CN102108350A - Preparation method of bromelin - Google Patents
Preparation method of bromelin Download PDFInfo
- Publication number
- CN102108350A CN102108350A CN2009102007555A CN200910200755A CN102108350A CN 102108350 A CN102108350 A CN 102108350A CN 2009102007555 A CN2009102007555 A CN 2009102007555A CN 200910200755 A CN200910200755 A CN 200910200755A CN 102108350 A CN102108350 A CN 102108350A
- Authority
- CN
- China
- Prior art keywords
- bromeline
- preparation
- bromelin
- concentration
- enzyme
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Enzymes And Modification Thereof (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention relates to a preparation method of bromelin, and the specific steps are as follows: 1) extracting the bromelin, adding an organic solvent into pineapple condensed juice after ultra-filtration by a PSA (pressure sensitive adhesive) film for precipitating an enzyme, and further performing drying under reduced pressure and other steps for getting a high-activity enzyme product; and 2) purifying the bromelin, (i) utilizing polyacrylic acid fixed by magnetic ferric oxide microspheres for adsorbing the bromelin, wherein the adsorption rate of the bromelin approaches 100%, the pH value is 6-8, 0.01-0.15mol/L of phosphoric acid buffer solution and KCl with the concentration of 0.6-2.0mol/L are adopted as eluent for performing elution, and the adsorption process and the desorption process are completed within 0.5-1.5 minutes; and (ii) using a double-water phase system of a copolymer of epoxy ethane and epoxypropane to separate and extract the high-purity bromelin. The preparation method has the advantages that the production is simple, and the large-scale industrial production can be performed; and the bromelin is derived from natural plants, thereby having the characteristics of safety, non-toxicity, wide application range and the like; and the application is wide.
Description
[technical field]
The present invention relates to the bromeline technical field, specifically, is a kind of preparation method of bromeline.
[background technology]
Bromeline (bromelain) is the general name of the proteinoid lytic enzyme that extracts from the pineapple plant, mainly is present in pineapple stem and the fruit, according to the difference of extract part, is divided into stem bromeline and fruit bromeline.Bromeline is vulnerable to the influence of temperature, pH, metal ion etc. as activated protein.The optimal reactive temperature of bromeline is between (55～60) ℃, and temperature is high again, and the heat inactivation of enzyme strengthens, and speed of response descends; The optimal pH of bromeline reaches maximum value 6.5～7.5 near 7.1, in sour environment, enzyme reaction rate descends fast, and the range of application of individual broad is arranged under alkalescence, enzyme than stable p H scope 3.5～4.5.
The application of bromeline has very long history, and along with the progress of extracting purification technique, it will have widely uses.China's pineapple aboundresources is produced pineapple per year more than 1,000,000 tons, and annual all have a large amount of pineapple peel, stem to be wasted.Simultaneously, China increases year by year to the demand of bromeline, particularly highly active bromeline.At field of medicaments, more and more higher to the purity requirement of bromeline, even to clearly play the concrete composition of function, and the enzyme of suitability for industrialized production is difficult to reach this standard at present, just requires to extract purification technique will constantly improve, to satisfy the demand in each field.Bromeline is as biologically active substance, determined influence that it is vulnerable to outside atmosphere and inactivation.Therefore, it is most important to keep required enzyme to live in production, storage, transportation, and this just requires the immobilization technology etc. of enzyme to have breakthrough.
The present situation of present domestic bromeline: the extracting and purifying method of bromeline is a lot, and the extracting method that is used for suitability for industrialized production mainly contains 3 kinds: kaolin absorption method, the tannin precipitator method, ultrafiltration and concentration method.In the above method, it is higher that kaolin absorption method products obtained therefrom enzyme is lived, but production technique and operate all more complicated, and required equipment is many, and raw materials consumption is also many, and the enzyme rate of recovery of living is low, contaminate environment, and environmental protection investments are big; The tannin precipitator method, production technique and operation are fairly simple, and the rate of recovery is low but enzyme is lived, and the activity of gained enzyme is not high yet, and added tannin causes certain pollution to environment.Produce the method that adopts ultrafiltration and concentration mostly on the domestic industry.This method is utilized the molecular retention principle, greater than removing by filter with most of material less than protease molecule, to reach the purpose that concentrates enzyme liquid, behind the pineapple juice ultrafiltration and concentration, carries out drying again and obtains enzyme preparation.The more preceding two kinds of method operation stepss of this method are few, and mild condition has reduced enzyme loss alive, produce and the environmental protection less investment, do not have the foreign protein of enzymic activity more but contain in the gained enzyme preparation, this type of molecular weight of albumen and bromeline are approximate, so ultra-filtration process can't be removed.
Simultaneously, China increases year by year to the demand of bromeline, particularly highly active bromeline.At field of medicaments, more and more higher to the purity requirement of bromeline, the enzyme of suitability for industrialized production is difficult to arrive this standard at present, and this just requires to extract purification technique and will constantly improve, to satisfy the demand in each field.
[summary of the invention]
The objective of the invention is to overcome the deficiencies in the prior art, a kind of preparation method of bromeline is provided.
The objective of the invention is to be achieved through the following technical solutions:
A kind of preparation method of bromeline, its concrete steps are:
One, the extraction of bromeline
The ultrafiltration and concentration method is improved, made the enzyme precipitation add organic solvent with the pineapple concentrated juice behind the PSA membrane ultrafiltration, obtain the high active enzyme goods through steps such as drying under reduced pressure again, enzyme work reaches 6.01 * 106U/g;
Two, the purifying of bromeline
(1) utilizes fixedly polyacrylic acid absorption bromeline of martial ethiops microballoon, the adsorption rate of bromeline is near 100%, in the pH value is 6～8, the KCl that is 0.6～2.0mol/L with phosphoric acid buffer and the concentration of 0.01～0.15mol/L carries out wash-out as elutriant, owing to there is not inside diffusional resistance, absorption and desorption process just can be finished in 0.5～1.5min, and the enzyme of the bromeline of this moment rate of recovery alive is 7.4%;
The concentration of described phosphoric acid buffer is preferably 0.1mol/L;
The concentration of described elutriant KCL is preferably 1～1.2mol/L;
Described absorption and desorb process time are preferably 1min;
(2) the highly purified bromeline of double-aqueous phase system separating and extracting of usefulness oxyethane and epoxy propane copolymer, separation condition is that the oxyethane mass percent is 5～15% for (content) in the multipolymer, the molecular weight of multipolymer is 2000g/mol, copolymer quality per-cent is 1～10%, temperature is 20～30 ℃, and the pH value is 5～7, and concentration of salt solution is when 5～20mmol/L, the enzyme rate of recovery alive can reach 94.2%, and enzyme work reaches 6.55 * 106U/g;
Ethylene oxide content is 10% in the described multipolymer;
The molecular weight of described multipolymer is 2000g/mol, and copolymer quality per-cent is 5%;
Described temperature is 25 ℃, and the pH value is 6.0, and concentration of salt solution is 15mmol/L.
The application of the bromeline of the present invention's preparation:
The bromeline that the present invention produces has important pharmaceutical use and has immunostimulation, studies show that, by to the hormesis of helper with to the direct inhibited reaction of T cell, can strengthen the immune response with suppressor T cell; Can prevent and treat cardiovascular disorder, the decomposition of its energy accelerating fibers proteinogen, allevating angina pectoris and apoplexy; In various tissues, oedema and inflammation can be treated effectively, and diarrhoea can be treated; Bromeline is used for decrustation after burn, can optionally remove Exfoliating Scrub, and normal skin is had no adverse effects, and decrustation is thorough, and is significant to the treatment burn; The bromelain endonuclease capable suppresses the growth of tumour cell, reduces toxin in conjunction with activity, and can suppress the secretion effect of heat-labile toxin and Toxins,exo-, cholera; Can promote the absorption of medicine, can improve its curative effect with various microbiotic couplings, and can promote the absorption of nutritive substance.
Compared with prior art, positively effect of the present invention is:
(1) bromeline production of the present invention is simple, but large-scale industrial production;
(2) bromeline of the present invention derives from natural phant, has spies such as safety non-toxic, sphere of action be wide;
(3) bromeline of the present invention's preparation has important pharmaceutical use and has immunostimulation; Be widely used.
[embodiment]
The preparation method's of a kind of bromeline of the present invention embodiment below is provided.
Embodiment 1
A kind of preparation method of bromeline, its concrete steps are:
One, the extraction of bromeline
The ultrafiltration and concentration method is improved, made the enzyme precipitation add organic solvent with the pineapple concentrated juice behind the PSA membrane ultrafiltration, obtain the high active enzyme goods through steps such as drying under reduced pressure again, enzyme work reaches 6.01 * 106U/g;
Two, the purifying of bromeline
(1) utilizes fixedly polyacrylic acid absorption bromeline of martial ethiops microballoon, the adsorption rate of bromeline is near 100%, in the pH value is 6, the KCl that is 0.6mol/L with phosphoric acid buffer and the concentration of 0.01mol/L carries out wash-out as elutriant, owing to there is not inside diffusional resistance, absorption and desorption process just can be finished in 0.5min, and the enzyme of the bromeline of this moment rate of recovery alive is 7.4%;
(2) the highly purified bromeline of double-aqueous phase system separating and extracting of usefulness oxyethane and epoxy propane copolymer, separation condition is that ethylene oxide content is 5% in the multipolymer, the molecular weight of multipolymer is 2000 for g/mol, copolymer quality per-cent is 1%, temperature is 20 ℃, and the pH value is 5, and concentration of salt solution is when 5mmol/L, the enzyme rate of recovery alive can reach 94.2%, and enzyme work reaches 6.55 * 106U/g.
Embodiment 2
A kind of preparation method of bromeline, its concrete steps are:
One, the extraction of bromeline
The ultrafiltration and concentration method is improved, made the enzyme precipitation add organic solvent with the pineapple concentrated juice behind the PSA membrane ultrafiltration, obtain the high active enzyme goods through steps such as drying under reduced pressure again, enzyme work reaches 6.01 * 106U/g;
Two, the purifying of bromeline
(1) utilizes fixedly polyacrylic acid absorption bromeline of martial ethiops microballoon, the adsorption rate of bromeline is near 100%, in the pH value is 6.5, the KCl that is 0.9mol/L with phosphoric acid buffer and the concentration of 0.05mol/L carries out wash-out as elutriant, owing to there is not inside diffusional resistance, absorption and desorption process just can be finished in 0.7min, and the enzyme of the bromeline of this moment rate of recovery alive is 7.4%;
(2) the highly purified bromeline of double-aqueous phase system separating and extracting of usefulness oxyethane and epoxy propane copolymer, separation condition is that ethylene oxide content is 8% in the multipolymer, the molecular weight of multipolymer is 2000 for g/mol, copolymer quality per-cent is 3%, temperature is 23 ℃, and the pH value is 5.5, and concentration of salt solution is when 8mmol/L, the enzyme rate of recovery alive can reach 94.2%, and enzyme work reaches 6.55 * 106U/g.
Embodiment 3
A kind of preparation method of bromeline, its concrete steps are:
One, the extraction of bromeline
The ultrafiltration and concentration method is improved, made the enzyme precipitation add organic solvent with the pineapple concentrated juice behind the PSA membrane ultrafiltration, obtain the high active enzyme goods through steps such as drying under reduced pressure again, enzyme work reaches 6.01 * 106U/g;
Two, the purifying of bromeline
(1) utilizes fixedly polyacrylic acid absorption bromeline of martial ethiops microballoon, the adsorption rate of bromeline is near 100%, in the pH value is 7, the KCl that is 1mol/L with phosphoric acid buffer and the concentration of 0.1mol/L carries out wash-out as elutriant, owing to there is not inside diffusional resistance, absorption and desorption process just can be finished in 1min, and the enzyme of the bromeline of this moment rate of recovery alive is 7.4%;
(2) the highly purified bromeline of double-aqueous phase system separating and extracting of usefulness oxyethane and epoxy propane copolymer, separation condition is an ethylene oxide content 10% in the multipolymer, the molecular weight of multipolymer is 2000g/mol, copolymer quality per-cent is 5%, temperature is 25 ℃, and the pH value is 6, and concentration of salt solution is when 15mmol/L, the enzyme rate of recovery alive can reach 94.2%, and enzyme work reaches 6.55 * 106U/g.
Embodiment 4
A kind of preparation method of bromeline, its concrete steps are:
One, the extraction of bromeline
The ultrafiltration and concentration method is improved, made the enzyme precipitation add organic solvent with the pineapple concentrated juice behind the PSA membrane ultrafiltration, obtain the high active enzyme goods through steps such as drying under reduced pressure again, enzyme work reaches 6.01 * 106U/g;
Two, the purifying of bromeline
(1) utilizes fixedly polyacrylic acid absorption bromeline of martial ethiops microballoon, the adsorption rate of bromeline is near 100%, in the pH value is 7.8, the KCl that is 1.6mol/L with phosphoric acid buffer and the concentration of 0.12mol/L carries out wash-out as elutriant, owing to there is not inside diffusional resistance, absorption and desorption process just can be finished in 1.2min, and the enzyme of the bromeline of this moment rate of recovery alive is 7.4%;
(2) the highly purified bromeline of double-aqueous phase system separating and extracting of usefulness oxyethane and epoxy propane copolymer, separation condition is that ethylene oxide content is 12% in the multipolymer, the molecular weight of multipolymer is 2000g/mol, copolymer quality per-cent is 8%, temperature is 27 ℃, and the pH value is 6.6, and concentration of salt solution is when 17mmol/L, the enzyme rate of recovery alive can reach 94.2%, and enzyme work reaches 6.55 * 106U/g.
Embodiment 5
A kind of preparation method of bromeline, its concrete steps are:
One, the extraction of bromeline
The ultrafiltration and concentration method is improved, made the enzyme precipitation add organic solvent with the pineapple concentrated juice behind the PSA membrane ultrafiltration, obtain the high active enzyme goods through steps such as drying under reduced pressure again, enzyme work reaches 6.01 * 106U/g;
Two, the purifying of bromeline
(1) utilizes fixedly polyacrylic acid absorption bromeline of martial ethiops microballoon, the adsorption rate of bromeline is near 100%, in the pH value is 8, the KCl that is 2.0mol/L with phosphoric acid buffer and the concentration of 0.15mol/L carries out wash-out as elutriant, owing to there is not inside diffusional resistance, absorption and desorption process just can be finished in 1.5min, and the enzyme of the bromeline of this moment rate of recovery alive is 7.4%;
(2) the highly purified bromeline of double-aqueous phase system separating and extracting of usefulness oxyethane and epoxy propane copolymer, separation condition is that ethylene oxide content is 15% in the multipolymer, the molecular weight of multipolymer is 2000g/mol, copolymer quality per-cent is 10%, temperature is 30 ℃, and the pH value is 7, and concentration of salt solution is when 20mmol/L, the enzyme rate of recovery alive can reach 94.2%, and enzyme work reaches 6.55 * 106U/g.
Bromeline production of the present invention is simple, but large-scale industrial production; Bromeline derives from natural phant, has spies such as safety non-toxic, sphere of action be wide; The bromeline of the present invention's preparation has important pharmaceutical use and has immunostimulation; Be widely used.
The above only is a preferred implementation of the present invention; should be pointed out that for those skilled in the art, without departing from the inventive concept of the premise; can also make some improvements and modifications, these improvements and modifications also should be considered within the scope of protection of the present invention.
Claims (8)
1. the preparation method of a bromeline is characterized in that, concrete steps are:
One, the extraction of bromeline
Add organic solvent with the pineapple concentrated juice behind the PSA membrane ultrafiltration and make the enzyme precipitation, obtain the high active enzyme goods through steps such as drying under reduced pressure again;
Two, the purifying of bromeline
(1) utilizes fixedly polyacrylic acid absorption bromeline of martial ethiops microballoon, the adsorption rate of bromeline is near 100%, in the pH value is 6～8, the KCl that is 0.6～2.0mol/L with phosphoric acid buffer and the concentration of 0.01～0.15mol/L carries out wash-out as elutriant, and absorption and desorption process are finished in 0.5～1.5min;
(2) the highly purified bromeline of double-aqueous phase system separating and extracting of usefulness oxyethane and epoxy propane copolymer, separation condition is that the oxyethane mass percent is 5～15% in the multipolymer, the molecular weight of multipolymer is 2000g/mol, copolymer quality per-cent is 1～10%, temperature is 20～30 ℃, the pH value is 5～7, and concentration of salt solution is 5～20mmol/L.
2. the preparation method of a kind of bromeline as claimed in claim 1 is characterized in that, in described step (), the concentration of described phosphoric acid buffer is 0.1mol/L.
3. the preparation method of a kind of bromeline as claimed in claim 1 is characterized in that, in described step (), the concentration of described elutriant KCL is 1～1.2mol/L.
4. the preparation method of a kind of bromeline as claimed in claim 1 is characterized in that, in described step (), described absorption and desorb process time are 1min.
5. the preparation method of a kind of bromeline as claimed in claim 1 is characterized in that, in described step (two), ethylene oxide content is 10% in the described multipolymer.
6. the preparation method of a kind of bromeline as claimed in claim 1 is characterized in that, in described step (two), the molecular weight of described multipolymer is 2000g/mol, and copolymer quality per-cent is 5%.
7. the preparation method of a kind of bromeline as claimed in claim 1 is characterized in that, in described step (two), described temperature is 25 ℃, and the pH value is 6.0, and concentration of salt solution is 15mmol/L.
8. the preparation method of a kind of bromeline as claimed in claim 1 is characterized in that, the application of described bromeline in medicine.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009102007555A CN102108350A (en) | 2009-12-25 | 2009-12-25 | Preparation method of bromelin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009102007555A CN102108350A (en) | 2009-12-25 | 2009-12-25 | Preparation method of bromelin |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102108350A true CN102108350A (en) | 2011-06-29 |
Family
ID=44172634
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2009102007555A Pending CN102108350A (en) | 2009-12-25 | 2009-12-25 | Preparation method of bromelin |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102108350A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103191581A (en) * | 2013-04-09 | 2013-07-10 | 江苏大学 | Method for separating/gathering trace ciprofloxacin in food |
| CN104531643A (en) * | 2015-01-21 | 2015-04-22 | 扬州大学 | Method for separating and purifying of bromelain from pineapple peels |
| CN104611316A (en) * | 2015-02-13 | 2015-05-13 | 广西壮族自治区分析测试研究中心 | Method for extracting bromelin |
| CN106119230A (en) * | 2016-08-11 | 2016-11-16 | 江苏大学 | A kind of method circulating the isolated and purified bromelain of double-aqueous phase system |
-
2009
- 2009-12-25 CN CN2009102007555A patent/CN102108350A/en active Pending
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103191581A (en) * | 2013-04-09 | 2013-07-10 | 江苏大学 | Method for separating/gathering trace ciprofloxacin in food |
| CN103191581B (en) * | 2013-04-09 | 2015-06-10 | 江苏大学 | Method for separating/gathering trace ciprofloxacin in food |
| CN104531643A (en) * | 2015-01-21 | 2015-04-22 | 扬州大学 | Method for separating and purifying of bromelain from pineapple peels |
| CN104611316A (en) * | 2015-02-13 | 2015-05-13 | 广西壮族自治区分析测试研究中心 | Method for extracting bromelin |
| CN106119230A (en) * | 2016-08-11 | 2016-11-16 | 江苏大学 | A kind of method circulating the isolated and purified bromelain of double-aqueous phase system |
| CN106119230B (en) * | 2016-08-11 | 2019-12-03 | 江苏大学 | A method of circulation double-aqueous phase system isolates and purifies bromelain |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN1273497C (en) | Process for preparing chondroitin sulfate | |
| CN104262413B (en) | Preparation method of trehalose anhydrous | |
| CN107217048A (en) | It is a kind of to catalyze and synthesize aminopeptidase of carnosine and its preparation method and application | |
| CN102108352A (en) | Immobilization method of bromelain | |
| CN102108350A (en) | Preparation method of bromelin | |
| CN107312807A (en) | The enzymolysis preparation of the brown alga function oligosaccharides in one main laminaria source | |
| WO2020042557A1 (en) | Method for extracting citrulline from watermelon | |
| CN110698525B (en) | Preparation method of cranberry extract | |
| CN101701069B (en) | Method for extracting epsilon-polylysine and salt thereof | |
| CN102228125A (en) | Preparation method of algal active peptide | |
| CN102249961A (en) | Method for extracting alliin from garlic | |
| US8173837B1 (en) | Process for the production of L-citrulline from watermelon flesh and rind | |
| CN102220390A (en) | Method for preparing citrulline by combining arginine fermentation and enzymatic conversion | |
| CN101434972A (en) | Process for preparing Lonicera edulis anthocyanin by solid state biological reaction | |
| CN104402976B (en) | A kind of method preparing enramycin fine powder | |
| CN101845089B (en) | Method for large-scale production of neurotoxin in cobra venin and reduction of neurotoxicity | |
| CN102071180A (en) | New process for preparing pineapple bromelain | |
| CN1408401A (en) | Cherry extract and its extracting method | |
| CN104232606A (en) | Improved beta-glucosidase as well as expression gene and application thereof | |
| CN114751825A (en) | Method for efficiently extracting chlorogenic acid from honeysuckle | |
| CN108147964A (en) | A kind of method with Integrated Membrane Technology separating chlorogenic acid | |
| CN102603450A (en) | Continuous extraction method of amino acid and pectin in watermelon peel | |
| CN104558212B (en) | Application of aminated bio-cellulose to preparation of moist dressing | |
| CN114249666A (en) | Method for extracting betaine from phoenix tree leaves | |
| Gu et al. | Production of recombinant β-glucosidase and simultaneous immobilization and purification using immobilized metal ion affinity membrane |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20110629 |