CN104212741B - Bacillus subtilis producing fermented chickpea having fibrinolysis and antioxidation functions, and its application - Google Patents
Bacillus subtilis producing fermented chickpea having fibrinolysis and antioxidation functions, and its application Download PDFInfo
- Publication number
- CN104212741B CN104212741B CN201410401745.9A CN201410401745A CN104212741B CN 104212741 B CN104212741 B CN 104212741B CN 201410401745 A CN201410401745 A CN 201410401745A CN 104212741 B CN104212741 B CN 104212741B
- Authority
- CN
- China
- Prior art keywords
- chickpea
- fermentation
- bacillus subtilises
- fibrinolytic
- fermented
- 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.)
- Expired - Fee Related
Links
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention discloses a Bacillus subtilis producing fermented chickpea having fibrinolysis and antioxidation functions, and its application. The Bacillus subtilis is Bacillus subtilis DC-Tx, and is preserved in China General Microbiological Culture Collection Center at No.3 Beichen West Road, Chaoyang District, Beijing on July 15, 2014 with the preservation number of CGMCC NO.9462. The Bacillus subtilis DC-Tx highly producing natto kinase is screened in the invention, and can produce fermented chickpea having fibrinolysis and antioxidation functions. Results show that the fibrinolysis activity of the fermented chickpea produced by the above strain can reach 3000FU/g, the .OH clearance of the fermented chickpea can reach 88.5%, and the Fe＜3+＞ reduction capability of the fermented chickpea is 3.33 times that of fresh chickpea. A fermented chickpea preparation method disclosed in the invention has the advantages of simple operation, low cost, short cycle, easy realization, abundant sources, easy obtaining and low cost of raw materials, no environmental pollution, low prices of used apparatuses and reagents, and easy realization.
Description
Technical field
The present invention relates to a kind of production has fibrinolytic and the bacillus subtilises of anti-oxidation function fermentation chickpea, also simultaneously
It is related to the application of this bacillus subtilis, belong to microbial technology field.
Background technology
In recent years, due to reasons such as dietary unbalance, ecological deterioration, inherited traits, blood vessel embolism class disease is significantly increased,
Seriously threaten health and the life security of the mankind, die from every year cerebral infarction, myocardial infarction patient up to millions of, occupy various
First of disease.Thrombus are the prefered methods treating this kind of disease, therefore, have the research and development day of Hyperfibrinolysis thrombolytic drug
Benefit is paid close attention to by people, has vast potential for future development.One of focus of thrombolytic drug research at present, is by bacterial fermentation
Produce orally active corroded rock mass food and health food nattokinase, but the nattokinase fibrinolytic of the method gained
How not fully up to expectations activity is.
On the other hand, reactive oxygen species and free radicals are the secondary metabolites of human metabolism's process, under normal circumstances always
It is in the dynamic equilibrium constantly producing and eliminating, but if its too high levels, dynamic equilibrium is destroyed, and will cause oxidation
Stress, thus the reduction of cause cancer, coronary heart disease, atherosclerosiss, diabetes, nervous system failure, immunity, joint
More than the 100 kind of common disease such as scorching.Its atherosclerosis can cause cardiovascular and cerebrovascular disease again, produces thrombosis, therefore, active oxygen
It is also one of cause of disease of cardiovascular and cerebrovascular disease with free radical.There are some researches show at present, mended by edible non-oxidizability functional food
Fill antioxidant, be one of prevention effective measures of multiple diseases including cardiovascular and cerebrovascular disease.
In sum, reactive oxygen species and free radicals can cause the multiple diseases including cardiovascular and cerebrovascular disease, and has anti-
The food of oxidative function and health food therefore can also have, with Scavenger of ROS and free radical, the work preventing cardiovascular and cerebrovascular disease
With；And the health food of Hyperfibrinolysis can dissolve the thrombosis having been formed, thus treating cardiovascular and cerebrovascular disease, if research and development one
Plant the food with fibrinolytic and anti-oxidation function, then both there is the pathogenetic function of prevention of cardiovascular disease, again can be in thrombosis
There is after generation therapeutic effect, thus reaching the purpose that prevention and treatment combine, but there is presently no the report of correlational study.
Chickpea (chickpea) scientific name cicer arietinum l., the various saccharides rich in necessary for human, plant egg
White and aminoacid, cellulose, vitamin and inorganic salt, have prophylaxis of hypertension and arteriosclerosis, reduction cholesterol and blood fat blood
Sugar, the effect such as diuresis, treatment insomnia.There are some researches show, the enzymatic hydrolysate (Chickpea short-peptide) of Chickpea Protein have significantly
Anti-oxidation function, also it is reported that, use bacillus natto to ferment chickpea, can obtain Hyperfibrinolysis natto swash
Enzyme, but yet there are no the report of the fermentation chickpea with fibrinolytic and anti-oxidation function.
Content of the invention
It is an object of the invention to provide a kind of production has fibrinolytic and the bacillus subtilis of anti-oxidation function fermentation chickpea
Bacterium and its application, produce fermentation chickpea using the bacillus subtilises of the present invention, can obtain with Hyperfibrinolysis and show
Write the chickpea tunning of antioxidation, and the method has simple to operate, low cost, cycle is short is excellent be easily achieved
Point, is that the industrialization of nattokinase provides technical support, and is its development and application as functional food and health food
Research provides new thinking.
In order to realize object above, the technical solution adopted in the present invention is to provide a kind of production to have fibrinolytic and antioxidation
The bacillus subtilises of function fermentation chickpea, described bacillus subtilises are bacillus subtilises (bacillus
Subtilis) dc-tx, depositary institution: China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation address:
Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, preserving number: cgmcc no.9462, preservation date: on July 15th, 2014.
The present invention filters out the bacillus subtilises dc-tx of high-yield nattokinase from Semen Sojae Preparatum, using the bacterial strain of the present invention,
The fermentation chickpea with Hyperfibrinolysis and notable anti-oxidation function can be produced.Test result indicate that, bacterial strain life of the present invention
The fermentation chickpea producing, up to 3000fu/g, hydroxyl radical free radical (oh) clearance rate is up to 88.5%, fe for its fibrinolytic3+Also
Proper energy power is 3.33 times of fresh chickpea.
The technical solution adopted in the present invention also resides in a kind of bacillus subtilises of offer and has fibrinolytic and antioxygen in production
Change the application of function fermentation chickpea aspect.
Bacillus subtilises of the present invention produce the method with fibrinolytic and anti-oxidation function fermentation chickpea, walk including following
Rapid:
(1) picking bacillus subtilises are inoculated in lb liquid seed culture medium, under the conditions of 28-40 DEG C, 120-180rpm
Shaking table constant temperature culture 12-18h, obtains bacillus subtilises seed liquor；
(2) by chickpea soaked overnight, drain away the water, autoclaving, after being cooled to room temperature, according to the inoculum concentration of 1-8%
Inoculation bacillus subtilises seed liquor, fermentation culture 36-72h under the conditions of 28-40 DEG C；
(3) by the chickpea fermentation culture product cryogenic vacuum lyophilization of step (2), pulverize, crossing 80-120 mesh sieve is
?.
Described lb liquid seed culture medium includes the raw material of following mass fraction: peptone 1%, yeast powder 0.5%, nacl
1%；Ph6.8,121 DEG C of autoclaving 20min.
The cryodesiccated condition of described cryogenic vacuum is: 50 DEG C of precooling temperature, vacuum 20-100pa, freeze temperature≤
45 DEG C, freeze-drying time 24-48h.In the present invention, lyophilization selects cryogenic vacuum lyophilization, and does not adopt spray drying, mesh
Be to preferably keep the fibrinolytic of gained tunning and anti-oxidation function.
The present invention has sending out of Hyperfibrinolysis and notable anti-oxidation function using bacillus subtilises dc-tx fermenting and producing
Ferment chickpea, the method has simple to operate, low cost, cycle is short and the advantage being easily achieved.Meanwhile, the present invention adopts olecranon
Bean is raw material, and abundance easily takes, low cost, non-environmental-pollution, and device therefor, reagent low price are easy to large-scale production.
Brief description
Below in conjunction with accompanying drawing, the specific embodiment of the present invention is described in further detail.
Fig. 1 is present invention fermentation chickpea fibrinolytic measurement result；
Fig. 2 is present invention fermentation chickpea oh clearance rate measurement result；Wherein,
A is gsh, and b is fermentation 36h chickpea, and c is fermentation 48h chickpea, and d is fermentation 72h chickpea, and e is fresh olecranon
Bean；
Fig. 3 is present invention fermentation chickpea fe3+Reducing power measurement result；Wherein,
A is gsh, and b is fermentation 36h chickpea, and c is fermentation 48h chickpea, and d is fermentation 72h chickpea, and e is fresh olecranon
Bean.
Specific embodiment
With reference to embodiments the specific embodiment of the present invention is described in further detail.
Embodiment 1
1st, the screening of nattokinase superior strain
The primary dcreening operation of 1.1 nattokinase superior strains takes commercially available Semen Sojae Preparatum, separates sample 10g, adds in 100ml sterilized water, fills
Divide vibration, 100 DEG C of boiling water boil 5min, in standing a moment, take upper strata dirty solution, after gradient dilution, take 100 μ l differences to dilute respectively
Degree solution, be spread evenly across on lb flat board, 37 DEG C of incubated overnight, choose single bacterium colony carry out streak culture, double.Picking lb
Single bacterium colony dibbling on separation flat board, on fibrin plate, cultivates 18h, measurement dissolving loop diameter for 37 DEG C, dissolving is enclosed larger
Strain liquid fermentation carry out secondary screening.
The bacterial strain that primary dcreening operation transparent circle is relatively large in diameter by the secondary screening of 1.2 nattokinase superior strains, is lived with lb seed culture medium
Change (37 DEG C, 150r/min, overnight), be respectively connected to liquid fermentation medium (in terms of mass fraction: 6% according to 5% inoculum concentration
Semen sojae atricolor powder, 2% glucose, 0.06%cacl2, 0.07%mgso4, ph6.5) in, 37 DEG C of fermentation 72h, its tunning is used
The phosphate buffer dilution of ph7.4, takes diluent 10 μ l, is loaded onto 37 DEG C of constant temperature culture 18h on fibrin plate, measure thoroughly
Bright loop diameter, chooses the maximum bacterial strain of dissolving circle, is named as dc-tx.
The feature of bacterial strain dc-tx is as follows:
Morphological characteristic:
37 DEG C of culture 24h on lb flat board, bacterial strain dc-tx bacterium colony is rounded, and edge is micro- lobate tooth, White-opalescent；Bacterium
Volume morphing is in little rod-short, and long 3.0-4.0 μm, wide 0.8-1.0 μm, Gram-positive is raw in spore, oval.
Physiological and biochemical property is shown in Table 1.
The physiological and biochemical property of table 1 bacterial strain dc-tx
16s rdna sequencing identification:
Extract the genome dna of bacterial strain dc-tx, with 16s rdna gene universal primer pcr amplification；The pcr product obtaining,
It is connected with pgm-t carrier, convert dh5 α competent cell；Screening positive clone, carries plasmid dna and carries out enzyme action identification；Just connect
True positive colony carries out 16s rdna sequencing, and its sequencing result is as follows:
Relatively find, the bacillus subtilises of the complete genome sequence of 16s rdna of bacterial strain dc-tx and online announcement, Xie Dian
Afnyloliquefaciens coincidence rate all reaches 99%.
The result of summary morphological characteristic, physiological and biochemical analysis and molecule sequencing identification, shows that dc-tx is one plant of hay
Bacillus cereuss (bacillus subtilis).
Embodiment 2
The present embodiment bacillus subtilises dc-tx produces the method with fibrinolytic and anti-oxidation function fermentation chickpea,
Comprise the following steps:
(1) picking bacillus subtilises dc-tx is inoculated in lb liquid seed culture medium, and under the conditions of 28 DEG C, 180rpm shakes
Bed constant temperature culture 15h, obtains bacillus subtilises dc-tx seed liquor；
(2) by chickpea soaked overnight, drain away the water, 121 DEG C of autoclaving 20min, after being cooled to room temperature, according to 8%
Inoculum concentration inoculation bacillus subtilises dc-tx seed liquor, fermentation culture 36h under the conditions of 28 DEG C；
(3) by the chickpea fermentation culture product cryogenic vacuum lyophilization of step (2), pulverize, cross 100 mesh sieves and obtain final product；
The cryodesiccated condition of cryogenic vacuum is: 50 DEG C of precooling temperature, vacuum 20pa, 50 DEG C of freeze temperature, freeze-drying time 24h.
Embodiment 3
The present embodiment bacillus subtilises dc-tx produces the method with fibrinolytic and anti-oxidation function fermentation chickpea,
Comprise the following steps:
(1) picking bacillus subtilises dc-tx is inoculated in lb liquid seed culture medium, and under the conditions of 34 DEG C, 120rpm shakes
Bed constant temperature culture 12h, obtains bacillus subtilises dc-tx seed liquor；
(2) by chickpea soaked overnight, drain away the water, 121 DEG C of autoclaving 20min, after being cooled to room temperature, according to 5%
Inoculum concentration inoculation bacillus subtilises dc-tx seed liquor, fermentation culture 48h under the conditions of 34 DEG C；
(3) by the chickpea fermentation culture product cryogenic vacuum lyophilization of step (2), pulverize, cross 80 mesh sieves and obtain final product；Low
The condition of warm vacuum lyophilization is: 50 DEG C of precooling temperature, vacuum 60pa, 55 DEG C of freeze temperature, freeze-drying time 48h.
Embodiment 4
The present embodiment bacillus subtilises dc-tx produces the method with fibrinolytic and anti-oxidation function fermentation chickpea,
Comprise the following steps:
(1) picking bacillus subtilises dc-tx is inoculated in lb liquid seed culture medium, and under the conditions of 40 DEG C, 150rpm shakes
Bed constant temperature culture 18h, obtains bacillus subtilises dc-tx seed liquor；
(2) by chickpea soaked overnight, drain away the water, 121 DEG C of autoclaving 20min, after being cooled to room temperature, according to 1%
Inoculum concentration inoculation bacillus subtilises dc-tx seed liquor, fermentation culture 72h under the conditions of 40 DEG C；
(3) by the chickpea fermentation culture product cryogenic vacuum lyophilization of step (2), pulverize, cross 120 mesh sieves and obtain final product；
The cryodesiccated condition of cryogenic vacuum is: 50 DEG C of precooling temperature, vacuum 100pa, 45 DEG C of freeze temperature, freeze-drying time 36h.
The mensure of experimental example, present invention fermentation chickpea fibrinolytic and anti-oxidation function
The preparation of sample: the fermentation olecranon Semen Glycines powder normal saline suspension of preparation mass fraction 2%, 4 DEG C of extraction 4h, 5000 ×
After g centrifugation 10min, after gained supernatant 15000 × g repeated centrifugation 10min, supernatant is used for fibrinolytic and the survey of anti-oxidation function
Fixed.
1st, dimension flat band method measures the fibrinolytic of fermentation chickpea
The preparation of fibrin plate: 1g agarose is dissolved in the sodium phosphate buffer 100ml of 0.01m ph7.4, takes
This agarose solution 7.5ml, 50 DEG C of water bath heat preservation 5min, add the 100bp/ml thrombin 225 μ l being dissolved in normal saline, mix
Even, 50 DEG C of water-bath 5min, take 7.5ml 3.6mg/ml bovine fibrinogen solution (to be dissolved in the sodium ascorbyl phosphate buffering of 0.1m ph7.4
Liquid), add in the thrombin agarose solution of insulation, rapid mixing, pour the culture dish of diameter 9cm into, wait to coagulate.
The drafting of standard curve: prepare 20,40,60,80 and 100iu/ml urokinase mark with sterile saline respectively
Quasi- product solution, respectively takes 10 μ l point samples on fibrin plate, 37 DEG C of insulation 18h, measures dissolving circle on fibrin plate
Diameter (see accompanying drawing 1), takes the average diameter of three tests, calculates dissolving circle area；With urokinase activity (iu/ml) for horizontal seat
Mark, dissolving circle area is vertical coordinate, draws urokinase standard curve.
The mensure of fibrinolytic: pipette 10 μ l fermentation olecranon Semen Glycines powder lixiviating solution with micro sample adding appliance, point sample is in fibrin
On flat board, 7 DEG C of insulation 18h, measurement fibrin plate dissolve the diameter of circle, calculate dissolving circle area.According to standard curve,
Calculate fibrinolytic.
Measurement result finds, before 48h, with the prolongation of fermentation time, fibrinolytic dramatically increases, after 48h, fibrinolytic
Prolongation in time is basically unchanged, and during fermentation 48h, tunning fibrinolytic can reach 3000fu/g.
2nd, fermentation chickpea removes the mensure of oh ability
50 μ l 3mm 1,10 phenanthrolene (being dissolved in the sodium phosphate buffer of 0.1m ph7.4) is taken to add 96 orifice plates
In, add 50 μ l samples, mix, plus 50 μ l feso4(3mm) aqueous solution, mixes, plus 50 μ l mass fraction 0.01% hydrogen peroxide
(h2o2) aqueous solution, close the lid, 37 DEG C of constant temperature 60min, measure 536nm place absorbance, this be sample sets (a1),
Control distilled water replaces sample (a0), and blank distilled water replaces sample and 0.01% hydrogen peroxide (a2).
With equation below calculate the clearance rate to oh for the sample: oh clearance rate (%)=[(a1-a0)/(a2-a0)] ×
100%
It was found that the oh clearance rate of the chickpea of fermentation 48h reaches 88.5%, it is that Glutathione (gsh) is removed
The nearly twice of oh ability (47.7%), and (measurement result is shown in attached to be significantly higher than the clearance rate (52.3%) to oh for the fresh chickpea
Fig. 2), illustrate that the fermentation chickpea that present invention fermentation obtains has very strong oh Scavenging activity.
3rd, fermentation chickpea reduction fe3+The mensure of ability
250 μ l samples are taken to mix with the potassium ferricyanide aqueous solution of 250 μ l mass fractions 1% (in compareing, not containing sample), 50
DEG C water bath with thermostatic control 20min, adds 250 μ l mass fraction 10% trichloroacetic acid solution afterwards, 5000 × g centrifugation 10min after mixing,
Take 100 μ l supernatant, add in 96 orifice plates, plus 20 μ l mass fraction 0.1%fecl3Aqueous solution, adds 80 μ l distilled water afterwards, mixes
Even, 25 DEG C of standing 10min of mixed liquor, measure the absorbance at supernatant 700nm.
Measurement result shows, ferment chickpea fe3+Reducing power the strongest, od when fermenting 72h700nmIt is worth for 0.40, though with
gsh od700nmValue (1.57) difference is larger, but fresh chickpea fe3+Reducing power (od700nmBe worth for 0.12) 3.33 times,
Show that fermentation chickpea has stronger fe3+Reducing power (see accompanying drawing 3).
Experimental result illustrates, present invention fermentation chickpea has very high fibrinolytic and stronger oxidation resistance.
The present invention is that the large-scale production of fermentation chickpea provides a kind of new approach, and for it as functional food and health care food
The development and application research of product provides new thinking.
Claims (5)
1. a kind of production have fibrinolytic and anti-oxidation function ferment chickpea bacillus subtilises it is characterised in that described withered
Careless bacillus cereuss are bacillus subtilises (bacillus subtilis) dc-tx, depositary institution: Chinese microorganism strain preservation
Administration committee's common micro-organisms center, preservation address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, preserving number: cgmcc
No.9462, preservation date: on July 15th, 2014.
2. a kind of bacillus subtilises as claimed in claim 1 have fibrinolytic and anti-oxidation function fermentation chickpea side in production
The application in face.
3. a kind of bacillus subtilises as claimed in claim 1 produce the side with fibrinolytic and anti-oxidation function fermentation chickpea
Method is it is characterised in that comprise the following steps:
(1) picking bacillus subtilises are inoculated in lb liquid seed culture medium, under the conditions of 28-40 DEG C, 120-180rpm shaking table
Constant temperature culture 12-18h, obtains bacillus subtilises seed liquor；
(2) by chickpea soaked overnight, drain away the water, autoclaving, after being cooled to room temperature, according to the inoculum concentration inoculation of 1-8%
Bacillus subtilises seed liquor, fermentation culture 36-72h under the conditions of 28-40 DEG C；
(3) by the chickpea fermentation culture product cryogenic vacuum lyophilization of step (2), pulverize, cross 80-120 mesh sieve and obtain final product.
4. bacillus subtilises according to claim 3 produce the side with fibrinolytic and anti-oxidation function fermentation chickpea
Method is it is characterised in that described lb liquid seed culture medium includes the raw material of following mass fraction: peptone 1%, yeast powder
0.5%, nacl 1%；Ph6.8,121 DEG C of autoclaving 20min.
5. bacillus subtilises according to claim 3 produce the side with fibrinolytic and anti-oxidation function fermentation chickpea
Method is it is characterised in that the cryodesiccated condition of described cryogenic vacuum is: 50 DEG C of precooling temperature, vacuum 20-100pa, lyophilizing
Temperature≤45 DEG C, freeze-drying time 24-48h.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410401745.9A CN104212741B (en) | 2014-08-15 | 2014-08-15 | Bacillus subtilis producing fermented chickpea having fibrinolysis and antioxidation functions, and its application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410401745.9A CN104212741B (en) | 2014-08-15 | 2014-08-15 | Bacillus subtilis producing fermented chickpea having fibrinolysis and antioxidation functions, and its application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104212741A CN104212741A (en) | 2014-12-17 |
| CN104212741B true CN104212741B (en) | 2017-01-18 |
Family
ID=52094651
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410401745.9A Expired - Fee Related CN104212741B (en) | 2014-08-15 | 2014-08-15 | Bacillus subtilis producing fermented chickpea having fibrinolysis and antioxidation functions, and its application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104212741B (en) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105087447B (en) * | 2015-09-16 | 2018-08-21 | 河南省科学院生物研究所有限责任公司 | One plant of resistance to microwave bacillus subtilis and its application in Nattokinase preparation |
| CN107688019A (en) * | 2017-08-29 | 2018-02-13 | 宝鸡文理学院 | A kind of method of micromethod detection ascorbic acid to hydroxy radical inhibiting rate |
| CN111676156B (en) * | 2020-06-03 | 2022-03-18 | 青岛农业大学 | Bacillus belgii MRS for improving reduction activity and fermentation product and application thereof |
| CN115317399A (en) * | 2022-07-07 | 2022-11-11 | 湖北真福医药有限公司 | Anti-aging composition and preparation method and application thereof |
| CN115005433A (en) * | 2022-07-19 | 2022-09-06 | 湖北真福医药有限公司 | Bacillus subtilis fibrinolytic enzyme composition with effects of maintaining beauty and keeping young, preparation method and application |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103243040A (en) * | 2012-02-13 | 2013-08-14 | 中国科学院过程工程研究所 | Bacillus subtilis LSSE-22 and application thereof |
-
2014
- 2014-08-15 CN CN201410401745.9A patent/CN104212741B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103243040A (en) * | 2012-02-13 | 2013-08-14 | 中国科学院过程工程研究所 | Bacillus subtilis LSSE-22 and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104212741A (en) | 2014-12-17 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104212741B (en) | Bacillus subtilis producing fermented chickpea having fibrinolysis and antioxidation functions, and its application | |
| CN104013657B (en) | A kind of American ginseng medicine extracts after saponin(e microbial fermentation extracting method again | |
| CN108034617A (en) | A kind of Bei Laisi bacillus and its separating screening method and application | |
| CN103243040B (en) | Bacillus subtilis LSSE-22 and application thereof | |
| CN103263448B (en) | Fermentation bacteria used for fermentation pretreatment to improve extraction of Ginkgo biloba L. leaf flavone and application | |
| CN105861381A (en) | Method for lactobacillus plantarum fermented ginseng liquid | |
| CN104187746B (en) | The technique that fermentation method prepares Testa Phaseoli radiati water soluble dietary fiber | |
| CN107828685A (en) | A kind of bacillus subtilis mutagenic strain of high-yield nattokinase and its application | |
| CN104905158B (en) | Marx-kluyveromyces fermented soybean dregs powder | |
| CN103205409B (en) | Method for preparing high-activity nattokinase by taking black beans as raw material | |
| CN102943060B (en) | Strain for nattokinase with high activity and thermal stability and fermented product thereof | |
| CN105918576B (en) | Fermented type coffee bean and preparation method thereof | |
| CN106085991B (en) | A kind of method that solid state fermentation prepares Nattokinase | |
| CN109259188A (en) | A kind of preparation method of the wheat bran lactic fermentation piece rich in viable bacteria | |
| CN105002249B (en) | A kind of method that monascus liquid state fermentation prepares ace inhibitory peptide | |
| CN104585828A (en) | Ganoderma lucidum fermented product and preparation method thereof | |
| CN107841473A (en) | A kind of bacillus subtilis of high-yield nattokinase and its application | |
| CN105087447B (en) | One plant of resistance to microwave bacillus subtilis and its application in Nattokinase preparation | |
| CN110507681A (en) | A kind of technique of higher value application flower of Panax ginseng | |
| CN107828686A (en) | A kind of bacillus subtilis subspecies of high-yield nattokinase and its application | |
| CN107475233A (en) | A kind of method that Nattokinase is produced with bean curd yellow pulp water | |
| CN101412993A (en) | Solid fermentation method of natto kinase | |
| CN108294224A (en) | A method of it is fermented using lactobacillus plantarum and removes heavy metal cadmium in rice | |
| CN108157933A (en) | The preparation method of soluble dietary fiber in a kind of raising citrus peel residue | |
| CN107801905A (en) | A kind of method of ferment Semen Coicis enrichment ligustrazine and fibrinolytic enzyme functional component |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20170118 Termination date: 20200815 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |