CN107459587A - A kind of method that water-soluble polysaccharide is extracted from fruit body of edible fungi - Google Patents
A kind of method that water-soluble polysaccharide is extracted from fruit body of edible fungi Download PDFInfo
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Abstract
The present invention relates to a kind of method that water-soluble polysaccharide is extracted from fruit body of edible fungi, belong to bioengineering and technical field.Its preparation process includes successively：Vacuum freeze drying, ultramicro grinding, NaCl extractions, water extraction, the removal of pigment, alcohol precipitation polysaccharide, the removal of protein and the preparation of refined polysaccharide.After the present invention crushes fruit body of edible fungi using ultramicro grinding method, extracted successively using NaCl and water, and cooperate with microwave ultrasound wave technology, significantly improve the recovery rate of edible fungus water-soluble polysaccharide, and gained polysaccharide has good anti-oxidant and Lipid-lowering activities；Secondly, obtained polysaccharide is removed method of protein using the resin decolorizations of AB 8, Sevag methods and carries out separating-purifying by the present invention, and gained edible fungi polysaccharide pigment removal rate reaches more than 67%, and polysaccharide retention rate reaches more than 65%.
Description
Technical field
The invention belongs to bioengineering and technical field, and in particular to one kind is extracted water-soluble more from fruit body of edible fungi
The method of sugar.
Background technology
Polysaccharide is the polymer that content is most abundant in nature.In recent years, with a large amount of reports, to point out that polysaccharide has special
The effect of, it has also become the focus studied both at home and abroad.Research shows, edible mushroom have anti-oxidant and blood fat reducing function activity into
Point, wherein the polysaccharide contained, particularly water-soluble polysaccharide are proved to containing anti-oxidant, reducing blood lipid and prevention of arterial atherosis
Function, certain booster action can be played to the treatment of angiocardiopathy.Therefore, edible fungi polysaccharide turn into domestic fungus resource exploitation with
An important branch in research, many people attempt the extraction polysaccharide from various edible mushrooms and analyzed and researched.
The Study on extraction of edible fungi polysaccharide mainly uses：Hot water extraction, microwave cooperating high-pressure water heating extraction, surpass
Sound wave synergic high-pressure water heating extraction, Subcritical water chromotagraphy method, alkalinity extraction, organic acid extraction, membrane separation process, enzymatic isolation method etc. carry
Taking technique.Wherein, water extract-alcohol precipitation is most simple and easy extracting method, but such extracting method can be simultaneously different polymerizations
Degree and different water-soluble substances such as protein, nucleic acid extract together, and extraction efficiency is low, and Subcritical water chromotagraphy method,
There is also the defect such as energy consumption is big, recovery rate is low for alkalinity extraction, organic acid extraction and membrane separation process；Microwave Extraction and ultrasonic wave skill
Although art reduces the big problem of energy consumption to a certain extent, the recovery rate of edible fungi polysaccharide is not significantly improved.Cause
This is, it is necessary to establish more efficient, energy-conservation, practicable edible fungi polysaccharide extracting method.
The content of the invention
In view of the shortcomings of the prior art, extracted it is an object of the invention to provide one kind from fruit body of edible fungi water-soluble more
The method of sugar, after the present invention crushes fruit body of edible fungi using ultramicro grinding method, extracted successively using NaCl and water, and cooperate with micro-
Ripple-ultrasonic technology, significantly improves the recovery rate of edible fungus water-soluble polysaccharide, and gained polysaccharide have it is good anti-oxidant and
Lipid-lowering activities；Secondly, obtained polysaccharide is removed method of protein and carried out by the present invention using AB-8 resin decolorizations, Sevag methods
Separating-purifying, gained edible fungi polysaccharide pigment removal rate reach more than 67%, and polysaccharide retention rate reaches more than 65%.
The present invention is achieved through the following technical solutions：
A kind of method that water-soluble polysaccharide is extracted from fruit body of edible fungi, comprises the following steps：
(1) pretreatment of raw material
A, vacuum freeze drying：Fresh food bacterium is selected, after rinsing 15~25min with clear water, then with clear water rinsed clean,
1~2cm, consistent bulk uniform in size are cut into, is put into refrigerator after pre-freeze certain time, puts it into vacuum freeze drying
In machine, start vavuum pump and vacuumize, it is 5~10% to dry to final moisture content；
B, ultramicro grinding：The edible mushroom that vacuum freeze drying is obtained, which is put into, shakes progress batch (-type) crushing in formula pulverizer,
Then powder is crossed into 80 mesh sieves, obtains edible mushroom coarse powder, then edible mushroom coarse powder is placed in micronizer and crushed, obtain edible mushroom
Ultramicro-powder；
(2) NaCl is extracted
Edible mushroom Ultramicro-powder made from step (1) is added in the NaCl aqueous solution, under certain conditions using ultrasonic wave
Microwave combination system extracts, and 6000r/min centrifuges 12min after the completion of extraction, obtains supernatant A and precipitation；
(3) water extracts
Precipitation obtained by step (2) is added in the distilled water of certain temperature, extraction, 6000r/min centrifugation 12min, obtained
Supernatant B；
(4) removal of pigment
A, the pretreatment of resin：It is 1 by weight:1.5 are immersed in AB-8 resins in 95% ethanol, and post water is filled after 24h
It is washed till no alcohol taste；
B, the removal of pigment：Supernatant A and supernatant B are well mixed, AB-8 resins is added and removes depigmentation, AB-8 resins
It is 3 with supernatant volume ratio:5；
(5) alcohol precipitation polysaccharide
After the supernatant for removing depigmentation is concentrated under reduced pressure into 1/5th of original volume with Rotary Evaporators, 3 times of bodies are added
95% long-pending ethanol, it is well mixed, the alcohol precipitation 10h in 4 DEG C of refrigerators, polysaccharide separates out in flocculent deposit, 5000r/min centrifugations
10min, precipitation is collected, obtains edible mushroom Thick many candies；
(6) removal of protein
Edible mushroom Thick many candies are redissolved in distilled water, the protein in solution is removed with Sevag methods；
(7) preparation of refined polysaccharide
95% ethanol that 3 times of volumes are added into the solution for removing isolating protein obtained by step (6) is precipitated, 5000r/
Min centrifuges 10min, collects precipitation, vacuum freeze drying, obtains edible mushroom refined polysaccharide, i.e., edible mushroom of the present invention is water-soluble
Property polysaccharide.
Preferably, refrigerator temperature is -20~-32 DEG C in the step (1), and the pre-freeze time is 1.5~12h.
Preferably, vacuum freeze drier condenser temperature is -30~-50 DEG C in the step (1).
Preferably, micronizer wind speed is 35.3Hz, rotating speed 35.57Hz in the step (1).
Preferably, the mass fraction of the NaCl aqueous solution is 0.9% in the step (2).
Preferably, edible mushroom Ultramicro-powder and NaCl aqueous solution solid-to-liquid ratio (kg/L) are 1 in the step (2):(25~35).
Preferably, ultrasonic microwave combined system extracting condition is in the step (2)：Ultrasonic power 200W, it is ultrasonic when
Between 30min, microwave power 400W, microwave time 30s, the ︰ 30g/mL of solid-liquid ratio 1.
Preferably, distillation coolant-temperature gage is 85~95 DEG C in the step (3).
Preferably, precipitation obtained by step (2) and distilled water solid-to-liquid ratio (kg/L) are 1 in the step (3):25~35, leaching
It is 1~3h to carry the time.
Edible fungi polysaccharide：Polysaccharide is the main energy sources and composition structural material of living organism, can strengthen organism
Immunoloregulation function, as broad immune accelerator；And with anti-infective, anticoagulation, hypoglycemic, reducing blood lipid, promote nucleic acid with
The biosynthesis effect of protein, controls cell division and differentiation, adjusts the functions such as growth and the decay of cell.He Qingti passes through
Research to edible fungi polysaccharide, it is indicated that edible fungi polysaccharide is a kind of by non-specific approach raising body fight original or microorganism
The material of specific reaction, it is mainly used in treatment immunologic function as caused by disease, clinical treatment and declines.
Beneficial effects of the present invention：
1st, ultramicro grinding can be by crushing material to 10~25 μm, and under the fineness, ordinary material cell-wall breaking ratio exceedes
95%, the active component in material is directly exposed, it is no longer necessary to it could be extracted by cell membrane or cell membrane, meanwhile,
Particle diminishes after material micronizing, and specific surface area sharply increases, so as to further improve the recovery rate of its active material.Therefore,
After the present invention crushes fruit body of edible fungi using ultramicro grinding method, extracted successively using NaCl and water, and cooperate with microwave-ultrasonic
Technology, can high degree improve the recovery rate of edible fungus water-soluble polysaccharide, and gained polysaccharide has good anti-oxidant and lipid-loweringing
Activity；
2nd, obtained polysaccharide is removed method of protein and separated by the present invention using AB-8 resin decolorizations, Sevag methods
Purification, gained edible fungi polysaccharide pigment removal rate reach more than 84.5%, and polysaccharide extract rate reaches more than 16.7%.
Embodiment
In order to make the purpose , technical scheme and advantage of the present invention be clearer, with reference to embodiments, to the present invention
It is further elaborated.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, it is not used to
Limit the present invention.
Embodiment 1
A kind of method that water-soluble polysaccharide is extracted from mushroom fruiting body, comprises the following steps：
(1) pretreatment of raw material
A, vacuum freeze drying：New fresh mushroom is selected, after rinsing 25min with clear water, then with clear water rinsed clean, is cut into
2cm, consistent bulk uniform in size, are put into pre-freeze 12h in -32 DEG C of refrigerators, then put it into vacuum freeze drier, start
Vavuum pump vacuumizes, and condenser temperature is -50 DEG C, and it is 5% to dry to final moisture content；
B, ultramicro grinding：The mushroom that vacuum freeze drying is obtained, which is put into, shakes progress batch (-type) crushing in formula pulverizer, so
Powder is crossed into 80 mesh sieves afterwards, obtains mushroom coarse powder, then mushroom coarse powder is placed in micronizer and crushed, micronizer wind speed
For 35.3Hz, rotating speed 35.57Hz, mushroom Ultramicro-powder is obtained；
(2) NaCl is extracted
Mushroom Ultramicro-powder made from step (1) is added in the NaCl aqueous solution that mass fraction is 0.9%, solid-to-liquid ratio (kg/
L it is) 1:30, extracted under certain conditions using ultrasonic microwave combined system, extracting condition is：Ultrasonic power 200W, surpass
Sound time 30min, microwave power 400W, microwave time 30s, the ︰ 30g/mL of solid-liquid ratio 1,6000r/min is centrifuged after the completion of extraction
12min, obtain supernatant A and precipitation；
(3) water extracts
It is 1 that precipitation obtained by step (2) is pressed into solid-to-liquid ratio (kg/L):30 add in the distilled water that temperature is 90 DEG C, extract 2h
Afterwards, 6000r/min centrifuges 12min, obtains supernatant B；
(4) removal of pigment
A, the pretreatment of resin：It is 1 by weight:1.5 are immersed in AB-8 resins in 95% ethanol, and post water is filled after 24h
It is washed till no alcohol taste；
B, the removal of pigment：Supernatant A and supernatant B are well mixed, AB-8 resins is added and removes depigmentation, AB-8 resins
It is 3 with supernatant volume ratio:5；
(5) alcohol precipitation polysaccharide
After the supernatant for removing depigmentation is concentrated under reduced pressure into 1/5th of original volume with Rotary Evaporators, 3 times of bodies are added
95% long-pending ethanol, it is well mixed, the alcohol precipitation 10h in 4 DEG C of refrigerators, polysaccharide separates out in flocculent deposit, 5000r/min centrifugations
10min, precipitation is collected, obtains mushroom Thick many candies；
(6) removal of protein
Mushroom Thick many candies are redissolved in distilled water, the protein in solution is removed with Sevag methods；
(7) preparation of refined polysaccharide
95% ethanol that 3 times of volumes are added into the solution for removing isolating protein obtained by step (6) is precipitated, 5000r/
Min centrifuges 10min, collects precipitation, vacuum freeze drying, obtains mushroom refined polysaccharide, i.e., mushroom of the present invention is water-soluble more
Sugar.
Embodiment 2
A kind of method that water-soluble polysaccharide is extracted from Tremella fructification, comprises the following steps：
(1) pretreatment of raw material
A, vacuum freeze drying：Fresh white fungus is selected, after rinsing 20min with clear water, then with clear water rinsed clean, is cut into
1cm, consistent bulk uniform in size, are put into pre-freeze 10h in -20 DEG C of refrigerators, then put it into vacuum freeze drier, start
Vavuum pump vacuumizes, and condenser temperature is -45 DEG C, and it is 7% to dry to final moisture content；
B, ultramicro grinding：The white fungus that vacuum freeze drying is obtained, which is put into, shakes progress batch (-type) crushing in formula pulverizer, so
Powder is crossed into 80 mesh sieves afterwards, obtains white fungus coarse powder, then white fungus coarse powder is placed in micronizer and crushed, micronizer wind speed
For 35.3Hz, rotating speed 35.57Hz, white fungus Ultramicro-powder is obtained；
(2) NaCl is extracted
White fungus Ultramicro-powder made from step (1) is added in the NaCl aqueous solution that mass fraction is 0.9%, solid-to-liquid ratio (kg/
L it is) 1:25, extracted under certain conditions using ultrasonic microwave combined system, extracting condition is：Ultrasonic power 200W, surpass
Sound time 30min, microwave power 400W, microwave time 30s, the ︰ 30g/mL of solid-liquid ratio 1,6000r/min is centrifuged after the completion of extraction
12min, obtain supernatant A and precipitation；
(3) water extracts
It is 1 that precipitation obtained by step (2) is pressed into solid-to-liquid ratio (kg/L):25 add in the distilled water that temperature is 95 DEG C, extract 1h
Afterwards, 6000r/min centrifuges 12min, obtains supernatant B；
(4) removal of pigment
A, the pretreatment of resin：It is 1 by weight:1.5 are immersed in AB-8 resins in 95% ethanol, and post water is filled after 24h
It is washed till no alcohol taste；
B, the removal of pigment：Supernatant A and supernatant B are well mixed, AB-8 resins is added and removes depigmentation, AB-8 resins
It is 3 with supernatant volume ratio:5；
(5) alcohol precipitation polysaccharide
After the supernatant for removing depigmentation is concentrated under reduced pressure into 1/5th of original volume with Rotary Evaporators, 3 times of bodies are added
95% long-pending ethanol, it is well mixed, the alcohol precipitation 10h in 4 DEG C of refrigerators, polysaccharide separates out in flocculent deposit, 5000r/min centrifugations
10min, precipitation is collected, obtains white fungus Thick many candies；
(6) removal of protein
White fungus Thick many candies are redissolved in distilled water, the protein in solution is removed with Sevag methods；
(7) preparation of refined polysaccharide
95% ethanol that 3 times of volumes are added into the solution for removing isolating protein obtained by step (6) is precipitated, 5000r/
Min centrifuges 10min, collects precipitation, vacuum freeze drying, obtains white fungus refined polysaccharide, i.e., white fungus of the present invention is water-soluble more
Sugar.
Embodiment 3
A kind of method that water-soluble polysaccharide is extracted from Tricholoma matsutake (lto et lmai) Singer sporophore, comprises the following steps：
(1) pretreatment of raw material
A, vacuum freeze drying：Fresh matsutake is selected, after rinsing 15min with clear water, then with clear water rinsed clean, is cut into
1cm, consistent bulk uniform in size, are put into pre-freeze 1.5h in -30 DEG C of refrigerators, then put it into vacuum freeze drier, open
Dynamic vavuum pump vacuumizes, and condenser temperature is -30 DEG C, and it is 10% to dry to final moisture content；
B, ultramicro grinding：The matsutake that vacuum freeze drying is obtained, which is put into, shakes progress batch (-type) crushing in formula pulverizer, so
Powder is crossed into 80 mesh sieves afterwards, obtains matsutake coarse powder, then matsutake coarse powder is placed in micronizer and crushed, micronizer wind speed
For 35.3Hz, rotating speed 35.57Hz, matsutake Ultramicro-powder is obtained；
(2) NaCl is extracted
Matsutake Ultramicro-powder made from step (1) is added in the NaCl aqueous solution that mass fraction is 0.9%, solid-to-liquid ratio (kg/
L it is) 1:35, extracted under certain conditions using ultrasonic microwave combined system, extracting condition is：Ultrasonic power 200W, surpass
Sound time 30min, microwave power 400W, microwave time 30s, the ︰ 30g/mL of solid-liquid ratio 1,6000r/min is centrifuged after the completion of extraction
12min, obtain supernatant A and precipitation；
(3) water extracts
It is 1 that precipitation obtained by step (2) is pressed into solid-to-liquid ratio (kg/L):35 add in the distilled water that temperature is 85 DEG C, extract 3h
Afterwards, 6000r/min centrifuges 12min, obtains supernatant B；
(4) removal of pigment
A, the pretreatment of resin：It is 1 by weight:1.5 are immersed in AB-8 resins in 95% ethanol, and post water is filled after 24h
It is washed till no alcohol taste；
B, the removal of pigment：Supernatant A and supernatant B are well mixed, AB-8 resins is added and removes depigmentation, AB-8 resins
It is 3 with supernatant volume ratio:5；
(5) alcohol precipitation polysaccharide
After the supernatant for removing depigmentation is concentrated under reduced pressure into 1/5th of original volume with Rotary Evaporators, 3 times of bodies are added
95% long-pending ethanol, it is well mixed, the alcohol precipitation 10h in 4 DEG C of refrigerators, polysaccharide separates out in flocculent deposit, 5000r/min centrifugations
10min, precipitation is collected, obtains matsutake Thick many candies；
(6) removal of protein
Matsutake Thick many candies are redissolved in distilled water, the protein in solution is removed with Sevag methods；
(7) preparation of refined polysaccharide
95% ethanol that 3 times of volumes are added into the solution for removing isolating protein obtained by step (6) is precipitated, 5000r/
Min centrifuges 10min, collects precipitation, vacuum freeze drying, obtains matsutake refined polysaccharide, i.e., matsutake of the present invention is water-soluble more
Sugar.
Comparative example 1
Tremella fructification shakes the crushing of formula pulverizer, not cooperateed with using microwave-ultrasonic in extraction process using spontaneously drying
Technology.
A kind of method that water-soluble polysaccharide is extracted from Tremella fructification, comprises the following steps：
(1) pretreatment of raw material
A, spontaneously dry：Select fresh white fungus, after rinsing 20min with clear water, then with clear water rinsed clean, be cut into 1cm,
Consistent bulk uniform in size, it is 7% to spontaneously dry to final moisture content；
B, pulverizer crushes：The step A white fungus being dried to obtain is put into and shakes progress batch (-type) crushing in formula pulverizer, then
80 mesh sieves are crossed, obtain white fungus coarse powder；
(2) NaCl is extracted
White fungus coarse powder made from step (1) is added in the NaCl aqueous solution that mass fraction is 0.9%, solid-to-liquid ratio (kg/L)
For 1:25,60 DEG C of dissolving 10h, 6000r/min centrifugation 12min, obtain supernatant A and precipitation；
(3) water extracts
It is 1 that precipitation obtained by step (2) is pressed into solid-to-liquid ratio (kg/L):25 add in the distilled water that temperature is 95 DEG C, extract 1h
Afterwards, 6000r/min centrifuges 12min, obtains supernatant B；
(4) removal of pigment
A, the pretreatment of resin：It is 1 by weight:1.5 are immersed in AB-8 resins in 95% ethanol, and post water is filled after 24h
It is washed till no alcohol taste；
B, the removal of pigment：Supernatant A and supernatant B are well mixed, AB-8 resins is added and removes depigmentation, AB-8 resins
It is 3 with supernatant volume ratio:5；
(5) alcohol precipitation polysaccharide
After the supernatant for removing depigmentation is concentrated under reduced pressure into 1/5th of original volume with Rotary Evaporators, 3 times of bodies are added
95% long-pending ethanol, it is well mixed, the alcohol precipitation 10h in 4 DEG C of refrigerators, polysaccharide separates out in flocculent deposit, 5000r/min centrifugations
10min, precipitation is collected, obtains white fungus Thick many candies；
(6) removal of protein
White fungus Thick many candies are redissolved in distilled water, the protein in solution is removed with Sevag methods；
(7) preparation of refined polysaccharide
95% ethanol that 3 times of volumes are added into the solution for removing isolating protein obtained by step (6) is precipitated, 5000r/
Min centrifuges 10min, collects precipitation, vacuum freeze drying, obtains white fungus refined polysaccharide, i.e., white fungus of the present invention is water-soluble more
Sugar.
Comparative example 2
A kind of method that water-soluble polysaccharide is extracted from Tricholoma matsutake (lto et lmai) Singer sporophore, comprises the following steps：
(1) pretreatment of raw material
A, vacuum freeze drying：Fresh matsutake is selected, after rinsing 15min with clear water, then with clear water rinsed clean, is cut into
1cm, consistent bulk uniform in size, are put into pre-freeze 1.5h in -30 DEG C of refrigerators, then put it into vacuum freeze drier, open
Dynamic vavuum pump vacuumizes, and condenser temperature is -30 DEG C, and it is 10% to dry to final moisture content；
B, ultramicro grinding：The matsutake that vacuum freeze drying is obtained, which is put into, shakes progress batch (-type) crushing in formula pulverizer, so
Powder is crossed into 80 mesh sieves afterwards, obtains matsutake coarse powder, then matsutake coarse powder is placed in micronizer and crushed, micronizer wind speed
For 35.3Hz, rotating speed 35.57Hz, matsutake Ultramicro-powder is obtained；
(2) NaCl is extracted
Matsutake Ultramicro-powder made from step (1) is added in the NaCl aqueous solution that mass fraction is 0.9%, solid-to-liquid ratio (kg/
L it is) 1:35, extracted under certain conditions using ultrasonic microwave combined system, extracting condition is：Ultrasonic power 200W, surpass
Sound time 30min, microwave power 400W, microwave time 30s, the ︰ 30g/mL of solid-liquid ratio 1,6000r/min is centrifuged after the completion of extraction
12min, obtain supernatant A and precipitation；
(3) water extracts
It is 1 that precipitation obtained by step (2) is pressed into solid-to-liquid ratio (kg/L):35 add in the distilled water that temperature is 85 DEG C, extract 3h
Afterwards, 6000r/min centrifuges 12min, obtains supernatant B；
(4) removal of pigment
Supernatant A and supernatant B are well mixed, 0.5% activated carbon is added, shakes up, constant temperature stands 30min, filters,
Take filtrate；
(5) alcohol precipitation polysaccharide
After the filtrate decompression for removing depigmentation is concentrated into 1/5th of original volume with Rotary Evaporators, 3 times of volumes are added
95% ethanol, be well mixed, the alcohol precipitation 10h in 4 DEG C of refrigerators, polysaccharide in flocculent deposit separate out, 5000r/min centrifuge 10min,
Precipitation is collected, obtains matsutake Thick many candies；
(6) removal of protein
Matsutake Thick many candies are redissolved in distilled water, is placed in the bag filter that combined closure system is 7000Da, is dialysed with distilled water,
1 water is changed per 12h, dialyse 2d；
(7) preparation of refined polysaccharide
95% ethanol that 3 times of volumes are added into the solution for removing isolating protein obtained by step (6) is precipitated, 5000r/
Min centrifuges 10min, collects precipitation, vacuum freeze drying, obtains matsutake refined polysaccharide, i.e., matsutake of the present invention is water-soluble more
Sugar.
Test example 1
Influence to edible fungus water-soluble polysaccharide pigment removal rate and polysaccharide extract rate
1st, test objective：The embodiment of the present invention 1~3 and the extracting method of comparative example 1~2 are detected to edible fungus water-soluble polysaccharide
The influence of pigment removal rate and polysaccharide extract rate.
2nd, detection method：Using spectrophotometry pigment removal rate, phend-sulphuric acid determines the recovery rate of polysaccharide.
3rd, testing result：Testing result is as shown in table 1 below.
Influence of the table 1 to edible fungus water-soluble polysaccharide pigment removal rate and polysaccharide extract rate
| Comparative example 1 | Comparative example 2 | Embodiment 1 | Embodiment 2 | Embodiment 3 | |
| Pigment removal rate (%) | 82.7 | 79.3 | 85.3 | 87.6 | 84.5 |
| Polysaccharide extract rate (%) | 5.61 | 12.7 | 16.9 | 17.0 | 16.7 |
4th, conclusion
After testing, compared with comparative example, polysaccharide pigment removal rate and polysaccharide extract rate in the embodiment of the present invention 1~3 show
Increase is write, wherein, the polysaccharide pigment removal rate of embodiment 2 and polysaccharide extract rate respectively reach 87.6% and 16.7%, for the present invention
Most preferred embodiment.
Test example 2
Influence to edible fungus water-soluble polysaccharide anti-oxidative activity
1st, test objective：The embodiment of the present invention 1~3 and the extracting method of comparative example 1~2 are detected to edible fungus water-soluble polysaccharide
The influence of FRAP oxidation resistances.
2nd, detection method：FRAP is a kind of the most frequently used assay method to sample total antioxidant activity, and principle is to be based on
The colorimetric method of redox reaction：In sour environment, the anti-oxidation active substance that is added in system is by Fe3+- TPTZ is reduced into
The Fe of blueness2+- TPTZ, the material have strong absorption at 593nm, and the big I of absorbance shows its Fe2+The content of ion,
That is its bigger oxidation resistance of absorbance is stronger.
3rd, testing result：Testing result is as shown in table 2 below.
Influence of the table 2 to edible fungus water-soluble polysaccharide anti-oxidative activity
| Comparative example 1 | Comparative example 2 | Embodiment 1 | Embodiment 2 | Embodiment 3 | |
| FRAP(mmol/L) | 0.46 | 0.75 | 0.81 | 0.84 | 0.79 |
4th, conclusion
After testing, compared with comparative example 1, the polysaccharide anti-oxidative activity in the embodiment of the present invention 1~3 is respectively increased
76.08%th, 82.61%, 71.74%；Compared with comparative example 2, the polysaccharide anti-oxidative activity in the embodiment of the present invention 1~3 is respectively
Improve 8.0%, 12.0%, 5.3%.Testing result shows that the edible fungus water-soluble polysaccharide prepared by extracting method of the present invention has
There is significant antioxidation activity.
Test example 3
Influence of the edible fungus water-soluble polysaccharide to lipids contents in rat body
1st, experimental animal：Male SD rat, cleaning grade, 100 ± 10g of body weight.
2nd, animal packet：Rat is randomly divided into 6 groups after being fed one week with basal feed, every group 8, and 6 groups are respectively：Base
Plinth feed control group, high lipid food group, positive controls and respectively add the embodiment of the present invention 1~3 prepare edible mushroom it is water-soluble
Property polysaccharide group.The specific compound method of each group rat institute feeding feed is as shown in table 3.
The rat feed formula (%) of table 3
Note："-" represents no added.
3rd, animal feeding and sampling：Zoopery is carried out six weeks altogether, before experiment starts, second week and experiment terminate (the
Six weeks) when docking carried out to rat respectively take blood, take Rat Fast 18h before blood.Taken blood centrifuges 15min in 6000r/min,
Serum is obtained, is preserved in -20 DEG C of refrigerators, to carry out the measure of related biochemical indicator.
4th, testing index：Triglycerides, T-CHOL, HDL-C, LDL-C.
5th, assay method：Triglycerides is determined using GPO-PAP methods, and T-CHOL is determined using CHOD-PAP methods, highly dense
Spend lipoprotein cholesterol to determine using direct measuring method, the calculation formula of LDL-C is as follows：
LDL-C (mmol/L)=T-CHOL-HDL-C-triglycerides/2.2.
6th, measurement result：Measurement result is as shown in table 4 below.
Influence of the table 4 to lipids contents in rat body
Measurement result shows, compared with high lipid food group, the edible mushroom prepared by the extracting method of the embodiment of the present invention 1~3
Water-soluble polysaccharide can significantly reduce Triglycerides in Serum, T-CHOL and LDL-C content, and make height
Density lipoprotein-cholesterol maintains normal level；Secondly, the edible mushroom water prepared by the extracting method of the embodiment of the present invention 1~3
Soluble polysaccharide lipid-lowering effect is suitable with positive controls.Result of the test shows that the edible fungus water-soluble prepared by the present invention is more
Sugar has lipid-lowering effect well.
Claims (9)
- A kind of 1. method that water-soluble polysaccharide is extracted from fruit body of edible fungi, it is characterised in that comprise the following steps：(1) pretreatment of raw materialA, vacuum freeze drying：Fresh food bacterium is selected, after rinsing 15~25min with clear water, then with clear water rinsed clean, cutting Into 1~2cm, consistent bulk uniform in size, it is put into refrigerator after pre-freeze certain time, puts it into vacuum freeze drier It is interior, start vavuum pump and vacuumize, it is 5~10% to dry to final moisture content；B, ultramicro grinding：The edible mushroom that vacuum freeze drying is obtained, which is put into, shakes progress batch (-type) crushing in formula pulverizer, then Powder is crossed into 80 mesh sieves, obtains edible mushroom coarse powder, then edible mushroom coarse powder is placed in micronizer and crushed, obtains edible mushroom ultra micro Powder；(2) NaCl is extractedEdible mushroom Ultramicro-powder made from step (1) is added in the NaCl aqueous solution, under certain conditions using ultrasonic microwave Combined system extracts, and 6000r/min centrifuges 12min after the completion of extraction, obtains supernatant A and precipitation；(3) water extractsPrecipitation obtained by step (2) is added in the distilled water of certain temperature, extraction, 6000r/min centrifugation 12min, obtain supernatant Liquid B；(4) removal of pigmentA, the pretreatment of resin：It is 1 by weight:1.5 are immersed in AB-8 resins in 95% ethanol, and post is filled after 24h and is washed to Without alcohol taste；B, the removal of pigment：Supernatant A and supernatant B are well mixed, AB-8 resins is added and removes depigmentation, AB-8 resins with it is upper Supernatant volume ratio is 3:5；(5) alcohol precipitation polysaccharideAfter the supernatant for removing depigmentation is concentrated under reduced pressure into 1/5th of original volume with Rotary Evaporators, 3 times of volumes of addition 95% ethanol, it is well mixed, the alcohol precipitation 10h in 4 DEG C of refrigerators, polysaccharide separates out in flocculent deposit, 5000r/min centrifugation 10min, receives Collection precipitation, obtains edible mushroom Thick many candies；(6) removal of proteinEdible mushroom Thick many candies are redissolved in distilled water, the protein in solution is removed with Sevag methods；(7) preparation of refined polysaccharide95% ethanol that 3 times of volumes are added into the solution for removing isolating protein obtained by step (6) is precipitated, 5000r/min 10min is centrifuged, precipitation is collected, vacuum freeze drying, obtains edible mushroom refined polysaccharide, i.e., edible fungus water-soluble of the present invention is more Sugar.
- A kind of 2. method that water-soluble polysaccharide is extracted from fruit body of edible fungi according to claim 1, it is characterised in that Refrigerator temperature is -20~-32 DEG C in the step (1), and the pre-freeze time is 1.5~12h.
- A kind of 3. method that water-soluble polysaccharide is extracted from fruit body of edible fungi according to claim 1, it is characterised in that Vacuum freeze drier condenser temperature is -30~-50 DEG C in the step (1).
- A kind of 4. method that water-soluble polysaccharide is extracted from fruit body of edible fungi according to claim 1, it is characterised in that Micronizer wind speed is 35.3Hz, rotating speed 35.57Hz in the step (1).
- A kind of 5. method that water-soluble polysaccharide is extracted from fruit body of edible fungi according to claim 1, it is characterised in that The mass fraction of the NaCl aqueous solution is 0.9% in the step (2).
- A kind of 6. method that water-soluble polysaccharide is extracted from fruit body of edible fungi according to claim 1, it is characterised in that Edible mushroom Ultramicro-powder and NaCl aqueous solution solid-to-liquid ratio (kg/L) are 1 in the step (2):(25~35).
- A kind of 7. method that water-soluble polysaccharide is extracted from fruit body of edible fungi according to claim 1, it is characterised in that Ultrasonic microwave combined system extracting condition is in the step (2)：Ultrasonic power 200W, ultrasonic time 30min, microwave power 400W, microwave time 30s, the ︰ 30g/mL of solid-liquid ratio 1.
- A kind of 8. method that water-soluble polysaccharide is extracted from fruit body of edible fungi according to claim 1, it is characterised in that Distillation coolant-temperature gage is 85~95 DEG C in the step (3).
- A kind of 9. method that water-soluble polysaccharide is extracted from fruit body of edible fungi according to claim 1, it is characterised in that Precipitation obtained by step (2) and distilled water solid-to-liquid ratio (kg/L) are 1 in the step (3):25~35, extraction time is 1~3h.
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| CN115144494A (en) * | 2022-06-28 | 2022-10-04 | 贵州大学 | Method for detecting oligosaccharide in mammal milk |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108077875A (en) * | 2017-12-29 | 2018-05-29 | 凤台县鼎足农业发展有限公司 | A kind of preparation method of mushroom-flavor compound edible mushroom baste |
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