CN106190866A - A kind of reuse method of Cordyceps fermented waste fluid - Google Patents
A kind of reuse method of Cordyceps fermented waste fluid Download PDFInfo
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- CN106190866A CN106190866A CN201610656157.9A CN201610656157A CN106190866A CN 106190866 A CN106190866 A CN 106190866A CN 201610656157 A CN201610656157 A CN 201610656157A CN 106190866 A CN106190866 A CN 106190866A
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- cordyceps
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
Abstract
The present invention with Cordyceps fungus through liquid submerged fermentation the waste liquid that produces after extracting mycelia as raw material, eliminate the inhibiting substances of suppression Cordyceps fungus growth after treatment, and remain the growth of Cordyceps fungus and breed favourable material, reach both to decrease the waste to resource, solve again the fermentation liquid problem to environment, and improve the comprehensive good result of the yield of Cordyceps mycelium.
Description
Technical field
The present invention relates to biological fermentation engineering field, in particular to the recycling of a kind of Cordyceps fermented waste fluid
Method.
Background technology
Cordyceps has another name called Cordyceps, Cordyceps, Cordyceps sinensis.Vespertilionidae is colonized in for Clavicipitaceae plant Cordyceps fungus
The complex of sclerotium is sat and in larva corpse be to son on insect larvae body.Main product in Sichuan, Qinghai, Tibet, the ground such as Yunnan.Summer
To front and back taking, dry or dry, raw use.Modern study shows, Cordyceps has important edible, medicinal and other is economical
It is worth, containing nutrient substance such as abundant cordycepic acid, cordycepin and each amino acids in Cordyceps, there is expansion trachea, calmness, resist
Each bacterioid, effect of blood pressure lowering.In recent years, the U.S., Japan etc. report that Cordyceps has raising human body and restrains tumor ability in succession,
Human body rejection effect can be strengthened.After liberation China also to manually the introducing a fine variety of edible fungi and Cordyceps, tame, cultivate, heredity, should
With etc. aspect carried out a lot of research.
Major part cultivated by China's Cordyceps at present is that artificial culture is cultivated, due to by geographical environment, weather, water and soil and worm
The effect of natural conditions of each side such as grass product kind, the quality of Cordyceps and yield extremely unstable, it is difficult to realize scale industrialization.
Use submerged fermentation to produce Cordyceps mycelium, on the one hand be easy to from culture fluid, separate and extract various active ingredient thing
Matter, such as, extract the utilities such as cordycepic acid, cordycepin, Cordyceps polysaccharide and have the Cordycepin of antitumaous effect and multiple ammonia
Base acid, cultivates on the other hand utilizing the fermentation tank of big volume and can improve yield, it is ensured that the material source of screening medication, alleviates
The pressure of worm grass resources.
During the artificial culture of Cordyceps, fermented after mycelium the most only account for the 3% left of fermented product total amount
The right side, and remaining fermentation liquid of 97% is discarded as waste liquid, find after fermentation broth contents is analyzed, the fermentation of Cordyceps polysaccharide
Waste liquid is contained within multiple proteins, aminoacid, vitamin, D-MANNOSE, ergosterol and trace element, if directly by worm
Grass fermented waste fluid discards, and the most not only wastes resource, will also result in environmental pollution simultaneously.Therefore, this area is needed one badly and can be returned
Receipts utilize Chinese caterpillar fungus fermentation waste liquid, reduce the wasting of resources and the method for environmental pollution.
The waste liquid that biofermentation has been produced by this area circulates as the ingredient of culture medium the most again
Use, but based on fermented bacterium and the difference of product, how to recycle biofermentation waste liquid, the most fixing method, also
Existing method can not be used for reference and directly apply mechanically, but need the concrete condition according to strain fermentation waste liquid, by constantly tasting
Examination just can draw.In the waste liquid that biofermentation produces in addition to comprising above-mentioned effective ingredient and some residual sugars, also include one
The material of growth of microorganism can be suppressed a bit, in order to realize recycling of fermented waste fluid, need these inhibiting substances are carried out
Process, eliminate the suppression to growth breeding.
Summary of the invention
For above-mentioned technical problem, the invention provides a kind of can efficient recovery and utilize Cordyceps fermented waste fluid
Method, will return, according to special ratios, the method that culture medium recycles by fermented waste fluid after adding specific nutrient substance.
Specifically comprise the following steps that
(1) collect Cordyceps fungus through liquid submerged fermentation the waste liquid that produces after extracting mycelia, use plate-and-frame filtration
Device filters, and collects filtrate A；
(2) with booster pump, filtrate A is pumped into three grades of mechanical accurate filters of deslagging to process, obtain liquor B；
(3) liquor B is concentrated into the 1/10 of its original volume, obtains concentrated broth C；
(4) concentrated broth C is carried out ultrafiltration, obtain ultrafiltration fermentation liquid D；
(5) in ultrafiltration fermentation liquid D, add new nutrient substance, obtain the fermentation liquid E of reusable edible；
(6) the fermentation liquid E of reusable edible is returned three-stage culture medium.
Specifically, the phorozoon that Cordyceps fungus is (Cordyceps the sinensis)-China's quilt in step (1)
Hair spore (Hirsutella sinensis Liu, Guo, Yu et Zeng)；Pressure 0.35-0.45Mpa during plate-and-frame filtration, warp
300 mesh filter-cloth filterings；
In step (2), the detailed process of use deslagging three grades mechanical accurate filter secondary filter filtrate A is, at deslagging
100 μm one-level accurate filters, two grades of accurate filters of 10 μm and two grades of essences of 1 μm are passed sequentially through in three grades of mechanical accurate filters
Close filter process；
In step (3), thickening temperature is 60-70 DEG C；
In step (4), the molecular cut off of the ultrafilter membrane that ultrafiltration uses is 500 dalton, intake pressure 1.2-1.3Mpa,
Ultrafiltrate temperature 20-30 DEG C, pH value is 6.2～6.8；
In step (5), the nutrient substance added by percent mass is as follows: Semen Viciae fabae powder 5.5-6%, Semen Maydis powder 4.5-5%, Portugal
Grape sugar 6-7%, yeast extract 2.0-2.5%, peptone 1.2-1.8%, magnesium sulfate 0.4-0.8%, dipotassium hydrogen phosphate 0.05-
0.1%.
In step (6), former three-stage culture medium is 3:1-5:1 with the volume ratio of the fermentation liquid E of addition.
The invention have the advantages that and good effect:
The present invention with Cordyceps fungus through liquid submerged fermentation the waste liquid that produces after extracting mycelia as raw material, through place
Eliminate the inhibiting substances of suppression Cordyceps fungus growth after reason, and remain the growth of Cordyceps fungus and breeding favourable
Material, be effectively utilized in Cordyceps fermented waste fluid the nutrient substance of remaining, and the fermentation liquid reclaimed by recycling,
It is found surprisingly that it can improve the yield of Cordyceps mycelium.With it, reached both to decrease the waste to resource,
Solve again the fermentation liquid problem to environment, and improve the comprehensive good effect of the yield of Cordyceps mycelium
Really, reduce the cost that enterprise produces simultaneously, improve the added value of industry, have broad application prospects.
Accompanying drawing explanation
Accompanying drawing 1 recycles the schematic flow sheet of Cordyceps fermented waste fluid for the present invention.
Detailed description of the invention
Embodiment 1
(1) collect Cordyceps fungus through liquid submerged fermentation the waste liquid that produces after extracting mycelia, use plate-and-frame filtration
Device filters, and collects filtrate A；Cordyceps fungus is the phorozoon-China pilose spore of (Cordyceps sinensis)
(Hirsutella sinensis Liu, Guo, Yu et Zeng)；Pressure 0.35Mpa during plate-and-frame filtration, through 300 mesh filter clothes
Filter；Concrete sweat can be found in the description of Chinese patent CN201210389147.5.
(2) with booster pump, filtrate A is pumped into three grades of mechanical accurate filters of deslagging to process, obtain liquor B, the row of use
The detailed process of slag three grades mechanical accurate filter secondary filter filtrate A is, in three grades of mechanical accurate filters of deslagging successively
Processed by 100 μm one-level accurate filters, two grades of accurate filters of 10 μm and two grades of accurate filters of 1 μm；
(3) liquor B being concentrated into the 1/10 of its original volume, obtain concentrated broth C, thickening temperature is 60 DEG C；
(4) concentrated broth C is carried out ultrafiltration, obtain ultrafiltration fermentation liquid D, the molecular cut off of the ultrafilter membrane that ultrafiltration uses
Being 500 dalton, intake pressure 1.3Mpa, ultrafiltrate temperature 20 DEG C, pH value is 6.2；
(5) in ultrafiltration fermentation liquid D, add new nutrient substance, obtain the fermentation liquid E of reusable edible, by percent mass
The nutrient substance added is as follows: Semen Viciae fabae powder 6%, Semen Maydis powder 4.5%, glucose 7%, yeast extract 2.0%, peptone 1.2%, sulfur
Acid magnesium 0.4%, dipotassium hydrogen phosphate 0.1%；
(6) the fermentation liquid E of reusable edible returning three-stage culture medium and continues fermentation, former three-stage culture medium is sent out with addition
The volume ratio of ferment liquid E is 3:1, separates and recovers the mycelium of generation after level Four is fermented, and dries, and pulverizes, measures mycelial
Yield and product quality.
Embodiment 2
(1) collect Cordyceps fungus through liquid submerged fermentation the waste liquid that produces after extracting mycelia, use plate-and-frame filtration
Device filters, and collects filtrate A；Cordyceps fungus is the phorozoon-China pilose spore of (Cordyceps sinensis)
(Hirsutella sinensis Liu, Guo, Yu et Zeng)；Pressure 0.4Mpa during plate-and-frame filtration, through 300 mesh filter cloth mistakes
Filter；Concrete sweat can be found in the description of Chinese patent CN201210389147.5.
(2) with booster pump, filtrate A is pumped into three grades of mechanical accurate filters of deslagging to process, obtain liquor B, the row of use
The detailed process of slag three grades mechanical accurate filter secondary filter filtrate A is, in three grades of mechanical accurate filters of deslagging successively
Processed by 100 μm one-level accurate filters, two grades of accurate filters of 10 μm and two grades of accurate filters of 1 μm；
(3) liquor B being concentrated into the 1/10 of its original volume, obtain concentrated broth C, thickening temperature is 65 DEG C；
(4) concentrated broth C is carried out ultrafiltration, obtain ultrafiltration fermentation liquid D, the molecular cut off of the ultrafilter membrane that ultrafiltration uses
Being 500 dalton, intake pressure 1.2Mpa, ultrafiltrate temperature 25 DEG C, pH value is 6.5；
(5) in ultrafiltration fermentation liquid D, add new nutrient substance, obtain the fermentation liquid E of reusable edible, by percent mass
The nutrient substance added is as follows: Semen Viciae fabae powder 5.8%, Semen Maydis powder 4.8%, glucose 6.5%, yeast extract 2.2%, peptone
1.5%, magnesium sulfate 0.6%, dipotassium hydrogen phosphate 0.07%；
(6) the fermentation liquid E of reusable edible returning three-stage culture medium and continues fermentation, former three-stage culture medium is sent out with addition
The volume ratio of ferment liquid E is 4:1, separates and recovers the mycelium of generation after level Four is fermented, and dries, and pulverizes, measures mycelial
Yield and product quality.
Embodiment 3
(1) collect Cordyceps fungus through liquid submerged fermentation the waste liquid that produces after extracting mycelia, use plate-and-frame filtration
Device filters, and collects filtrate A；Cordyceps fungus is the phorozoon-China pilose spore of (Cordyceps sinensis)
(Hirsutella sinensis Liu, Guo, Yu et Zeng)；Pressure 0.45Mpa during plate-and-frame filtration, through 300 mesh filter clothes
Filter；Concrete sweat can be found in the description of Chinese patent CN201210389147.5.
(2) with booster pump, filtrate A is pumped into three grades of mechanical accurate filters of deslagging to process, obtain liquor B, the row of use
The detailed process of slag three grades mechanical accurate filter secondary filter filtrate A is, in three grades of mechanical accurate filters of deslagging successively
Processed by 100 μm one-level accurate filters, two grades of accurate filters of 10 μm and two grades of accurate filters of 1 μm；
(3) liquor B being concentrated into the 1/10 of its original volume, obtain concentrated broth C, thickening temperature is 70 DEG C；
(4) concentrated broth C is carried out ultrafiltration, obtain ultrafiltration fermentation liquid D, the molecular cut off of the ultrafilter membrane that ultrafiltration uses
Being 500 dalton, intake pressure 1.3Mpa, ultrafiltrate temperature 30 DEG C, pH value is 6.8；
(5) in ultrafiltration fermentation liquid D, add new nutrient substance, obtain the fermentation liquid E of reusable edible, by percent mass
The nutrient substance added is as follows: Semen Viciae fabae powder 5.5%, Semen Maydis powder 5%, glucose 6%, yeast extract 2.5%, peptone 1.8%, sulfur
Acid magnesium 0.8%, dipotassium hydrogen phosphate 0.05%；
(6) the fermentation liquid E of reusable edible returning three-stage culture medium and continues fermentation, former three-stage culture medium is sent out with addition
The volume ratio of ferment liquid E is 5:1, separates and recovers the mycelium of generation after level Four is fermented, and dries, and pulverizes, measures mycelial
Yield and product quality.
Comparative example 1
Phorozoon-the China pilose spore three grades of configuration Cordyceps fungus adds culture fluid, and concrete composition is (by percentage matter
Amount): Semen Viciae fabae powder 6%, Semen Maydis powder 4.5%, glucose 7%, yeast extract 2.0%, peptone 1.2%, magnesium sulfate 0.4%, phosphoric acid
Hydrogen dipotassium 0.1%, remaining is water；This is added culture fluid and adds three-stage culture medium continuation fermentation, former three-stage culture medium and addition
The volume ratio adding culture fluid be 3:1, after level Four fermented, separate and recover the mycelium of generation, dry, pulverize, measure bacterium
The yield of filament and product quality.
Comparative example 2
Phorozoon-China pilose spore third stage culture the liquid of configuration Cordyceps fungus, concrete composition is (by percent mass):
Semen Viciae fabae powder 5.8%, Semen Maydis powder 4.8%, glucose 6.5%, yeast extract 2.2%, peptone 1.5%, magnesium sulfate 0.6%, phosphoric acid
Hydrogen dipotassium 0.07%, remaining is water；This is added culture fluid and adds three-stage culture medium continuation fermentation, former three-stage culture medium and addition
The volume ratio adding culture fluid be 4:1, after level Four fermented, separate and recover the mycelium of generation, dry, pulverize, measure bacterium
The yield of filament and product quality.
Comparative example 3
Phorozoon-China pilose spore third stage culture the liquid of configuration Cordyceps fungus, concrete composition is (by percent mass):
Semen Viciae fabae powder 5.5%, Semen Maydis powder 5%, glucose 6%, yeast extract 2.5%, peptone 1.8%, magnesium sulfate 0.8%, phosphoric acid hydrogen two
Potassium 0.05%, remaining is water；This is added culture fluid and adds three-stage culture medium continuation fermentation, former three-stage culture medium and the benefit of addition
The volume ratio adding culture fluid is 5:1, separates and recovers the mycelium of generation after level Four is fermented, and dries, and pulverizes, and measures mycelium
Yield and product quality.
Comparative example 4
Ferment according to the method in CN201210389147.5 embodiment 5, when three grade fermemtation without culture medium,
After level Four is fermented, separate and recover the mycelium of generation, dry, pulverize, measure mycelial yield and product quality.
By assay method described in " Pharmacopoeia of People's Republic of China version in 2005 ", obtained by embodiment 1-3
Product carries out contrast test with the product obtained by comparative example 1-4, and result see table:
By said determination result it is found that fermentation liquid returns interpolation after treatment in three-stage culture medium, both reduced
Waste to resource, solves again the fermentation liquid problem to environment, is the most also found surprisingly that and can improve the winter
The mycelial yield of worm summer grass fermentation gained and quality, thus it is speculated that the fermented waste fluid that reason is Cordyceps of this effect occurs
In containing to the growth of Cordyceps fungus and breeding or to producing the favourable material composition of effective metabolite.
Claims (9)
1. the method reclaimed and utilize Cordyceps fermented waste fluid, it is characterised in that it is useless that described method includes collecting fermentation
Liquid, filters, fine straining, concentrates, ultrafiltration, adds nutrient substance, returns culture medium.
A kind of method reclaiming and utilize Cordyceps fermented waste fluid the most according to claim 1, it is characterised in that described
Specifically comprising the following steps that of method
(1) collect Cordyceps fungus through liquid submerged fermentation the waste liquid that produces after extracting mycelia, use plate filter to enter
Row filters, and collects filtrate A；
(2) with booster pump, filtrate A is pumped into three grades of mechanical accurate filters of deslagging to process, obtain liquor B；
(3) liquor B is concentrated into the 1/10 of its original volume, obtains concentrated broth C；
(4) concentrated broth C is carried out ultrafiltration, obtain ultrafiltration fermentation liquid D；
(5) in ultrafiltration fermentation liquid D, add new nutrient substance, obtain the fermentation liquid E of reusable edible；
(6) the fermentation liquid E of reusable edible is returned three-stage culture medium.
A kind of method reclaiming and utilize Cordyceps fermented waste fluid the most according to claim 2, it is characterised in that: step
(1) the Cordyceps fungus in is the phorozoon-China pilose spore (Hirsutella of (Cordyceps sinensis)
Sinensis Liu, Guo, Yu et Zeng).
A kind of method reclaiming and utilize Cordyceps fermented waste fluid the most according to claim 2, it is characterised in that: step
(1) pressure 0.35-0.45Mpa during plate-and-frame filtration in, through 300 mesh filter-cloth filterings.
A kind of method reclaiming and utilize Cordyceps fermented waste fluid the most according to claim 2, it is characterised in that: step
(2), in, the detailed process of use deslagging three grades mechanical accurate filter secondary filter filtrate A is, accurate at deslagging three grades machinery
Pass sequentially through in filter at 100 μm one-level accurate filters, two grades of accurate filters of 10 μm and two grades of accurate filters of 1 μm
Reason.
A kind of method reclaiming and utilize Cordyceps fermented waste fluid the most according to claim 2, it is characterised in that: step
(3), in, thickening temperature is 60-70 DEG C.
A kind of method reclaiming and utilize Cordyceps fermented waste fluid the most according to claim 2, it is characterised in that: step
Suddenly, in (4), the molecular cut off of the ultrafilter membrane that ultrafiltration uses is 500 dalton, intake pressure 1.2-1.3Mpa, ultrafiltrate temperature
20-30 DEG C, pH value is 6.2～6.8.
A kind of method reclaiming and utilize Cordyceps fermented waste fluid the most according to claim 2, it is characterised in that: step
(5) in, the nutrient substance added by percent mass is as follows: Semen Viciae fabae powder 5.5-6%, Semen Maydis powder 4.5-5%, glucose 6-7%, ferment
Female cream 2.0-2.5%, peptone 1.2-1.8%, magnesium sulfate 0.4-0.8%, dipotassium hydrogen phosphate 0.05-0.1%.
A kind of method reclaiming and utilize Cordyceps fermented waste fluid the most according to claim 2, it is characterised in that: step
(6), in, former three-stage culture medium is 3:1-5:1 with the volume ratio of the fermentation liquid E of addition.
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Cited By (4)
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| CN107586728A (en) * | 2017-10-26 | 2018-01-16 | 青海珠峰冬虫夏草工程技术研究有限公司 | A kind of cordyceps sinensis nutrient solution and preparation method thereof |
| CN108676729A (en) * | 2018-05-28 | 2018-10-19 | 杭州雪域生物技术有限公司 | A kind of culture media composition containing fermentation mother liquor and preparation method thereof and fermentation process |
| CN112586454A (en) * | 2020-12-15 | 2021-04-02 | 青海珠峰冬虫夏草工程技术研究有限公司 | Large-scale breeding method for cordyceps sinensis host hepialus armoricanus larvae |
| CN114591878A (en) * | 2022-05-10 | 2022-06-07 | 齐鲁工业大学 | Process for preparing mariculture tail water treatment microbial inoculum and tail water treatment method |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107586728A (en) * | 2017-10-26 | 2018-01-16 | 青海珠峰冬虫夏草工程技术研究有限公司 | A kind of cordyceps sinensis nutrient solution and preparation method thereof |
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| CN112586454A (en) * | 2020-12-15 | 2021-04-02 | 青海珠峰冬虫夏草工程技术研究有限公司 | Large-scale breeding method for cordyceps sinensis host hepialus armoricanus larvae |
| CN114591878A (en) * | 2022-05-10 | 2022-06-07 | 齐鲁工业大学 | Process for preparing mariculture tail water treatment microbial inoculum and tail water treatment method |
| CN114591878B (en) * | 2022-05-10 | 2022-08-19 | 齐鲁工业大学 | Process for preparing mariculture tail water treatment microbial inoculum and tail water treatment method |
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| CN106190866B (en) | 2019-09-20 |
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