CN110055237A - A kind of extracting method of alkaline phosphatase - Google Patents
A kind of extracting method of alkaline phosphatase Download PDFInfo
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- CN110055237A CN110055237A CN201910394507.2A CN201910394507A CN110055237A CN 110055237 A CN110055237 A CN 110055237A CN 201910394507 A CN201910394507 A CN 201910394507A CN 110055237 A CN110055237 A CN 110055237A
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- 238000000034 method Methods 0.000 title claims abstract description 25
- 102000002260 Alkaline Phosphatase Human genes 0.000 title claims abstract description 21
- 108020004774 Alkaline Phosphatase Proteins 0.000 title claims abstract description 21
- 210000004347 intestinal mucosa Anatomy 0.000 claims abstract description 28
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims abstract description 20
- 238000000605 extraction Methods 0.000 claims abstract description 20
- 238000004108 freeze drying Methods 0.000 claims abstract description 12
- 239000000843 powder Substances 0.000 claims abstract description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 10
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims abstract description 8
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims abstract description 8
- 235000011130 ammonium sulphate Nutrition 0.000 claims abstract description 8
- 239000006228 supernatant Substances 0.000 claims abstract description 7
- 239000013049 sediment Substances 0.000 claims abstract description 4
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 9
- 244000309466 calf Species 0.000 claims description 3
- 230000000968 intestinal effect Effects 0.000 claims description 3
- 150000002576 ketones Chemical class 0.000 claims description 3
- 238000000926 separation method Methods 0.000 claims description 3
- 230000000694 effects Effects 0.000 abstract description 10
- 230000000844 anti-bacterial effect Effects 0.000 abstract description 4
- 238000005119 centrifugation Methods 0.000 abstract description 4
- 210000000813 small intestine Anatomy 0.000 abstract description 2
- 102000004190 Enzymes Human genes 0.000 description 13
- 108090000790 Enzymes Proteins 0.000 description 13
- 102100024319 Intestinal-type alkaline phosphatase Human genes 0.000 description 6
- 101710184243 Intestinal-type alkaline phosphatase Proteins 0.000 description 6
- 239000012530 fluid Substances 0.000 description 6
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 4
- 230000004888 barrier function Effects 0.000 description 4
- 210000001035 gastrointestinal tract Anatomy 0.000 description 4
- 230000008859 change Effects 0.000 description 3
- 239000000284 extract Substances 0.000 description 3
- 210000004185 liver Anatomy 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 230000028327 secretion Effects 0.000 description 3
- 102100025683 Alkaline phosphatase, tissue-nonspecific isozyme Human genes 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- 101000574445 Homo sapiens Alkaline phosphatase, tissue-nonspecific isozyme Proteins 0.000 description 2
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 2
- 210000000988 bone and bone Anatomy 0.000 description 2
- 230000003197 catalytic effect Effects 0.000 description 2
- 238000006555 catalytic reaction Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 210000001842 enterocyte Anatomy 0.000 description 2
- 238000006911 enzymatic reaction Methods 0.000 description 2
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 2
- 210000000232 gallbladder Anatomy 0.000 description 2
- 210000004837 gut-associated lymphoid tissue Anatomy 0.000 description 2
- 210000002490 intestinal epithelial cell Anatomy 0.000 description 2
- 210000003734 kidney Anatomy 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000012423 maintenance Methods 0.000 description 2
- 239000003960 organic solvent Substances 0.000 description 2
- 229910052698 phosphorus Inorganic materials 0.000 description 2
- 239000011574 phosphorus Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- ASWBNKHCZGQVJV-UHFFFAOYSA-N (3-hexadecanoyloxy-2-hydroxypropyl) 2-(trimethylazaniumyl)ethyl phosphate Chemical compound CCCCCCCCCCCCCCCC(=O)OCC(O)COP([O-])(=O)OCC[N+](C)(C)C ASWBNKHCZGQVJV-UHFFFAOYSA-N 0.000 description 1
- IIZPXYDJLKNOIY-JXPKJXOSSA-N 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCC\C=C/C\C=C/C\C=C/C\C=C/CCCCC IIZPXYDJLKNOIY-JXPKJXOSSA-N 0.000 description 1
- 102100025677 Alkaline phosphatase, germ cell type Human genes 0.000 description 1
- 102000003886 Glycoproteins Human genes 0.000 description 1
- 108090000288 Glycoproteins Proteins 0.000 description 1
- 101000574440 Homo sapiens Alkaline phosphatase, germ cell type Proteins 0.000 description 1
- 101001068480 Homo sapiens Guanylyl cyclase-activating protein 1 Proteins 0.000 description 1
- 108010044467 Isoenzymes Proteins 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- 102100037611 Lysophospholipase Human genes 0.000 description 1
- 108010058864 Phospholipases A2 Proteins 0.000 description 1
- 102000004160 Phosphoric Monoester Hydrolases Human genes 0.000 description 1
- 108090000608 Phosphoric Monoester Hydrolases Proteins 0.000 description 1
- 102000000591 Tight Junction Proteins Human genes 0.000 description 1
- 108010002321 Tight Junction Proteins Proteins 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000011149 active material Substances 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 239000012431 aqueous reaction media Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 230000001079 digestive effect Effects 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 230000002496 gastric effect Effects 0.000 description 1
- 229930004094 glycosylphosphatidylinositol Natural products 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 239000000710 homodimer Substances 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 230000003053 immunization Effects 0.000 description 1
- 238000002649 immunization Methods 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000007358 intestinal barrier function Effects 0.000 description 1
- 210000000936 intestine Anatomy 0.000 description 1
- 229940067606 lecithin Drugs 0.000 description 1
- 235000010445 lecithin Nutrition 0.000 description 1
- 239000000787 lecithin Substances 0.000 description 1
- 235000012054 meals Nutrition 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 210000003097 mucus Anatomy 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 125000002467 phosphate group Chemical group [H]OP(=O)(O[H])O[*] 0.000 description 1
- 230000000704 physical effect Effects 0.000 description 1
- 230000004962 physiological condition Effects 0.000 description 1
- 210000002826 placenta Anatomy 0.000 description 1
- 230000003169 placental effect Effects 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000012429 reaction media Substances 0.000 description 1
- 230000009257 reactivity Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000007790 scraping Methods 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 238000013517 stratification Methods 0.000 description 1
- 210000001578 tight junction Anatomy 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y301/00—Hydrolases acting on ester bonds (3.1)
- C12Y301/03—Phosphoric monoester hydrolases (3.1.3)
- C12Y301/03001—Alkaline phosphatase (3.1.3.1)
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Enzymes And Modification Thereof (AREA)
Abstract
A kind of extracting method of alkaline phosphatase, belongs to zymetology field.The method is as follows: 1, taking fresh pig (ox) small intestine, scrape intestinal mucosa.2, the uniform scumbling of the intestinal mucosa scraped down is lyophilized in freeze dryer in plate, grinds after freeze-drying.3, intestinal mucosa is packed into supercritical CO2In the feed chamber of extraction equipment, extraction retains kettle midgutmucosa coarse powder after the completion.4, water is added into intestinal mucosa coarse powder, it is homogeneous, pH6.5 is adjusted, ammonium sulfate and acetone are added, 20min is centrifuged in 4 DEG C of standings 40min, 9000r/min, takes supernatant that acetone is added, stand, centrifugation takes final sediment to be lyophilized.The invention has the advantages that intestinal mucosa gets rid of extra moisture, then through supercritical CO after freeze-drying2Extraction, gets rid of extra fat, while realizing the interference for excluding moisture and fat, and also has certain bactericidal effect, compared with the extraction effect that traditional handicraft will have better alkaline phosphatase.
Description
Technical field
The invention belongs to zymetology fields, and in particular to a kind of extracting method of alkaline phosphatase.
Background technique
Alkaline phosphatase (ALP or AKP) is a kind of monoester phosphohydrolase, be distributed widely in human liver, bone, intestines,
The tissue such as kidney and placenta, is finally all discharged to outside gallbladder through liver, belongs to homodimer enzyme.This enzyme energy catalytic nucleic acid molecule is de-
Fall 5' phosphate group, so that the end 5'-P of DNA or RNA segment be made to be converted into the end 5'-OH.But it is not single enzyme, and
It is one group of isoenzymes, i.e. tissue non-specific alkaline phosphatase (TNAP) (being mainly expressed in liver, bone and kidney), human placental alkaline phosphorus
Sour enzyme (PLAP), reproduction cell alkaline phosphatase (GCAP) and intestinal alkaline phosphatase (IAP).IAP is a kind of through glycosyl-phosphatidyl
Inositol key is anchored to the glycoprotein on enterocyte teleblem.Under physiological conditions, the pancreatic phospholipase a2 secreted after the meal hydrolyzes gallbladder
Lecithin lysolecithin generated specific can accelerate IAP to be released into enteric cavity through enterocyte teleblem in juice.
Gut barrie r is made of four parts, i.e. the mechanical barrier of Intestinal epithelial cells and intercellular tight junction composition；
The compositions such as the antibacterial substance that the mucus of Intestinal epithelial cells secretion, the digestive juice of gastro-intestinal secretion and commensal gut bacterium generate
Chemical barrier；The immunization barrier that gut associated lymphoid tissue (GALT) and its immunologic active material of secretion are constituted；Commensal gut
The biological barrier for interdepending, interacting that bacterium and host's microenvironment are formed.The maintenance of intestinal barrier function depends on its four
Partial integrality, the damage of any one part can have an adverse effect to body, threaten the life and health of body.Phase
It closes result of study and shows that intestinal alkaline phosphatase (IAP) rises in the normal performance of the integrality and function of maintenance Gut barrie r structure
To certain effect.
With the development of bioengineering, enzymic catalytic reaction is increasingly valued by people.However, due in exoenzyme
Activity reduce and its unstability, significantly limit its application industrially.To find out its cause, mainly enzyme extract and
The change of structure in purification process.Studies have shown that the performance of enzymatic activity is largely dependent upon its molecular conformation, conformation is risen
Variation, activity or function also become therewith.Therefore people in depth inquire into enzyme kinetics and influence enzymatic reaction
Factor, and by the way that the external environment of enzyme reaction is modified or changed to enzyme molecule to improve the activity and stabilization of enzyme in vitro
Property.
Supercritical fluid (supercritical fluid, SCF) refers to that temperature and pressure are in critical-temperature and critical pressure
Fluid more than power is the fluid during a kind of liquid gaseous state is in homogeneous.The advantages of this fluid has liquids and gases concurrently, viscosity
It is small, diffusion coefficient is big, density is big, there are good dissolubility and mass transfer performances, and in Near The Critical Point fluid to pressure and temperature
Variation it is very sensitive, be both a kind of good separating medium and a kind of good reaction medium.Further, since SCF has
The physical property of consecutive variations can control reactivity and selectivity by influencing its local action.As a kind of non-aqueous reaction
Medium, SCF is with unique property by more and more extensive concern in enzyme catalysis field.So enzyme is placed in supercritical state
Its catalytic kinetics can also be made to be improved under state.With supercritical CO2For, CO under high pressure2It changes in enzyme microenvironment
PH value, the free amino of covalent modification protein surface simultaneously form carbaminate, and it is residual that these carbaminates change lysine
The charge of base, to change the activity of enzyme.
Alkaline phosphatase extractive technique route more commonly used at present is as follows: fresh pig (ox) small intestine → scraping intestinal mucosa →
Add water, it is homogeneous → to add n-butanol, homogeneous → centrifugation (9000r/min, 20min) → stratification, phase supernatant → tune of fetching water
PH4.9 → centrifugation (9000r/min, 20min) → takes supernatant, adds NaOH tune pH6.5, adds 5% ammonium sulfate, adds acetone → standing
40min(4 DEG C), centrifugation (9000r/min, 20min) → supernatant is taken to add acetone → 40min(4 DEG C of standing), it is centrifuged (9000r/
Min, 20min) → precipitating → freeze-drying.This method needs a large amount of organic solvent, in the process for extracting fat using n-butanol
Alkaline phosphatase can also partially remain in organic solvent, and extraction effect is not good enough.
Summary of the invention
The purpose of the present invention is to solve the bad problems of current alkaline phosphatase extraction effect, provide a kind of alkaline phosphorus
The extracting method of sour enzyme, i.e. supercritical CO2Extraction.First the intestinal mucosa scraped down is lyophilized before experiment for this method,
Pass through supercritical CO again2Extraction removes the fat in intestinal mucosa, and this method effectively eliminates moisture extra in intestinal mucosa
With the interference of fat, and supercritical CO2Extraction also has certain bactericidal effect, is more advantageous to subsequent to alkaline phosphatase
Extraction.
To achieve the above object, the technical solution adopted by the present invention is as follows:
A kind of extracting method of alkaline phosphatase, specific step is as follows for the method:
Step 1: taking fresh chitterlings or calf intestinal, clean up, and scrapes intestinal mucosa；
Step 2: the intestinal mucosa scraped down being uniformly coated in plate and is lyophilized in freeze dryer, with a thickness of 3 ~ 5mm,
Grinds after freeze-drying, granularity are no more than 100 mesh；
Step 3: the intestinal mucosa of above-mentioned grinds is packed into supercritical CO2In the feed chamber of extraction equipment, setting extracting pressure is
20MPa, separating pressure 8MPa, extraction and separation carry out simultaneously, and total time is 1.5 ~ 2h, obtain intestinal mucosa coarse powder；
Step 4: intestinal mucosa coarse powder, water, the ammonium sulfate and third for taking step 3 to obtain according to the volume ratio of 1:0.8:0.05:0.47
Then water is added in ketone into intestinal mucosa coarse powder, homogeneous, and adjusting pH is 6.5, and ammonium sulfate and acetone is added, in 4 DEG C of standing 40min,
9000r/min is centrifuged 20min, takes supernatant additionally to add the acetone of its 1.07 volume, in 4 DEG C of standings 40min, 9000r/
Min is centrifuged 20min, and final sediment is taken to be lyophilized.
The beneficial effect of the present invention compared with the existing technology is: intestinal mucosa gets rid of extra water after freeze-drying
Point, then through supercritical CO2Extraction, gets rid of extra fat, while realizing the interference for excluding moisture and fat, and also have
There is certain bactericidal effect, there is better extraction effect compared with traditional handicraft.
Specific embodiment
Further description of the technical solution of the present invention below, and however, it is not limited to this, all to the technology of the present invention
Scheme is modified or replaced equivalently, and without departing from the spirit and scope of the technical solution of the present invention, should all be covered in the present invention
Protection scope in.
Specific embodiment 1: present embodiment record be a kind of alkaline phosphatase extracting method, the method
By freeze-drying, supercritical CO2Extraction extracts, alkaline phosphatase extracts three parts composition, and specific step is as follows for the method:
Step 1: taking fresh chitterlings or calf intestinal, clean up, and scrapes intestinal mucosa；
Step 2: the intestinal mucosa scraped down being uniformly coated in plate and is lyophilized in freeze dryer, with a thickness of 3 ~ 5mm,
Grinds after freeze-drying, granularity are no more than 100 mesh；
Step 3: the intestinal mucosa of above-mentioned grinds is packed into supercritical CO2In the feed chamber of extraction equipment, setting extracting pressure is
20MPa, separating pressure 8MPa, extraction and separation carry out simultaneously, and total time is 1.5 ~ 2h, obtain intestinal mucosa coarse powder；
Step 4: intestinal mucosa coarse powder, water, the ammonium sulfate and third for taking step 3 to obtain according to the volume ratio of 1:0.8:0.05:0.47
Then water is added in ketone into intestinal mucosa coarse powder, homogeneous, and adjusting pH is 6.5, and ammonium sulfate and acetone is added, in 4 DEG C of standing 40min,
9000r/min is centrifuged 20min, takes supernatant additionally to add the acetone of its 1.07 volume, in 4 DEG C of standings 40min, 9000r/
Min is centrifuged 20min, and final sediment is taken to be lyophilized.
The supercritical CO2Extraction equipment is purchased from Jiangsu Huaan Science Research Instrument Co., Ltd, model HA121-50-
01。
Specific embodiment 2: a kind of extracting method of alkaline phosphatase described in specific embodiment one, in step 2,
The freeze-drying specifically: after -40 DEG C of pre-freeze 40min, -40 DEG C of operation 120min, -35 DEG C of operation 120min, -30 DEG C are run
360min, -25 DEG C of operation 360min, -15 DEG C of operation 360min, 0 DEG C of operation 120min, 10 DEG C of operation 120min, 20 DEG C run
360min。
Specific embodiment 3: a kind of extracting method of alkaline phosphatase described in specific embodiment one, in step 4,
PH is adjusted using the NaOH of 1M.
Specific embodiment 4: a kind of extracting method of alkaline phosphatase described in specific embodiment one, in step 4,
The freeze-drying specifically: after -40 DEG C of pre-freeze 40min, -40 DEG C of operation 120min, -35 DEG C of operation 120min, -30 DEG C are run
360min, -25 DEG C of operation 360min, -15 DEG C of operation 360min, 0 DEG C of operation 120min, 10 DEG C of operation 120min, 20 DEG C run
360min。
Claims (4)
1. a kind of extracting method of alkaline phosphatase, it is characterised in that: specific step is as follows for the method:
Step 1: taking fresh chitterlings or calf intestinal, clean up, and scrapes intestinal mucosa；
Step 2: the intestinal mucosa scraped down being uniformly coated in plate and is lyophilized in freeze dryer, with a thickness of 3 ~ 5mm,
Grinds after freeze-drying, granularity are no more than 100 mesh；
Step 3: the intestinal mucosa of above-mentioned grinds is packed into supercritical CO2In the feed chamber of extraction equipment, setting extracting pressure is
20MPa, separating pressure 8MPa, extraction and separation carry out simultaneously, and total time is 1.5 ~ 2h, obtain intestinal mucosa coarse powder；
Step 4: intestinal mucosa coarse powder, water, the ammonium sulfate and third for taking step 3 to obtain according to the volume ratio of 1:0.8:0.05:0.47
Then water is added in ketone into intestinal mucosa coarse powder, homogeneous, and adjusting pH is 6.5, and ammonium sulfate and acetone is added, in 4 DEG C of standing 40min,
9000r/min is centrifuged 20min, takes supernatant additionally to add the acetone of its 1.07 volume, in 4 DEG C of standings 40min, 9000r/
Min is centrifuged 20min, and final sediment is taken to be lyophilized.
2. a kind of extracting method of alkaline phosphatase according to claim 1, it is characterised in that: described in step 2
Freeze-drying specifically: after -40 DEG C of pre-freeze 40min, -40 DEG C of operations 120min, -35 DEG C of operations 120min, -30 DEG C of operation 360min, -
25 DEG C of operations 360min, -15 DEG C of operations 360min, 0 DEG C of operation 120min, 10 DEG C of operations 120min, 20 DEG C of operation 360min.
3. a kind of extracting method of alkaline phosphatase according to claim 1, it is characterised in that: in step 4, use 1M
NaOH adjust pH.
4. a kind of extracting method of alkaline phosphatase according to claim 1, it is characterised in that: described in step 4
Freeze-drying specifically: after -40 DEG C of pre-freeze 40min, -40 DEG C of operations 120min, -35 DEG C of operations 120min, -30 DEG C of operation 360min, -
25 DEG C of operations 360min, -15 DEG C of operations 360min, 0 DEG C of operation 120min, 10 DEG C of operations 120min, 20 DEG C of operation 360min.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117050969A (en) * | 2023-08-18 | 2023-11-14 | 北京中检葆泰生物技术有限公司 | Preparation method of alkaline phosphatase |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101182495A (en) * | 2007-11-21 | 2008-05-21 | 郑州市通源生物技术有限公司 | Joint production process joint production producing alkaline phosphatase and heparin sodium with pig small intestine as raw material |
| CN102283904A (en) * | 2011-09-08 | 2011-12-21 | 南京泽朗医药科技有限公司 | Preparation method for rumex japonicus houtt tannin |
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2019
- 2019-05-13 CN CN201910394507.2A patent/CN110055237A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101182495A (en) * | 2007-11-21 | 2008-05-21 | 郑州市通源生物技术有限公司 | Joint production process joint production producing alkaline phosphatase and heparin sodium with pig small intestine as raw material |
| CN102283904A (en) * | 2011-09-08 | 2011-12-21 | 南京泽朗医药科技有限公司 | Preparation method for rumex japonicus houtt tannin |
Non-Patent Citations (2)
| Title |
|---|
| KIL-YOON KANG等: "Separation of protein and fatty acids from tuna viscera using supercritical carbon dioxide", 《BIOTECHNOLOGY AND BIOPROCESS ENGINEERING》 * |
| 蒋益众等: "高活性牛肠碱性磷酸酶的制备", 《应用科学学报》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117050969A (en) * | 2023-08-18 | 2023-11-14 | 北京中检葆泰生物技术有限公司 | Preparation method of alkaline phosphatase |
| CN117050969B (en) * | 2023-08-18 | 2024-04-02 | 北京中检葆泰生物技术有限公司 | Preparation method of alkaline phosphatase |
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