CN105769938A - Oudemansiella canarii and application thereof - Google Patents
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Abstract
The invention discloses Oudemansiella canarii and application thereof. The strain number of Oudemansiella canarii is JZB2115055, and the accession serial number of Oudemansiella canarii in the China General Microbiological Culture Collection Center is CGMCC No.6171. The IC50 of the scavenging rate of Oudemansiella canarii flavone extract on DPPH free radicals is 1.73 micrograms/ml, the IC50 of the scavenging rate of Oudemansiella canarii polysaccharide extract on DPPH free radicals is 593.66 micrograms/ml, the IC50 of the total antioxidant capacity of Oudemansiella canarii flavone extract is 24.74 micrograms/ml, and the IC50 of the total antioxidant capacity of Oudemansiella canarii polysaccharide extract is 240.18 micrograms/ml. Oudemansiella canarii flavone extract and Oudemansiella canarii polysaccharide extract can be developed into an antioxidant or a free radical scavenger.
Description
Technical field
The present invention relates to strain light brown Aode mushroom and an application thereof in biological field.
Background technology
Free radical and oxidation reaction and the multiple disease of induction that human body produces are closely related, and can accelerate human senility, antioxidant (antioxidant) can effective scavenging free radicals, delaying human body caducity and prevent disease.At present, using more in medical treatment and field of food is synthesis antioxidant, such as BHT (di-t-butyl hydroxyl cresol), BHA (butylhydroxy anisole), TBHQ (tert-butylhydroquinone) etc..Research shows, these synthetic antioxidants exist safety issue, have many side effect, as human liver, spleen, lung are all had adverse effect, can bring out malignant tumor etc..Therefore, develop biological anti-oxidant and become the development trend of current Food Science.The Natural antioxidant developed at present is many from plant, such as spice extract, tea polyphenols, natural flavonoid, vitamins, protein and enzyme (SOD), carotenoid, Chinese herbal medicine extract etc..
Light brown Aode mushroom (Oudemansiellacanarii (Jungh.)), it is again Oudemansiella canarii, belongs to Agaricales (Agaricales), swollen coral Cordycepps (Physalacriaceae), Genus Oudeman Siella Spec (Oudemansiella).This bacterium has that mycelial growth rate is fast, the fruiting cycle is short, yield advantages of higher, does not also see the report of the light brown Aode mushroom with antioxidant activity at present.
Summary of the invention
The technical problem to be solved is how antioxidation.
For solving above technical problem, the invention provides a strain light brown Aode mushroom.
Light brown Aode mushroom provided by the present invention, its bacterial strain number is JZB2115055, it is numbered CGMCCNo.6171 in registering on the books of China Committee for Culture Collection of Microorganisms's common micro-organisms center, hereinafter referred to as light brown Aode mushroom JZB2115055 or JZB2115055.
Above-mentioned light brown Aode mushroom JZB2115055 can sporophore, mycelia and/or spore form exist.
The culture of above-mentioned light brown Aode mushroom JZB2115055 falls within protection scope of the present invention.
The culture of light brown Aode mushroom JZB2115055 provided by the present invention, being that light brown Aode mushroom JZB2115055 is cultivated the material in the culture vessel obtained in microbiological culture media, described material includes described light brown Aode mushroom JZB2115055 and the metabolite of described light brown Aode mushroom JZB2115055.
In the culture of above-mentioned light brown Aode mushroom JZB2115055, described microbiological culture media can be solid medium or fluid medium.
The metabolite of above-mentioned light brown Aode mushroom JZB2115055 can obtain from the fermentation liquid of described light brown Aode mushroom JZB2115055.The metabolite of described light brown Aode mushroom JZB2115055 specifically can be prepared as follows, liquid medium within is cultivated described light brown Aode mushroom JZB2115055, removes the light brown Aode mushroom JZB2115055 in liquid culture (fermentation liquid) and namely obtain the metabolite of described light brown Aode mushroom JZB2115055.
For solving above technical problem, the invention provides the application in preparing antioxidant or free radical scavenger of the light brown Aode mushroom extract, described light brown Aode mushroom extract can be light brown Aode mushroom flavone extract or light brown Aode mushroom polyoses extract；
Described light brown Aode mushroom flavone extract can be prepared according to the method comprised the steps: extracts light brown Aode mushroom JZB2115055 with ethanol water, collects extracting solution and removes the second alcohol and water in described extracting solution, obtaining light brown Aode mushroom flavone extract；
Described light brown Aode mushroom polyoses extract can be prepared according to the method comprised the steps: precipitates described light brown Aode mushroom JZB2115055 Aqueous extracts with ethanol, collects precipitation, obtains light brown Aode mushroom polyoses extract；Described light brown Aode mushroom JZB2115055 Aqueous extracts is the water-soluble substances extracted from described light brown Aode mushroom JZB2115055 with water；
Described light brown Aode mushroom JZB2115055 is numbered CGMCCNo.6171 registering on the books of China Committee for Culture Collection of Microorganisms's common micro-organisms center.
Above-mentioned light brown Aode mushroom JZB2115055 can be the sporophore of light brown Aode mushroom JZB2115055, mycelia and/or spore.
The preparation method of above-mentioned light brown Aode mushroom flavone extract falls within protection scope of the present invention.
The preparation method of light brown Aode mushroom flavone extract provided by the invention, comprise the steps: to extract light brown Aode mushroom JZB2115055 with ethanol water, collect extracting solution and remove the second alcohol and water in described extracting solution, obtaining light brown Aode mushroom flavone extract, described light brown Aode mushroom flavone extract is light brown Aode mushroom extract；
Described light brown Aode mushroom JZB2115055 is numbered CGMCCNo.6171 registering on the books of China Committee for Culture Collection of Microorganisms's common micro-organisms center.
Above, in described ethanol water, the concentration expressed in percentage by volume of ethanol can be 50%.
Above, extract light brown Aode mushroom JZB2115055 with ethanol water can for extract light brown Aode mushroom JZB2115055 with the ethanol water that ethanol concentration expressed in percentage by volume is 50% when microwave intermittent radiation, described microwave intermittent radiation carries out 3 microwave radiations for interval, 1st time microwave irradiation time is 3 minutes, interval carries out the 2nd microwave radiation after 1 minute, radiated time is 1.5 minutes, then 1 minute, interval again, carry out the 3rd microwave radiation, radiated time is 1.5 minutes, and the power of each microwave radiation in 3 microwave radiations is 700W.
The light brown Aode mushroom flavone extract prepared by above-mentioned preparation method falls within protection scope of the present invention.
Active component is the antioxidant of above-mentioned light brown Aode mushroom flavone extract or free radical scavenger falls within protection scope of the present invention.
The preparation method of light brown Aode mushroom polyoses extract provided by the invention, comprises the steps: to precipitate described light brown Aode mushroom JZB2115055 Aqueous extracts with ethanol, collects precipitation, obtain light brown Aode mushroom polyoses extract；Described light brown Aode mushroom JZB2115055 Aqueous extracts is the water-soluble substances extracted from described light brown Aode mushroom JZB2115055 with water；
Described light brown Aode mushroom JZB2115055 is numbered CGMCCNo.6171 registering on the books of China Committee for Culture Collection of Microorganisms's common micro-organisms center.
In said method, described precipitation drying obtains described light brown Aode mushroom polyoses extract.Described drying can carry out at 60-70 DEG C.
The light brown Aode mushroom polyoses extract prepared by above-mentioned preparation method falls within protection scope of the present invention.
Active component is the antioxidant of above-mentioned light brown Aode mushroom polyoses extract or free radical scavenger falls within protection scope of the present invention.
Light brown Aode mushroom JZB2115055 application in preparing antioxidant or free radical scavenger falls within protection scope of the present invention.
Above-mentioned free radical can be DPPH free radical.
Experiment proves, the IC50 of the clearance rate of DPPH free radical is 1.73 μ g/ml by described light brown Aode mushroom flavone extract, the IC50 of DPPH free radical scavenging activity is 593.66 μ g/ml by described light brown Aode mushroom polyoses extract, the IC50 of described light brown Aode mushroom flavone extract total antioxidant capacity is 24.74 μ g/ml, the IC50=240.18 μ g/ml of described light brown Aode mushroom polyoses extract total antioxidant capacity.Described light brown Aode mushroom flavone extract and described light brown Aode mushroom polyoses extract can be developed as antioxidant or free radical scavenger.
Preservation explanation
Strain name: light brown Aode mushroom (Oudemansiellacanarii)
Strain number: JZB2115055
Preservation mechanism: China Committee for Culture Collection of Microorganisms's common micro-organisms center
Preservation mechanism is called for short: CGMCC
Address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Preservation date: on May 28th, 2012
Register on the books numbering: CGMCCNo.6171 at preservation center
Accompanying drawing explanation
Fig. 1 is the sporophore of JZB2115055.
Fig. 2 is the basidiospore form of JZB2115055.
Fig. 3 is the standard curve that flavone concentration measures.
Fig. 4 is the standard curve that polysaccharide concentration measures.
Fig. 5 is FeSO4Standard curve.
Detailed description of the invention
Below in conjunction with detailed description of the invention, the present invention being further described in detail, the embodiment provided is only for illustrating the present invention, rather than in order to limit the scope of the present invention.Experimental technique in following embodiment, if no special instructions, is conventional method.Material used in following embodiment, reagent etc., if no special instructions, all commercially obtain.
Embodiment 1, light brown Aode mushroom JZB2115055 Isolation and identification
1, the separation and Culture of light brown Aode mushroom JZB2115055
From the great Du Gang withered tree of Yunnan, gather fungus, adopt comprehensive PDA culture medium (to be made up of following raw material for every liter: Rhizoma Solani tuber osi 200g, agar 20g, glucose 20g, soy peptone 5g, KH2PO43g、MgSO41.5g, vitamin B110mg, surplus water.121 DEG C of autoclavings 30 minutes) separate tissue, obtain the pure culture (mycelia) that strain number is JZB2115055, the access of the pure culture of JZB2115055 is added rich Optimal Medium and (is made up of following raw material for every liter: Rhizoma Solani tuber osi 200g, glucose 20g, soy peptone 4g, KH2PO41g、MgSO40.5g, vitamin C 10mg agar powder 16g, surplus water, pH8.5) at 28 DEG C constant temperature culture 7-10 days, be transferred in second-generation culture medivm, at 24 DEG C-26 DEG C, lucifuge is cultivated 14-16 days, and mycelia covers with culture medium, obtains the second class inoculum of JZB2115055.Second-generation culture medivm is prepared as follows: it is 65%-70% (weight/mass percentage composition), the natural second-generation culture medivm of pH that apple branch wood flour, Testa Tritici, sucrose, Calx and water are mixed water content, and wherein the mass ratio of apple branch wood flour, Testa Tritici, sucrose and Calx is 80:18:1:1 (with dry weight basis).Second-generation culture medivm is sub-packed in vial, 121 DEG C of autoclavings 120 minutes.
With water, cotton seed hulls, Testa Tritici, apple branch wood flour and Calx are mixed and made into water content according to the mass ratio of 65:18:15:2 is 65%-70% (weight/mass percentage composition), the natural culture medium of pH.Culture medium is sub-packed in the high pressure resistant plastic bag of 17cm × 33cm × 0.04cm, every packed siccative (in sporophore culture medium other components) in addition to water 380g.121 DEG C of autoclavings 2 hours, obtain the bacterium bag equipped with sporophore culture medium.
The second class inoculum of JZB2115055 uniformly accesses the bacterium bag equipped with above-mentioned sporophore culture medium, and every bag of inoculum concentration is 3.14 × 10-5m3.Setup Experiments three repetition, each repetition cultivates 100 bacterium bags, every bag culture medium dry weight 380g.
Being 24 DEG C-26 DEG C in ambient temperature after inoculation, relative air humidity is 50%-60%, well-ventilated, and lucifuge is cultivated, and after 21-23 days, mycelia covers with culture medium.Bacterium bag is transferred under-5 DEG C of-5 DEG C of conditions and processes 3-5 days, complete low temperature stimulation；It is 23 DEG C-26 DEG C that bacterium bag is transferred to ambient temperature, relative air humidity 80%-90%, intensity of illumination 300Lux, well-ventilated, cultivates 5-7 days and formed to fruit body primordium when gas concentration lwevel 1500ppm；After continuing cultivation 3-5 days, sporophore stem length is to about 10cm, and cap launches, but when spore not yet launches, the sporophore of the JZB2115055 that gathers.
2, the qualification of light brown Aode mushroom JZB2115055
The Morphological Identification of JZB2115055: observe the mycelium shape of bacterium colony, size, color, taste, vitality etc. on solid medium.Optical microscope inspection basidiospore form, and adopt flat board oblique cutting coverslip method, observation mycelia clamp connection structure, cell size, form etc..
The Molecular Identification of JZB2115055: utilize ITS1 and ITS4 universal primer that JZB2115055 is carried out ITS-PCR qualification.
Result shows the bacteria cover diameter 4-12cm of JZB2115055, and first flat hemispherical, there is mucus on surface；Initial stage central authorities' brown, edge are gradually shallow, and the later stage becomes greyish white to Slate grey；Bacterial context white, colloid；Stem often bends, long 5-16cm, thick 5-15mm, and dirty white, there are dark brown cilium and vertical stripe (Fig. 1) in surface；Lamella white is to dirty white, rarer, and Length discrepancy is directly given birth to；Spore print white, spore is spherical, diameter 14-16 μ m 16-18 μm (Fig. 2).
Sequence 1 in the ITS sequence of JZB2115055 such as sequence table, reaches 99% with the light brown Aode mushroom bacterial strain homology that accession number is AF321476.Therefore combining form qualification result, is accredited as light brown Aode mushroom by JZB2115055.
Light brown Aode mushroom JZB2115055 is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on May 28th, 2012, and preserving number is CGMCCNo.6171.
Embodiment 2, the preparation of light brown Aode mushroom JZB2115055 extract and antioxidant activity research thereof
1, the preparation of light brown Aode mushroom JZB2115055 flavone extract
1.1, preparation light brown Aode mushroom JZB2115055 flavone extract
The sporophore of the JZB2115055 of embodiment 1 is obtained 20-25 DEG C of dry in the sun the sporophore of the JZB2115055 dried.The sporophore of dry JZB2115055 is put into the omnipotent disintegrating machine of high speed.Broken 4 times repeatedly, each 20 seconds, make the 100 homogeneous JZB2115055 fruit body powders of purpose.Take 10gJZB2115055 fruit body powder in beaker, with deionized water and the ethanol water 450mL that dehydrated alcohol preparation ethanol concentration expressed in percentage by volume is 50%, inject beaker, after stirring evenly, put in microwave oven, carry out microwave intermittent radiation extraction, this microwave intermittent radiation carries out 3 microwave radiations for interval, 1st time microwave irradiation time is 3 minutes, interval carries out the 2nd microwave radiation after 1 minute, radiated time is 1.5 minutes, then 1 minute, interval again, carry out the 3rd microwave radiation, radiated time is 1.5 minutes, the microwave power of each microwave radiation in 3 microwave radiations is 700W.Then carrying out the centrifugal 100min of 8000r/min and collect supernatant, measure volume lyophilizing, grind and obtain powder, this powder is light brown Aode mushroom JZB2115055 flavone extract.
1.2, the drafting of the standard curve that flavone concentration measures:
Adopt sodium nitrite-aluminum nitrate colorimetry.Precision weighs and adds 30% ethanol water through 115 DEG C of freeze-day with constant temperature to constant weight rutin standard substance 10mg, is settled to 100mL, obtains rutin titer.Take rutin titer 0 μ l, 80 μ l, 320 μ l, 640 μ l, 1000 μ l, in 5 2ml centrifuge tubes, add 30% ethanol water and are supplemented to 1mL, add 100 μ l5% sodium nitrite solutions, shake up, stand 5min, add 100 μ l10% aluminum nitrate solutions, stand 6min, add 400 μ l4.3%NaOH solution mixings, manage (adding 0 μ l rutin titer) reagent solution with first and make blank, by microplate reader at 510nm place survey absorbance.With light absorption value for vertical coordinate, rutin concentration is that abscissa does linear graph drawing standard curve.
Being obtained linear equation by standard curve is y=0.0039x-0.0075, R2=0.9989, when the content of total flavones is within the scope of 8ug/mL-100ug/mL, the flavone concentration of solution and 510nm place light absorption value present once linear relationship.When making testing sample, it is necessary to make sample concentration in linear interval, the accuracy of guarantee determination data.
1.3, flavone concentration measures:
The light brown Aode mushroom JZB2115055 flavone extract weighed in 0.5000g step 1.1 adds 6mL deionized water dissolving, obtain light brown Aode mushroom flavone extract solution, light brown Aode mushroom flavone extract solution is diluted to different concentration with 30% ethanol water respectively, obtains flavonoids solution sample.Take 1ml flavonoids solution sample respectively as in 2ml centrifuge tube, add 100ul5% sodium nitrite solution, shake up, stand 5min, add 100 μ l10% aluminum nitrate solutions, stand 6min, add 400 μ l4.3%NaOH solution mixings, blank is made, by microplate reader at 510nm place survey absorbance with corresponding solvent.Standard curve according to step 1.2, obtains the general flavone content in light brown Aode mushroom flavone extract solution.
2, the preparation of light brown Aode mushroom JZB2115055 polyoses extract
2.1, preparation light brown Aode mushroom JZB2115055 polyoses extract
The sporophore of the JZB2115055 of embodiment 1 is obtained 20-25 DEG C of dry in the sun the sporophore of the JZB2115055 dried.The sporophore of dry JZB2115055 is put into the omnipotent disintegrating machine of high speed.Broken 4 times repeatedly, each 20 seconds, make the 100 homogeneous JZB2115055 fruit body powders of purpose.In JZB2115055 fruit body powder, add the deionized water of the 25 times of quality being JZB2115055 fruit body powder, obtain JZB2115055 sporophore mixed liquor.This JZB2115055 sporophore mixed liquor is placed after 8 hours at the refrigerator of 4 DEG C and carries out water-bath.Shake up before water-bath, sealing, with shaking bath, JZB2115055 sporophore mixed liquor carried out 90 DEG C, the high temperature bath of 4h.After water-bath, JZB2115055 sporophore mixed liquor carrying out the centrifugal 30min of 6000r/min and collects supernatant (water-soluble substances), this supernatant is JZB2115055 sporophore Aqueous extracts.Measure this supernatant volume, add in this supernatant is that the dehydrated alcohol of these 4 times of volumes of supernatant makes polysaccharide precipitate out, stir masking foil on bonnet, stand and treat solid-liquid separation in 12 hours, 6000g collects precipitation in centrifugal 20 minutes, precipitation being put to the baking oven of 60 DEG C drying until constant mass, be ground into powder, this powder is light brown Aode mushroom JZB2115055 polyoses extract.
The Specification Curve of Increasing that 2.2 polysaccharide concentrations measure
The making of standard curve: glucose 105 DEG C is dried to constant weight, the glucose solution of variable concentrations (10,20,30,40,50,60,70,80,90,100 μ g/mL) it is configured to distilled water, draw the various glucose solution of 2ml respectively in 20ml tool plug test tube, separately take 2ml distilled water and make blank.Respectively adding 6% phenol (heavily smoked) solution 1.0mL, jolting mixes, and adds sulphuric acid 5.0mL, and rapid jolting mixes, and in left at room temperature 15min, is placed in 100 DEG C of water-bath 30min, and mixing cools down rapidly, then mixes.Trap is measured respectively in 492nm wavelength place with ultraviolet spectrophotometer.With the concentration C (mg/mL) of glucose solution for abscissa, trap A is vertical coordinate, drawing standard curve.Being obtained linear equation by standard curve is y=0.0135x+0.0543, R2=0.9988, when the content of total glucose is within the scope of 0ug/mL-60ug/mL, the polysaccharide concentration of solution and 492nm place light absorption value present once linear relationship.When making testing sample, it is necessary to make sample concentration in linear interval, the accuracy of guarantee determination data.
2.3 polysaccharide concentrations measure
Weigh the light brown Aode mushroom JZB2115055 polyoses extract 0.0100g of 2.1, add 3ml deionized water, it is made to dissolve with vibrating machine is broken, obtain light brown Aode mushroom polyoses extract solution, light brown Aode mushroom polyoses extract solution deionized water is diluted to different concentration, obtains polysaccharide solution sample.
Pipetting 250ul polysaccharide solution sample to put in 2mL centrifuge tube, be separately added into 6% phenol (heavily smoked) solution 1.0mL, jolting mixes, and adds sulphuric acid 5.0mL, rapid jolting mixes, and in left at room temperature 15min, is placed in 100 DEG C of water-bath 30min, mixing, cools down rapidly, then mixes.Trap is measured respectively in 492nm wavelength place with ultraviolet spectrophotometer.Standard curve according to step 2.2, obtains the polyoses content in light brown Aode mushroom polyoses extract solution.
3, DPPH antioxidant activity detection
The DPPH stock solution of 1mg/mL is diluted to by 1:4 with 50% ethanol water the DPPH diluent of 0.2mg/mL, standby.Respectively the light brown Aode mushroom flavone extract solution of 1.3 and the light brown Aode mushroom polyoses extract solution of 2.3 with the ethanol water of 50%, are diluted to desired concn and obtain testing sample diluent, testing sample diluent is mixed with equal-volume 50% ethanol water and shakes up, stand 30min, under 517nm, measure light absorption value (Aj)；Testing sample diluent mixes with equal-volume DPPH diluent and shakes up, and stands 30min, measures light absorption value (Ai) under 517nm；DPPH diluent mixes with equal-volume 50% ethanol water and shakes up, and stands 30min, measures light absorption value (A0) under 517nm.Each 3 times of operation repetitive, takes its meansigma methods.Calculate clearance rate as follows: W (%)=(A0-Ai+Aj)/A0 × 100
The table 1. variable concentrations light brown Aode mushroom flavone extract solution clearance rate to DPPH free radical
| General flavone content μ g/ml | 5 | 10 | 15 | 20 | 25 |
| Clearance rate W (100%) | 67.00±0.97b | 84.05±0.55a | 85.71±1.62a | 85.74±0.76a | 87.46±3.64a |
Note: in table, numeral is mean+SD (n=3)；Different English lower case represent 5% significant level (p < 0.05).
As can be seen from Table 1, DPPH free radical is respectively provided with certain clearance rate by the light brown Aode mushroom flavone extract solution of variable concentrations, and wherein, when concentration is 25 μ g/ml, clearance rate is maximum, is 87.46 ± 3.64%；When concentration is 5 μ g/ml, clearance rate is minimum, is 67.00 ± 0.97%, and and other process between significant difference.It is 10-25 μ g/ml in concentration, between each process, the clearance rate difference of DPPH free radical is not notable.The light brown Aode mushroom JZB2115055 flavone extract IC50=1.73 μ g/ml to DPPH free radical scavenging activity.
The table 2. variable concentrations light brown Aode mushroom polyoses extract solution clearance rate to DPPH free radical
| Polysaccharide concentration μ g/ml | 10 | 20 | 30 | 40 | 50 |
| Clearance rate W (100%) | 21.69±0.56d | 23.94±0.94c | 27.86±0.69b | 30.47±0.81a | 30.71±1.14a |
Note: in table, numeral is mean+SD (n=3)；Different English lower case represent 5% significant level (p < 0.05).
As can be seen from Table 2, DPPH free radical is respectively provided with certain clearance rate by the light brown Aode mushroom polyoses extract solution of variable concentrations, and wherein, when concentration is 50 μ g/ml, clearance rate is maximum, is 30.71 ± 1.14%；Next is 40 μ g/ml, is 30.47 ± 0.81%, and difference is not notable between the two；When concentration is 10 μ g/ml, clearance rate is minimum, is 21.69 ± 0.56%, and and other process between significant difference.The light brown Aode mushroom JZB2115055 polyoses extract IC50=593.66 μ g/ml to DPPH free radical scavenging activity.
4, FRAP oxidation resistance detection
According to total antioxidant capacity detection kit (FRAP method) (green skies biotechnology research institute), preparation FeSO is described4Solution concentration respectively 0.1,0.2,0.4,0.8,1.6mmol/L, make standard curve.
Respectively the light brown Aode mushroom flavone extract solution of 1.3 and the light brown Aode mushroom polyoses extract solution of 2.3 with the ethanol water of 50%, are diluted to desired concn and obtain testing sample diluent, replace FeSO with testing sample diluent4Solution carries out reduction reaction, mixes gently, hatches the absorbance measuring 593nm place after 3-5min, judges the oxidation resistance of testing sample according to standard curve for 37 DEG C.
With FeSO4Solution is standard curve such as Fig. 5 of standard preparation.Fe2+ concentration light absorption value with 593nm place in the scope of 0-1.2mmol/L be once linear relationship, illustrate that testing sample concentration should control within the specific limits, just meet conversion requirement.
The total antioxidant capacity of table 3. variable concentrations light brown Aode mushroom flavone extract solution
| General flavone content μ g/ml | 52.67 | 39.51 | 31.60 | 26.34 | 22.57 |
| Antioxidation (mM) | 1.06±0.04a | 0.78±0.02b | 0.62±0.04c | 0.53±0.01d | 0.46±0.01e |
Note: in table, numeral is mean+SD (n=3)；Different English lower case represent 5% significant level (p < 0.05).
As can be seen from Table 3, there is difference in the light brown Aode mushroom flavone extract solution total antioxidant capacity of variable concentrations, wherein, when concentration is 52.67 μ g/ml, oxidation resistance is maximum, is 1.06 ± 0.04mM；Next is 39.51 μ g/ml, is 0.78 ± 0.02mM, between the two significant difference；When concentration is 22.57 μ g/ml, oxidation resistance is minimum, is 0.46 ± 0.01mM, significant difference between each process.The IC50=24.74 μ g/ml of light brown Aode mushroom JZB2115055 flavone extract total antioxidant capacity.
The total antioxidant capacity of table 4 variable concentrations light brown Aode mushroom polyoses extract solution
| Polysaccharide concentration μ g/ml | 430 | 215 | 143 | 108 | 86 |
| Antioxidation (mM) | 0.77±0.00a | 0.39±0.00b | 0.26±0.05c | 0.22±0.00d | 0.19±0.01d |
Note: in table, numeral is mean+SD (n=3)；Different English lower case represent 5% significant level (p < 0.05).
As can be seen from Table 4, there is difference in the total antioxidant capacity of variable concentrations light brown Aode mushroom polyoses extract solution, wherein, when concentration is 430 μ g/ml, oxidation resistance is maximum, is 0.77 ± 0.00mM；Next is 215 μ g/ml, is 0.39 ± 0.00mM, between the two significant difference；When concentration is 108 μ g/ml and 86 μ g/ml, oxidation resistance is minimum, respectively 0.22 ± 0.00mM and 0.19 ± 0.01mM, and difference is notable between the two, and significant difference between other process.The IC50=240.18 μ g/ml of light brown Aode mushroom JZB2115055 polyoses extract total antioxidant capacity.
Can be seen that from above experimental result, DPPH free radical is had certain scavenging action by light brown Aode mushroom JZB2115055 flavone extract and light brown Aode mushroom JZB2115055 polyoses extract, but the elimination effect of DPPH free radical is substantially better than light brown Aode mushroom JZB2115055 polyoses extract by light brown Aode mushroom JZB2115055 flavone extract.Wherein light brown Aode mushroom JZB2115055 flavone extract removes the optimum concentration of DPPH free radical is 10 μ g/ml, and it is 40 μ g/ml that light brown Aode mushroom JZB2115055 polyoses extract removes the optimum concentration of DPPH free radical；Under both the above concentration, the clearance rate of DPPH is 2.76 times of light brown Aode mushroom JZB2115055 polyoses extract by light brown Aode mushroom JZB2115055 flavone extract；In addition, the light brown Aode mushroom JZB2115055 flavone extract IC50=1.73 μ g/ml to DPPH free radical scavenging activity, the light brown Aode mushroom JZB2115055 polyoses extract IC50=593.66 μ g/ml to DPPH free radical scavenging activity, compares it appeared that DPPH free radical scavenging activity is significantly stronger than light brown Aode mushroom JZB2115055 polyoses extract by light brown Aode mushroom JZB2115055 flavone extract.Light brown Aode mushroom JZB2115055 flavone extract and light brown Aode mushroom JZB2115055 polyoses extract are respectively provided with oxidation resistance, but the oxidation resistance effect of light brown Aode mushroom JZB2115055 flavone extract is substantially better than light brown Aode mushroom JZB2115055 polyoses extract.The wherein IC50=24.74 μ g/ml of light brown Aode mushroom JZB2115055 flavone extract total antioxidant capacity, the IC50=240.18 μ g/ml of light brown Aode mushroom JZB2115055 polyoses extract total antioxidant capacity, the IC50 of the total antioxidant capacity of light brown Aode mushroom JZB2115055 flavone extract are the 1/10 of light brown Aode mushroom JZB2115055 polyoses extract；Therefore, in oxidation resistance, light brown Aode mushroom JZB2115055 flavone extract is also significantly stronger than light brown Aode mushroom JZB2115055 polyoses extract.
Claims (10)
1. light brown Aode mushroom extract application in preparing antioxidant or free radical scavenger, described light brown Aode mushroom extract is light brown Aode mushroom flavone extract or light brown Aode mushroom polyoses extract；
Described light brown Aode mushroom flavone extract is prepared according to the method comprised the steps: extract light brown Aode mushroom JZB2115055 with ethanol water, collects extracting solution and removes the second alcohol and water in described extracting solution, obtaining light brown Aode mushroom flavone extract；
Described light brown Aode mushroom polyoses extract is prepared according to the method comprised the steps: precipitate described light brown Aode mushroom JZB2115055 Aqueous extracts with ethanol, collects precipitation, obtains light brown Aode mushroom polyoses extract；Described light brown Aode mushroom JZB2115055 Aqueous extracts is the water-soluble substances extracted from described light brown Aode mushroom JZB2115055 with water；
Described light brown Aode mushroom JZB2115055 is numbered CGMCCNo.6171 registering on the books of China Committee for Culture Collection of Microorganisms's common micro-organisms center.
2. the preparation method of light brown Aode mushroom extract, comprise the steps: to extract light brown Aode mushroom JZB2115055 with ethanol water, collect extracting solution and remove the second alcohol and water in described extracting solution, obtaining light brown Aode mushroom flavone extract, described light brown Aode mushroom flavone extract is a kind of light brown Aode mushroom extract；
Described light brown Aode mushroom JZB2115055 is numbered CGMCCNo.6171 registering on the books of China Committee for Culture Collection of Microorganisms's common micro-organisms center.
3. the method described in application according to claim 1 or claim 2, it is characterised in that: in described ethanol water, the concentration expressed in percentage by volume of ethanol is 50%.
4. the light brown Aode mushroom extract that prepared by the preparation method described in claim 2.
5. antioxidant or free radical scavenger, its active component is the light brown Aode mushroom extract described in claim 4.
6. the preparation method of light brown Aode mushroom extract, comprise the steps: to precipitate described light brown Aode mushroom JZB2115055 Aqueous extracts with ethanol, collecting precipitation, obtain light brown Aode mushroom polyoses extract, described light brown Aode mushroom polyoses extract is a kind of light brown Aode mushroom extract；Described light brown Aode mushroom JZB2115055 Aqueous extracts is the water-soluble substances extracted from described light brown Aode mushroom JZB2115055 with water；
Described light brown Aode mushroom JZB2115055 is numbered CGMCCNo.6171 registering on the books of China Committee for Culture Collection of Microorganisms's common micro-organisms center.
7. the light brown Aode mushroom extract that prepared by the preparation method described in claim 6.
8. antioxidant or free radical scavenger, its active component is the light brown Aode mushroom extract described in claim 7.
9. light brown Aode mushroom or its culture, it is characterised in that: the bacterial strain number of described light brown Aode mushroom is JZB2115055, and it is numbered CGMCCNo.6171 in registering on the books of China Committee for Culture Collection of Microorganisms's common micro-organisms center.
10. the application in preparing antioxidant or free radical scavenger of the light brown Aode mushroom described in claim 9.
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| CN106258999A (en) * | 2016-08-11 | 2017-01-04 | 广东省微生物研究所 | A kind of thick pleat Aode mushroom novel bacterial, cultural method and application thereof |
| CN108456742A (en) * | 2018-04-02 | 2018-08-28 | 北京市农林科学院 | The molecular labeling of light brown Aode mushroom JZB2115055 and its application |
| CN111937680A (en) * | 2020-06-17 | 2020-11-17 | 广东省微生物研究所（广东省微生物分析检测中心） | New spawn of oospore oudemansiella mucida, artificial cultivation method and application thereof |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106258999A (en) * | 2016-08-11 | 2017-01-04 | 广东省微生物研究所 | A kind of thick pleat Aode mushroom novel bacterial, cultural method and application thereof |
| CN108456742A (en) * | 2018-04-02 | 2018-08-28 | 北京市农林科学院 | The molecular labeling of light brown Aode mushroom JZB2115055 and its application |
| CN111937680A (en) * | 2020-06-17 | 2020-11-17 | 广东省微生物研究所（广东省微生物分析检测中心） | New spawn of oospore oudemansiella mucida, artificial cultivation method and application thereof |
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