%) respectively, micro-filter dialyzate 35.6Kg, 35.7Kg, 35.8Kg and 99Kg, the transmittance of dialyzate is 83-87%, COD is 40-60mg/L. Micro-filtration filter mould material used is aperture is the ceramic membrane of 50nm, and concrete processing condition are: service temperature is 50～60 DEG C, enters pressure for 3.0bar, goes out pressure for 1.0bar, and pressure difference is 2.0bar.
3) cryodrying: 4.4kg, 4.3kg, 4.2kg, 11kg China pilose spore wet mycelium step 2 obtained enters vacuum drying oven dries, obtain the Hirsutella sinensis mycelium of 1.135kg, 1.108kg, 1.08kg and 2.842kg moisture content about 3%, this mycelium adenosine is 0.12-0.16%, mannitol content 10-13%, aminoacids content is at 35-40%. According to WS3-181 (Z-60)-2006 (Z)-2010 standard method is checked. Concrete drying process condition is: drying temperature 65 DEG C, and vacuum tightness is-0.085Mpa.
Traditional processing technology and the embodiment of the present invention 2 handicraft product Yield and quality contrast table
Can find out from above table and adopt traditional technology fermentation in 64 days 2 batches, mycelium 5.522kg can be obtained; And adopt the total fermentation period of technique of the present invention to be 55 days, and the mycelium 6.165kg of 4 batches can be obtained, fermentation period is traditional 86%, and to obtain mycelium production be traditional 111.6%, and mycelium quality product is higher than traditional technology produces. Therefore adopt technique of the present invention both can shorten fermentation period, it is to increase seed output and quality, reduce production cost.
Claims (7)
1. a method for rapid deep zone liquid state fermentation production hirsutella sinensis fungal powder, is characterized in that the method comprises the steps:
1) China pilose spore semicontinuous fermentation liquid preparation: the fermention medium after China pilose spore seed liquor is inoculated into sterilizing carries out deep layer liquid state fermentation and obtains China pilose spore fermentation culture, inoculum size is the 10～20% of volume content, ventilating ratio is 1:0.5-1.5vvm, tank pressure 0.02～0.05Mpa, at 15～20 DEG C after fermentation culture 10-30 days, release the fermented liquid of the 10-50% of incubator volume, fill into the fermention medium after the sterilizing of equal volume, continue fermentation 1-10 days;So constantly putting material, feed supplement, each feed supplement continues fermentation 1-10 days, repeated multiple times, obtains China pilose spore fermented liquid;
2) China pilose spore fermented liquid ceramic membrane filter: by step 1) in the China pilose spore fermented liquid of gained obtained micro-filter dialyzate and the mycelium of fermented liquid by microfiltration membrane, collect mycelium;
3) cryogenic vacuum is dried: by step 2) in the mycelium of gained be-0.065～-0.095Mpa in vacuum tightness, be dried under 70～85 DEG C of conditions.
2. the method for rapid deep zone liquid state fermentation production hirsutella sinensis fungal powder according to claim 1, is characterized in that step 1) described in fermentation condition be ventilating ratio be 1:0.5vvm, tank pressure 0.04Mpa, fermentation culture 25 days at 20 DEG C.
3. the method for rapid deep zone liquid state fermentation production hirsutella sinensis fungal powder according to claim 1, is characterized in that described China pilose spore seed liquor prepares by the following method: China pilose spore seed culture medium consists of glucose 1-3%, peptone 0.5-2%, milk powder 0.1-1%, wheat bran 1-3%, potassium primary phosphate 0.1-0.5%, magnesium sulfate heptahydrate 0.1-0.5%, defoamer 0.01-0.05%; Adjust ph 5.5-6.5; Culture condition is: ventilating ratio is 1:0.5-1.5vvm, tank pressure 0.02～0.05Mpa, culture temperature 15～20 DEG C, culture cycle 5-15 days.
4. the method for rapid deep zone liquid state fermentation production hirsutella sinensis fungal powder according to claim 3, it is characterized in that described China pilose spore seed liquor prepares by the following method: activated shaking flask hirsutella sinensis fungal bacterial classification is accessed seed culture medium, and China pilose spore seed culture medium consists of glucose 2%, peptone 0.8%, milk powder 0.5%, wheat bran 1.5%, potassium primary phosphate 0.3%, magnesium sulfate heptahydrate 0.3%, defoamer 0.02-0.03%; Adjust ph 5.5-6.5; Culture condition is: ventilating ratio is 1:0.5-1vvm, tank pressure 0.03～0.04Mpa, culture temperature 18～20 DEG C, culture cycle 5-10 days.
5. the method for rapid deep zone liquid state fermentation production hirsutella sinensis fungal powder according to claim 1, is characterized in that China pilose spore fermention medium consists of glucose 1-3%, Semen Maydis powder 1-3%, peptone 0.5-2%, yeast powder 0.1-1%, wheat bran 1-3%, potassium primary phosphate 0.1-0.5%, magnesium sulfate heptahydrate 0.1-0.5%, defoamer 0.01-0.05%; Adjust ph 5.5-6.5.
6. the method for rapid deep zone liquid state fermentation production hirsutella sinensis fungal powder according to claim 1, it is characterized in that the microfiltration membrane described in the method is ceramic membrane or organic membrane, its aperture is at 0.05～0.5 μm, service temperature is 50～60 DEG C, operating pressure: entering pressure is 2～4bar, going out pressure is 0～5bar.
7. the method for rapid deep zone liquid state fermentation production hirsutella sinensis fungal powder according to claim 6, it is characterized in that the microfiltration membrane described in the method is ceramic membrane or organic membrane, its aperture is at 50nm, and service temperature is 50～60 DEG C, operating pressure: enter pressure for 3bar, goes out pressure for 1bar.
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| CN112795493B (en) * | 2021-04-02 | 2023-07-07 | 青海珠峰冬虫夏草原料有限公司 | Method for improving yield of fermented cordyceps sinensis mycelia |
| CN116333963A (en) * | 2021-12-23 | 2023-06-27 | 江苏神华药业有限公司 | Rapid submerged liquid fermentation culture method for hirsutella sinensis |
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