CN105802855A - Method for rapidly producing antrodia comphorata powder through deep liquid state fermentation - Google Patents
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Abstract
The invention discloses a method for rapidly producing antrodia comphorata powder through deep liquid state fermentation.The method includes the steps that fermentation liquor which has rich and massive mycelia is rapidly acquired from an antrodia comphorata strain through a semi-continuous fermentation process, and then the mycelia of antrodia comphorata are obtained after being separated from the fermentation liquor through micro-filtration membrane treatment.A traditional antrodia comphorata intermittent batch fermentation process is changed, and micro-filtration of an efficient ceramic membrane is adopted for replacing a centrifuge separation mode on the aspect of separation of the mycelia.The defects that links of the traditional batch fermentation process is abundant, the middle interval time is long, and equipment efficiency is low are overcome through the semi-continuous fermentation process.The fermentation liquor is filtered through the efficient ceramic membrane, the mycelia entering filtrate can be reduced, the yield of the mycelia can be increased, COD of the filtrate is reduced, the sewage treatment amount is reduced, cells are not damaged, and it is guaranteed that all effective ingredients in the cells are recycled.Compared with a traditional process, the method has remarkable economic benefits, social benefits and environmental protection benefits.
Description
Technical field
The invention belongs to microbial technology field, be specifically related to a kind of method quickly utilizing deep layer liquid fermentation technology to produce fungal mycelium, a kind of method that rapid deep zone liquid fermentation produces Antrodia camphorata mycopowder 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 the operation such as every batch fermentation is required for carrying out feeding, sterilizing, inoculation, blowing, cleaning, operation is loaded down with trivial details, and fermentation period is longer, and production efficiency is relatively low；Though fed-batch fermentation can pass through feed supplement supplements the deficiency of nutrient or precursor, but due to the continuous accumulation of unwanted metabolic products, Product formation is finally checked unavoidably；Continuous fermentation relatively batch fermentation and fed-batch fermentation produce intensity and are greatly improved, but are subject to the pollution of miscellaneous bacteria, and strain is easily degenerated, and equipment investment is relatively big, and Fermentation Substance Concentration is relatively low；In semicontinuous fermentation process, fill into fresh culture again by bleeding off Partial fermentation liquid, be possible not only to supplement nutrient and precursor, and metabolism nuisance is diluted, thus being conducive to the continuation of product to synthesize.
The application of semicontinuous fermentation process can play the effect alleviated Product inhibiton and avoid metabolic by-product to accumulate, improve the culture environment of microorganism, help to maintain stablizing of thalline vigor, and for some secondary metabolite, its highest throughput rate only can be only achieved under some transient condition.Adopting semicontinuous fermentation process to be possible not only to make this transient condition repeatedly occur, but also can improve the utilization rate of equipment, therefore semicontinuous fermentation process has important application in the production of secondary metabolite.
Antrodia camphorata has another name called the raw Antrodia of Camphor tree, Antrodia camphorata, Antrodia Camphorata, red Antrodia camphorata, Camphor tree mushroom etc., is subordinate to Eumycota, and Basidiomycotina, Hymenomycetes, Polyporaceae, Antrodia belongs to perennial Mushrooms, is the peculiar one of TaiWan, China famous and precious pharmaceutical fungi and edible fungi rare, still undeveloped.The heartwood inwall that to be only grown between TaiWan, China mountain area height above sea level 450～2000 meters distinctive Cinnamomum kanahirai hay tree trunk rotten, or the surface of the Cinnamomum kanahirai hay timber darkness humidity of withered lodging, it has been found that really not easily.Antrodia camphorata is as the distinctive strain of TaiWan, China, and in natural environment, Cinnamomum kanahirai hay tree is its unique host, and general fungus can not grow thereon.Owing to Antrodia camphorata can cause Cinnamomum kanahirai hay tree central cavity rotten, add that people is to Cinnamomum kanahirai hay tree excessive and random lumbering, cause Cinnamomum kanahirai hay tree quantity quite rare, be listed in one-level child care seeds at present.In the last hundred years, Cinnamomum kanahirai hay tree is rare to see, and the Cinnamomum kanahirai hay tree that can grow Antrodia Camphorata is then less, and it results in wild Antrodia camphorata resource critical shortage.Therefore Antrodia camphorata is very precious, is referred to as " Ganoderma " in Hong Kong, Macao and Taiwan, and TaiWan, China then title among the people Antrodia camphorata is " ruby in forest ".
Containing multiple physiologically active ingredient in Antrodia camphorata, such as polysaccharide, triterpenoid compound, sudismase (SOD) adenosine, protein (containing immune protein), multivitamin, trace element (calcium, phosphorus, paper mulberry), nucleic acid, agglutinin, aminoacid, cholesterol, lignin, blood pressure stabilization material etc..It physiological active functions possessed has antitumor, enhancing immunity, antiviral, antiallergic, resisting hypertension, suppression platelet aggregation, blood pressure lowering, cholesterol reducing, antibacterium, the liver protecting etc..
Tradition Antrodia camphorata liquid-state fermentation technology mostly is the batch process of intermittence, and the human and material resources consumption of batch process is relatively big, the operation such as every batch fermentation is required for carrying out feeding, sterilizing, inoculation, blowing, cleaning, operation is loaded down with trivial details, the non-fermented cycle is longer, and production efficiency is relatively low；And follow-up separating technology tradition multiplace centrifuge is easily separated, which is because being limited by the impact of centrifuge separation factor, in separation process, mycelium suffers that shearing force is destroyed, part effective ingredient is lost in filtrate and causes that product quality and yield decline, separating obtained filtrate is more muddy, the inside is containing more broken mycelia, cause that outer row liquor C OD is higher, if this filtrate reaches sewage drainage standard, need through complicated sewage disposal, sewage disposal system workload is too big, brings serious financial burden to enterprise.
Summary of the invention
The invention aims to solve the deficiency that above-mentioned Antrodia camphorata mycopowder produces, thus devising a kind of rapid deep zone liquid fermentation to produce the novel environment-friendly process of Antrodia camphorata mycopowder, this technique adopts semicontinuous fermentation process can decrease process procedure, shorten the non-fermented time, improve utilization rate of equipment and installations, reduce production cost；Adopt again efficient ceramic membrane filter simultaneously, improve mycelial yield, reduce the mycelium entering filtrate, reduce the COD of filtrate, reduce sewage load；Do not destroy again cell itself, it is ensured that in born of the same parents, the whole of effective ingredient reclaim.This invention not only can quickly obtain the mycelium of the Antrodia camphorata of high-quality, improves again mycelial yield in separation process, adds economic benefit；Reduce again COD in waste water, alleviate sewage disposal system pressure, add social benefit.
The object of the invention realizes mainly by the following manner:
A kind of rapid deep zone liquid fermentation produces the method for Antrodia camphorata mycopowder, and the method carries out as steps described below:
1) prepared by Antrodia camphorata semicontinuous fermentation liquid:: the fermentation medium after Antrodia camphorata seed liquor is inoculated into sterilizing carries out deep layer liquid fermentation and obtains Antrodia camphorata fermentation culture, inoculum concentration is the 10～20% of volume content, ventilating ratio is 1:0.1-1.0vvm, tank pressure 0.02～0.05Mpa, after 25～30 DEG C of bottom fermentations are cultivated 4-8 days, release the fermentation liquid of the 10-50% of incubator volume, fill into the fermentation medium after the sterilizing of equal volume, continue fermentation 1-3 days；So constantly blowing, feed supplement, each feed supplement continues fermentation 1-3 days, repeated multiple times, obtains Antrodia camphorata fermentation liquid, and the operation cycle can be adjusted to 6-20 days according to number of times；Antrodia camphorata seed liquor is prepared after Antrodia camphorata bacterial strain can be activated in conventional manner before fermentation medium after seed liquor is inoculated into sterilizing；Preferred fermentation condition is ventilating ratio is 1:0.5vvm, tank pressure 0.04Mpa, cultivates 7 days at 27 DEG C of bottom fermentations；
2) Antrodia camphorata fermentation liquid ceramic membrane filter: by step 1) in the Antrodia camphorata fermentation liquid of gained obtained microfiltration dialysis solution and the mycelium of fermentation liquid by micro-filtration 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, be dried under 70～85 DEG C of conditions, gained dried object is Antrodia Camphorata mycelium.
Described Antrodia camphorata seed liquor prepares by the following method: the Antrodia camphorata seed culture medium of employing consists of glucose 1-3%, peptone 0.1-1%, yeast powder 0.5-2%, potassium dihydrogen phosphate 0.1-0.5%, Magnesium sulfate heptahydrate 0.1-0.5%, Oleum Glycines 0.01-0.05%；Regulate pH value 4.0-5.0；Condition of culture is: ventilating ratio is 1:0.1-1.0vvm, tank pressure 0.02～0.05Mpa, cultivation temperature 25～30 DEG C, cultivation cycle 4-8 days.
Preferred Antrodia camphorata seed liquor prepares by the following method: the Antrodia camphorata seed culture medium of employing consists of glucose 2%, peptone 0.3-0.4%, yeast powder 1.3-1.8%, potassium dihydrogen phosphate 0.3%, Magnesium sulfate heptahydrate 0.15-0.2%, Oleum Glycines 0.02%；Regulate pH value 4.8-5.0；Condition of culture is: ventilating ratio is 1:0.4vvm, tank pressure 0.03Mpa, cultivation temperature 26～28 DEG C, cultivation cycle 7 days.
Described fermentation medium consists of glucose 1-3%, peptone 0.1-1%, yeast powder 0.5-2%, potassium dihydrogen phosphate 0.1-0.5%, Magnesium sulfate heptahydrate 0.1-0.5%, Oleum Glycines 0.01-0.05%；Regulate pH value 4.0-5.0.Preferred fermentation medium consists of glucose 2.5%, peptone 0.6-1%, yeast powder 1.8-2%, potassium dihydrogen phosphate 0.3-0.5%, Magnesium sulfate heptahydrate 0.2-0.5%, Oleum Glycines 0.03-0.05%；Regulate pH value 4.7-4.8.
Micro-filtration membrane described in the method is ceramic membrane or organic membrane, and its aperture is at 0.05～0.5 μm, and operation temperature is 50～60 DEG C, and operating pressure: entrance pressure is 2～4bar, going out pressure is 0～5bar.Preferred micro-filtration membrane is ceramic membrane or organic membrane, and its aperture is at 50nm, and operation temperature is 50～60 DEG C, operating pressure: entrance pressure is 3bar, goes out pressure for 1bar.
The present invention is applicable to any Antrodia camphorata strain (Antrodiacamphorata), all can use as being preserved in institute of microbiology of the Chinese Academy of Sciences (strain coding CGMCCNO.0543) etc..
Antrodia camphorata 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 fermentation production Antrodia camphorata mycopowder provides a kind of new method.The method has following prominent advantage:
1, fermentation period is short: traditional batch process, the operations such as every batch fermentation is required for carrying out feeding, sterilizing, inoculation, blowing, cleaning, operation is loaded down with trivial details, the non-fermented cycle is long, and adopt semicontinuous fermentation process to decrease process procedure, shorten non-fermented time and fermentation time, improve utilization rate of equipment and installations and ferment strength, reduce production cost.
2, yield is improved: adopt 0.05～10 μm of micro-filtration membrane aperture, significantly improve mycelia yield.
3, product quality is improved: in conventional centrifuge separation process, cephalosporium sinensis filament suffers that shearing force is destroyed, part effective ingredient is lost in filtrate and causes that mycelium effective ingredient reduces, this technique adopts micro-filtrate membrane filtration, operating process will not produce shearing force, be effectively ensured mycelium effective ingredient.
3, filter liquor COD is reduced, alleviate environmental protection pressure: the sleeping spiral shell of tradition separates owing to mycelium rejection is low, and part mycelium enters filtrate, causes that in filtrate, COD content increases, this method separation factor is high, mycelium rejection is greatly increased, and filtrate is clarified, and in filtrate, mycelium is few, the dialysis solution COD obtained is substantially reduced, its COD index can reach direct discharge standard, only sewage disposal every day just reduces by 30 tons for the treatment of capacities for enterprise, greatly reduces sewage disposal expense.
4, cleaning produces continuously, and energy consumption is low: the inventive method can produce continuously, film and corollary equipment long service life thereof, convenience for device maintenance, small investment, and operating process noise-less pollution is easy to clean.
Accompanying drawing explanation
Fig. 1 is conventional processing routes figure described in embodiment.
Detailed description of the invention
The present invention is further illustrated below by way of specific embodiment.But the detail of embodiment is only used for explaining the present invention, should not be construed as limited overall technical solution.
Embodiment 1
1) Antrodia camphorata bacterial strain (Antrodiacamphorata) is preserved in institute of microbiology of the Chinese Academy of Sciences (strain coding CGMCCNO.0543).By cultured Antrodia camphorata bacterial strain seed liquor kind to seed culture medium, inoculum concentration is 10%, and seed culture medium consists of glucose 2%, peptone 0.3%, yeast powder 1.3%, potassium dihydrogen phosphate 0.3%, Magnesium sulfate heptahydrate 0.15%, Oleum Glycines 0.02%；Regulate pH value 4.8.Condition of culture is: ventilating ratio is 1:0.4vvm, tank pressure 0.03Mpa, cultivation temperature 26-28 DEG C, cultivates and can obtain seed liquor in about 7 days.Being inoculated into 50L fermentation medium after sterilizing by having cultivated seed liquor, inoculum concentration 15%, fermentation medium consists of glucose 2.5%, peptone 0.5%, yeast powder 1.5%, potassium dihydrogen phosphate 0.3%, Magnesium sulfate heptahydrate 0.2%, Oleum Glycines 0.03%；Regulate pH value 4.8；Ventilating ratio is 1:0.5vvm, tank pressure 0.04Mpa, after 27 DEG C of bottom fermentations are cultivated 7 days, releases the fermentation liquid of 20L, fills into the fermentation medium after 20L sterilizing, continues fermentation 2.5 days；So constantly blowing, feed supplement, 3 times repeatedly, fermentation ends can obtain fermentation liquid 115L, and fermentation period is 14.5 days.
2) microfiltration filters: Antrodia camphorata fermentation liquid 20L, 20L, 20L and 55L after fermenting four times respectively are filtered by microfiltration, respectively obtain Antrodia camphorata wet mycelium 0.83kg, 0.828kg, 0.824kg, 2.288kg (water content about 76%), microfiltration dialysis solution 19.17Kg, 19.172Kg, 19.176Kg and 52.712Kg, the light transmittance of dialysis solution is 84-87%, COD is 30-60mg/L.Membrane material used by microfiltration filtration is aperture is the ceramic membrane of 50nm, and concrete technology condition is: operation temperature is 50～60 DEG C, and entrance pressure is 3.0bar, goes out pressure for 1.0bar, and pressure differential is 2.0bar.
3) cold drying: 0.83kg, 0.828kg, 0.824kg, 2.288kg Antrodia camphorata wet mycelium step 2 obtained enters vacuum drying oven and dries, obtain the Antrodia Camphorata mycelium of 0.206kg, 0.2kg, 0.194kg and 0.564kg moisture about 3.8%, this mycelium polysaccharides content 3-6%, total triterpene contents 2-4%.According to Q/JSFF0012S-2013 standard method inspection.Concrete drying process condition is: baking temperature 65 DEG C, and vacuum is-0.085Mpa.
Traditional processing technology and the embodiment of the present invention 1 handicraft product Yield and quality contrast table
Can be seen that employing traditional handicraft is fermented 2 batches for 16 days from above table, mycelium 1.048kg can be obtained；And adopting the total fermentation period of present invention process is 14.5 days, the mycelium 1.164kg of 4 batches can be obtained, it is thus achieved that mycelium production is traditional 110.1%, and mycelium product quality is higher than conventionally produced.Therefore adopt present invention process both can shorten fermentation period, improve yield and quality, reduce production cost.
Embodiment 2
1) Antrodia camphorata bacterial strain (Antrodiacamphorata) is preserved in institute of microbiology of the Chinese Academy of Sciences (strain coding CGMCCNO.0543).By cultured Antrodia camphorata bacterial strain seed liquor kind to seed culture medium, inoculum concentration is 10%, and seed culture medium consists of glucose 2.0%, peptone 0.4%, yeast powder 1.8%, potassium dihydrogen phosphate 0.3%, Magnesium sulfate heptahydrate 0.2%, Oleum Glycines 0.02%；Regulate pH value 4.8.Condition of culture is: ventilating ratio is 1:0.4vvm, tank pressure 0.03Mpa, cultivation temperature 26-28 DEG C, cultivates and can obtain seed liquor in about 7 days.Being inoculated into 100L fermentation medium after sterilizing by having cultivated seed liquor, inoculum concentration 15%, fermentation medium consists of glucose 2.5%, peptone 0.6%, yeast powder 1.8%, potassium dihydrogen phosphate 0.3%, Magnesium sulfate heptahydrate 0.2%, Oleum Glycines 0.03%；Regulate pH value 4.7；Ventilating ratio is 1:0.5vvm, tank pressure 0.04Mpa, after 27 DEG C of bottom fermentations are cultivated 7 days, releases the fermentation liquid of 20L, fills into the fermentation medium after 40L sterilizing, continues fermentation 2.5 days；So constantly blowing, feed supplement, 3 times repeatedly, fermentation ends can obtain fermentation liquid 232L, and fermentation period is 14.5 days.
2) microfiltration filters: Antrodia camphorata fermentation liquid 40L, 40L, 40L and 112L after fermenting four times respectively are filtered by microfiltration, respectively obtain Antrodia camphorata wet mycelium 2.0kg, 1.96kg, 1.92kg, 5.58kg (water content about 80%), microfiltration dialysis solution 38Kg, 38.04Kg, 38.08Kg and 106.42Kg, the light transmittance of dialysis solution is 82-85%, COD is 25-60mg/L.Membrane material used by microfiltration filtration is aperture is the ceramic membrane of 50nm, and concrete technology condition is: operation temperature is 50～60 DEG C, and entrance pressure is 3.0bar, goes out pressure for 1.0bar, and pressure differential is 2.0bar.
3) cold drying: 2.0kg, 1.96kg, 1.92kg, 5.58kg Antrodia camphorata wet mycelium step 2 obtained enters vacuum drying oven and dries, obtain the Antrodia Camphorata mycelium of 0.410kg, 0.406kg, 0.402kg and 1.152kg moisture about 3.5%, this mycelium polysaccharides content 3-6%, total triterpene contents 2-4%.According to Q/JSFF0012S-2013 standard method inspection.Concrete drying process condition is: baking temperature 65 DEG C, and vacuum is-0.085Mpa.
Traditional processing technology and the embodiment of the present invention 2 handicraft product Yield and quality contrast table
Can be seen that employing traditional handicraft is fermented 2 batches for 16 days from above table, mycelium 2.038kg can be obtained；And adopting the total fermentation period of present invention process is 14.5 days, the mycelium 2.37kg of 4 batches can be obtained, it is thus achieved that mycelium production is the 116.3% of traditional handicraft, and mycelium product quality is higher than conventionally produced.Therefore adopt present invention process both can shorten fermentation period, improve yield and quality, reduce production cost.
Claims (8)
1. the method that rapid deep zone liquid fermentation produces Antrodia camphorata mycopowder, is characterized in that the method comprises the steps:
1) prepared by Antrodia camphorata semicontinuous fermentation liquid: the fermentation medium after Antrodia camphorata seed liquor is inoculated into sterilizing carries out deep layer liquid fermentation and obtains Antrodia camphorata fermentation culture, inoculum concentration is the 10～20% of volume content, ventilating ratio is 1:0.1-1.0vvm, tank pressure 0.02～0.05Mpa, after 25～30 DEG C of bottom fermentations are cultivated 4-8 days, release the fermentation liquid of the 10-50% of incubator volume, fill into the fermentation medium after the sterilizing of equal volume, continue fermentation 1-3 days；So constantly blowing, feed supplement, each feed supplement continues fermentation 1-3 days, repeated multiple times, obtains Antrodia camphorata fermentation liquid；
2) Antrodia camphorata fermentation liquid ceramic membrane filter: by step 1) in the Antrodia camphorata fermentation liquid of gained obtained microfiltration dialysis solution and the mycelium of fermentation liquid by micro-filtration 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, be dried under 70～85 DEG C of conditions.
2. the method that rapid deep zone liquid fermentation according to claim 1 produces Antrodia camphorata mycopowder, is characterized in that described Antrodia camphorata seed liquor prepares by the following method: the Antrodia camphorata seed culture medium of employing consists of glucose 1-3%, peptone 0.1-1%, yeast powder 0.5-2%, potassium dihydrogen phosphate 0.1-0.5%, Magnesium sulfate heptahydrate 0.1-0.5%, Oleum Glycines 0.01-0.05%；Regulate pH value 4.0-5.0；Condition of culture is: ventilating ratio is 1:0.1-1.0vvm, tank pressure 0.02～0.05Mpa, cultivation temperature 25～30 DEG C, cultivation cycle 4-8 days.
3. the method that rapid deep zone liquid fermentation according to claim 2 produces Antrodia camphorata mycopowder, is characterized in that described Antrodia camphorata seed liquor prepares by the following method: the Antrodia camphorata seed culture medium of employing consists of glucose 2%, peptone 0.3-0.4%, yeast powder 1.3-1.8%, potassium dihydrogen phosphate 0.3%, Magnesium sulfate heptahydrate 0.15-0.2%, Oleum Glycines 0.02%；Regulate pH value 4.8-5.0；Condition of culture is: ventilating ratio is 1:0.4vvm, tank pressure 0.03Mpa, cultivation temperature 26～28 DEG C, cultivation cycle 7 days.
4. the method that rapid deep zone liquid fermentation according to claim 1 produces Antrodia camphorata mycopowder, is characterized in that described fermentation medium consists of glucose 1-3%, peptone 0.1-1%, yeast powder 0.5-2%, potassium dihydrogen phosphate 0.1-0.5%, Magnesium sulfate heptahydrate 0.1-0.5%, Oleum Glycines 0.01-0.05%；Regulate pH value 4.0-5.0.
5. the method that rapid deep zone liquid fermentation according to claim 4 produces Antrodia camphorata mycopowder, is characterized in that described fermentation medium consists of glucose 2.5%, peptone 0.6-1%, yeast powder 1.8-2%, potassium dihydrogen phosphate 0.3-0.5%, Magnesium sulfate heptahydrate 0.2-0.5%, Oleum Glycines 0.03-0.05%；Regulate pH value 4.7-4.8.
6. the method that rapid deep zone liquid fermentation according to claim 1 produces Antrodia camphorata mycopowder, is characterized in that described fermentation condition be ventilating ratio is 1:0.5vvm, and tank pressure 0.04Mpa cultivates 7 days at 27 DEG C of bottom fermentations.
7. the method that rapid deep zone liquid fermentation according to claim 1 produces Antrodia camphorata mycopowder, it is characterized in that the micro-filtration membrane described in the method is ceramic membrane or organic membrane, its aperture is at 0.05～0.5 μm, and operation temperature is 50～60 DEG C, operating pressure: entrance pressure is 2～4bar, going out pressure is 0～5bar.
8. the method that rapid deep zone liquid fermentation according to claim 7 produces Antrodia camphorata mycopowder, it is characterized in that the micro-filtration membrane described in the method is ceramic membrane or organic membrane, its aperture is at 50nm, and operation temperature is 50～60 DEG C, operating pressure: entrance pressure is 3bar, goes out pressure for 1bar.
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| CN106865804A (en) * | 2017-03-23 | 2017-06-20 | 福建凯立生物制品有限公司 | A kind of Zhongshengmycin mother medicine clean preparation method |
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