CN101942423A - Catalase and extraction method thereof - Google Patents
Catalase and extraction method thereof Download PDFInfo
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- CN101942423A CN101942423A CN 201010265399 CN201010265399A CN101942423A CN 101942423 A CN101942423 A CN 101942423A CN 201010265399 CN201010265399 CN 201010265399 CN 201010265399 A CN201010265399 A CN 201010265399A CN 101942423 A CN101942423 A CN 101942423A
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- catalase
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- animal livers
- slurries
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Abstract
The invention discloses catalase and an extraction method thereof. The catalase is extracted by taking animal livers as raw materials and adopting a supercritical CO2 extraction method. The extraction method has the advantages of short production period, little wastewater discharge, safety, energy saving and high production yield. The product concentration of the produced catalase (CAT) reaches 211824.5U/ml proteins as well as exceeds and reaches that of a similar product, the shelf life of the produced catalase is 180 days, and the total colony count of the produced catalase is no more than 1000/ml.
Description
Technical field the present invention relates to a kind of catalase and extracting method thereof.
(Catalase is a kind of efficient, safety, environmental protection, energy-conservation biological catalyst CAT) to the background technology catalase, can be water and oxygen with hydrogen peroxide decomposition rapidly.Thereby fresh-keeping at food antiseptic, the bleachinging and dyeing of textile printing and dyeing industry, paper industry bleaching are handled and waste water control, the chemical rubber moulding, be used widely in aspects such as plastics binder.
The method of home and abroad hydrogen peroxidase extracting has two kinds usually at present; A kind of is the Production by Microorganism Fermentation catalase; Another kind is to be the raw material hydrogen peroxidase extracting with the animal livers.Preceding a kind of extraction process still is in the experimental study stage at present; Belong to short run, also do not have large-scale industrial production; Second kind is raw material with the animal livers, at home, report is all arranged as U.S. Pat 2992167 etc. outward, the Chinese patent ZL03152957.7 " a kind of method of extraction method hydrogen peroxidase extracting " of Southern Yangtze University application and number of patent application are 02110792.0 the method for hydrogen peroxidase extracting " a kind of from animal livers ", these two kinds of methods all are to be to extract raw material with the animal livers, needing to add various chemical compositions such as ammonium sulfate etc. in leaching process precipitates, with an organic solvent dissolve, lixiviate is got the time more than 20 hours, the lixiviate slag, liquid separates, parting liquid adds ammonium sulfate precipitation and promptly gets product.Need to use a large amount of organic solvents in its production process, the product stability of its extraction is poor, the easy inactivation of producing of enzyme causes certain pollution to environment, produces long in time limit, and the production yield is low, and be that product is easy to separate and purifying with fermentative Production catalase advantage, but complex manufacturing, it is high that manufacturing condition requires, investment is big, and three wastes treatment capacity is big.
The catalatic enterprise of production and supply mainly contains Denmark Novozymes Company on the domestic market at present, and it is maximum in the world enzyme product production exploitation enterprise, be provided with how tame branch office at home, but it produces hydrogen peroxide enzyme product, market price height; Also have the Chinese patent application 200910027116.3 " a kind of is the technology of raw material while hydrogen peroxidase extracting and hepatocyte growth-promoting liquid with the animal livers " of Jiangsu Engineering Research Center for Bioactive Products Processing Technology's application to explore and when with the animal livers being raw material hydrogen peroxidase extracting product, produce other biochemical industry product, can significantly reduce like this and produce the catalase product price, but existing also still at the experimental stage, and the production process three wastes are handled complicated use strong acid, alkaline solution is big for environment pollution.The catalatic technical matters of existing production from the home and abroad, yet there are no with the animal livers is raw material, adopts supercritical carbon dioxide extraction catalase technical scheme.
Summary of the invention the present invention will provide a kind of catalase and extracting method thereof exactly.It is that raw material adopts supercritical CO with the animal livers
2The extraction process hydrogen peroxidase extracting, with short production cycle, discharge of wastewater is few, energy-saving safe, production yield height.
Catalase extracting method of the present invention is raw material with the animal livers, adopts supercritical CO
2The extraction process hydrogen peroxidase extracting.
Catalase extracting method of the present invention, described supercritical carbon dioxide extraction method hydrogen peroxidase extracting, undertaken by following processing step:
(1) raw material is prepared the animal livers processing animal livers pulverization process is made slurries;
(2) dissolving mixes above-mentioned slurries and supercritical CO
2Fluid, thorough mixing in mixing device makes mixed stream;
(3) separate removal of impurities mixed stream is placed tripping device, separate and remove insoluble sludge and coarse particles in the mixed stream, get and just divide mixed stream;
(4) extraction preparing hydrogen peroxide enzyme will just divide mixed stream to carry out the gas-liquid CO that is separated with tripping device
2, the liquid phase that stays is the hydrogen peroxide enzyme product.
Described catalase extracting method, preferably undertaken by following processing step:
(1) raw material prepare animal livers handle with animal livers clean, mowing, removal of impurities, soak pulverization process and make slurries;
(2) dissolving mixes: with above-mentioned slurries and supercritical CO
2Fluid places the mixing device of pressurized vessel, carries out thorough mixing and makes the component that contains solubility in the slurries dissolve in supercritical CO fully
2Form mixed stream in the fluid solvent;
(3) separate removal of impurities mixed stream placed tripping device, through once or the separation of some levels time remove insoluble sludge and coarse particles in the mixed stream, make the first minute mixed stream that is full of the catalase solute;
(4) extraction preparing hydrogen peroxide enzyme will just divide the mixed stream tripping device, carry out once or several times extracting gas liquid phase separation CO
2, supercritical CO
2Become gas phase by liquid phase and reclaim, stay liquid phase and be the hydrogen peroxide enzyme product.
Mixing device of the present invention is an extraction tower, and described tripping device is the pressurized vessel separator.
Supercritical CO of the present invention
2Be equipped with the high-pressure pump suction in the mixing device of animal livers slurries, make supercritical CO
2Fluid fully dissolves with the animal livers slurries and mixes, and the solubility active ingredient dissolves in supercritical CO fully in the animal livers
2In the fluid, form mixed stream.
Described catalase extracting method is a supercritical CO
2The gas phase CO that in leaching process, produces
2, be recycled to CO by compressor
2In the gas-holder.
Animal livers of the present invention is that pig, ox, sheep, horse, chicken, duck, all kinds of animal liverss of goose are raw materials for production.
According to catalase extracting method of the present invention extract catalase.
The present invention is supercritical CO
2Be equipped with the high-pressure pump suction in the extraction tower of mixing device of animal livers slurries and make supercritical CO
2Fluid fully dissolves with the animal livers slurries and mixes, and the solubility active ingredient dissolves in supercritical CO fully in the animal livers
2In the fluid, form mixed stream; Tripping device separates removes animal livers slurries residue and coarse particles, forms and just divides mixed stream, just divides mixed stream to make CO through gas-liquid separator separates
2Be divided into gas-liquid two-phase with catalase, the supercritical CO that separating and extracting is finished
2Fluid is recycled to CO through high-pressure pump
2Gas-holder, and gasification part compressed machine is recycled to CO after pressurized liquefied
2Gas-holder, the liquid phase that stays is the hydrogen oxide enzyme product.
The catalatic production principle of the present invention is: according to the molecular weight size, molecular conformation etc. are passed through supercritical CO
2Extraction obtains the hydrogen peroxide enzyme product of high vigor.
The technological process of production of the present invention is as follows:
Animal livers → break process slurries → separator separates resolves → removes slag → hydrogen peroxide enzyme product → tail gas CO
2Recovery → slag liquid pollution-free is handled and is reclaimed.
The present invention uses supercritical CO
2The method of extraction of hydrogen peroxide enzyme, except that explanation is disclosed, other technology, step condition and the existing supercritical CO of using
2The technology condition and the operating process that extract other biological products are identical.
The technical process of traditional with the animal livers is raw material extracts CAT is: animal livers → cutting and crushing → make stay starch → add buffered soln → cytoclasis thing → accent PH → stirring soak → leave standstill → centrifugation → saltout → leave standstill → filtering separation (membrane sepn) → dissolving → desalination → foam that disappears is anticorrosion → the hydrogen peroxide enzyme product, the tediously long complexity of technology, and yield is lower, the production cost height.
The present invention compared with prior art has following technical superiority: use supercritical CO
2Extraction process directly extracts the animal livers extracting solution, saves traditional technology ammonium sulfate precipitation step, greatly reduces production cost, and product yield is high by 60% than current technology, and procedure is simple, and quality product is easy to control.
Technical target of the product of the present invention: WD-JX-20 〉=2000U/ml, WD-JX-30 〉=3000U/ml, WD-JX-50 〉=5000U/ml, high density: 211824.5/ml albumen surpasses and reaches like product. the quality guaranteed period: 180 days.Total number of bacterial colony :≤1000/ml.Also use supercritical CO in other biological products leaching process of domestic extraction
2Abstraction technique still belongs to initiative but be applied to hydrogen peroxidase extracting.Use supercritical CO
2Extraction, low at cost, easy and simple to handle, under the condition of temperature gentleness, can selective extraction catalase and maintenance biological activity.Have advantages such as product yield and steady quality simultaneously.
Extraction and isolation integral, technology is simple; Extraction can be finished on a production line with separating, as the CO that contains solute
2Supercutical fluid makes CO through separator because pressure descends
2Form gas-liquid two-phase with CAT and separate immediately, separation efficiency height and less energy consumption, working cost is low, CO
2Low price, the purity height obtains easily, and recycles in process of production, thereby reduces production costs.
In the extraction production process, do not have organic solvent residual, do not need complicated subtractive process.Because whole process is without organic solvent, so CAT product noresidue solvent, prevented that leaching process is to the murder by poisoning of human body with to the pollution of environment.CO
2Be a kind of inactive gas, chemical reaction and tasteless can not take place during extraction, non-toxic and safe is good.
Heavy metal in raw materials for production, inorganics, dust etc. are difficult for by CO
2Extraction is taken quality product out of and is controlled easily.
Extraction is carried out in airtight high-pressure system, but kill bacteria, and product is directly gone into a minute packaging container internal reaction water closed circulation, non-wastewater discharge from the separator belt pressure.
Adopt low-temperature extraction and separate, do not destroy biological activity, at nearly room temperature 35-40 ℃ and CO
2Gas extraction prevents oxidation and the loss of CAT to thermo-responsive thing effectively, therefore keeps the natural radioactivity of product to greatest extent.
The present invention adopts supercritical CO
2The extraction of hydrogen peroxide enzyme product detects through China Guangzhou Analysis ﹠. Test Center, and quality product is better than the state quality standard requirement, sees Table 1
Name of product: catalase;
State: aqua; Type of detection: censorship.
Table 1 analyzing and testing result
Catalase (CAT) product that adopts the inventive method to produce, quality standard is put on record through Yichun City Department of Qulity Supervision of National Quality ﹠ Technology Inspection Bureau.
Product of the present invention is divided into 4 kinds of WD-JX-20, WD-JX-30, WD-JX-50 and high densitys by catalase activity
Table 2 physical and chemical index
Description of drawings:
Fig. 1 is a supercritical CO of the present invention
2The extraction process flow synoptic diagram.
1, CO
2Storage tank, 2, high-pressure pump, 3, extraction tower, 4, separator, 5, the tail gas storage tank, 6, compressor.
Embodiment: following embodiment is an example with pig, cattle liver only, and the inventive method and product are described, and other animal livers is similar to aforesaid method, does not explain at this.
Embodiment 1
1) raw material prepares to get pork liver 50kg, fresh or does not freezingly limit, and prunes away to remove the white tissue and cut into pieces, and adds suitable quantity of water and pulverizes to grind and make the pork liver slurries;
2) will go up the pork liver slurries made of step places mixing device extraction tower 3 CO
2Supercritical CO in the storage tank 1
2Fluid adds in the extraction tower 3 with high-pressure pump 3 makes pork liver slurries and CO
2Soluble component in the fluid thorough mixing, pork liver slurries comprises that active pharmaceutical ingredients promptly is added to pork liver slurries and supercritical CO fully
2Promptly make mixed stream in the mixed stream that fluid solvent forms, finish above-mentioned steps and can carry out several times in extraction tower 3, this example divides secondary to carry out.
3) separate removal of impurities, the mixed stream that the last step was made joins in the separator separator 4, and by the pressure and temperature in the control separator 4, its pressure and temperature is same as the prior art; Make mixed stream in separator 4, carry out the inferior separation of some levels, remove insoluble sludge and the coarse particles in the pork liver slurries in the mixed stream first the separation, dissolve in the fluid that contains active ingredient in the mixed stream, enter next stage time separator 4 and separate again, through making the first minute mixed stream that is full of CAT after time separation of some levels that is being the overcritical critical CO that is dissolved with CAT
2Fluid.Pressure and temperature in the separator 4 is identical with other biological products scheme of extraction in the prior art, and this example divides secondary to remove residue and coarse particles.
4) extraction preparing hydrogen peroxide enzyme CAT product
The critical CO that the first minute mixed stream that is full of CAT promptly is full of CAT
2Fluid places the separator 4 of gas-liquid separation device to carry out gas-liquid and is separated, and pressure makes CO in the reduction separator 4
2Be separated into gas-liquid two-phase rapidly with CAT and separate CO immediately
2The liquid phase that changes the gas phase remainder into is the cat catalase product, the supercritical CO that extraction finishes
2Be back to CO by air compressor 6 transportation flows
2In the gas-holder 1, the vaporization part is collected through collector tail gas storage tank 5, and same compressed machine 6 reclaims and is delivered to CO
2In the storage tank 1; So just finished the technological process of a supercritical extraction CAT, this example is carried out the reextraction leaching process, gets hydrogen peroxide enzyme product 100kg.
The specific activity that product of the present invention detects CAT through China Guangzhou Analysis ﹠. Test Center reaches 211824.5 υ/mG albumen.
Adopt the existing Technology production cost of this technology to reduce more than 80%, product yield improves more than 60%.
Embodiment 2
With the beef liver be raw material to get the 50kg beef liver be that raw material is produced, other manufacturing condition extracts to such an extent that enzyme lives that 〉=60000 υ/the proteic catalase of mL is more than the 100kg with example 1, protein content 〉=95%, all other technical indicators all meet table 2 requirement.
The present invention adopts supercritical CO
2The method of extraction of hydrogen peroxide enzyme is removed existing disclosed technical scheme, other and application supercritical CO
2It is identical to extract other biological products technology.Separating removal of impurities and extraction preparing hydrogen peroxide enzyme CAT product can divide and carry out for 3-4 time.
Claims (8)
1. a catalase extracting method is a raw material with the animal livers, it is characterized in that adopting supercritical CO
2The extraction process hydrogen peroxidase extracting.
2. according to the described catalase extracting method of claim 1, it is characterized in that described supercritical carbon dioxide extraction method hydrogen peroxidase extracting, undertaken by following processing step:
(1) raw material is prepared the animal livers processing animal livers pulverization process is made slurries;
(2) dissolving mixes above-mentioned slurries and supercritical CO
2Fluid, thorough mixing in mixing device makes mixed stream;
(3) separate removal of impurities mixed stream is placed tripping device, separate and remove insoluble sludge and coarse particles in the mixed stream, make and just divide mixed stream;
(4) extraction preparing hydrogen peroxide enzyme will just divide the mixed stream tripping device, carry out the gas-liquid CO that is separated
2, the liquid phase that stays is the hydrogen peroxide enzyme product.
3. according to claim 1 or 2 described catalase extracting method, it is characterized in that being undertaken by following processing step:
(1) raw material prepare animal livers handle with animal livers clean, mowing, removal of impurities, soak pulverization process and make slurries;
(2) dissolving mixes: with above-mentioned slurries and supercritical CO
2Fluid places the mixing device of pressurized vessel, carries out thorough mixing and makes the component that contains solubility in the slurries dissolve in supercritical CO fully
2Form mixed stream in the fluid solvent;
(3) separate removal of impurities mixed stream placed tripping device, through once or the separation of some levels time remove insoluble sludge and coarse particles in the mixed stream, make the first minute mixed stream that is full of the catalase solute;
(4) extraction preparing hydrogen peroxide enzyme will just divide the mixed stream tripping device, carry out once or several times extracting gas liquid phase separation CO
2, supercritical CO
2Become gas phase by liquid phase and reclaim, stay liquid phase and be the hydrogen peroxide enzyme product.
4. according to the described catalase extracting method of claim 2, it is characterized in that described mixing device is an extraction tower, described tripping device is the pressurized vessel separator.
5. according to claim 1 or 2 described catalase extracting method, the described supercritical CO of its feature
2Supercritical CO is housed with the high-pressure pump suction in the mixing device of animal livers slurries
2Fluid fully dissolves with the animal livers slurries and mixes, and the solubility active ingredient dissolves in supercritical CO fully in the animal livers
2In the fluid, form mixed stream.
6. according to the described catalase extracting method of claim 2, it is characterized in that supercritical CO
2The gas phase CO that in leaching process, produces
2, be recycled to CO by compressor
2In the gas-holder.
7. according to claim 1 or 2 described catalase extracting method, it is characterized in that described animal livers is that pig, ox, sheep, horse, chicken, duck, all kinds of animal liverss of goose are raw materials for production.
8. a catalase extracts and obtains according to claim 1, one of 2 or 3 described extracting method.
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|---|---|---|---|
| CN 201010265399 CN101942423A (en) | 2010-08-22 | 2010-08-22 | Catalase and extraction method thereof |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201010265399 CN101942423A (en) | 2010-08-22 | 2010-08-22 | Catalase and extraction method thereof |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1493688A (en) * | 2003-09-04 | 2004-05-05 | 江南大学 | Method of preparing catalase using extraction method |
| KR20090039470A (en) * | 2007-10-18 | 2009-04-22 | 대한민국(관리부서 : 산림청 국립산림과학원장) | Saccharification of woody biomasses using supercritical water in presence of acid catalysts |
| CN101445795A (en) * | 2008-11-24 | 2009-06-03 | 江苏省江大绿康生物工程技术研究有限公司 | Clean production technology for separation and extraction of catalase |
-
2010
- 2010-08-22 CN CN 201010265399 patent/CN101942423A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1493688A (en) * | 2003-09-04 | 2004-05-05 | 江南大学 | Method of preparing catalase using extraction method |
| KR20090039470A (en) * | 2007-10-18 | 2009-04-22 | 대한민국(관리부서 : 산림청 국립산림과학원장) | Saccharification of woody biomasses using supercritical water in presence of acid catalysts |
| CN101445795A (en) * | 2008-11-24 | 2009-06-03 | 江苏省江大绿康生物工程技术研究有限公司 | Clean production technology for separation and extraction of catalase |
Non-Patent Citations (3)
| Title |
|---|
| 《Surg Endosc.》 20020131 Sare M et al "Effects of carbon dioxide pneumoperitoneum on free radical formation in lung and liver tissues." 第16卷, 第1期 * |
| 《化学与生物工程》 20080131 沈国强 等 "牛肝过氧化氢酶提取工艺的研究" 第25卷, 第1期 * |
| 《生物技术通讯》 20050131 黄永洪 等 "猪肝过氧化氢酶提取条件的研究" 第16卷, 第1期 * |
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Application publication date: 20110112 |