CN110256599A - A kind of enzyme assisted extraction lycium ruthenicum polysaccharide process - Google Patents
A kind of enzyme assisted extraction lycium ruthenicum polysaccharide process Download PDFInfo
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- CN110256599A CN110256599A CN201910652403.7A CN201910652403A CN110256599A CN 110256599 A CN110256599 A CN 110256599A CN 201910652403 A CN201910652403 A CN 201910652403A CN 110256599 A CN110256599 A CN 110256599A
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- lycium ruthenicum
- enzyme
- assisted extraction
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- polysaccharide process
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
Abstract
The invention discloses a kind of enzyme assisted extraction lycium ruthenicum polysaccharide process, belong to the biological extraction and separation technology field in phytochemicals production.A kind of enzyme assisted extraction lycium ruthenicum polysaccharide process, including the following steps: S1. cleans lycium ruthenicum three times with pure water, is then dried, crushes, crossing 40 meshes, obtaining lycium ruthenicum powder；S2. enzyme is added in the lycium ruthenicum powder, then plus water is digested, and is centrifuged, is obtained supernatant；S3. it after adjusting supernatant pH to 3 with 2mol/L hydrochloric acid, stands overnight, is then filtered, obtain filtrate, be then concentrated, obtain concentrate；S4. it is stayed overnight after the dehydrated alcohol of 4 times of volumes is added into the concentrate in 4 DEG C of low temperature, is then centrifuged, 50 DEG C of vacuum drying, obtains lycium ruthenicum polyoses extract.Equipment needed for this process is simple, and operation is easy and lycium ruthenicum polysaccharide yield is high.
Description
Technical field
The invention belongs to the biological extraction and separation technology fields in phytochemicals production, are specifically related to a kind of enzyme assisted extraction
Lycium ruthenicum polysaccharide process.
Background technique
Lycium ruthenicum polysaccharide is primary bioactivity substance in lycium ruthenicum, is important health-care components, be by many identical or
Different monosaccharide has immunological regulation, antitumor, anti-oxidant, anti-aging, drop blood with compound composed by glucosides key connection
Rouge, protection liver, promotes the multiple pharmacological effects such as development, prevention and treatment cataract of old people at hypoglycemic, anti-radiation, anti-bacteria and anti-virus
And physiological function.Currently, fructus lycii used in food is mostly fructus lycii powder and full fructus lycii, it is difficult to after consumer is edible to therein
Active constituent is completely absorbed, so that fructus lycii product health-care effect is unobvious.Therefore, it is necessary to a kind of extraction processes to carry out
The extraction of lycium ruthenicum polysaccharide, to meet the needs of the majority of consumers.Currently, lycium ruthenicum extraction method of polysaccharides is mostly water extraction, acid
More or less there is some lack in alkali extraction method, ultrasonic extraction, microwave loss mechanisms, organic solvent extraction, but these methods
It falls into, for example recovery rate is low, required equipment is expensive, has radiation etc..
Summary of the invention
In view of the above problems, the present invention provides a kind of enzyme assisted extraction lycium ruthenicum polysaccharide process, this methods
Required equipment is simple, and operation is easy and lycium ruthenicum polysaccharide yield is high.
The technical solution of the present invention is as follows:
A kind of enzyme assisted extraction lycium ruthenicum polysaccharide process, including the following steps:
S1. lycium ruthenicum is cleaned three times with pure water, is then dried, crushes, crossing 40 meshes, obtains lycium ruthenicum powder；
S2. enzyme is added in the lycium ruthenicum powder, then plus water is digested, and is centrifuged, is obtained supernatant；
S3. it after adjusting supernatant pH to 3 with 2mol/L hydrochloric acid, stands overnight, is then filtered, obtain filtrate, it is then dense
Contracting, obtains concentrate；
S4. it stays overnight, is then centrifuged, 50 in 4 DEG C of low temperature after the dehydrated alcohol of 4 times of volumes is added into the concentrate
DEG C vacuum drying, obtain lycium ruthenicum polyoses extract.
Preferably, condition dry described in the S1 are as follows: temperature 60 C, drying time 4h.
Preferably, the enzyme is pectase.
Preferably, the mass ratio of lycium ruthenicum powder and enzyme described in the S2 is 1:0.01~0.05.
Preferably, lycium ruthenicum silty amount described in the S2 and water volume ratio are 1:30~40.
Preferably, the condition of enzymatic hydrolysis described in the S2 are as follows: 30 DEG C of hydrolysis temperature, 45~75min of enzymolysis time.
Preferably, concentrate described in the S3 and filtrate volume ratio are 1:14~15.
Preferably, centrifugal condition described in the S4 are as follows: centrifugation rate 2000r/min, centrifugation time 10min.
The invention has the benefit that a kind of enzyme assisted extraction lycium ruthenicum polysaccharide process of the invention, lycium ruthenicum is cleaned,
It dry, pulverize, be sieved, then extracted with enzyme extraction process, be concentrated, polysaccharide can be extracted from lycium ruthenicum to greatest extent, mentioned
High lycium ruthenicum polysaccharide yield；Equipment needed for simultaneously is simple, concise in technology, and operation is easy.
Specific embodiment
Embodiment of the present invention can be replaced by the difference of specific range based on the above technical solution,
Available numerous embodiment, therefore, several embodiments as described below are only merely the more excellent reality in numerous embodiment
Example is applied, any technology replacement done in above-mentioned technical proposal all belongs to the scope of protection of the present invention.
Embodiment 1
A kind of enzyme assisted extraction lycium ruthenicum polysaccharide process, including the following steps:
S1. it takes the lycium ruthenicum of 5 parts by weight to be cleaned three times with pure water, then carries out 60 DEG C of 4h dryings, crushing, crosses 40 mesh
Sieve, obtains lycium ruthenicum powder；
S2. 0.15 parts by weight pectase is added in the lycium ruthenicum powder, the water that 150 parts by volume are then added carries out 30 DEG C
60min enzymatic hydrolysis, centrifugation obtain supernatant；
S3. it after adjusting supernatant pH to 3 with 2mol/L hydrochloric acid, stands overnight, is then filtered, by filtrate rotary evaporation
Obtain 1/15 concentrate of extracting liquid volume；
S4. it is stayed overnight after the dehydrated alcohol of 4 times of volumes is added into the concentrate in 4 DEG C of low temperature, then in 2000r/min
Lower centrifugation 10min will then be deposited at 50 DEG C and be dried in vacuo, and obtain lycium ruthenicum polyoses extract.
Embodiment 2
A kind of enzyme assisted extraction lycium ruthenicum polysaccharide process, including the following steps:
S1. it takes the lycium ruthenicum of 5 parts by weight to be cleaned three times with pure water, then carries out 60 DEG C of 4h dryings, crushing, crosses 40 mesh
Sieve, obtains lycium ruthenicum powder；
S2. 0.15 parts by weight pectase is added in the lycium ruthenicum powder, the water that 200 parts by volume are then added carries out 30 DEG C
75min enzymatic hydrolysis, centrifugation obtain supernatant；
S3. it after adjusting supernatant pH to 3 with 2mol/L hydrochloric acid, stands overnight, is then filtered, by filtrate rotary evaporation
Obtain 1/14 concentrate of extracting liquid volume；
S4. it is stayed overnight after the dehydrated alcohol of 4 times of volumes is added into the concentrate in 4 DEG C of low temperature, then in 2000r/min
Lower centrifugation 10min will then be deposited at 50 DEG C and be dried in vacuo, and obtain lycium ruthenicum polyoses extract.
Embodiment 3
A kind of enzyme assisted extraction lycium ruthenicum polysaccharide process, including the following steps:
S1. it takes the lycium ruthenicum of 5 parts by weight to be cleaned three times with pure water, then carries out 60 DEG C of 4h dryings, crushing, crosses 40 mesh
Sieve, obtains lycium ruthenicum powder；
S2. 0.05 parts by weight pectase is added in the lycium ruthenicum powder, the water that 175 parts by volume are then added carries out 30 DEG C
45min enzymatic hydrolysis, centrifugation obtain supernatant；
S3. it after adjusting supernatant pH to 3 with 2mol/L hydrochloric acid, stands overnight, is then filtered, by filtrate rotary evaporation
Obtain 1/15 concentrate of extracting liquid volume；
S4. it is stayed overnight after the dehydrated alcohol of 4 times of volumes is added into the concentrate in 4 DEG C of low temperature, then in 2000r/min
Lower centrifugation 10min will then be deposited at 50 DEG C and be dried in vacuo, and obtain lycium ruthenicum polyoses extract.
Embodiment 4
A kind of enzyme assisted extraction lycium ruthenicum polysaccharide process, including the following steps:
S1. it takes the lycium ruthenicum of 5 parts by weight to be cleaned three times with pure water, then carries out 60 DEG C of 4h dryings, crushing, crosses 40 mesh
Sieve, obtains lycium ruthenicum powder；
S2. 0.25 parts by weight pectase is added in the lycium ruthenicum powder, the water that 150 parts by volume are then added carries out 30 DEG C
50min enzymatic hydrolysis, centrifugation obtain supernatant；
S3. it after adjusting supernatant pH to 3 with 2mol/L hydrochloric acid, stands overnight, is then filtered, by filtrate rotary evaporation
Obtain 1/13 concentrate of extracting liquid volume；
S4. it is stayed overnight after the dehydrated alcohol of 4 times of volumes is added into the concentrate in 4 DEG C of low temperature, then in 2000r/min
Lower centrifugation 10min will then be deposited at 50 DEG C and be dried in vacuo, and obtain lycium ruthenicum polyoses extract.
Lycium ruthenicum measurement of the polysaccharide content is carried out to lycium ruthenicum polyoses extract obtained by Examples 1 to 3, measuring method is anthracene
Ketone-sulfuric acid process, specifically: carbohydrate meets concentrated sulfuric acid dehydration and generates alditol or derivatives thereof, can be condensed with anthrone reagent and generate color
Substance, solution is in blue-green after reaction, there is absorption maximum at 620nm, is developed the color in a linear relationship with polyoses content.
Make standard curve:
Dextran solution 0.05mL, 0.10mL, 0.20mL, 0.30mL, 0.40mL, the 0.50mL of 0.1g/L are taken respectively,
0.80mL is separately added into 4.00mL anthrone reagent with distilled water constant volume to 1.00mL, is dipped in ice-water bath cooling, each pipe rapidly
It is dipped in boiling water bath together after adding, nozzle cover glass ball from water-bath is boiled again, accurately boils 10min with vaporization prevention
It takes out, it is cooling with tap water, 10 minutes or so are placed at room temperature for, in 620nm colorimetric.Using the double distilled water equally handled as blank,
Carry out colorimetric.Using optical density as ordinate, sugared content micrograms are abscissa, obtain standard curve: y=5.1169x-
0.0795, R2=0.9997
Lycium ruthenicum measurement of the polysaccharide content:
It is lycium ruthenicum polyoses extract made from Examples 1 to 3 is soluble in water and constant volume is in 100mL volumetric flask, from capacity
It after drawing 25 times of 1mL dilution in bottle, takes 1mL dilution in test tube, 4.00mL anthrone reagent is added, is dipped in ice-water bath rapidly
Cooling, each pipe is dipped in boiling water bath together after adding, nozzle cover glass ball, with vaporization prevention, from water-bath is boiled again, accurately
10min taking-up is boiled, it is cooling with tap water, it is placed at room temperature for 10min or so, in 620nm colorimetric.And contain according to standard curve calculating
Amount calculates lycium ruthenicum polysaccharide yield.
The yield of 1. lycium ruthenicum polysaccharide of table
| Embodiment | Embodiment 1 | Embodiment 2 | Embodiment 3 | Embodiment 4 |
| Lycium ruthenicum polysaccharide yield % | 5.05 | 4.8 | 5.46 | 4.7 |
The invention is not limited to specific embodiments above-mentioned.The present invention, which expands to, any in the present specification to be disclosed
New feature or any new combination, and disclose any new method or process the step of or any new combination.
Claims (8)
1. a kind of enzyme assisted extraction lycium ruthenicum polysaccharide process, characterized in that it comprises the following steps:
S1. lycium ruthenicum is cleaned three times with pure water, is then dried, crushes, crossing 40 meshes, obtains lycium ruthenicum powder；
S2. enzyme is added in the lycium ruthenicum powder, then plus water is digested, and is centrifuged, is obtained supernatant；
S3. it after adjusting supernatant pH to 3 with 2mol/L hydrochloric acid, stands overnight, is then filtered, obtain filtrate, be then concentrated, obtain
Concentrate；
S4. stayed overnight after the dehydrated alcohol of 4 times of volumes is added into the concentrate in 4 DEG C of low temperature, be then centrifuged, 50 DEG C it is true
Sky is dry, obtains lycium ruthenicum polyoses extract.
2. a kind of enzyme assisted extraction lycium ruthenicum polysaccharide process according to claim 1, which is characterized in that described in the S1
Dry condition are as follows: temperature 60 C, drying time 4h.
3. a kind of enzyme assisted extraction lycium ruthenicum polysaccharide process according to claim 1, which is characterized in that the enzyme is pectin
Enzyme.
4. a kind of enzyme assisted extraction lycium ruthenicum polysaccharide process according to claim 1, which is characterized in that described in the S2
The mass ratio of lycium ruthenicum powder and enzyme is 1:0.01~0.05.
5. a kind of enzyme assisted extraction lycium ruthenicum polysaccharide process according to claim 1, which is characterized in that described in the S2
Lycium ruthenicum silty amount and water volume ratio are 1:30~40.
6. a kind of enzyme assisted extraction lycium ruthenicum polysaccharide process according to claim 1, which is characterized in that described in the S2
The condition of enzymatic hydrolysis are as follows: 30 DEG C of hydrolysis temperature, 45~75min of enzymolysis time.
7. a kind of enzyme assisted extraction lycium ruthenicum polysaccharide process according to claim 1, which is characterized in that described in the S3
Concentrate and filtrate volume ratio are 1:14~15.
8. a kind of enzyme assisted extraction lycium ruthenicum polysaccharide process according to claim 1, which is characterized in that described in the S4
Centrifugal condition are as follows: centrifugation rate 2000r/min, centrifugation time 10min.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103333268A (en) * | 2013-07-23 | 2013-10-02 | 草本创想生物科技（成都）有限公司 | Method for preparing lycium ruthenicum polysaccharide |
| CN108329399A (en) * | 2018-05-15 | 2018-07-27 | 重庆市洲仨科技发展有限公司 | A kind of extraction process of polysaccharides |
| CN108497378A (en) * | 2018-03-02 | 2018-09-07 | 中国科学院过程工程研究所 | A kind of black fruit fructus lycii instant powder and its preparation method and application |
| US20190388447A1 (en) * | 2008-04-08 | 2019-12-26 | B&H Biotechnologies | Glycan-based drugs, therapies and biomarkers |
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2019
- 2019-07-18 CN CN201910652403.7A patent/CN110256599A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20190388447A1 (en) * | 2008-04-08 | 2019-12-26 | B&H Biotechnologies | Glycan-based drugs, therapies and biomarkers |
| CN103333268A (en) * | 2013-07-23 | 2013-10-02 | 草本创想生物科技（成都）有限公司 | Method for preparing lycium ruthenicum polysaccharide |
| CN108497378A (en) * | 2018-03-02 | 2018-09-07 | 中国科学院过程工程研究所 | A kind of black fruit fructus lycii instant powder and its preparation method and application |
| CN108329399A (en) * | 2018-05-15 | 2018-07-27 | 重庆市洲仨科技发展有限公司 | A kind of extraction process of polysaccharides |
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Application publication date: 20190920 |