CN108936320A - A kind of natto extracts the preparation method of freeze-dried powder - Google Patents
A kind of natto extracts the preparation method of freeze-dried powder Download PDFInfo
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- CN108936320A CN108936320A CN201810914866.1A CN201810914866A CN108936320A CN 108936320 A CN108936320 A CN 108936320A CN 201810914866 A CN201810914866 A CN 201810914866A CN 108936320 A CN108936320 A CN 108936320A
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- 235000013557 nattō Nutrition 0.000 title claims abstract description 68
- 239000000843 powder Substances 0.000 title claims abstract description 33
- 239000000284 extract Substances 0.000 title claims abstract description 30
- 238000002360 preparation method Methods 0.000 title claims abstract description 28
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 20
- 230000000694 effects Effects 0.000 claims abstract description 12
- 239000000706 filtrate Substances 0.000 claims abstract description 12
- 229910000831 Steel Inorganic materials 0.000 claims abstract description 11
- 239000010959 steel Substances 0.000 claims abstract description 11
- 108091000080 Phosphotransferase Proteins 0.000 claims abstract description 7
- 102000020233 phosphotransferase Human genes 0.000 claims abstract description 7
- 239000007787 solid Substances 0.000 claims abstract description 7
- 238000001035 drying Methods 0.000 claims abstract description 6
- 238000007873 sieving Methods 0.000 claims abstract description 6
- 238000000855 fermentation Methods 0.000 claims description 28
- 230000004151 fermentation Effects 0.000 claims description 28
- 244000046052 Phaseolus vulgaris Species 0.000 claims description 15
- 235000010627 Phaseolus vulgaris Nutrition 0.000 claims description 15
- 244000068988 Glycine max Species 0.000 claims description 12
- 235000010469 Glycine max Nutrition 0.000 claims description 12
- 239000000853 adhesive Substances 0.000 claims description 8
- 230000001070 adhesive effect Effects 0.000 claims description 8
- 238000004108 freeze drying Methods 0.000 claims description 8
- 238000000034 method Methods 0.000 claims description 6
- 235000013305 food Nutrition 0.000 claims description 5
- 230000001954 sterilising effect Effects 0.000 claims description 4
- 238000001514 detection method Methods 0.000 claims description 3
- 244000039328 opportunistic pathogen Species 0.000 claims description 3
- 238000004821 distillation Methods 0.000 claims description 2
- 230000009261 transgenic effect Effects 0.000 claims description 2
- 108010073682 nattokinase Proteins 0.000 description 12
- 229940086319 nattokinase Drugs 0.000 description 11
- 208000007536 Thrombosis Diseases 0.000 description 8
- 239000000047 product Substances 0.000 description 6
- 239000004615 ingredient Substances 0.000 description 5
- 239000000463 material Substances 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- 229960000103 thrombolytic agent Drugs 0.000 description 5
- 102000009123 Fibrin Human genes 0.000 description 4
- 108010073385 Fibrin Proteins 0.000 description 4
- BWGVNKXGVNDBDI-UHFFFAOYSA-N Fibrin monomer Chemical compound CNC(=O)CNC(=O)CN BWGVNKXGVNDBDI-UHFFFAOYSA-N 0.000 description 4
- 238000000605 extraction Methods 0.000 description 4
- 229950003499 fibrin Drugs 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 244000063299 Bacillus subtilis Species 0.000 description 3
- 235000014469 Bacillus subtilis Nutrition 0.000 description 3
- 108010049003 Fibrinogen Proteins 0.000 description 3
- 102000008946 Fibrinogen Human genes 0.000 description 3
- 239000012792 core layer Substances 0.000 description 3
- 238000004090 dissolution Methods 0.000 description 3
- 229940012952 fibrinogen Drugs 0.000 description 3
- 239000003527 fibrinolytic agent Substances 0.000 description 3
- 239000008363 phosphate buffer Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 230000002537 thrombolytic effect Effects 0.000 description 3
- 238000009777 vacuum freeze-drying Methods 0.000 description 3
- 238000005303 weighing Methods 0.000 description 3
- 229920000936 Agarose Polymers 0.000 description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- 208000024172 Cardiovascular disease Diseases 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 108010022999 Serine Proteases Proteins 0.000 description 2
- 102000012479 Serine Proteases Human genes 0.000 description 2
- 102000003990 Urokinase-type plasminogen activator Human genes 0.000 description 2
- 108090000435 Urokinase-type plasminogen activator Proteins 0.000 description 2
- 238000004364 calculation method Methods 0.000 description 2
- 208000026106 cerebrovascular disease Diseases 0.000 description 2
- 230000007812 deficiency Effects 0.000 description 2
- 230000002526 effect on cardiovascular system Effects 0.000 description 2
- 229940088598 enzyme Drugs 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 229960005356 urokinase Drugs 0.000 description 2
- 238000009423 ventilation Methods 0.000 description 2
- 239000012224 working solution Substances 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
- 206010008132 Cerebral thrombosis Diseases 0.000 description 1
- 206010008190 Cerebrovascular accident Diseases 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 101710145505 Fiber protein Proteins 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 201000001429 Intracranial Thrombosis Diseases 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 102000001253 Protein Kinase Human genes 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 108010023197 Streptokinase Proteins 0.000 description 1
- 208000006011 Stroke Diseases 0.000 description 1
- 108090000190 Thrombin Proteins 0.000 description 1
- 102000003978 Tissue Plasminogen Activator Human genes 0.000 description 1
- 108090000373 Tissue Plasminogen Activator Proteins 0.000 description 1
- 108010092464 Urate Oxidase Proteins 0.000 description 1
- 206010000891 acute myocardial infarction Diseases 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 235000013527 bean curd Nutrition 0.000 description 1
- 230000023555 blood coagulation Effects 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000010102 embolization Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- XLYOFNOQVPJJNP-ZSJDYOACSA-N heavy water Substances [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 description 1
- GOMNOOKGLZYEJT-UHFFFAOYSA-N isoflavone Chemical compound C=1OC2=CC=CC=C2C(=O)C=1C1=CC=CC=C1 GOMNOOKGLZYEJT-UHFFFAOYSA-N 0.000 description 1
- CJWQYWQDLBZGPD-UHFFFAOYSA-N isoflavone Natural products C1=C(OC)C(OC)=CC(OC)=C1C1=COC2=C(C=CC(C)(C)O3)C3=C(OC)C=C2C1=O CJWQYWQDLBZGPD-UHFFFAOYSA-N 0.000 description 1
- 235000008696 isoflavones Nutrition 0.000 description 1
- 238000012792 lyophilization process Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 229910000403 monosodium phosphate Inorganic materials 0.000 description 1
- 235000019799 monosodium phosphate Nutrition 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 108060006633 protein kinase Proteins 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 229960005202 streptokinase Drugs 0.000 description 1
- 229960004072 thrombin Drugs 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 230000002792 vascular Effects 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L11/00—Pulses, i.e. fruits of leguminous plants, for production of food; Products from legumes; Preparation or treatment thereof
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L11/00—Pulses, i.e. fruits of leguminous plants, for production of food; Products from legumes; Preparation or treatment thereof
- A23L11/30—Removing undesirable substances, e.g. bitter substances
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
- A23L3/36—Freezing; Subsequent thawing; Cooling
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Agronomy & Crop Science (AREA)
- Botany (AREA)
- Beans For Foods Or Fodder (AREA)
Abstract
The invention discloses the preparation methods that a kind of natto extracts freeze-dried powder, and solving fresh natto has micro- stink and need to refrigerate, be not easy the technical issues of transporting and carrying, and belong to natto extract preparation technical field.Then the present invention is added water according to the mass ratio 1:1.2~2.0 of fresh natto and water, is sufficiently stirred under room temperature the following steps are included: (1) weighs fresh natto pours into container；(2) solid content is removed with 8~12 mesh stainless (steel) wire sievings again, obtains filtrate；(3) gained filtrate was reduced the temperature at 1~2 hour -18 DEG C hereinafter, progress drying in 32~35 hours, obtains dry matter of the moisture content less than 5%；(4) pulverizer through 80 meshes crushes again, and finally obtained natto extracts freeze-dried powder.Preparation method of the invention not only remains the excellent activity of original various bioactivators and kinases in fresh natto to the greatest extent, eliminates micro- stink, and can store and transport under normal temperature conditions.
Description
Technical field
The present invention relates to the preparation sides that natto extract preparation technical field more particularly to a kind of natto extract freeze-dried powder
Method.
Background technique
The intracorporal thrombus of people often causes vascular embolization, cerebral thrombosis apoplexy, the serious cardiovascular and cerebrovascular disease such as acute myocardial infarction
Disease causes great harm to the health of the mankind especially middle-aged and the old.Current Thrombolytic agent, such as urokinase (SK) and
Streptokinase (UK) is used as first generation thrombolytics, lacks Fibrin selectivity, and side effect is big；Second generation thrombolytics is as recombinated
Tissue-type plasminogen activator, but there is price height to be difficult for the shortcomings that masses receive.And contain in fresh natto it is special
Substance --- Nattokinase, it is a kind of isoelectric point (pI) that the Bacillus natto being seeded on beans carrier generates during the fermentation
It is 8.6 ± 0.3, serine protease of the molecular weight between 20000~30000Da, this protease has very strong fiber
Protein dissolution ability, and the effectiveness with activatable elastic enzyme and uricase have significantly to cardiovascular and cerebrovascular disease is prevented and treated
Effect.This kinase source in food, have the characteristics that thrombolysis ability it is strong, it is safe and reliable, have no toxic side effect with it is at low cost, therefore
With extensive development prospect.
The 1980s, Japanese Fu Qi medical university must see that foreign firm professor did such test, he is brain
The thrombolytic agent of thrombosis patients' informal dress is put on the artificial thrombus of certain deal, and the artificial thrombus of 2 centimetres of dissolution needs nearly 2 days
Time could complete, and changing the substance extracted from natto in the same way then only needs 3 hours that can complete, also
It is to say that the speed of natto fermentation object thrombus is as many as 19 times of urokinase.Then just this strong thrombus-dissolving object of natto is ordered
Entitled Nattokinase Nattokinase, abbreviation NK, here it is " afternoons famous on world-shaking thrombolytic agent research history
Half past two " experiment.The thrombus dissolving ability that this test sufficiently demonstrates Nattokinase be more than be currently known it is most molten
Thrombus drug.
Nattokinase (NK) is a kind of Proteinkinase, is in soybean isoflavone by natto strain by Bacillus subtilis natto from Traditional Japanese Food
A kind of serine protease that (Bacillus subtilis natto) is generated, is effect ingredient in natto.But it is received due to fresh
The fermenting step of beans in process of production can generate a kind of particular matter " ammonia ", it is made to contain certain micro- stink, this taste
Cause many people to be unwilling to receive, and fresh natto needs to refrigerate, be not easy to transport and carry.
Summary of the invention
The technical issues of present invention has micro- stink and need to refrigerate, be not easy transport and carry for fresh natto, provides one
Kind natto extracts the preparation method of freeze-dried powder, revolutionizes many deficiencies of fresh natto.
To achieve the goals above, the invention provides the following technical scheme:
A kind of natto extracts the preparation method of freeze-dried powder, comprising the following steps:
(1) it weighs fresh natto to pour into container, water then is added according to the mass ratio 1:1.2~2.0 of fresh natto and water, often
Beans adhesive surface object is stirred well under temperature to be all dissolved into water；
(2) solid content is removed with 8~12 mesh stainless (steel) wire sievings again, obtains filtrate；
(3) gained filtrate is reduced the temperature in 1~2 hour -18 DEG C hereinafter, progress drying in 32~35 hours, obtains
Dry matter of the moisture content less than 5%；
(4) pulverizer through 80 meshes crushes again, and finally obtained natto extracts freeze-dried powder.
Further, the drying process of step (3) includes lyophilization and parsing-desiccation, wherein the time of lyophilization
It is 24~30 hours, the time of parsing-desiccation is 5~8 hours.
Further, the time of the lyophilization of step (3) is 28 hours, and the time of parsing-desiccation is 6 hours.
Further, the rustless steel container that container sterilizes using prior process in step (1).
It further, further include the step of natto extraction freeze-dried powder kinase activity detection after natto is made and extracts freeze-dried powder
Suddenly.
Further, fresh natto the preparation method comprises the following steps: three will introduced first from Japanese crown prince's food industry Co., Ltd.
Generation improvement natto opportunistic pathogen is inoculated on the organic Small Grain Soybean of cooked non-transgenic, and the diameter of soya bean is 4~5mm, the shape of soya bean
Shape is ball-type or spheroid shape；Ferment under ventilation condition to soya bean, wherein wind speed be 3~5m/s, wind pressure be 100 ±
10Pa, fermentation temperature are 37~42 DEG C, and fermentation time is 18~22 hours.
Further, the method for the fermentation is fermented using multisection type, and first stage fermentation temperature is 38 DEG C, fermentation time
It is 5 hours；Second stage fermentation temperature is 40 DEG C, and fermentation time is 3 hours；Phase III fermentation temperature is 42 DEG C, when fermentation
Between be 3~4 hours；Fourth stage fermentation temperature is 37 DEG C, fermentation time 7~10 hours.
Compared with prior art, the invention has the benefit that
Natto provided by the invention extracts the preparation method of freeze-dried powder, revolutionizes many deficiencies of fresh natto, it is
By the biology washing lift-off technology of original creation, after the effect of being attached to natto grain surface ingredient " Nattokinase " removing will be wrapped up in, beans
The solid contents such as grain, skin of beancurd filter out, and exaltation goes out the Nattokinase aqueous solution of high-purity, after quick-freezing plant freezes, then use
Moisture content in material is directly become gaseous state (vapor) from the solid-state (ice) freezed by vacuum freeze, finally using catching
Storage is thoroughly taken out moisture content, in drying process, since the presence of ice in material locates material all in entire lyophilization process
Under cryogenic, the fine powder of corresponding mesh number is made according to the grinding of different requirements for the material after drying.It is this to utilize vacuum
Natto of the moisture content that Freeze Drying Technique dries out under vacuum and cryogenic conditions less than 5% extracts freeze-dried powder, not only most
The excellent activity for remaining original various bioactivators and kinases in natto of big degree, eliminates micro- stink, and
It can store and transport under normal temperature conditions.
Specific embodiment
In order to make those skilled in the art more fully understand technical solution of the present invention, below in conjunction with embodiment to this
Invention is further detailed.Raw materials and reagents of the present invention are except product when specified otherwise is.
The preparation of the fresh natto of embodiment 1
The three generations introduced from Japanese crown prince's food industry Co., Ltd. improvement natto opportunistic pathogen is inoculated into first cooked non-
On the organic Small Grain Soybean of transgenosis, the diameter of soya bean is 4~5mm, and the shape of soya bean is ball-type or spheroid shape；It recycles advanced
Multistage program control type ferment at constant temperature system, makes material submerged fermentation at optimal temperature and ventilation condition, wherein wind speed be 3~
5m/s, wind pressure are 100 ± 10Pa, and first stage temperature is 38 DEG C, and fermentation time is 5 hours；Second stage temperature is 40 DEG C, hair
The ferment time is 3 hours；Phase III temperature is 42 DEG C, and fermentation time is 3~4 hours；Fourth stage temperature is 37 DEG C, when fermentation
Between 7~10 hours.Length of string is made after fermentation up to 2 meters or more of the fresh natto of high-quality.
The preparation of 2 natto of embodiment extraction freeze-dried powder
The fresh natto for weighing 100kg, pouring into advance is 0.5m by sterilizing volume3Rustless steel container in, then according to
Water is added in the mass ratio 1:1.2 of fresh natto and water, is stirred well to beans adhesive surface object under room temperature and is all dissolved into water, beans
Grain adhesive surface object includes Nattokinase ingredient；The solid contents such as beans are removed with 8 mesh stainless (steel) wire sievings again, obtain filtrate；It is small with 2
When gained filtrate core layer temperature dropped to -18 DEG C hereinafter, being sent to the lyophilization and 8 carried out in vacuum freeze drying tank 24 hours
The parsing-desiccation of hour obtains dry matter of the moisture content less than 5%；The pulverizer through 80 meshes crushes again, and natto is finally made
Extract freeze-dried powder product.
The preparation of 3 natto of embodiment extraction freeze-dried powder
The fresh natto for weighing 120kg, pouring into advance is 0.5m by sterilizing volume3Rustless steel container in, then according to
Water is added in the mass ratio 1:1.5 of fresh natto and water, is stirred well to beans adhesive surface object under room temperature and is all dissolved into water, beans
Grain adhesive surface object includes Nattokinase ingredient；The solid contents such as beans are removed with 10 mesh stainless (steel) wire sievings again, obtain filtrate；With
Gained filtrate core layer temperature -18 DEG C were dropped in 1.5 hours to do hereinafter, being sent to the distillation carried out 28 hours in vacuum freeze drying tank
Dry and 6 hours parsing-desiccations obtain dry matter of the moisture content less than 5%；The pulverizer through 80 meshes crushes again, final to be made
Natto extracts freeze-dried powder product.
The preparation of 4 natto of embodiment extraction freeze-dried powder
The fresh natto for weighing 150kg, pouring into advance is 0.6m by sterilizing volume3Rustless steel container in, then according to
Water is added in the mass ratio 1:2.0 of fresh natto and water, is stirred well to beans adhesive surface object under room temperature and is all dissolved into water, beans
Grain adhesive surface object includes Nattokinase ingredient；The solid contents such as beans are removed with 12 mesh stainless (steel) wire sievings again, obtain filtrate；With 1
Gained filtrate core layer temperature is dropped to -18 DEG C hereinafter, being sent to the lyophilization carried out in vacuum freeze drying tank 30 hours by hour
With 5 hours parsing-desiccations, dry matter of the moisture content less than 5% is obtained；The pulverizer through 80 meshes crushes again, and final be made is received
Beans extract freeze-dried powder product.
5 natto of embodiment extracts the detection of freeze-dried powder kinase activity
1, the preparation of phosphate buffer (0.01mol/L, pH7.75):
A liquid: disodium hydrogen phosphate (Na is taken2HPO4·12H2O) 3.58g adds water to 1000ml solution；
B liquid: sodium dihydrogen phosphate (NaH is taken2PO4·2H2O) 0.78g adds water to 500ml solution；
A liquid: B liquid=1000ml:190.5ml, adjust pH7.75, obtain phosphate buffer (0.01mol/L,
pH7.75)。
2, the preparation of working solution:
Phosphate buffer (0.01mol/L) and 0.9%NaCl solution is taken to prepare by 1:17.
3, the preparation of 1% agar liquid glucose:
It takes 162mg agarose that 16.2ml working solution is added, is put into 45 DEG C of water-baths after dissolving by heating (to fully transparent)
Inside holding.
4, the preparation of fibrinogen:
Take fibrinogen appropriate: the i.e. each required fibrin commercial weight of (gross weight-tare weight)/content × 48.6mg, later
Add water 16.2ml wiring solution-forming.
5, the preparation of thrombin solution:
Take fibrin ferment appropriate: 0.9% physiological saline is added in the i.e. each blood coagulation enzyme amount in (gross weight-tare weight)/content × 1.5
1.3ml。
6, bed board:
Quickly fibrin ferment is poured into fibrinogen and is shaken up, then the two poured into agarose shake up after bed board, put
Set 20~30 minutes etc. it is to be solidified.16 holes will be made a call to after 30 minutes in plate, as the position that sample is added；
The 3 groups of nattos respectively prepared by embodiment 2~4 extract freeze-dried powder, 4 concentration of every group of dilution, and each concentration corresponds to
One hole separately sets 4 blank wells, and 10 μ l are injected with microsyringe in every hole, covers to take out after 37 DEG C of incubator constant temperature 18h and put down
Plate, the vertical two diameters product value that plate dissolution circle is measured with micrometer is log area value.
Calculation formula: (a+blog area value)/antilogarithm × 20
A=-4.3602
B=2.7292
Two diameter product value of log area value-sample
The inverse of 20-sample concentrations
1 natto of table extracts freeze-dried powder kinase activity testing result
It is obtained by calculation, natto prepared by embodiment 2~4 extracts freeze-dried powder, and natto kinase activity entirely reaches
12000FU/g or more.
It is above that certain exemplary embodiments of the invention are only described by way of explanation, undoubtedly, for ability
The those of ordinary skill in domain without departing from the spirit and scope of the present invention can be with a variety of different modes to institute
The embodiment of description is modified.Therefore, foregoing description is regarded as illustrative in nature, and should not be construed as wanting right of the present invention
Ask the limitation of protection scope.
Claims (7)
1. the preparation method that a kind of natto extracts freeze-dried powder, which comprises the following steps:
(1) it weighs fresh natto to pour into container, water then is added according to the mass ratio 1:1.2~2.0 of fresh natto and water, under room temperature
Beans adhesive surface object is stirred well to all to be dissolved into water；
(2) solid content is removed with 8~12 mesh stainless (steel) wire sievings again, obtains filtrate；
(3) gained filtrate is reduced the temperature in 1~2 hour -18 DEG C hereinafter, carrying out drying in 32~35 hours, acquisition is aqueous
Dry matter of the rate less than 5%；
(4) pulverizer through 80 meshes crushes again, and finally obtained natto extracts freeze-dried powder.
2. the preparation method that natto according to claim 1 extracts freeze-dried powder, which is characterized in that step (3) it is dried
Journey includes lyophilization and parsing-desiccation, wherein the time of lyophilization is 24~30 hours, and the time of parsing-desiccation is 5~8
Hour.
3. the preparation method that natto according to claim 2 extracts freeze-dried powder, which is characterized in that the distillation of step (3) is dry
The dry time is 28 hours, and the time of parsing-desiccation is 6 hours.
4. the preparation method that natto according to claim 1 extracts freeze-dried powder, which is characterized in that container is adopted in step (1)
It is the prior rustless steel container by sterilizing.
5. the preparation method that natto according to claim 1 extracts freeze-dried powder, which is characterized in that extract and freeze in obtained natto
Further include the steps that natto extracts the detection of freeze-dried powder kinase activity after dry powder.
6. described in any item nattos extract the preparation method of freeze-dried powder according to claim 1~5, which is characterized in that fresh natto
The preparation method comprises the following steps: first by the three generations introduced from Japanese crown prince's food industry Co., Ltd. improvement natto opportunistic pathogen be inoculated into it is cooked
The organic Small Grain Soybean of non-transgenic on, the diameter of soya bean is 4~5mm, and the shape of soya bean is ball-type or spheroid shape；In air duct slats
It ferments under part to soya bean, wherein wind speed is 3~5m/s, and wind pressure is 100 ± 10Pa, and fermentation temperature is 37~42 DEG C, fermentation
Time is 18~22 hours.
7. the preparation method that natto according to claim 6 extracts freeze-dried powder, which is characterized in that the method for the fermentation is adopted
It is fermented with multisection type, first stage fermentation temperature is 38 DEG C, and fermentation time is 5 hours；Second stage fermentation temperature is 40 DEG C, hair
The ferment time is 3 hours；Phase III fermentation temperature is 42 DEG C, and fermentation time is 3~4 hours；Fourth stage fermentation temperature is 37
DEG C, fermentation time 7~10 hours.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0884569A (en) * | 1994-09-14 | 1996-04-02 | Takeo Ochi | Utilization of nattou, viscous material, viscous material-free nattou and their applications |
| CN104799201A (en) * | 2014-01-24 | 2015-07-29 | 辽宁天润生物技术有限公司 | Anti-oxidation natto freeze-drying powder preparation method |
| CN107343605A (en) * | 2017-05-15 | 2017-11-14 | 蚌埠市星光豆制品厂 | A kind of instant natto of flavor |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0884569A (en) * | 1994-09-14 | 1996-04-02 | Takeo Ochi | Utilization of nattou, viscous material, viscous material-free nattou and their applications |
| CN104799201A (en) * | 2014-01-24 | 2015-07-29 | 辽宁天润生物技术有限公司 | Anti-oxidation natto freeze-drying powder preparation method |
| CN107343605A (en) * | 2017-05-15 | 2017-11-14 | 蚌埠市星光豆制品厂 | A kind of instant natto of flavor |
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