CN105316260A - Bacillus, method for producing plasmin by virtue of bacillus and application of plasmin in thrombolytic drugs - Google Patents
Bacillus, method for producing plasmin by virtue of bacillus and application of plasmin in thrombolytic drugs Download PDFInfo
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Abstract
The invention aims at providing bacillus, a preparation method of plasmin and application of the plasmin. The bacillus is named as bacillus tequilensis, which is preserved with preservation number of CGMCC No. 11462; the method for producing the plasmin comprises steps of conducting fermentation culture on the bacillus tequilensis (with preservation number of CGMCC No. 11462), and implementing separation and purification on a fermentation product so as to obtain the bacillus plasmin; and in thrombolytic drugs, the plasmin, prepared by the method, is used for activating plasminogen in blood so as to degrade fibrin. The invention is applicable to the fields of production and processing of food raw materials.
Description
Technical field
The invention belongs to fermentation engineering and biological pharmacy technical field, specifically, relate to a kind of subtilis, utilize this bacterium to prepare method and the application of plasmin in thrombolytic drug of plasmin.
Background technology
Thrombus disease is the disease of a class serious harm human health and life, according to estimates, there is thrombus disease patient about 1,500 ten thousand people in the whole world, wherein die from every year heart and brain thrombus disease just up to 1,200 ten thousand people, add up according to the World Health Organization (WHO), the death caused due to the thrombotic diseases such as myocardial infarction, Intracerebral hemorrhage has accounted for 51% of the total death toll in the whole world, in China, along with people's living standard and living-pattern preservation, the excess ingestion of higher fatty acid in meals, high protein, thrombotic diseases has exceeded cancer and diabetes become first Health Killer.The medicine for the treatment of thrombus disease can be divided into according to its different mechanism of action: one, plasminogen activator, as urokinase (UK) tissue type plasminogen activator (tPA) etc. passes through to activate the original fibrin degradation of plasmin in blood; Two, Taka-proteinase, as Lumbrukinase, multiple snake venom antithrombotic enzyme etc., the scleroproein of directly degrading in blood.Microorganism always is the important sources of plasmin, especially bacillus, tens of kinds of dissimilar plasmins are separated to, as being separated the NK from BacillusSubtilis, be separated the DC4 etc. from Bacillusamyloliquefaciens, other are as BacillussubtilisinCarlsberg, the bacteriums such as Bacilluscereus all have report to be separated to plasmin (at Peng, Yong, XiaojuanYang, andYizhengZhang. " Microbialfibrinolyticenzymes:anoverviewofsource, production, properties, andthrombolyticactivityinvivo. " Appliedmicrobiologyandbiotechnology69.2 (2005): 126-132 has report), but utilize Bacillustequilensis to be separated and prepare plasmin and have no report always.
Summary of the invention
Technical problem to be solved by this invention overcomes the deficiencies in the prior art, aims to provide a kind of novel genus bacillus and utilize this genus bacillus to carry out the method for plasmin production, also comprising this plasmin in the application in thrombolytic drug.
The technical scheme that genus bacillus of the present invention adopts is, this Strain Designation is
bacillustequilensis, its deposit number is: CGMCCNo.11462.
The method utilizing above-mentioned genus bacillus to carry out plasmin production comprises the following steps: by bacterial classification genus bacillus (
bacillustequilensis) (its deposit number is: CGMCCNo.11462) carry out fermentation culture, from tunning, separation and purification obtains Fibrinolytic Enzyme from Bacillus.
Further, described genus bacillus is separated from soil, after completing fermentation, carries out following steps:
(1) the complete protein precipitation of 75% ammonium sulfate is added in fermented liquid or solid fermentation product elutriant;
(2) dissolution precipitation, removes remaining ammonium sulfate molecule through dialysis;
(3) dialysis terminate after with MonoQ strong anion chromatography, use NaCl wash-out, collect active part;
(4) the activated plasmin of gained tool is through Sepcryal-100 gel filtration chromatography, collect Peak Activity, obtain after freeze-drying purity higher than 90% plasmin, SDS-PAGE electrophoresis is single band.
Further, during the fermentation, employing be liquid nutrient medium, this liquid nutrient medium is: take soybean and MgCl in proportion
2, add distilled water, be placed in 200ml beaker, 121 DEG C, 20min sterilizing, substratum to be cooled, soybean wherein, MgCl
2and the mass/mass/volume ratio of distilled water is w/w/v=10:1:5; Or take peptone: yeast powder: sodium-chlor: the weight of magnesium chloride (2:1:2:1) is dissolved in the distilled water of 100 times of volumes (w/v), and packing is in the triangular flask of 200ml, 1/3 liquid amount, 121 DEG C, 20min sterilizing, substratum to be cooled.
The application of the plasmin utilizing aforesaid method to prepare in thrombolytic drug, described plasmin originally explained scleroproein for the plasmin activated in blood in thrombolytic drug.
The invention has the beneficial effects as follows: the enzyme that the inventive method prepares can directly, rapid fibrin degradation, there is good thrombolytic effect, molecular weight is little compared with urokinase, causes Human immune responses risk lower, has the important value being developed to clinical medicine.Meanwhile, the production abundant raw materials of this enzyme, with low cost, preparation technology is simple, and easily amplifying and realize suitability for industrialized production, is a kind of important thrombolysis composition that can be used for dietary supplements and medicine.
Accompanying drawing explanation
Fig. 1 is for preparing gained high purity enzyme after SDS-PAGE electrophoresis in single band;
Fig. 2 measures enzymic activity, the fibrinolytic circle that this plasmin produces after 2h for using fiber flat band method.
Embodiment
Below, with specific embodiment, the present invention is further described.
(1) the separation preparation of genus bacillus
The pedotheque of Zhuhai City, Guangdong Province will be picked up from, dilute through sterilized water, leave standstill and get supernatant, (often liter of substratum is containing peptone: 5g to adopt coated plate method scleroproein substratum to carry out screening and separating after dilution, yeast powder: 2g, technology agar: 18g, skimmed milk: 250ml), the bacterial classification that can form transparent circle on fibrin plate chooses separately further cultivation.
The bacterium cultivated for the first step adopts fibrin plate (Fibrinogen: 5g, ammonium sulfate: 2g, CaCl after improving
2: 1g, K
2hPO
4: 0.1g, KH
2pO
4: 0.1g, MgSO
4: 0.2g) carry out multiple sieve, preserve for cultivation can be carried out at the bacterial strain of secondary formation transparent circle.
(2) choose the bacterial strain forming transparent circle largest diameter, extract its 16srDNA and check order, sequencing result is compared in ncbi database, and 16srDNA information is the Main Basis of this identification of strains.Wherein, in order-checking, forward primer is 27f:5'-AGAGTTTGATCCTGGCTCAG-3'; Reverse primer 1492r:5'-GGTTACCTTGTTACGACTT-3'.
(3) preparation of plasmin
Embodiment one:
Take 20 grams of soybean and 2gMgCl
2, add distilled water 10ml, be placed in 200ml beaker, 121 DEG C, 20min sterilizing, after cooling, inoculate a ring Bacillustequilensis bacterial classification.32 DEG C, 72h ferments.
After having fermented, 200ml sterilized water is added substratum, stir 0.5h, 4 layers of filtered through gauze removing thalline and insoluble impurities, all the other impurity can use 11500rpm, the centrifugal removing of 20min.Slowly add 112g solid ammonium sulfate, stir, 4 DEG C of standing 1h, treat that enzyme precipitates completely.
Precipitation adds 45mlNa
2cO
3/ NaHCO
3(30mMpH9.2) solution dissolves completely, and the dialysis tubing that use molecular weight is 5kDa or 8kDa is at Na
2cO
3/ NaHCO
3(30mMpH9.2) dialyse in solution, every 8h changes a dialyzate, totally three times, removes residual ammonium sulfate molecule.
With 2.5ml/min speed distilled water balance MonoQ prepacked column (1.6*2.5cmGEHealthcare Sweden) 10 times of column volumes, then changing moving phase is Na
2cO
3/ NaHCO
3(30mMpH9.2 mobile phase A), with identical flow velocity balance columns 16 times of column volumes, crude enzyme liquid sample obtained by previous step is carried on pillar, 6 times of column volumes are rinsed with mobile phase A, then increase Mobile phase B (mobile phase A is containing 1MNaCl) to 100% wash-out 3 column volume internal linear, collect and have fibrinolytic part.
The target enzyme liquid above-mentioned collection obtained uses and is carried on Sephcryal-100 gel chromatography column, uses moving phase Na
2cO
3/ NaHCO
3(30mMpH9.2) rinse with the speed of 0.5ml/min, collect Peak Activity, after desalination lyophilize, obtain high purity plasmin.
Embodiment two:
Take peptone: yeast powder: sodium-chlor: the weight of magnesium chloride (2:1:2:1) is dissolved in the distilled water of 100 times of volumes (w/v), packing is in the triangular flask of 200ml, 1/3 liquid amount, 121 DEG C, 20min sterilizing, after cooling, every bottle graft kind one ring Bacillustequilensis bacterial classification.35 DEG C, 72h ferments.
By fermentation 4 layers of filtered through gauze removing thalline and insoluble impurities after having fermented, all the other impurity can use 11500rpm, the centrifugal removing of 20min.Slowly add often liter of fermented liquid after cooling and add 560g anhydrous slufuric acid ammonium powder, stir, 4 DEG C of standing 4h, treat that enzyme precipitates completely.
The a small amount of as far as possible Na of precipitation
2cO
3/ NaHCO
3(30mMpH9.2) solution dissolves completely, and 1L fermented liquid produces precipitation 15ml solubilize.The dialysis tubing that use molecular weight is 5kDa or 8kDa is at Na
2cO
3/ NaHCO
3(30mMpH9.2) dialyse in solution, every 8h changes a dialyzate, totally three times, removes residual ammonium sulfate molecule.
With 2.5ml/min speed distilled water balance MonoQ prepacked column (1.6*2.5cmGEHealthcare Sweden) 10 times of column volumes, then changing moving phase is Na
2cO
3/ NaHCO
3(30mMpH9.2 mobile phase A), with identical flow velocity balance columns 16 times of column volumes, crude enzyme liquid sample obtained by previous step is carried on pillar, 6 times of column volumes are rinsed with mobile phase A, then increase Mobile phase B (mobile phase A is containing 1MNaCl) to 100% wash-out 3 column volume internal linear, collect and have fibrinolytic part.
The target enzyme liquid above-mentioned collection obtained uses and is carried on Sephcryal-100 gel chromatography column, uses moving phase Na
2cO
3/ NaHCO
3(30mMpH9.2) rinse with the speed of 0.5ml/min, collect Peak Activity, after desalination lyophilize, obtain high purity plasmin.
Used by pure for gained enzyme Edman edman degradation Edman to measure N terminal amino acid residue sequence, result shows front 10 aminoacid sequences of this enzyme N end and is: AQSVPYGISQ, and this plasmin sequence and other genus bacillus are produced plasmin sequence and contrasts, obtaining result is:
table 1 different sources plasmin N terminal sequence compares
Live measuring its enzyme after pure for gained enzyme interpolation different sorts and concentration metal ion and proteinase inhibitor.
table 2 proteinase inhibitor affects this enzymic activity
table 3 metal ion affects this enzyme
As shown in Table 2, the fibrinolytic of plasmin can be suppressed by PMSF, EDTA and beta-mercaptoethanol.K as shown in Table 3
+, Mg
2+, Mn
2+and Ca
2+all have stronger promoter action to this plasmin activity, this result is consistent with the plasmin result that other genus bacillus produce.
Plasmin of the present invention has the fibrinolytic of the direct fibrin degradation of energy, there are good thrombolysis and the remarkable effect suppressing venous thrombosis, and molecular weight, simultaneously, the production abundant raw materials of this enzyme, production technique is easy, with low cost, the existing clinical medicines such as urokinase can be made up in the deficiency of producing and in treatment, there is great Development volue.
The present invention can be applicable to fermentation engineering and biological pharmacy technical field.
SEQUENCELISTING
The pungent hero of <110>, Leibo, Cai ancestor reed
<120> genus bacillus, the method utilizing genus bacillus production plasmin and the application of plasmin in thrombolytic drug
<130>
<160>3
<170>PatentInversion3.3
<210>1
<211>20
<212>DNA
<213> artificial sequence
<220>
<221>misc_feature
<222>(1)…(20)
<223> forward primer 27f
<400>1
agagtttgatcctggctcag20
<210>2
<211>19
<212>DNA
<213> artificial sequence
<220>
<221>misc_feature
<222>(1)…(19)
<223> reverse primer 1492r
<400>2
ggttaccttgttacgactt19
<210>3
<211>1293
<212>DNA
<213> artificial sequence
<220>
<221>misc_feature
<222>(1)…(1293)
<223> Bacillus strain 16srDNA sequence
<400>3
ggcgatgcgcgtgctatacatgcagtcgagcggacagatgggagcttgctccctgatgtt60
agcggcggacgggtgagtaacacgtgggtaacctgcctgtaagactgggataactccggg120
aaaccggggctaataccggatggttgtttgaaccgcatggttcaaacataaaaggtggct180
tcggctaccacttacagatggacccgcggcgcattagctagttggtgaggtaatggctca240
ccaaggcaacgatgcgtagccgacctgagagggtgatcggccacactgggactgagacac300
ggcccagactcctacgggaggcagcagtagggaatcttccgcaatggacgaaagtctgac360
ggagcaacgccgcgtgagtgatgaaggttttcggatcgtaaagctctgttgttagggaag420
aacaagtaccgttcgaatagggcggtaccttgacggtacctaaccagaaagccacggcta480
actacgtgccagcagccgcggtaatacgtaggtggcaagcgttgtccggaattattgggc540
gtaaagggctcgcaggcggtttcttaagtctgatgtgaaagcccccggctcaaccgggga600
gggtcattggaaactggggaacttgagtgcagaagaggagagtggaattccacgtgtagc660
ggtgaaatgcgtagagatgtggaggaacaccagtggcgaaggcgactctctggtctgtaa720
ctgacgctgaggagcgaaagcgtggggagcgaacaggattagataccctggtagtccacg780
ccgtaaacgatgagtgctaagtgttagggggtttccgccccttagtgctgcagctaacgc840
attaagcactccgcctggggagtacggtcgcaagactgaaactcaaaggaattgacgggg900
gcccgcacaagcggtggagcatgtggtttaattcgaagcaacgcgaagaaccttaccagg960
tcttgacatcctctgacaatcctagagataggacgtccccttcgggggcagagtgacagg1020
tggtgcatggttgtcgtcagctcgtgtcgtgagatgttgggttaagtcccgcaacgagcg1080
caacccttgatcttagttgccagcattcagttgggcactctaaggtgactgccggtgaca1140
aaccggaggaaaggtggggatgacgtcaaatcatcatgccccttatgacctgggctaaca1200
ccttgctacaatgggacagaacaaaggggcagcgaaacccgcgaggttaacccaatccac1260
aaatttgtttcaattttggatccattttggaac1293
Claims (5)
1. a genus bacillus, is characterized in that: this Strain Designation is
bacillustequilensis, its deposit number is: CGMCCNo.11462.
2. utilize genus bacillus as claimed in claim 1 to produce a method for plasmin, it is characterized in that: bacterial classification genus bacillus is carried out fermentation culture, from tunning, separation and purification obtains Fibrinolytic Enzyme from Bacillus.
3. method according to claim 2, is characterized in that, described genus bacillus is separated from soil, after completing fermentation, carries out following steps:
(1) the complete protein precipitation of 75% ammonium sulfate is added in fermented liquid or solid fermentation product elutriant;
(2) dissolution precipitation, removes remaining ammonium sulfate molecule through dialysis;
(3) dialysis terminate after with MonoQ strong anion chromatography, use NaCl wash-out, collect active part;
(4) the activated plasmin of gained tool is through Sepcryal-100 gel filtration chromatography, collect Peak Activity, obtain after freeze-drying purity higher than 90% plasmin, SDS-PAGE electrophoresis is single band.
4. method according to claim 3, is characterized in that, during the fermentation, employing be liquid nutrient medium, this liquid nutrient medium is: take soybean and MgCl in proportion
2, add distilled water, be placed in 200ml beaker, 121 DEG C, 20min sterilizing, substratum to be cooled, soybean wherein, MgCl
2and the mass/mass/volume ratio of distilled water is w/w/v=10:1:5; Or take peptone: yeast powder: sodium-chlor: the weight of magnesium chloride (2:1:2:1) is dissolved in the distilled water of 100 times of volumes (w/v), and packing is in the triangular flask of 200ml, 1/3 liquid amount, 121 DEG C, 20min sterilizing, substratum to be cooled.
5. the application of plasmin in thrombolytic drug utilizing the method described in claim 2 to prepare, described plasmin originally explained scleroproein for the plasmin activated in blood in thrombolytic drug.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109706092A (en) * | 2018-10-08 | 2019-05-03 | 四川农业大学 | The preparation method of one plant of bacillus coagulans for producing fibrinolysin and fibrinolysin and viable bacteria tablet |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101113422A (en) * | 2007-06-28 | 2008-01-30 | 浙江大学 | Bacillus licheniformis fibrinolytic enzyme and production method thereof |
| CN101117631A (en) * | 2006-08-03 | 2008-02-06 | 人宇生物科技股份有限公司 | Method for manufacturing natto kinase |
| CN101451130A (en) * | 2007-12-07 | 2009-06-10 | 江苏梁丰食品集团有限公司 | Method for preparing fibrinolysis enzyme |
| CN102533576A (en) * | 2010-12-08 | 2012-07-04 | 中国科学院过程工程研究所 | Bacillus amyloliquefaciens LSSE-62 and application thereof |
| US20130156884A1 (en) * | 2011-12-19 | 2013-06-20 | Triarco Industries | Protease enzymes for increased protein digestion rate and absorption and methods of using the same |
| CN104877936A (en) * | 2015-05-15 | 2015-09-02 | 江西师范大学 | Fibrinolytic-enzyme-producing Bacillus amyloliquefaciens strain separated from traditional fermented black beans |
-
2015
- 2015-11-03 CN CN201510735562.5A patent/CN105316260B/en not_active Expired - Fee Related
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101117631A (en) * | 2006-08-03 | 2008-02-06 | 人宇生物科技股份有限公司 | Method for manufacturing natto kinase |
| CN101113422A (en) * | 2007-06-28 | 2008-01-30 | 浙江大学 | Bacillus licheniformis fibrinolytic enzyme and production method thereof |
| CN101451130A (en) * | 2007-12-07 | 2009-06-10 | 江苏梁丰食品集团有限公司 | Method for preparing fibrinolysis enzyme |
| CN102533576A (en) * | 2010-12-08 | 2012-07-04 | 中国科学院过程工程研究所 | Bacillus amyloliquefaciens LSSE-62 and application thereof |
| US20130156884A1 (en) * | 2011-12-19 | 2013-06-20 | Triarco Industries | Protease enzymes for increased protein digestion rate and absorption and methods of using the same |
| CN104877936A (en) * | 2015-05-15 | 2015-09-02 | 江西师范大学 | Fibrinolytic-enzyme-producing Bacillus amyloliquefaciens strain separated from traditional fermented black beans |
Non-Patent Citations (7)
| Title |
|---|
| ABDHUL,K.等: "ACCESSION：JX847616，Bacillus tequilensis strain MML2 16S ribosomal RNA gene, partial sequence", 《GENBANK》 * |
| NADIA ZARAÎ JAOUADIA等: "A novel keratinase from Bacillus tequilensis strain Q7 with promisingpotential for the leather bating process", 《INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES》 * |
| SEON-JU JEONG等: "Characterization of AprE176, a Fibrinolytic Enzyme from Bacillus subtilis HK176", 《J. MICROBIOL. BIOTECHNOL.》 * |
| SHARMILA THOKCHOM AND SANTA RAM JOS: "Screening of Fibrinolytic Enzymes from Lactic Acid Bacterial Isolates Associated with Traditional Fermented Soybean Foods", 《FOOD SCI. BIOTECHNOL.》 * |
| 吕凤霞等: "内生多黏类芽孢杆菌纤溶酶的纯化及其体外溶栓作用", 《生物工程》 * |
| 王光利等: "芽孢杆菌纤溶酶的纯化及其生物活性研究", 《西南农业大学学报》 * |
| 贾楠等: "纤溶酶分离纯化的研究进展", 《食品科技》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109706092A (en) * | 2018-10-08 | 2019-05-03 | 四川农业大学 | The preparation method of one plant of bacillus coagulans for producing fibrinolysin and fibrinolysin and viable bacteria tablet |
| CN109706092B (en) * | 2018-10-08 | 2020-11-06 | 四川农业大学 | Preparation method of plasmin-producing bacillus coagulans, plasmin and live bacterium tablet |
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