CN107475230A - Containing sesame, lily, Chinese yam composition high-yield nattokinase liquid culture medium - Google Patents
Containing sesame, lily, Chinese yam composition high-yield nattokinase liquid culture medium Download PDFInfo
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- CN107475230A CN107475230A CN201710830142.4A CN201710830142A CN107475230A CN 107475230 A CN107475230 A CN 107475230A CN 201710830142 A CN201710830142 A CN 201710830142A CN 107475230 A CN107475230 A CN 107475230A
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- bacillus subtilis
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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
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- C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
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- C12N9/52—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from bacteria or Archaea
- C12N9/54—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from bacteria or Archaea bacteria being Bacillus
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- C12Y—ENZYMES
- C12Y304/00—Hydrolases acting on peptide bonds, i.e. peptidases (3.4)
- C12Y304/21—Serine endopeptidases (3.4.21)
- C12Y304/21062—Subtilisin (3.4.21.62)
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Abstract
The invention discloses containing sesame, lily, Chinese yam composition high-yield nattokinase liquid culture medium, belong to biological technical field.Fermentation medium includes following components：Navy bean powder 3.0%, white sesameseed powder 3.0%, yam flour 2.0%, lily root flour 2.0%, glucose 4.0%, sodium chloride 1.5%, dipotassium hydrogen phosphate 0.1%, potassium dihydrogen phosphate 0.1%, pH value 7.0.Compared with prior art, the fermentation medium of the present invention contains white sesameseed, Chinese yam, the raw material of three kinds of integration of drinking and medicinal herbs of lily, wherein more beneficiating ingredients are discharged while high-yield nattokinase after microbial fermentation, are preferably absorbed beneficial to human body, zymotic fluid natto kinase activity reaches 33250u/ml.
Description
Technical field
The invention belongs to biological technical field, be related to it is a kind of containing sesame, lily, Chinese yam composition high-yield nattokinase liquid
State culture medium.
Background technology
Thrombotic diseases are the high morbidities of mid-aged population, are the second largest illnesss for being only second to cancer now, to person in middle and old age
People has very big death threats.By the end of 2016, more than the 60 years old elderly population in China were 2.2 hundred million people, account for total population
16.1%, China has been enter into aging society.At present, thrombus is the important means for treating this kind of disease, is used for now
The medicine of clinical treatment includes streptokinase, urokinase, tissue-type plasminogen activator, the work of single chain urokinase type plasminogen activator type plasminogen
Agent, the streptokinase activator complex of fibrinolysin one etc. of chemical modification, but obvious deficiency all be present in these medicines：It is systemic
Bleeding, drug half-life be short, expensive etc., therefore, develop new thrombolytic agent turned into current important subject it
One.
Nattokinase (Nattokinase, NK) is a kind of Proteinkinase, is by receiving in soybean isoflavone by natto strain
A kind of serine protease caused by beans hay bacillus, have thrombus, reduce blood viscosity, improve blood circulation, softening and
Increase the effect such as blood vessel elasticity.Nattokinase thrombolysis mechanism is：Nattokinase is by stimulating vascular endothelial cell to produce tectotype
Plasminogen activation is fibrinolysin by plasminogen activator (t-PA), t-PA, solution fibrin, direct thrombus；Together
When by the prourokinase activation in human body be urokinase, urokinase and t-PA together plasminogen activation, indirect thrombus.Receive
The main substance fibrin of beans kinases thrombus, does not hydrolyze plasma fibrinogen, so will not trigger the danger of bleeding
Danger, simultaneously, moreover it is possible to the thrombolysis ability of activation enhancing human body itself, so as to reach the thrombolytic effect of long lasting and stable.
The technique that bacillus subtilis solid state fermentation contains Nattokinase product easily infects miscellaneous bacteria, and radiating and recovery have
It is difficult；And the research in terms of liquid state fermentation also focuses mostly in the seed selection of excellent species, the optimization of zymotechnique and the separation of product
In terms of purifying, the research to liquid culture medium composition selection is less, wherein screening Nattokinase unit enzyme activity is high, and has
The research of the culture medium of other food therapy health effects is few.
Lily is the bulb of liliaceous plant lily, containing a variety of alkaloids such as colchicin and starch, protein and
The nutritional ingredients such as reduced sugar, there is the effect of clearing away the heart fire and tranquillizing, clinic is used for restlessness of asrhenia type and insomnia, wandering etc...The drying of Chinese yam
Rhizome is included in pharmacopeia, and its is sweet, mild-natured, returns spleen lung kidney channel, has tonifying spleen nourishing the stomach, engender liquid and benefic lung, the puckery smart work(of kidney tonifying
Can, for spleen eating less, the diarrhea just disease such as the pool, leukorrhea；The effect of in Chinese yam composition Thick many candies, soap it is general etc. all to promoting machine
The balance of body microscopic system, the reaction of immunocyte, there is good positive facilitation in immune system.Sesame is rich in a variety of battalion
Form point, containing abundant unrighted acid, vitamin and protein, wherein methionine, arginine and tryptophane shows
Work is higher than other plant, also all universal in world many countries containing special physiologically active ingredient sesame lignan, sesame
As the indispensable raw material of dietary food product, it is one of kind of anticancer food that the U.S. announces Chinese sesame in recent years.
Nattokinase product containing other healthcare functions in the market, most of is in later stage and natto powder or natto
Capsule or other health foods are made in the pure powder of kinases after being simply mixed, this technique hinders macromolecular in raw material, micro member
The absorption and utilization of the active ingredients such as element.Further culture medium raw material of the exploitation with a variety of integration of drinking and medicinal herbs combines, high obtaining
While producing Nattokinase, the more beneficiating ingredients of integration of drinking and medicinal herbs raw material in culture medium are discharged by microbial fermentation, are beneficial to
Human body preferably absorbs, and the exploitation for Nattokinase product is significant.
The content of the invention
It is an object of the invention to provide a kind of fermentation of bacillus subtilis culture medium, the fermentation medium contain white sesameseed,
Chinese yam, the raw material of three kinds of integration of drinking and medicinal herbs of lily, being discharged after microbial fermentation while high-yield nattokinase wherein more has
Beneficial composition, preferably absorbed beneficial to human body.
The technical scheme is that：
A kind of Nattokinase liquid state fermentation culture medium of integration of drinking and medicinal herbs, contains following components：Navy bean powder 3.0-10.0%,
White sesameseed powder 2.0-5.0%, yam flour 2.0-5.0%, lily root flour 2.0-5.0%, glucose 3.0-8.0%, sodium chloride 1.0-
3.0%, dipotassium hydrogen phosphate 0.05-0.2%, potassium dihydrogen phosphate 0.05-0.2%, pH value 7.0-7.4, it is w/v.
Preferable culture medium, which forms, is：Navy bean powder 3.0%, white sesameseed powder 3.0%, yam flour 2.0%, lily root flour
2.0%, glucose 4.0%, sodium chloride 1.5%, dipotassium hydrogen phosphate 0.1%, potassium dihydrogen phosphate 0.1%, pH value 7.0, it is w/
v。
For verifying the bacillus subtilis bacterial strain Bacillus subtilis subsp.UJS-1 of above-mentioned culture medium, now protect
It is hidden in China Committee for Culture Collection of Microorganisms's common micro-organisms of city of BeiJing, China Chaoyang District North Star West Road 1 institute 3
Center (CGMCC), deposit number are CGMCC No.14434, and preservation date is on 07 17th, 2017, it is proposed that Classification And Nomenclature
For：Bacillus subtilis Bacillus subtilis.
Fermentation medium provided by the invention contains white sesameseed, Chinese yam, the raw material of three kinds of integration of drinking and medicinal herbs of lily, through microorganism
Wherein more beneficiating ingredients are discharged after fermentation while high-yield nattokinase, are preferably absorbed beneficial to human body.
Embodiment
With reference to specific embodiment, the present invention is described in detail
The present invention is used for the bacillus subtilis strain Bacillus subtilis for verifying embodiment culture medium
Subsp.UJS-1, now it is preserved in the Chinese microorganism strain preservation management of city of BeiJing, China Chaoyang District North Star West Road 1 institute 3
Committee's common micro-organisms center (CGMCC), deposit number are CGMCC No.14434, and preservation date is 07 month 2017 17
Day, it is proposed that Classification And Nomenclature be：Bacillus subtilis Bacillus subtilis.
Embodiment 1
Culture medium forms：Navy bean powder 3.0%, glucose 4.0%, sodium chloride 1.5%, dipotassium hydrogen phosphate 0.1%, phosphorus
Acid dihydride potassium 0.1%, pH value 7.0, it is w/v.
The culture medium being made into above-mentioned composition proportion carries out bacillus subtilis (Bacillus subtilis
Subsp.UJS-1, CGMCC No.14434) liquid state fermentation, 37 DEG C, 225rpm, ferment 48h, pass through agarose-fibrin
It is 29170u/ml that flat band method, which measures zymotic fluid Nattokinase enzymatic activity,.
Embodiment 2
Culture medium forms：Navy bean powder 4.0%, glucose 4.0%, sodium chloride 1.5%, dipotassium hydrogen phosphate 0.1%, phosphorus
Acid dihydride potassium 0.1%, pH value 7.0, it is w/v.
The culture medium being made into above-mentioned composition proportion carries out bacillus subtilis (Bacillus subtilis
Subsp.UJS-1, CGMCC No.14434) liquid state fermentation, 37 DEG C, 225rpm, ferment 48h, pass through agarose-fibrin
Flat band method (with reference to Xie Pinghui etc., 2010, Nattokinase vigor in fibrin plate method measure health food) measure zymotic fluid
Nattokinase enzymatic activity is 26650u/ml.
Embodiment 3
Culture medium forms：Navy bean powder 5.0%, glucose 4.0%, sodium chloride 1.5%, dipotassium hydrogen phosphate 0.1%, phosphorus
Acid dihydride potassium 0.1%, pH value 7.0, it is w/v.
The culture medium being made into above-mentioned composition proportion carries out bacillus subtilis (Bacillus subtilis
Subsp.UJS-1, CGMCC No.14434) liquid state fermentation, 37 DEG C, 225rpm, ferment 48h, pass through agarose-fibrin
It is 27580u/ml that flat band method, which measures zymotic fluid Nattokinase enzymatic activity,.
Embodiment 4
Culture medium forms：Navy bean 3.0%, white sesameseed powder 3.0%, glucose 4.0%, sodium chloride 1.5%, phosphoric acid hydrogen
Dipotassium 0.1%, potassium dihydrogen phosphate 0.1%, pH value 7.0, it is w/v.
The culture medium being made into above-mentioned composition proportion carries out bacillus subtilis (Bacillus subtilis
Subsp.UJS-1, CGMCC No.14434) liquid state fermentation, 37 DEG C, 225rpm, ferment 48h, pass through agarose-fibrin
It is 25010u/ml that flat band method, which measures zymotic fluid Nattokinase enzymatic activity,
Embodiment 5
Culture medium forms：Navy bean powder 3.0%, white sesameseed powder 4.0%, glucose 4.0%, sodium chloride 1.5%, phosphoric acid
Hydrogen dipotassium 0.1%, potassium dihydrogen phosphate 0.1%, pH value 7.0, it is w/v.
The culture medium being made into above-mentioned composition proportion carries out bacillus subtilis (Bacillus subtilis
Subsp.UJS-1, CGMCC No.14434) liquid state fermentation, 37 DEG C, 225rpm, ferment 48h, pass through agarose-fibrin
It is 25450u/ml that flat band method, which measures zymotic fluid Nattokinase enzymatic activity,.
Embodiment 6
Culture medium forms：Navy bean powder 3.0%, white sesameseed powder 5.0%, glucose 4.0%, sodium chloride 1.5%, phosphoric acid
Hydrogen dipotassium 0.1%, potassium dihydrogen phosphate 0.1%, pH value 7.0, it is w/v.
The culture medium being made into above-mentioned composition proportion carries out bacillus subtilis (Bacillus subtilis
Subsp.UJS-1, CGMCC No.14434) liquid state fermentation, 37 DEG C, 225rpm, ferment 48h, pass through agarose-fibrin
It is 26680u/ml that flat band method, which measures zymotic fluid Nattokinase enzymatic activity,.
Embodiment 7
Culture medium forms：Navy bean powder 3.0%, yam flour 2.0%, glucose 4.0%, sodium chloride 1.5%, phosphoric acid hydrogen
Dipotassium 0.1%, potassium dihydrogen phosphate 0.1%, pH value 7.0, it is w/v.
The culture medium being made into above-mentioned composition proportion carries out bacillus subtilis (Bacillus subtilis
Subsp.UJS-1, CGMCC No.14434) liquid state fermentation, 37 DEG C, 225rpm, ferment 48h, pass through agarose-fibrin
It is 35630u/ml that flat band method, which measures zymotic fluid Nattokinase enzymatic activity,
Embodiment 8
Culture medium forms：Navy bean powder 3.0%, yam flour 3.0%, glucose 4.0%, sodium chloride 1.5%, phosphoric acid hydrogen
Dipotassium 0.1%, potassium dihydrogen phosphate 0.1%, pH value 7.0, it is w/v.
The culture medium being made into above-mentioned composition proportion carries out bacillus subtilis (Bacillus subtilis
Subsp.UJS-1, CGMCC No.14434) liquid state fermentation, 37 DEG C, 225rpm, ferment 48h, pass through agarose-fibrin
It is 32840u/ml that flat band method, which measures zymotic fluid Nattokinase enzymatic activity,.
Embodiment 9
Culture medium forms：Navy bean powder 3.0%, yam flour 4.0%, glucose 4.0%, sodium chloride 1.5%, phosphoric acid hydrogen
Dipotassium 0.1%, potassium dihydrogen phosphate 0.1%, pH value 7.0, it is w/v.
The culture medium being made into above-mentioned composition proportion carries out bacillus subtilis (Bacillus subtilis
Subsp.UJS-1, CGMCC No.14434) liquid state fermentation, 37 DEG C, 225rpm, ferment 48h, pass through agarose-fibrin
It is 33690u/ml that flat band method, which measures zymotic fluid Nattokinase enzymatic activity,.
Embodiment 10
Culture medium forms：Navy bean powder 3.0%, lily root flour 2.0%, glucose 4.0%, sodium chloride 1.5%, phosphoric acid hydrogen
Dipotassium 0.1%, potassium dihydrogen phosphate 0.1%, pH value 7.0, it is w/v.
The culture medium being made into above-mentioned composition proportion carries out bacillus subtilis (Bacillus subtilis
Subsp.UJS-1, CGMCC No.14434) liquid state fermentation, 37 DEG C, 225rpm, ferment 48h, pass through agarose-fibrin
It is 33540u/ml that flat band method, which measures zymotic fluid Nattokinase enzymatic activity,
Embodiment 11
Culture medium forms：Navy bean powder 3.0%, lily root flour 3.0%, glucose 4.0%, sodium chloride 1.5%, phosphoric acid hydrogen
Dipotassium 0.1%, potassium dihydrogen phosphate 0.1%, pH value 7.0, it is w/v.
The culture medium being made into above-mentioned composition proportion carries out bacillus subtilis (Bacillus subtilis
Subsp.UJS-1, CGMCC No.14434) liquid state fermentation, 37 DEG C, 225rpm, ferment 48h, pass through agarose-fibrin
It is 30910u/ml that flat band method, which measures zymotic fluid Nattokinase enzymatic activity,.
Embodiment 12
Culture medium forms：Navy bean powder 3.0%, lily root flour 2.0%, glucose 4.0%, sodium chloride 1.5%, phosphoric acid hydrogen
Dipotassium 0.1%, potassium dihydrogen phosphate 0.1%, pH value 7.0, it is w/v.
The culture medium being made into above-mentioned composition proportion carries out bacillus subtilis (Bacillus subtilis
Subsp.UJS-1, CGMCC No.14434) liquid state fermentation, 37 DEG C, 225rpm, ferment 48h, pass through agarose-fibrin
It is 28920u/ml that flat band method, which measures zymotic fluid Nattokinase enzymatic activity,.
Embodiment 13
Culture medium forms：Navy bean powder 3.0%, white sesameseed powder 3.0%, yam flour 2.0%, lily root flour 2.0%, grape
Sugar 4.0%, sodium chloride 1.5%, dipotassium hydrogen phosphate 0.1%, potassium dihydrogen phosphate 0.1%, pH value 7.0, is w/v.
The culture medium being made into above-mentioned composition proportion carries out bacillus subtilis (Bacillus subtilis
Subsp.UJS-1, CGMCC No.14434) liquid state fermentation, 37 DEG C, 225rpm, ferment 48h, pass through agarose-fibrin
It is 33250u/ml that flat band method, which measures zymotic fluid Nattokinase enzymatic activity,.
Comparative example
Culture medium forms：Analysis for soybean powder 4.0%, glucose 4.0%, sodium chloride 1.5%, dipotassium hydrogen phosphate 0.1%, phosphoric acid
Potassium dihydrogen 0.1%, pH value 7.0, it is w/v.
The culture medium being made into above-mentioned composition proportion carries out bacillus subtilis (Bacillus subtilis
Subsp.UJS-1, CGMCC No.14434) liquid state fermentation, 37 DEG C, 225rpm, ferment 48h, pass through agarose-fibrin
It is 9830u/ml that flat band method, which measures zymotic fluid Nattokinase enzymatic activity,.
Claims (3)
1. it is a kind of containing sesame, lily, Chinese yam composition high-yield nattokinase liquid culture medium, it is characterised in that containing with the following group
Point：Navy bean powder 3.0-10.0%, white sesameseed powder 2.0-5.0%, yam flour 2.0-5.0%, lily root flour 2.0-5.0%, grape
Sugared 3.0-8.0%, sodium chloride 1.0-3.0%, dipotassium hydrogen phosphate 0.05-0.2%, potassium dihydrogen phosphate 0.05-0.2%, pH value
7.0-7.4 it is w/v.
2. it is according to claim 1 it is a kind of containing sesame, lily, Chinese yam composition high-yield nattokinase liquid culture medium,
It is characterized in that preferable culture medium composition is：Navy bean powder 3.0%, white sesameseed powder 3.0%, yam flour 2.0%, lily root flour
2.0%, glucose 4.0%, sodium chloride 1.5%, dipotassium hydrogen phosphate 0.1%, potassium dihydrogen phosphate 0.1%, pH value 7.0, it is w/
v。
A kind of 3. high-yield nattokinase liquid culture containing sesame, lily, Chinese yam composition according to claim 1 or 2
Base, it is characterised in that one plant of Nattokinase superior strain Bacillus subtilis subsp.UJS-1 that culture medium uses, protect
Hide numbering be CGMCC No.14434, it is proposed that Classification And Nomenclature be：Bacillus subtilis Bacillus subtilis.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108949733A (en) * | 2018-08-09 | 2018-12-07 | 江苏大学 | A kind of Nattokinase extracting solution and its application |
Citations (2)
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| CN101597192A (en) * | 2008-06-05 | 2009-12-09 | 姚淑先 | Chinese herb culture medium, its preparation method and culturing edible fungi |
| CN105725008A (en) * | 2016-02-20 | 2016-07-06 | 三株福尔制药有限公司 | Natto bacillus subtilis fermented milkvetch root composition and preparation method and application thereof |
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2017
- 2017-09-08 CN CN201710830142.4A patent/CN107475230A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101597192A (en) * | 2008-06-05 | 2009-12-09 | 姚淑先 | Chinese herb culture medium, its preparation method and culturing edible fungi |
| CN105725008A (en) * | 2016-02-20 | 2016-07-06 | 三株福尔制药有限公司 | Natto bacillus subtilis fermented milkvetch root composition and preparation method and application thereof |
Non-Patent Citations (1)
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108949733A (en) * | 2018-08-09 | 2018-12-07 | 江苏大学 | A kind of Nattokinase extracting solution and its application |
| CN108949733B (en) * | 2018-08-09 | 2021-09-24 | 江苏大学 | Nattokinase extracting solution and application thereof |
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