CN107383242A - A kind of ultrasonic enzymolysis and extraction liquaemin technology - Google Patents
A kind of ultrasonic enzymolysis and extraction liquaemin technology Download PDFInfo
- Publication number
- CN107383242A CN107383242A CN201710694173.1A CN201710694173A CN107383242A CN 107383242 A CN107383242 A CN 107383242A CN 201710694173 A CN201710694173 A CN 201710694173A CN 107383242 A CN107383242 A CN 107383242A
- Authority
- CN
- China
- Prior art keywords
- liquaemin
- technology
- enzymolysis
- extraction
- ultrasonic
- 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
Classifications
-
- 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
- C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
- C08B37/0063—Glycosaminoglycans or mucopolysaccharides, e.g. keratan sulfate; Derivatives thereof, e.g. fucoidan
- C08B37/0075—Heparin; Heparan sulfate; Derivatives thereof, e.g. heparosan; Purification or extraction methods thereof
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Materials Engineering (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicinal Chemistry (AREA)
- Polymers & Plastics (AREA)
- Organic Chemistry (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The present invention relates to a kind of liquaemin technology, especially a kind of ultrasonic enzymolysis and extraction liquaemin technology, enzymatic extraction heparin sodium is carried out using double enzyme groups, by selecting ratio, the intensity of ultrasonic field that the species of enzyme, enzyme combine, controlling the time and liquaemin is made in other several procedures.The present invention compared with prior art, has the following advantages that：1st, the time of whole process reduces general 33%；2nd, the yield of product improves 29.6%；3rd, titer plateaus (domestic advanced technology product potency 90 " 0) more than 120.
Description
The present invention relates to a kind of liquaemin technology, especially a kind of ultrasonic enzymolysis and extraction liquaemin technology.
Background technology
The almost all of China's production at present uses chitterlings as raw material, with salt solution, alkaline hydrolysiss in conventional reactor
Several with enzymatic isolation method etc., wherein salt solution application is the most common, and its production process is：In raw material (intestinal mucosa), conventional reactor
Salt solution extraction, filtering, resin adsorption, elution, ethanol precipitation, dehydration, drying, heparin sodium crude, above method production liquaemin
Technical process it is although fairly simple be easily achieved, but the existing exquisiteness for being disadvantageous in that process, because crude heparin sodium
Handled through soda acid repeatedly, the yield of liquaemin can not only reduced, and the bioactivity of liquaemin also influenceed very greatly, especially
Its potency is extremely difficult to more than 90USPu/mg, and the time is longer.
The content of the invention
The purpose of the present invention is to be to provide a kind of ultrasonic enzymolysis and extraction liquaemin technology, and carrying out enzymatic using double enzyme groups carries
Taking heparin is received, by selecting ratio, the intensity of ultrasonic field that the species of enzyme, enzyme combine, controlling time and other a few road works
Liquaemin is made in sequence.
This method uses following steps：
A kind of ultrasonic enzymolysis and extraction liquaemin technology, it is characterised in that this method comprises the following steps：
(a), the conventional enzymatic vessel of mucous membrane of small intestine suction made will be scraped, it is 2.3%- that weight ratio is first added before heating
2.8% refined salt, pH value is adjusted to 9.0-9.2；The mucous membrane of small intestine suction multiple-frequency supersonic retort made, multiple-frequency supersonic condition will be scraped
For 20+40kHZ, ultrasound intensity is selected in 0.1W/m2 to 0.9W/m2；
(b) trypsase and lipase are added when the solution in step (a), being warmed into 40 DEG C -45 DEG C, and makes its ratio
Example is 10：0.5 to 10：Between 5, double enzymes weight ratio of solution in step (a) is 0.3%-0.4%, continues to be warmed to 50
DEG C -55 DEG C, insulation enzymolysis 2h-5h, then proceed to be warmed to 80 DEG C -85 DEG C, filtered after being incubated 10-20 minutes；
(c), the filtering solution in step (b) is adsorbed, washed, is eluted, is precipitated, dehydrate conventional steps after obtain
Obtain heparin sodium product.
In described step (b), selected trypsase and lipase are preferably in a proportion of 10：2.
The present invention compared with prior art, has the following advantages that：
1st, the time of whole process reduces general 33%；
2nd, the yield of product improves 29.6%；
3rd, titer plateaus (domestic advanced technology product potency 90- " 0) more than 120.
Embodiment
Embodiment 1
Mucous membrane of small intestine (10 secondary small intestine) processed, every small intestine mixed with water after about 4-6 kilograms, will scrape the small intestine that makes be pumped into it is normal
Enzymatic vessel is advised, the refined salt of 2.3-2.8% (scale of construction) is first added before heating, adjusts pH value 9.0-9.2：Tryptose is added at 42 DEG C
Enzyme and lipase (ratio 10：2) total solution weight ratio, is accounted for as 0.4%；Continue to be heated up to 52 DEG C, insulation carries out enzyme after 3 hours
Solution, then proceeding to be warmed to 80 DEG C -85, ' C, insulation are filtered after 15 minutes, the filtering solution in above-mentioned steps are adsorbed：Inhale
Attached water is filtered, and resin adsorption is added when being cooled to 58 ' C；Washing：Resin is collected, clear water rinses for several times, then heats 58- with clear water
60 ' C, adjust PH to 7.5, untill resin wash clean, with 1.2N saline solutions, adjust PH to 8 to add in resin, be warmed to 60 DEG C,
0.5h is washed, place to go wash solution, is filtered dry resin；Elution：After being eluted 2 times with 4M NaCl solution, solution conjunction will be eluted twice
And filter：Precipitation：By first and second eluent with 85% ethanol precipitation, volume ratio is 1：1.5 or so, the alcohol after converting
Mixed liquid concentration is about 30-40 degree, and heparin sodium product is dehydrated to obtain after precipitation.
Claims (4)
1. a kind of ultrasonic enzymolysis and extraction liquaemin technology, it is characterised in that this method comprises the following steps：
(a), the conventional enzymatic vessel of mucous membrane of small intestine suction made will be scraped, weight is first added before heating than for 2.3%-2.8%'s
Refined salt, pH value is adjusted to 9.0-9.2；The mucous membrane of small intestine suction multiple-frequency supersonic retort made will be scraped, multiple-frequency supersonic condition is 20+
40kHZ, ultrasound intensity is selected in 0.1W/m2 to 0.9W/m2；
(b) trypsase and lipase are added when the solution in step (a), being warmed into 40 DEG C -45 DEG C, and its ratio is existed
10：0.5 to 10：Between 5, double enzymes weight ratio of solution in step (a) is 0.3%-0.4%, continues to be warmed to 50 DEG C -55
DEG C, insulation enzymolysis 2h-5h, then proceed to be warmed to 80 DEG C -85 DEG C, filtered after being incubated 10-20 minutes；
(c), the filtering solution in step (b) is adsorbed, washed, is eluted, is precipitated, dehydrate conventional steps after obtain liver
Plain sodium product.
A kind of 2. ultrasonic enzymolysis and extraction liquaemin technology according to claim l, it is characterised in that：Described step (b)
In, selected trypsase and lipase are preferably in a proportion of 10：2.
A kind of 3. ultrasonic enzymolysis and extraction liquaemin technology according to claim 1 or 2, it is characterised in that：Described step
(b) in, the preferred time of selected insulation enzymolysis is in 2h.
A kind of 4. ultrasonic enzymolysis and extraction liquaemin technology according to claim 1, it is characterised in that：Multiple-frequency supersonic retort
Selected ultrasound intensity is preferably 0.3W/m2 to 0.7W/m2.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710694173.1A CN107383242A (en) | 2017-08-10 | 2017-08-10 | A kind of ultrasonic enzymolysis and extraction liquaemin technology |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710694173.1A CN107383242A (en) | 2017-08-10 | 2017-08-10 | A kind of ultrasonic enzymolysis and extraction liquaemin technology |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107383242A true CN107383242A (en) | 2017-11-24 |
Family
ID=60355792
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710694173.1A Pending CN107383242A (en) | 2017-08-10 | 2017-08-10 | A kind of ultrasonic enzymolysis and extraction liquaemin technology |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107383242A (en) |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101343645A (en) * | 2008-08-29 | 2009-01-14 | 罗时通 | Compound frequently combined ultrasonic reinforced double-enzyme heparin sodium extracting technique |
| CN102633907A (en) * | 2012-05-02 | 2012-08-15 | 如皋市永兴肠衣有限公司 | Process for using intestine casing to extract sodium heparin |
| CN102633904A (en) * | 2011-03-21 | 2012-08-15 | 如皋市坝新肠衣有限公司 | Hydrolysis technology for protease of heparin sodium |
| CN102993335A (en) * | 2011-09-09 | 2013-03-27 | 谭科 | Heparin sodium balance extraction method |
| CN103122040A (en) * | 2013-03-19 | 2013-05-29 | 如皋市永兴肠衣有限公司 | Method for preparing heparin sodium through ultrasound enzymolysis |
| CN105175578A (en) * | 2015-10-23 | 2015-12-23 | 南通仁寿食品有限公司 | Extraction technology of heparin sodium |
| CN105693886A (en) * | 2016-04-19 | 2016-06-22 | 常州市蓝勖化工有限公司 | Preparation method of heparin sodium |
-
2017
- 2017-08-10 CN CN201710694173.1A patent/CN107383242A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101343645A (en) * | 2008-08-29 | 2009-01-14 | 罗时通 | Compound frequently combined ultrasonic reinforced double-enzyme heparin sodium extracting technique |
| CN102633904A (en) * | 2011-03-21 | 2012-08-15 | 如皋市坝新肠衣有限公司 | Hydrolysis technology for protease of heparin sodium |
| CN102993335A (en) * | 2011-09-09 | 2013-03-27 | 谭科 | Heparin sodium balance extraction method |
| CN102633907A (en) * | 2012-05-02 | 2012-08-15 | 如皋市永兴肠衣有限公司 | Process for using intestine casing to extract sodium heparin |
| CN103122040A (en) * | 2013-03-19 | 2013-05-29 | 如皋市永兴肠衣有限公司 | Method for preparing heparin sodium through ultrasound enzymolysis |
| CN105175578A (en) * | 2015-10-23 | 2015-12-23 | 南通仁寿食品有限公司 | Extraction technology of heparin sodium |
| CN105693886A (en) * | 2016-04-19 | 2016-06-22 | 常州市蓝勖化工有限公司 | Preparation method of heparin sodium |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106011199B (en) | Pretreatment method of crop straws | |
| CN101343645B (en) | Compound frequently combined ultrasonic reinforced double-enzyme heparin sodium extracting technique | |
| CN103665192B (en) | A kind of method extracting sodium heparin and co-producing protein powder from chitterlings | |
| CN100497647C (en) | Method for adsorbing and decomposing stalk cellulose using cellulase | |
| CN111004827B (en) | Preparation method of xylo-oligosaccharide | |
| CN101659714B (en) | Method for extracting sodium heparin and co-producing amino acid | |
| KR20180081805A (en) | Soluble fiber and process for making same | |
| CN103320486A (en) | Method for producing chondroitin sulfate with coproduction of hydrolyzed collagen by employing fish cartilage | |
| CN105754015A (en) | Method for extracting chitin from oyster shell | |
| CN103130904A (en) | High-valued utilization method for patinopecten yessoensis offal | |
| CN108314749A (en) | A kind of method of driven Object Extraction heparin sodium | |
| CN109576324A (en) | A kind of astragalus polyose and its biological extraction method | |
| KR101170685B1 (en) | Method for producing a water-soluble dietary fiber from a rice by-products | |
| CN102732592A (en) | Method for preparing freshwater fish bone gelatin by enzyme process | |
| CN103122040A (en) | Method for preparing heparin sodium through ultrasound enzymolysis | |
| JP5408999B2 (en) | Soluble low calorie fiber composition and process for preparing the same | |
| CN110074418B (en) | Extraction method of seaweed dietary fibers | |
| CN102071180A (en) | New process for preparing pineapple bromelain | |
| CN107383242A (en) | A kind of ultrasonic enzymolysis and extraction liquaemin technology | |
| CN105859916A (en) | Preparing method for NO.9 south jerusalem artichoke powder | |
| CN107280018B (en) | Preparation method of rice bran insoluble dietary fiber | |
| CN102775511A (en) | Method for extracting pepper polysaccharide from pepper residue | |
| CN107312111A (en) | A variety of enzymes are combined intestinal mucosa technology for hydrolyzing | |
| CN107267572A (en) | A kind of combined-enzyme method heparin sodium extracting technique | |
| CN105504098B (en) | A kind of technique that Heparan sulfate is extracted from duodenum |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20171124 |