CN110250496A - Cordyceps sinensis powder and preparation method thereof - Google Patents
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Abstract
The present invention provides a kind of cordyceps sinensis powder and preparation method thereof, belong to Cordyceps militaris effective component extractive technique field.Specifically, the described method comprises the following steps: A, which will activate Cordyceps militaris and be inoculated into the container containing fermentation medium, to ferment, and is then separated by solid-liquid separation and obtains supernatant;Then the PH to 4~6 for the supernatant that B regulating step A is obtained is added complex enzyme and is digested to obtain enzymolysis liquid, the complex enzyme is the compound of dextranase, cellulase and hemicellulase;Enzymolysis liquid filtering that C obtains step B, it is dry after up to cordyceps sinensis powder.For cordycepin content in cordyceps sinensis powder of the invention up to 30wt% or more, functional oligosaccharide content is higher than 20wt%, and effective component is much higher than presently commercially available level, may be used in liquid health care product.
Description
Technical field
The invention belongs to Cordyceps militaris effective component extractive technique fields, and in particular to a kind of cordyceps sinensis powder and preparation method thereof.
Background technique
Cordyceps militaris also known as northern Chinese caterpillar Fungus, Cordceps militaris etc. belong to xenogenesis with cordyceps sinensis, are Cordyceps kinds, belong to ascus
Bacterium subphylum Cordyceps is that it colonizes in answering for the stroma that is grown on Insecta Lepidoptera noctuid pupal cell and ossified pupal cell
It is fit.Years of researches show that Cordyceps militaris has a variety of pharmacological activity such as immunological regulation is antitumor, antiviral, anti-infective, extensively
For tumour, immunologic hypofunction including HIV infection etc. in a variety of diseases.
It is related studies have shown that riched Cordyceps Militaris is containing various bioactive components, such as cordycepin, Cordyceps sinensis polysaccharide, adenosine etc..Its
Middle cordycepin is first nucleoside antibiotic separated from fungi.In terms of Chinese medicine medical knowledge angle, cordycepin have and
The same yin-yang of cordyceps sinensis is the same as the function of mending with bidirectional modulation total balance of the body;In terms of doctor trained in Western medicine medical knowledge angle, cordycepin has anti-
Tumour, antibacterial, antiviral, immunological regulation, improves metabolism, removes the multiple pharmacological effects such as free radical anti-aging, has good
Potential applicability in clinical practice.Cordyceps sinensis polysaccharide can adjust immune function of human body, enhance disease resistance, have anticancer anticancer activity, right
The reduction of leucocyte caused by various diseases has good auxiliary therapeutic action, also has antidiabetic function.But due to cordyceps sinensis
Polysaccharide is not soluble in water, and meeting ethyl alcohol can settle, and cordycepin is in contrast soluble easily in water, so, extract worm simultaneously from Cordyceps militaris
Careless element and Cordyceps sinensis polysaccharide have certain difficulty.
In recent years, domestic more as the drug of raw material and health food development using Cordyceps militaris, it is most commonly seen with cordyceps sinensis powder, but
It is that cordycepin content, simultaneously because the dissolubility of Cordyceps sinensis polysaccharide is very poor, limits worm generally between 3%-10% in cordyceps sinensis powder
Application of the grass meal in liquid health care product.
Summary of the invention
The purpose of the present invention is to provide a kind of preparation method of cordyceps sinensis powder, this method can with high efficiency extraction cordycepin and
Functional oligosaccharide, simple process, it is easy to accomplish industrial automation production.
The present invention provides a kind of preparation methods of cordyceps sinensis powder, comprising the following steps:
A, which will activate Cordyceps militaris and be inoculated into the container containing fermentation medium, to ferment, and is then separated by solid-liquid separation and obtains supernatant;
Then the PH to 4~6 for the supernatant that B regulating step A is obtained is added complex enzyme and is digested to obtain enzymolysis liquid, institute
State the compound that complex enzyme is dextranase, cellulase and hemicellulase;
Enzymolysis liquid filtering that C obtains step B, it is dry after up to cordyceps sinensis powder.
Wherein, the preparation method of above-mentioned cordyceps sinensis powder, in step A, the Cordyceps militaris is classified as Cordyceps militaris Cordyceps
Militaris, number are CICC 14014;
The activation Cordyceps militaris is to be inoculated into Cordyceps militaris on slant medium to carry out Spore cultivation, cultivation temperature be 24~
27 DEG C, incubation time is 5~7 days, and the slant medium is made as follows: removing 200~250g of potato of skin, cuts
Block, adds water 1000mL to boil 20~40min, and 5~6 layers of gauze filter off potato ball, filtrate is complemented to 1000mL, adds glucose
15~20g, KH2PO41~5g, MgSO4·7H20.5~5g of O, vitaminB10 .1~0.8g, 10~25g of agar divide after fusing
Dress, sterilizing.
Wherein, the preparation method of above-mentioned cordyceps sinensis powder, in step A, the inoculum concentration of the activation Cordyceps militaris is 5~30%v/v.
Wherein, the preparation method of above-mentioned cordyceps sinensis powder, in step A, the fermentative medium formula is as follows: glucose 30~
60g/L, 2~10g/L of yeast extract, 2~10g/L of corn pulp, 1~5g/L of soybean protein, KH2PO40.5~5g/L, K2HPO4·
3H20.5~5g/L of O, MgSO4·7H20.5~5g/L of O, solvent are water, and pH is 5~7;
The condition of the fermentation are as follows: 24~28 DEG C of fermentation temperature, fermentation time is 3~4 weeks;
The step of separation of solid and liquid, is as follows: removing the residue of fermentation liquid using 2~8 layers of filtered through gauze, then will obtain
Crude liquid 10~20min is centrifuged at 4000~10000r/min, sterilize.
Wherein, the preparation method of above-mentioned cordyceps sinensis powder, in step B, the pH of the supernatant is 4.5~6;
The additive amount of the dextranase is 10~100U/g Cordyceps sinensis polysaccharide, and the additive amount of cellulase is 10~100U/g
Cordyceps sinensis polysaccharide, the additive amount of hemicellulase are 10~100U/g Cordyceps sinensis polysaccharide;
Preferably, the additive amount of the dextranase is 20~50U/g Cordyceps sinensis polysaccharide, the additive amount of cellulase is 20~
50U/g Cordyceps sinensis polysaccharide, the additive amount of hemicellulase are 20~50U/g Cordyceps sinensis polysaccharide.
Wherein, the preparation method of above-mentioned cordyceps sinensis powder, the enzymatic hydrolysis meet at least one of following:
The temperature of the enzymatic hydrolysis is 35~60 DEG C;Preferably, the temperature of the enzymatic hydrolysis is 50~55 DEG C;
The time of the enzymatic hydrolysis is 1~4h;Preferably, 1~2h of time of the enzymatic hydrolysis;
The revolving speed of the enzymatic hydrolysis is 50~200r/min;Preferably, 100~200r/min of revolving speed of the enzymatic hydrolysis.
Wherein, the preparation method of above-mentioned cordyceps sinensis powder, in step C, the nanofiltration after being filtered into ultrafiltration;What the ultrafiltration used
The molecular cut off of film is 2000~3000D, and ultrafiltration pressure is 0.5~3MPa, and ultrafiltration flow velocity is 30~90L/h;The nanofiltration
The molecular cut off of the film used is 150~200D, and nanofiltration pressure is 0.5~3MPa, and nanofiltration flow velocity is 10~40L/h.
Wherein, the preparation method of above-mentioned cordyceps sinensis powder, in step C, the ultrafiltration pressure is 1.5~3MPa, and ultrafiltration flow velocity is
40~75L/h;Nanofiltration pressure is 2~3MPa, and nanofiltration flow velocity is 10~30L/h.
Wherein, the preparation method of above-mentioned cordyceps sinensis powder, in step C, the drying is concentration drying or spray drying.
The present invention also provides the cordyceps sinensis powder that the preparation method using above-mentioned cordyceps sinensis powder is prepared.
The beneficial effects of the present invention are:
It is few that insoluble Cordyceps sinensis polysaccharide in fermentation liquid can be hydrolyzed to soluble functionality by the complex enzyme that the present invention uses
Sugar, percent hydrolysis is up to 90% or more, and the molecular weight of oligosaccharides is controllably in 500~900D;The separating technology of product of the present invention is simple, easily
In realization industrial automation production;Cordycepin content in cordyceps sinensis powder of the present invention is up to 30wt% or more, functional oligosaccharide content
Higher than 20wt%, effective component is much higher than presently commercially available level, may be used in liquid health care product.
Specific embodiment
Specifically, a kind of preparation method of cordyceps sinensis powder, comprising the following steps: A is inoculated into Cordyceps militaris is activated containing fermentation
It ferments in the container of culture medium, is then separated by solid-liquid separation and obtains supernatant;The PH to 4~6 for the supernatant that B regulating step A is obtained, so
Complex enzyme is added afterwards to be digested to obtain enzymolysis liquid, the complex enzyme is answering for dextranase, cellulase and hemicellulase
Close object;Enzymolysis liquid filtering that C obtains step B, it is dry after up to cordyceps sinensis powder.
In the present invention, the enzyme activity of dextranase is defined as under the conditions of 50 DEG C, 4.8 pH, and 1min hydrolyzes barley beta-glucan
Enzyme amount needed for generating 1 μm of ol reduced sugar is 1 enzyme-activity unit.The enzyme activity of cellulase is defined as in 50 DEG C, 4.8 condition of pH
Under, enzyme amount needed for 1min hydrocellulose generates 1 μm of ol reduced sugar is 1 enzyme-activity unit.The enzyme activity of hemicellulase defines
For under the conditions of 50 DEG C, 4.8 pH, enzyme amount needed for 1min hydrolyzed hemicellulose generates 1 μm of ol reduced sugar is 1 enzyme-activity unit.
In the present invention, the concentration furnace drying method are as follows: with revolving evaporator at 50~80 DEG C of temperature, vacuum degree 0.05~
Separating liquid is concentrated into sticky shape under 0.1MPa, then under the conditions of 60~95 DEG C, drying to constant weight, then it is crushed to 60~
100 mesh.The spray drying process are as follows: by feed liquid spray dryer, under the conditions of 85~120 DEG C of temperature, 1~4L/h of flow velocity
Carry out spray drying.
Below by test example and embodiment, the present invention is described in further detail, but does not therefore protect the present invention
Scope limitation is protected among the embodiment described range.
Embodiment 1
Cordyceps militaris activation: Cordyceps militaris being inoculated on slant medium and carries out Spore cultivation, and 25 DEG C are cultivated 6 days, the work
Change culture medium to be prepared as follows: removing the potato 230g of skin, be cut into 1cm square fritter, water 1000mL is added to boil
30min, 4 layers of gauze filter off potato ball, filtrate are complemented to 1000mL, adds 20g glucose, 3g KH2PO4, 1.5g
MgSO4·7H2O, 0.4g vitamin B1,15g agar dispense after fusing, 121 DEG C of 20~30min of sterilizing.
Fermentation: scraping inclined-plane Cordyceps militaris spore is placed in the shaking flask equipped with fermentation medium, wherein fermentative medium formula
Are as follows: glucose 35g/L, yeast extract 5g/L, corn pulp 8g/L, soybean protein 2g/L, KH2PO4 1g/L、K2HPO4·3H2O 1g/
L、MgSO4·7H2O 0.5g/L, pH5.5;25 DEG C stationary culture 3 weeks, 121 DEG C of sterilizing 20min.
Filtering: fermentation liquid residue is filtered out with 4 layers of gauze;Clear liquid 5000r/min is centrifuged 10min again, takes supernatant fraction;
Enzyme hydrolysis: the pH to 4.5 of supernatant is adjusted before hydrolysis with the HCL aqueous solution of 3M;Then add dextranase, fibre respectively
Plain enzyme and hemicellulase: 20U/g, 20U/g and 20U/g are tieed up, must be hydrolyzed after hydrolyzing 1h under the conditions of revolving speed 100r/min, 50 DEG C
Liquid.
UF membrane: removing the macromolecular substances such as albumen, nucleic acid in hydrolyzate with ultrafiltration membrane (molecular cut off 3000D), then
The small-molecule substances such as inorganic salts are removed with nanofiltration membrane (molecular cut off 150D), wherein ultra-filtration conditions are as follows: pressure 2MPa, flow
60L/h;Nanofiltration condition are as follows: pressure 3MPa, flow 30L/h, the feed liquid after collecting separation.
Dry powder processed: under the conditions of 55 DEG C of temperature, vacuum degree 0.08Mpa that the feed liquid after UF membrane is dense with revolving evaporator
It is reduced to sticky shape, then drying to constant weight under the conditions of 80 DEG C, is finally crushed to 80 mesh with pulverizer up to cordyceps sinensis powder.
Cordycepin content 25wt% in the cordyceps sinensis powder of acquisition, oligosaccharide content are not less than 20wt%, and oligosaccharide molecular amount is 900D,
And cordyceps sinensis powder is completely solvable.
Embodiment 2
Cordyceps militaris activation and fermentation step are the same as embodiment 1.
Filtering: fermentation liquid residue is filtered out with 6 layers of gauze;Clear liquid 8000r/min is centrifuged 5min again, takes supernatant fraction;
Enzyme hydrolysis: the pH to 5.5 of supernatant is adjusted before hydrolysis with the HCL aqueous solution of 2M;Then add dextranase, fibre respectively
Plain enzyme and hemicellulase: 30U/g, 30U/g and 40U/g are tieed up, obtains water after 1.5h is hydrolyzed under the conditions of revolving speed 200r/min, 55 DEG C
Solve liquid.
UF membrane: removing the macromolecular substances such as albumen, nucleic acid in hydrolyzate with ultrafiltration membrane (molecular cut off 2000D), then
The small-molecule substances such as inorganic salts are removed with nanofiltration membrane (molecular cut off 200D), wherein ultra-filtration conditions are as follows: pressure 3MPa, flow
50L/h;Nanofiltration condition are as follows: pressure 2MPa, flow 15L/h, the feed liquid after collecting separation.
Dry powder processed: under the conditions of 75 DEG C of temperature, vacuum degree 0.1MPa that the feed liquid after UF membrane is dense with revolving evaporator
It is reduced to sticky shape, then drying to constant weight under the conditions of 95 DEG C, is finally crushed to 60 mesh with pulverizer up to cordyceps sinensis powder.
Cordycepin content 30wt% in the cordyceps sinensis powder of acquisition, oligosaccharide content are not less than 25wt%, molecular weight 540D, and worm
Grass meal is completely solvable.
Embodiment 3
Cordyceps militaris activation, fermentation and filtration step are the same as embodiment 1.
Enzyme hydrolysis: the pH to 5 of supernatant is adjusted before hydrolysis with the HCL aqueous solution of 2M;Then add dextranase, fiber respectively
Plain enzyme and hemicellulase: 40U/g, 50U/g and 50U/g must be hydrolyzed after hydrolyzing 1h under the conditions of revolving speed 150r/min, 50 DEG C
Liquid.
UF membrane: removing the macromolecular substances such as albumen, nucleic acid in hydrolyzate with ultrafiltration membrane (molecular cut off 2500D), then
The small-molecule substances such as inorganic salts are removed with nanofiltration membrane (molecular cut off 150D), wherein ultra-filtration conditions are as follows: pressure 3MPa, flow
75L/h;Nanofiltration condition are as follows: pressure 2.5MPa, flow 30L/h, the feed liquid after collecting separation.
Dry powder processed: by the feed liquid spray dryer after UF membrane, under the conditions of 105 DEG C of temperature, flow velocity 1.5L/h into
Row spray drying is up to cordyceps sinensis powder.
Cordycepin content 32wt% in the cordyceps sinensis powder of acquisition, oligosaccharide content be not less than 20wt%, molecular weight be 500~
900D, and cordyceps sinensis powder is completely solvable.
Embodiment 4
Cordyceps militaris activation, fermentation and filtration step are the same as embodiment 1.
Enzyme hydrolysis: the pH to 6 of supernatant is adjusted before hydrolysis with the HCL aqueous solution of 2M;Then add dextranase, fiber respectively
Plain enzyme and hemicellulase: 50U/g, 30U/g and 20U/g hydrolyze the hydrolyzate of 2h under the conditions of revolving speed 120r/min, 55 DEG C;
UF membrane: removing the macromolecular substances such as albumen, nucleic acid in hydrolyzate with ultrafiltration membrane (molecular cut off 2000D), then
The small-molecule substances such as inorganic salts are removed with nanofiltration membrane (molecular cut off 150D), wherein ultra-filtration conditions are as follows: pressure 1.5MPa, stream
Measure 40L/h;Nanofiltration condition are as follows: pressure 2MPa, flow 10L/h, the feed liquid after collecting separation.
Dry powder processed: by the feed liquid spray dryer after UF membrane, under the conditions of 115 DEG C of temperature, flow velocity 2.5L/h into
Row spray drying is up to cordyceps sinensis powder.
Cordycepin content 36wt% in the cordyceps sinensis powder of acquisition, oligosaccharide content be not less than 20wt%, molecular weight be 500~
900D, and cordyceps sinensis powder is completely solvable.
Embodiment 5
Method with embodiment 1, unlike:
Fermentation liquid residue is filtered out using 2 layers of gauze in filtration step;
The pH to 4 of supernatant is adjusted in enzyme hydrolysis step, the dosage of dextranase, cellulase and hemicellulase is
100U/g, 100U/g, 100U/g hydrolyze 4h at 35 DEG C;
Ultra-filtration conditions in UF membrane step are as follows: pressure 0.5MPa, flow 30L/h;Nanofiltration condition are as follows: pressure 0.5MPa, stream
10L/h is measured, the feed liquid after collecting separation.
Embodiment 6
Method with embodiment 1, unlike:
Fermentation liquid residue is filtered out using 8 layers of gauze in filtration step;
The pH to 5.5 of supernatant is adjusted in enzyme hydrolysis step, the dosage of dextranase, cellulase and hemicellulase is
50U/g, 30U/g, 20U/g hydrolyze 1h at 60 DEG C;
Ultra-filtration conditions in UF membrane step are as follows: pressure 3MPa, flow 90L/h;Nanofiltration condition are as follows: pressure 3MPa, flow
40L/h, the feed liquid after collecting separation.
Claims (10)
1. the preparation method of cordyceps sinensis powder, which comprises the following steps:
A, which will activate Cordyceps militaris and be inoculated into the container containing fermentation medium, to ferment, and is then separated by solid-liquid separation and obtains supernatant;
Then the PH to 4~6 for the supernatant that B regulating step A is obtained is added complex enzyme and is digested to obtain enzymolysis liquid, described multiple
Synthase is the compound of dextranase, cellulase and hemicellulase;
Enzymolysis liquid filtering that C obtains step B, it is dry after up to cordyceps sinensis powder.
2. the preparation method of cordyceps sinensis powder according to claim 1, which is characterized in that in step A, the Cordyceps militaris is classified as
Cordyceps militaris Cordyceps militaris, number are CICC 14014;
The activation Cordyceps militaris is to be inoculated into Cordyceps militaris on slant medium to carry out Spore cultivation, and cultivation temperature is 24~27
DEG C, incubation time is 5~7 days, and the slant medium is made as follows: removing 200~250g of potato of skin, cuts
Block, adds water 1000mL to boil 20~40min, and 5~6 layers of gauze filter off potato ball, filtrate is complemented to 1000mL, adds glucose
15~20g, KH2PO41~5g, MgSO4·7H20.5~5g of O, 0.1~0.8g of vitamin B1,10~25g of agar, after fusing
Packing, sterilizing.
3. the preparation method of cordyceps sinensis powder according to claim 1, which is characterized in that in step A, the activation Cordyceps militaris is connect
Kind amount is 5~30%v/v.
4. the preparation method of cordyceps sinensis powder according to claim 1, which is characterized in that in step A, the fermentative medium formula
It is as follows: 30~60g/L of glucose, 2~10g/L of yeast extract, 2~10g/L of corn pulp, 1~5g/L of soybean protein, KH2PO4 0.5
~5g/L, K2HPO4·3H20.5~5g/L of O, MgSO4·7H20.5~5g/L of O, solvent are water, and pH is 5~7;
The condition of the fermentation are as follows: 24~28 DEG C of fermentation temperature, fermentation time is 3~4 weeks;
The step of separation of solid and liquid, is as follows: the residue of fermentation liquid is removed using 2~8 layers of filtered through gauze, it is then thick by what is obtained
Liquid is centrifuged 10~20min at 4000~10000r/min, sterilizing.
5. the preparation method of cordyceps sinensis powder according to claim 1, which is characterized in that in step B, the pH of the supernatant is
4.5~6;
The additive amount of the dextranase is 10~100U/g Cordyceps sinensis polysaccharide, and the additive amount of cellulase is 10~100U/g cordyceps sinensis
Polysaccharide, the additive amount of hemicellulase are 10~100U/g Cordyceps sinensis polysaccharide;
Preferably, the additive amount of the dextranase is 20~50U/g Cordyceps sinensis polysaccharide, and the additive amount of cellulase is 20~50U/
G Cordyceps sinensis polysaccharide, the additive amount of hemicellulase are 20~50U/g Cordyceps sinensis polysaccharide.
6. the preparation method of cordyceps sinensis powder according to claim 1, which is characterized in that the enzymatic hydrolysis meets at least one of following:
The temperature of the enzymatic hydrolysis is 35~60 DEG C;Preferably, the temperature of the enzymatic hydrolysis is 50~55 DEG C;
The time of the enzymatic hydrolysis is 1~4h;Preferably, 1~2h of time of the enzymatic hydrolysis;
The revolving speed of the enzymatic hydrolysis is 50~200r/min;Preferably, 100~200r/min of revolving speed of the enzymatic hydrolysis.
7. the preparation method of cordyceps sinensis powder according to claim 1, which is characterized in that in step C, it is described be filtered into ultrafiltration after receive
Filter;The molecular cut off for the film that the ultrafiltration uses is 2000~3000D, and ultrafiltration pressure is 0.5~3MPa, and ultrafiltration flow velocity is 30
~90L/h;The molecular cut off for the film that the nanofiltration uses is 150~200D, and nanofiltration pressure is 0.5~3MPa, nanofiltration flow velocity
For 10~40L/h.
8. the preparation method of cordyceps sinensis powder according to claim 6, which is characterized in that in step C, the ultrafiltration pressure is 1.5
~3MPa, ultrafiltration flow velocity are 40~75L/h;Nanofiltration pressure is 2~3MPa, and nanofiltration flow velocity is 10~30L/h.
9. the preparation method of cordyceps sinensis powder according to claim 1, which is characterized in that in step C, the drying is concentration drying
Or spray drying.
10. the cordyceps sinensis powder that the preparation method of any one of the claim 1~9 cordyceps sinensis powder is prepared.
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