CN109280091A - A kind of biological enzyme extraction process of bitter melon polysaccharide - Google Patents
A kind of biological enzyme extraction process of bitter melon polysaccharide Download PDFInfo
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- 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 present invention provides a kind of biological enzyme extraction processes of bitter melon polysaccharide, comprising the following steps: (1) balsam pear is cleaned slice, water is added and is homogenized；(2) the mixing enzyme solutions being made of cellulase, pectase and papain are added into balsam pear homogenate, mechanical stirring uniformly adjusts pH extraction, separating filtrate and residue afterwards；(3) residue is extracted again, merges and extracts filtrate twice and be concentrated, added active carbon processing and obtain filtrate；(4) 80% ethyl alcohol is added into the filtrate of step (3), bitter melon polysaccharide is precipitated, then it is handled through infra-red drying.The present invention does solvent with water; edible stage pectase and cellulase make catalyst and extract bitter melon polysaccharide; papain deproteinized; active carbon is except electrification and foreign pigment; the quality for improving bitter melon polysaccharide reduces organic solvent use, avoids the pollution to environment; the extraction process is easy to operate, operating procedure is few, at low cost, opens the new way of bitter melon polysaccharide large-scale production.
Description
Technical field
The invention belongs to technical field of natural product extraction, and in particular to a kind of biological enzyme extraction process of bitter melon polysaccharide.
Background technique
In recent years, its drug effect in disease treatment of the polysaccharide component in animals and plants has obtained extensive verifying, polysaccharide
Ingredient receives the extensive concern and research of pharmaceutical field, becomes the focus and emphasis of recent pharmacy research.It is bitter according to records
Melon has effects that relieve inflammation or internal heat improving eyesight, removing toxic substances, hypoglycemic, nourishing, QI invigorating, has certain disease treatment effect, illustrates to contain in balsam pear
Have bioactive ingredients abundant, wherein having a kind of bioactive ingredients is polysaccharide, also known as bitter melon polysaccharide, it have improve it is immune
Power, it is hypoglycemic, antibacterial, anti-oxidant the effects of, bitter melon polysaccharide (momordic acharantia polysaccharide, referred to as
MCP) it is one of polysaccharide, is a kind of heteroglycan being made of rhamnose, arabinose, mannose, glucose and galactolipin,
Bitter melon polysaccharide is frequently utilized for the clinical diseases such as hypoglycemic, anti-oxidant, antitumor and antibacterial, so, the extraction of bitter melon polysaccharide is in hardship
It is had a decisive role in the clinical application of melon polysaccharide.
The common extracting method of bitter melon polysaccharide has Hot water extraction, ultrasonic wave assisted extraction method, microwave abstracting, double-enzyme hydrolysis
Method, ultrasonic extraction etc..The existing extraction process of every kind of extracting method is as follows: 1, conventional method (Hot water extraction), in routine
The optimum condition of optimization technique in method extraction process: 100 DEG C of extraction temperature, extraction time 1 hour, alcohol precipitation concentration 85%, material-water ratio
1:20 (m:v), is tested, and the purity of bitter melon polysaccharide crude product is 36.1% at this time；2, ultrasonic wave assisted extraction method, in ultrasonic wave
The optimum condition of optimization technique in assisted extraction: 70 DEG C of extraction temperature, extraction time 25 minutes, 150 watts of ultrasonic power, alcohol precipitation
Concentration 85%, material-water ratio 1:20 (m:v), is tested, and the purity of bitter melon polysaccharide crude product is 80.77% at this time；3, microwave extraction method,
Optimize the optimum condition of technique during microwave abstracting are as follows: material-water ratio is 1:20 (m:v), and extraction time is 25 minutes, microwave
Power is 600 watts, and alcohol precipitation concentration 85% is tested, and the purity of bitter melon polysaccharide crude product is 56.81% at this time；4, dual-enzyme hydrolysis method,
The optimum condition of optimization technique in double-enzyme hydrolysis extraction process: cellulase, 50 DEG C of Extracting temperature, pH value 6.0, enzyme dosage
5%；Neutral proteinase, hydrolysis time 48 hours, pH value 7.0, enzyme dosage 10%, alcohol precipitation concentration 85% tested, bitter melon polysaccharide
The purity of crude product is 94.01%；5, ultrasonic extraction optimizes the optimum condition of technique during ultrasonic wave extraction: extraction temperature
55 DEG C of degree, extraction time 35 minutes, 350 watts of ultrasonic power, material-water ratio 1:30(m:V), alcohol precipitation concentration 85% is tested, hardship
The purity of melon polysaccharide crude is 82.51%.Pass through the bitter melon polysaccharide crude product purity obtained to above five kinds of bitter melon polysaccharide extracting methods
It is compared and learns, the bitter melon polysaccharide crude product purity highest that dual-enzyme hydrolysis method obtains.Therefore, bitter melon polysaccharide extracting method is determined as
Dual-enzyme hydrolysis method.However existing dual-enzyme hydrolysis method is lower to the recovery rate of bitter melon polysaccharide, so that the waste of big content of starting materials is caused,
High production cost, production efficiency are low.Therefore DNA purity and recovery rate in order to guarantee bitter melon polysaccharide simultaneously, it needs further to grind
The extracting method of bitter melon polysaccharide is studied carefully, to meet the dual requirements of bitter melon polysaccharide high-purity, high extraction.
Summary of the invention
The purpose of the present invention is to provide a kind of biological enzyme extraction processes for improving bitter melon polysaccharide recovery rate and purity, reduce
Production cost simplifies processing step, to overcome existing bitter melon polysaccharide extraction process that cannot meet high extraction and height simultaneously
The demand of purity.
To achieve the goals above, the present invention the following technical schemes are provided:
A kind of biological enzyme extraction process of bitter melon polysaccharide, comprising the following steps:
(1) it selects full balsam pear deseeding and removes flesh, clean slice, water and balsam pear are homogenized by the ratio of water to material of 1-10:1；
(2) homogenate of 1000g balsam pear is taken to be added in round-bottomed flask, what addition was made of cellulase, pectase and papain
Enzyme solutions are mixed, after mechanical stirring is uniform, pH value of solution is adjusted, solution is placed under 50-120 DEG C of bath temperature and extracts 1-7h,
It is cooled to room temperature after extraction, collects filtrate and residue respectively by filtering；
(3) water is added with the ratio of water to material of 1:1 into residue, is extracted again according to step (2), filtered after extraction by collected by suction
Liquid simultaneously merges with the filtrate of step (2) acquisition, and filtrate is concentrated, active carbon is then added thereto, mechanical stirring is uniformly taken out afterwards
Filtrate is collected in filter；
(4) 80% ethyl alcohol of 5 times of filtrate weight parts is added into the filtrate of step (3) under normal temperature condition, shakes up sealing and places
In 5 DEG C of refrigerator stand at low temperature, collected by suction sediment, by sediment with 80% ethanol washing 2 times, then by sediment with infrared dry
It is dry, obtain bitter melon polysaccharide.
Preferably, the ratio of water to material of water and balsam pear is 1:1 in the step (1).
Preferably, the dosage of the step (2) cellulase, pectase and papain is respectively as follows: 10,000 U/g
Cellulase 3g, 10,000 U/g pectase 1g, papain 1g.
Preferably, pH value of solution is 5.0 in the step (2).
Preferably, the extraction temperature in the step (2) is 80 DEG C.
Preferably, the extraction time in the step (2) is 1h.
Preferably, filtrate is concentrated into 100mL in the step (3), and the additive amount of active carbon is 3g.
Preferably, churned mechanically revolving speed is 80r/min in the extraction process.
Preferably, the balsam pear that the extraction process uses is informal voucher balsam pear.
The utility model has the advantages that doing solvent, food grade fruit the present invention provides a kind of biological enzyme extraction process of bitter melon polysaccharide with water
Glue enzyme and cellulase make catalyst and extract bitter melon polysaccharide, papain deproteinized, and active carbon is mentioned except electrification and foreign pigment
The high quality of bitter melon polysaccharide, reduces organic solvent use, avoids the pollution to environment, while having investigated the choosing of balsam pear
It selects, the condition of storage of fresh bitter, the technique for optimizing the extraction of bitter melon polysaccharide biological enzyme, reduces the extraction step of bitter melon polysaccharide
Suddenly, production link is simplified, production cost has been saved, opens the new way of bitter melon polysaccharide large-scale production.Therefore, the extraction
Technique is suitable for large-scale industrial production.
Detailed description of the invention
Fig. 1 is the extraction process flow chart of the bitter melon polysaccharide in embodiment 1.
Fig. 2 is bitter melon polysaccharide yield and extraction temperature relational graph in embodiment 1.
Fig. 3 is bitter melon polysaccharide yield and extraction time relational graph in embodiment 1.
Fig. 4 is bitter melon polysaccharide yield and ratio of water to material relational graph in embodiment 1.
Fig. 5 is the canonical plotting of absorbance and sugared content.
Fig. 6 is the extraction optimization technological process figure of the bitter melon polysaccharide of Preliminary design in embodiment 3.
Fig. 7 is the extraction optimization technological process figure of the bitter melon polysaccharide in embodiment 3.
Specific embodiment
The determination of 1 bitter melon polysaccharide extraction process of embodiment
1 instrument and reagent
1.1 instrument
SHZ-D circulating water type vacuum pump (Ying Yu Yu Hua instrument plant of Henan Gongyi City)；(Shanghai Soviet Union is up in fact for 112GKC temperature controlled water bath pot
Test Instrument Ltd.)；MS-DS10A1 soy bean milk making machine (fine work electric appliance Manufacturing Co., Ltd of Guangdong Province)；TU-1800 UV, visible light point
Light photometer (Beijing Puxi General Instrument Co., Ltd).
1.2 materials and reagent
Balsam pear: it is commercially available, come from the Suzhou vegetables market of farm produce.Dehydrated alcohol, common water, glucose, the concentrated sulfuric acid and anthrone reagent.
2 methods
The process of 2.1 extraction processes
It selects full balsam pear deseeding and removes flesh, weigh balsam pear 100g respectively after cleaning slice, a certain amount of water is added by requirement of experiment
It is homogenized, balsam pear homogenate is poured into the round-bottomed flask of different size, 3g cellulase and 1g pectase is separately added into, shakes
It is even, it extracts and cools down after a certain period of time, until room temperature, filters, residue continues plus water 10mL keeps old terms extraction 2 times, again mistake
Filter, merging filtrate, and it is concentrated into about 50mL, after cooling, papain 1g is added, 45 DEG C of constant temperature are dialysed 1 hour after shaking up, cold
But the dehydrated alcohol of 5 times of volumes is added to room temperature, is placed refrigerator 48 hours after shaking up sealing, collected by suction precipitating is dried at 50 DEG C
It is placed 12 hours in case, obtains bitter melon polysaccharide, weighed, finally carry out the assay of bitter melon polysaccharide.Extraction process flow chart is such as
Shown in Fig. 1.
2.2 factors influencing
Using balsam pear homogenate as raw material, bitter melon polysaccharide is extracted using the method for hot water extraction, alcohol chromatography, influences bitter melon polysaccharide extraction
Principal element have: extraction temperature, time, ratio of water to material etc..Therefore, this experiment is with extraction temperature, ratio of water to material, extraction time 3
A investigation factor by the single factor test changing rule of comprehensive extraction efficiency and multifactor interferes with each other the experiment final with determination
Condition.
2.2.1 the influence of extraction temperature
Condition: fixed ratio of water to material is 1:1, extraction time is 3 hours (h), and extraction temperature is respectively with 60 DEG C, 80 DEG C, 100 DEG C, 120
It DEG C is tested, as a result as shown in Figure 2.
2.2.2 the influence of extraction time
Condition: fixed temperature is 60 DEG C, ratio of water to material 1:1, extraction time are respectively with 1 h(hours), 3h, 5h, 7h tested,
As a result as shown in Figure 3.
2.2.3 the influence of ratio of water to material
Condition: fixed extraction temperature is 60 DEG C, extraction time is 3 hours (h), and ratio of water to material is respectively with 1:1,1:5,1:10,1:20
It is tested, as a result as shown in Figure 4.
By Fig. 2, Fig. 3 and Fig. 4 as it can be seen that bitter melon polysaccharide yield and extraction temperature, extraction time and ratio of water to material have it is close
Relationship can be seen that yield is increased with extraction temperature by the changing rule of curve and first increase then gradually gentle and have downward to become
Gesture；Yield has the tendency that gradually gentle after first reducing and has slightly upward with the increase of extraction time, but changes little；And yield
It tends to be steady again with first gentle slightly downward rear rise of the increase of ratio of water to material.So according to the reality of the influence of single factor test and experiment
The feasibility angle of border operation is set out, and selects three factors: extraction temperature, extraction time and ratio of water to material else in addition to blank column, while each
3 levels are taken, L is carried out9(34) orthogonal test[4], in which: extraction temperature takes 60 DEG C, 80 DEG C, 100 DEG C；Ratio of water to material take 1:1,5:1,
10:1；Extraction time takes 1h, 2h, 3h；The recovery rate and polyoses content of determining bitter melon polysaccharide is determined by experiment.Specific design is shown in
Table 1.
1 experimental factor level design table of table
2.3 experimental procedure
It is tested respectively according to experimental factor level design table according to the process of extraction process after optimization.
3 experimental results
The yield of 3.1 bitter melon polysaccharides
Pass through L9(34) orthogonal test designs table tested according to the process of extraction process after optimization, experimental result is shown in Table 2.
2 positive quadraturing design test result of table
3.2 bitter melon polysaccharide assays
3.2.1 principle and method
The assay of bitter melon polysaccharide usually relies on the characteristic color reaction of glucide.There are many common reagent, just react
For chemical process, it may be summarized to be two different types.(1) carbohydrate is made to be dehydrated shape with non-oxidizing acid (sulfuric acid or hydrochloric acid)
At furans aldehyde derivative, coloring matter further is generated with phenols or nitrogenous base condensation.(2) alkali process carbohydrate is used, is generated multiple
Miscellaneous cracking derivative acts on with certain reagents, special color is presented.Currently used Methods in Determination of Polysaccaride Content is
Anthrone-sulfuricacid method is to be hydrolyzed into monosaccharide under concentrated sulfuric acid effect using polysaccharide, monosaccharide is further based on first kind principle
It is dehydrated into alditol, alditol and anthrone, which are bound directly, generates blue-green derivative, which absorption maximum occurs at 620nm.
The operation has many advantages, such as that simple, quick, experimental phenomena is stable, reproducibility is preferable.Using glucose as standard items in this experiment,
The content of measurement bitter melon polysaccharide is analyzed using Anthrone-sulfuricacid method.
3.2.2 glucose standard curve is drawn
9 band plug rub oral examination tubes are taken, are that 0.5g/L glucose standards solution prepares sugar content (mg/mL) respectively and is with concentration
0.06,0.08,0.10,0.12,0.14,0.16,0.18,0.20,0.22 glucose solution.0.33g anthrone is weighed, is added
The 100mL concentrated sulfuric acid, is placed in brown bottle, shakes up.1mL series glucose standards solution is pipetted respectively in rub oral examination tube, simultaneously
Make blank with 1mL distilled water, every pipe is added 4.00mL Anthrone Sulphuric acid reagent, shakes up, be placed in ice-water bath immediately, is subsequently placed in
10min is heated in boiling water bath, it is cooling with tap water, 10 minutes are placed at room temperature for, absorbance is measured at 620nm.And with extinction
Degree is ordinate, and the content of glucose is abscissa, draws standard curve, as shown in Figure 5.
3.2.3 bitter melon polysaccharide assay and analysis
It takes 1mg extraction product difference label quantitative to 1mL respectively, measures its absorbance；The results are shown in Table 3:
The relationship of 3 polyoses content of table, polysaccharide amount and absorbance
By the bitter melon polysaccharide product volume and bitter melon polysaccharide content substitution L in table 39(34) orthogonal test table analyzed, it derives
Optimize the optimum experimental condition in technique out.
4 positive quadraturing design test result of table
5 L of table9(34) orthogonal test analysis of variance table
Pass through L9(34) orthogonal test table result and analysis of variance table determine the factor B(extraction temperature for influencing polysaccharide extract rate),
Factor C(ratio of water to material), factor D(extraction time) primary-slave relation, it is as a result as follows: 1, by very poor R value size analysis result be RD>
RC >RB, that is, influence the factor primary and secondary sequence of product yield: D > C > B.2, according to conspicuousness: FD>FC>FB Product is influenced to produce
The factor primary and secondary sequence of rate: D > C > B.It can be seen that all obtaining influence product yield according to Orthogonal experiment results and variance analysis
Factor primary and secondary sequence be D > C > B.Meanwhile each factor is respectively to the influence degree of polysaccharide extract rate on different level
D1>D2>D3, C3>C1>C2, B2>B3>B1, so, the optimal level collocation of bitter melon polysaccharide recovery rate is D1C3B2, i.e. the time is 1 small
When, ratio of water to material 10:1, extraction temperature be 80 DEG C.Bitter melon polysaccharide recovery rate at this time be 3.1%, purity 92.8%, this also with
Experimental result is consistent.
The determination of 2 balsam pear type of embodiment
Classification experiments are carried out according to different type balsam pear in different time periods, first according to different balsam pear type bar shaped balsam pears, oval
Shape balsam pear, short round balsam pear are tested, and optimal balsam pear type are determined by experiment, then in different time periods white (old)
Balsam pear and green (tender) balsam pear are tested, with the determination optimal period.Finally determine optimal balsam pear source.
Bitter melon polysaccharide is extracted using extracting method same as Example 1, is mentioned according to the optimum extraction condition of optimization
It takes: extraction time 1 hour, ratio of water to material 10:1,80 DEG C of extraction temperature.
Influence of the separate sources balsam pear to bitter melon polysaccharide recovery rate: with dual-enzyme hydrolysis method, to different type balsam pear, (bar shaped is bitter
Melon, oblong balsam pear, short round balsam pear) bitter melon polysaccharide extraction experiment is carried out, and bitter melon polysaccharide crude product amount and polyoses content are measured,
It the results are shown in Table 6.
The percentage composition of 6 different type balsam pear dual-enzyme hydrolysis method of table extraction bitter melon polysaccharide
The result shows that: by experiment measure different type balsam pear on momordica polysaccharide crude product and polyoses content, bar shaped balsam pear compared with
Short round and oblong balsam pear recovery rate with higher.
Influence of the balsam pear in different time periods to bitter melon polysaccharide recovery rate: selection bar shaped balsam pear, to hardship in different time periods
Melon (white (old) bar shaped balsam pear, green (tender) bar shaped balsam pear) carries out bitter melon polysaccharide again and extracts experiment, and measures bitter melon polysaccharide crude product
Amount and polyoses content, the results are shown in Table 7.
The recovery rate of bitter melon polysaccharide in white (old) the bar shaped balsam pear of table 7 and green (tender) bar shaped balsam pear
The result shows that: it is white on momordica polysaccharide crude product and polyoses content that bar shaped balsam pear in different time periods is measured by experiment
Greener (tender) the bar shaped balsam pear of (old) bar shaped balsam pear is good.
To sum up conclude that selection is white (old) bar shaped balsam pear in bitter melon polysaccharide extraction process, what bitter melon polysaccharide extracted
Method is the optimum extraction condition that dual-enzyme hydrolysis method and dual-enzyme hydrolysis method extract bitter melon polysaccharide are as follows: extraction time 1 hour, water
Material is than 10:1, and 80 DEG C of extraction temperature.
3 bitter melon polysaccharide biological enzyme extraction process of embodiment advanced optimizes
In conjunction with the embodiments 1 and embodiment 2 experimental data, balsam pear selects white (old) bar shaped balsam pear, the extraction preliminary process obtained
(see figure 6) are as follows: extraction time is 1 hour, ratio of water to material 10:1, and extraction temperature is 80 DEG C.But active, the extensive life of desmoenzyme
The physical condition of production, the factors such as raw material own characteristic, on the basis of original extraction process, the process conditions that advanced optimize
Are as follows: under conditions of ratio of water to material is 8) 1:1(is shown in Table, 10,000 U/g cellulase 3g, 10,000 U/g fruits are added in the homogenate of 1000g balsam pear
9) glue enzyme 1g and papain 1g(are shown in Table, mechanical stirring (80r/min), and pH value is adjusted to 5, and water bath heating temperature is 80 DEG C,
Extraction time is 1 hour.
Ratio of water to material determines: in order to fully consider reality, now carrying out further check experiment to ratio of water to material 10:1 and 1:1.
The different ratio of water to material of table 8 do the percentage composition that check experiment extracts bitter melon polysaccharide
The determination of enzyme dosage: original 1/5 is reduced into enzyme dosage and does the influence that check experiment determines enzyme dosage.Cellulase is 1
Ten thousand U/g, pectase are 10,000 U/g and papain.Amount ratio is dosage after 3g:1g:1g(optimizes by 1000g balsam pear slurry) and
Dosage before 15g:5g:5g(optimizes), water and balsam pear are by 1:1(mL:g) control experiment is carried out respectively.
9 enzyme different amounts of table do the percentage composition that check experiment extracts bitter melon polysaccharide
The determination of the storage requirement of fresh bitter: since balsam pear is to belong to fresh seasonal vegetable, period of storage is short, to avoid storing
The improper loss for being easy to cause balsam pear ingredient is deposited, therefore, the balsam pear and room temperature balsam pear for freezing 48 hours by p- 25 DEG C compare
Test, in the identical situation of experiment condition, it is determined that the low tempertaure storage of fresh bitter influences less the loss of balsam pear ingredient,
So -25 DEG C of refrigerated storages and spare can be carried out to balsam pear during balsam pear lists.Concrete outcome is shown in Table 10.
10-25 DEG C of the table balsam pears for freezing 48 hours and room temperature balsam pear do the percentage composition that check experiment extracts bitter melon polysaccharide
Specific optimal extraction technology are as follows: selection informal voucher balsam pear, -25 DEG C of low temperature refrigerator freeze 48 hours, take out and thaw, and deseeding is gone
Flesh cleans slice, and balsam pear and water are homogenized by 1:1, takes the homogenate of 1000g balsam pear to be added in round-bottomed flask, it is fine that 10,000 U/g are added
Plain enzyme 3g, 10,000 U/g pectase 1g and papain 1g, mechanical stirring (80r/min) are tieed up, pH value is adjusted to 5, heating water bath
Temperature is 80 DEG C (foundation: such as Fig. 2), and extraction time is 1 hour, is cooled to room temperature after extraction, is filtered, and residue 1:1 adds water
Extracted again by old terms, after merging filtrate, be concentrated into about 100mL, be added active carbon 3g, mechanical stirring 20 minutes
(80r/min) is filtered, and 5 times of 80% ethyl alcohol of amount are added in filtrate room temperature, is shaken up sealing and is placed 5 DEG C of refrigerators, 48 hours stand at low temperature, takes out
Filter, is washed 2 times, sediment infra-red drying obtains bitter melon polysaccharide with 80% ethyl alcohol.Optimized Extraction Process process is as shown in Figure 7.
As a result: the amount for bitter melon polysaccharide of weighing is that 14.6g(balsam pear amount is 500g, show that recovery rate is 2.92%, connects very much
Closely by the nutritional ingredient and content to fresh bitter point of Chinese Academy Of Preventive Medicine Research Institute Of Nutrition And Food Hygiene's publication
Carbohydrate percentage composition (2.60-3.50%) in analysis), by bitter melon polysaccharide preparing standard solution obtained measure absorbance A=
4.404, substitute into the standard curve of Fig. 5 absorbance and sugared content, it was therefore concluded that: the bitter melon polysaccharide purity extracted is
94.6%。
Claims (9)
1. a kind of biological enzyme extraction process of bitter melon polysaccharide, which comprises the following steps:
(1) it selects full balsam pear deseeding and removes flesh, clean slice, water and balsam pear are homogenized by the ratio of water to material mL/g of 1-10:1；
(2) homogenate of 1000g balsam pear is taken to be added in round-bottomed flask, what addition was made of cellulase, pectase and papain
Enzyme solutions are mixed, after mechanical stirring is uniform, pH value of solution is adjusted, solution is placed under 50-120 DEG C of bath temperature and extracts 1-7h,
It is cooled to room temperature after extraction, collects filtrate and residue respectively by filtering；
(3) water is added with the ratio of water to material of 1:1 into residue, is extracted again according to step (2), filtered after extraction by collected by suction
Liquid simultaneously merges with the filtrate of step (2) acquisition, and filtrate is concentrated, active carbon is then added thereto, mechanical stirring is uniformly taken out afterwards
Filtrate is collected in filter；
(4) 80% ethyl alcohol of 5 times of filtrate weight parts is added into the filtrate of step (3) under normal temperature condition, shakes up sealing and places
In 5 DEG C of refrigerator stand at low temperature, collected by suction sediment, by sediment with 80% ethanol washing 2 times, then by sediment with infrared dry
It is dry, obtain bitter melon polysaccharide.
2. a kind of biological enzyme extraction process of bitter melon polysaccharide according to claim 1, which is characterized in that the step
(1) ratio of water to material of water and balsam pear is 1:1 in.
3. a kind of biological enzyme extraction process of bitter melon polysaccharide according to claim 1, which is characterized in that the step
(2) dosage of cellulase, pectase and papain be respectively as follows: 10,000 U/g cellulase 3g, 10,000 U/g pectase 1g,
Papain 1g.
4. a kind of biological enzyme extraction process of bitter melon polysaccharide according to claim 1, which is characterized in that the step
(2) pH value of solution is 5.0 in.
5. a kind of biological enzyme extraction process of bitter melon polysaccharide according to claim 1, which is characterized in that the step
(2) extraction temperature in is 80 DEG C.
6. a kind of biological enzyme extraction process of bitter melon polysaccharide according to claim 1, which is characterized in that the step
(2) extraction time in is 1h.
7. a kind of biological enzyme extraction process of bitter melon polysaccharide according to claim 1, which is characterized in that the step
(3) filtrate is concentrated into 100mL in, and the additive amount of active carbon is 3g.
8. a kind of biological enzyme extraction process of bitter melon polysaccharide according to claim 1, which is characterized in that the extraction process
In churned mechanically revolving speed be 80r/min.
9. a kind of biological enzyme extraction process of bitter melon polysaccharide according to claim 1, which is characterized in that the extraction process
The balsam pear used is informal voucher balsam pear.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110845633A (en) * | 2019-11-11 | 2020-02-28 | 江苏大学 | Normal-temperature method for extracting active polysaccharide from fresh bitter gourds |
| RU2806822C2 (en) * | 2021-10-05 | 2023-11-07 | Евгений Михайлович Герасимов | Method for processing gourds with obtaining pectin-containing compositions |
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Application publication date: 20190129 |