CN106473065A - The production method of natto fermentation pollen - Google Patents
The production method of natto fermentation pollen Download PDFInfo
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- CN106473065A CN106473065A CN201610905724.XA CN201610905724A CN106473065A CN 106473065 A CN106473065 A CN 106473065A CN 201610905724 A CN201610905724 A CN 201610905724A CN 106473065 A CN106473065 A CN 106473065A
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Classifications
-
- 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
- Beans For Foods Or Fodder (AREA)
Abstract
The present invention relates to a kind of production method of natto fermentation pollen, it carries out amplification cultivation as nutrient raw material to bafillus natto with soya bean, and adding pollen to carry out pollen fermentation based on this culture, its fermentation process includes solid fermentation method and two kinds of solution fermentation.Pollen large protein is hydrolyzed into by the micromolecule polypeptide beneficial to absorption of human body by fermentation, while producing thrombolytic Nattokinase, treating osteoporotic farnoquinone and a large amount of vitamine B groups beneficial to human body, soluble dietary fiber and Bacillus Subtilis Natto of Probiotics viable bacteria etc., the nutritional advantages of collection pollen and natto are organically combined, improve and enhance the nutrient content of soya bean and pollen, give the new function of soya bean and pollen and curative effect.
Description
Technical field
Present invention belongs to the preparing technical field of biochemical article, is related to a kind of production method of natto fermentation pollen.
Background technology
Pollen is the Male reproductive cell of plant, similar to the sperm of animal.Though pollen is little, can be multiplied into newly by it
Plant, so claim pollen be plant " Source of life ", it has concentrated the elite of whole plant, containing abundant nutrition
Composition and medicinal ingredient, reputation is complete cell nutritious element in the world.It not only carries the hereditary information of life, and includes
And breed complete nutrients matter necessary to new life, be plant produce a male heir to continue the family line basic, containing abundant cytology level
Nutrient content, such as protein, amino acid, vitamin, trace element, organized enzyme, flavone compound, lipid, nucleic acid, rape
Element, phytic acid etc., wherein amino acid content and ratio are closest to the amino acid pattern that FAO (Food and Agriculture Organization of the United Nation) (FAO) is recommended, this
Extremely rare in wholefood.What pollen was gathered by honeybee is referred to as Bee Pollen, because it is not only pollen, also includes honeybee
The special glandular secretion thing added in gatherer process, thus with unique natural health effect and medical treatment and beauty valency
Value.
Pollen is the guarantee of honeybee source of nutrition, and royal jelly is that pollen obtains " nutrition elite after honeybee conversion in the body
Crystallization ".However, the yield of royal jelly is limited, so how artificial by pollen transformation so as to which nutrition is more enriched simultaneously
The nutritive value for being similar to royal jelly is produced, is also always the problem that those skilled in the art endeavour to solve for a long time.Microorganism
Fermentation is exactly a large amount of enzymes (ferment) using generation in microorganism growth process, and the technology converted by nutriment is micro-
Biofermentation can be turned the foul and rotten into the rare and ethereal, and bland rice, face, transformation of soybean are that wine, vinegar, sauce etc. are delicious and nutrition is easy to
The novel substance of absorption.The common wine in living on ordinary days of people, vinegar, sauce, fermented bean curd etc. are all fermented products.Pollen is in honeybee body
Royal jelly is converted in the presence of enzyme, conventional people are exactly to combine using different microorganisms, pollen is turned by enzyme needed for producing
Turn to " the class emperor slurry " of the effect of similar royal jelly.Be characterized in, 1, vitamin content increase, particularly human body can not oneself synthesis
Farnoquinone blood calcium can be converted into bone calcium, prevent and treat senile osteoporosis;2nd, high molecular weight protein degrades to be formed greatly
The active peptides of amount, helpful to step-down, hypoglycemic, reducing blood lipid;3rd, Flavone content increases, hypotensive, blood fat, antiviral energy
Power strengthens;4th, some peculiar polysaccharide are formed, stimulates immunologic function to strengthen;In addition they can also produce some special enzymes, such as receive
Beans kinases, can prevent heart infarction, dredging and softening blood vessel, prevention of cardiovascular disease.Therefore, " the class emperor formed by fermented pollen
Health care sage's product that the slurry " Johnson & Johnson that can yet be regarded as is healthy and strong.
The bacterial strain that can be used to ferment is numerous, and wherein bafillus natto is one of current study hotspot.Natto Japan
Through there is the history of more than 1000 year, which is made of initially using the fermentation of bacillus subtilis soya bean on the straw of kitchen, is vegetarian diet monk
The important foodstuffs source of nutrition of companions, then also gradually comes into vogue in imperial family.Japan has started whole people's food natto within 1996
Upsurge.Japanese government is disposed as special project in its " plan educated by food " special secondary school goalkeeper's natto that carries out, i.e., from little culture children
Teenager eats natto.The Japanese whole people ate natto less than 10 years, just became the long-lived big country of first for generally acknowledging in the world.Soya bean
Under bacillus natto to ferment effect, in the natto of 100g, just include different amino acid (16g), Aneurine
(0.07mg), Verdoflavin (0.56mg), Vitamin B6 (0.24mg), fiber (6.7g) etc., the natto of generation of particularly fermenting
Kinases has corroded rock mass, can understand thrombus with oral absorption, for coronary heart disease, the treatment of cerebral thrombus and artery sclerosis.
At present, the domestic health-care natto food that national Bureau of Drugs Supervision has been approved by has 14 kinds, and its function is strengthen immunity
With auxiliary reducing blood lipid.Japan, the country such as Korea also have developed multiple products with Nattokinase as primary efficacy in recent years, only
The annual marketing volume of the nutriment of US and European and functional food just reaches 500,000,000,000 dollars, and Nattokinase is eaten as feature
Quickly growing for product and health products, gathers around and holds out broad prospects.
Although pollen itself is nutritious, often with a small amount of peculiar smell (as fishy smell), part population is not easily accepted by, and
Pollen is changed into through honeybee after royal jelly, and nutrition is more comprehensive and is easy to absorb, and whether we artificially can simulate honeybee, will flower
Where powder transforms into " class bee milk " through fermentable?The purpose of the present invention is exactly the peculiar smell for eliminating pollen, while by pollen
Middle nutrient content is changed into more nutritious to human body and is more easy to the composition that is absorbed by the body.Though human use's fermentable
The history of more than one thousand years is so had, but as pollen source is limited, is belonged to and treasures article, the past, never someone was once luxuriously using flower
Powder is fermented for raw material.Only bee-keeping scale now, Bee Pollen wide material sources, quantity have guarantee, and people just open successively
Begin the research of this respect.And though natto is the food of fashionable Japan, simply just starts within nearly 2 years China is introduced, receive at home
Beans are not yet received by most crowds, so pollen and natto are organically combined by not yet someone's consideration so far, Gu Shi
At present also still without this kind of product on field.
Content of the invention
It is an object of the invention to solving to the problem that prior art is present, one kind is provided with soya bean and pollen as original
Material makes the production method of natto fermentation pollen, on the basis of the original nutrient of pollen is played, produces in conjunction with fermenting bacillus natto
The Nattokinase of thrombus dissolving and promote the farnoquinone of bone calcium uptake, thus give the new function of pollen and glamour.
Technical solution for realizing object above is as described below.
A kind of production method of natto fermentation pollen, it is characterised in that bafillus natto is entered as nutrient raw material with soya bean
Row amplification cultivation, and based on this culture, add pollen to carry out pollen fermentation, specifically include solid fermentation method and liquid is sent out
Two kinds of ferment method:
1) solid fermentation method, comprises the steps:
1a, the preparation of bafillus natto nutrient solution (natto bacterium solution) choose the natto raw material of multiple brands respectively,
Various brands natto raw material is isolated and purified with General nutrition agar plate respectively multiple natto bacterial strains accordingly are obtained, use fiber egg
White flat band method determines the ability that each natto bacterial strain produces Nattokinase respectively, selects and produces the most strong individuality of Nattokinase ability, and -70
DEG C Refrigerator store bacterial classification, used as production bacterial classification;Production bacterial classification is inoculated in LB liquid nutrient media, 35 DEG C~40 DEG C, per point
150~200 turns of clock shaking table culture 16~24 hours overnight, obtains bafillus natto kind nutrient solution;
1b, soya bean is soaked in water 12~24 hours overnight, 115 DEG C~130 DEG C autoclavings 20~30 minutes, with high pressure
Afterwards soya bean soft glutinous be defined, be cooled to 40 DEG C;
1c, the mass ratio after high pressure on soft glutinous soya bean by 2%~10% access Liquid Culture natto gemma overnight
Bacillus, after stirring, cultivates 8~24 hours, and cultivation temperature is 20 DEG C~40 DEG C, has canescence bacterium to soya bean superficial growth
Film;
1d, by pollen Steam by water bath 30 minutes, be cooled to room temperature, then the pollen after cooling and superficial growth had canescence
The soya bean of mycoderm presses 1:1~10 mass ratio is mixed and stirred, and continues fermentation 24~48 hours;
The mixture of 1e, the soya bean of collection fermentation and pollen, vacuum are freezed dried 36~48 hours, and vacuum requirement≤
10Pa, vacuum freeze dried initial temperature for -30~-50 DEG C, after vacuum freezes dried 12 hours, heat up 5 DEG C per hour, temperature
Maintain to parching completely after rising to 25 DEG C, obtain and dry thalline;
1f, the fine powder of 50~100 mesh thalline will be dry will be ground into SAPMAC method pulverizer, packaging, product is obtained;
2) solution fermentation, comprises the steps:
2a, the preparation of bafillus natto nutrient solution (natto bacterium solution) choose the natto raw material of multiple brands respectively,
Various brands natto raw material is isolated and purified with General nutrition agar plate respectively multiple natto bacterial strains accordingly are obtained, use fiber egg
White flat band method determines the ability that each natto bacterial strain produces Nattokinase respectively, selects and produces the most strong individuality of Nattokinase ability, and -70
DEG C Refrigerator store bacterial classification, used as production bacterial classification;Production bacterial classification is inoculated in LB liquid nutrient media, 35 DEG C~40 DEG C, per point
150~200 turns of clock shaking table culture 16~24 hours overnight, obtains bafillus natto kind nutrient solution;
2b, soya bean and water are pressed 1:2~5 mass ratio soaks 16~24 hours, and soak is homogenized or defibrination, with total matter
1%~5% interpolation glycerine or glucose or wheat bran of amount, 115 DEG C~130 DEG C autoclavings 20~30 minutes, it is cooled to 40 DEG C;
2c, the mass ratio after high pressure in soya bean slurries by 2%~5% access Liquid Culture natto gemma bar overnight
Bacterium, is passed through filtrated air, and dissolved oxygen amount is more than 10%, with 100~200 revs/min of agitated liquid to after uniform, cultivates 6~12 hours,
Cultivation temperature is 30 DEG C~42 DEG C;
2d, pollen is placed in water and is boiled 15~25 minutes with 100 DEG C, pollen boil liquid is added to and above-mentioned has cultivated 6~12
In the soya bean slurries of hour, the 10%~60% of total amount of liquid is accounted for pollen boil liquid, continue fermentation 24 hours;
2e, collection zymotic fluid, vacuum are freezed dried 36~48 hours, and vacuum requirement≤10Pa, vacuum freeze dried starting
Temperature is -30~-50 DEG C, after vacuum freezes dried 12 hours, heats up 5 DEG C per hour, and temperature is maintained after rising to 25 DEG C to complete
Till parching, obtain and dry thalline;
2f, the fine powder of 50~100 mesh thalline will be dry will be ground into SAPMAC method pulverizer, packaging, product is obtained.
In the production method of above-mentioned natto fermentation pollen, in step 1a and step 2a, selected natto raw material is produced for Japan
" natto life " board natto, the natto of " the big delicious food in mountain " board and minimum grain natto, BeiJing, China " Japanese flavor Beijing " for producing
Board natto.
In the production method of above-mentioned natto fermentation pollen, described pollen is rape pollen, pollen pini, camellia pollen, jujube flower
Powder, pollen of Semen Fagopyri Esculenti, zasiokaurin, lotus pollen, Pollen Codonopsis, motherwort pollen, corn poppy's pollen, sesame pollen, dandelion
Pollen, pollen powder of chrysanthemun flower, Chinese chestnut pollen, fruit of Chinese magnoliavine pollen, sunflower Bee Pollen, peach blossom pollen, "Hami" melon pollen, acacia flower powder, mountain
One or more combination in short, bristly hair or beard flower pollen, wild rose pollen.
In the production method of above-mentioned natto fermentation pollen, the vacuum of tunning is freezed dried and freezes dried machine using vacuum
Carry out, the initial temperature that vacuum is freezed dried is -50 DEG C, vacuumizes after reaching vacuum, after being gradually heating to 20 DEG C~35 DEG C
Maintain to parching.
In the production method of above-mentioned natto fermentation pollen, in step 1b and step 2b, optimal autoclave temperature used is
121℃.
In the production method of above-mentioned natto fermentation pollen, bafillus natto refers to produce bacterial classification used by natto, in micro- life
Bafillus natto subspecies in the bacillus subtilis bacterial classification belonged in the classification of thing bacteriology, abbreviation bafillus natto.
In the production method of above-mentioned natto fermentation pollen, alleged fermentation refers to using traditional solid fermentation or fermentation tank liquid
Body ferments, and specifically includes pure pollen fermentation or adds the fermentation of the additive grown beneficial to bafillus natto, wherein described
Additive includes that one or several use cooperatively in soya bean, glucose, glycerine, flour, wheat bran.
In state of the art, the assay method of kinase activity ability is relatively more, such as fibrin plate method, tetrapeptide bottom
Thing method, serum plate method etc., adopt fibrin plate method in step 1a of the present invention and step 2a, that is, prepare artificial thrombus flat board,
The sample point sample of Nattokinase on flat board, the area of the dissolving circle which occurs just is become with the molten fiber-reactive of Nattokinase
Than.Concrete assay method is comprised the following steps:
A) fibrin plate is prepared --- first 0.20g BFG is mixed with the Tris-HlC buffer solution of 20ml,
50 DEG C of water-bath 20min, make fibrinogen solution standby;Then by the 1.5% of 0.3g agar and the Tris-HlC of 20ml
Buffer solution mixes, and is cooled to and soon solidifies, make fibrin ferment (20U/ml) after boiling;Off-the-shelf fibrinogen solution
Mix with fibrin ferment;The mixed solution of each flat-plate inverted 20ml, after cooling, is evenly distributed and makes a call to 6 holes and (can be respectively used to four
The Nattokinase of natto brand, positive control and negative control is determined;Or the Nattokinase of different condition fermented pollen is determined),
Standby;
B) detected sample suspension is prepared --- and natto, the fermented pollen for weighing different brands respectively is each 1 gram, uses respectively
The 10ml Tris-HlC buffer solution of 0.05mol/l is made into sample suspension, standby;
C) determination of activity and result observation --- on fibrin plate, different holes are separately added into the natto of different brands or send out
Ferment pollen samples suspension, does positive control with Nattokinase sterling, and the 10ml Tris-HlC buffer solution of 0.05mol/l is used as the moon
Property control.37 DEG C of incubators stand overnight, and observe result;Appearance transparent circle is positive findings, measures transparent loop diameter, transparent circle
Diameter is bigger, and natto kinase activity is higher;
D) the quantitative calculating of Nattokinase --- tested with the standard Nattokinase of different enzyme-activity units as stated above, survey
The diameter of the transparent circle formed by fixed difference enzyme-activity unit.With a diameter of ordinate of transparent circle, made with enzyme-activity unit as abscissa
Figure, obtains natto kinase activity quantitation curves;According in test, the Nattokinase of different brands, different fermentations pollen
Transparent loop diameter, determines corresponding unit of enzyme activity on natto kinase activity quantitation curves;
E) according to the testing result of upper method, in detached difference natto bacterial strain, choose and produce the high bacterium of natto kinase activity
Strain is production bacterial strain;In different pollen fermentation conditions, same selection produces the high method of natto kinase activity.The present invention is produced
In method, select in step 1a and step 2a and produce the individual work in fermented pollen of the most strong bafillus natto of Nattokinase ability
Bacterium number is no less than 106CFU/ gram, natto kinase activity is more than 800U/ gram.
The present invention is fermented using traditional solid-state microorganism fermentation and modern liquid microbe, with pollen and soya bean as original
Material, is fermented with bafillus natto, pollen large protein is hydrolyzed into the micromolecule polypeptide (protein hydrolysate beneficial to absorption of human body
Polypeptide), while producing Nattokinase, farnoquinone, substantial amounts of vitamine B group, soluble dietary fiber and probio natto bud
Spore bacillus viable bacteria etc..Which is with high content of technology, improves and enhance the nutrient content of soya bean and pollen, gives soya bean and pollen is new
Function and curative effect.Flavone compound in pollen and soya bean, rue is remained through natto fermentation pollen obtained in the production method
" selenium " in tongue element, phytic acid, trace element such as rape pollen etc., while by fermentation, produces thrombolytic Nattokinase, controls
Osteoporotic farnoquinone, a large amount of B family vitamins, dietary fiber and a large amount of probios beneficial to human body is treated, makes natto
Fermented pollen has incorporated the thrombolysis of natto in addition to original health-care effect, again and has promoted bone calcium absorption effect, collects pollen and receives
The nutritional advantages of beans are organically combined, and pollen transformation is " class royal jelly ".The present invention extends the health care scope of pollen, except pollen
Outside original health-care effect, also there are good pharmacological action or trophic function to coronary heart disease and sufferers of osteoporosis face.
Specific embodiment
Embodiment 1
A kind of production method of natto fermentation pollen, using solid fermentation method, its production stage is:
1st, the preparation of bafillus natto nutrient solution (natto bacterium solution) is chosen following brand natto respectively (Japan enters
" the big delicious food natto in mountain ", " minimum grain natto " of " the natto life " of mouth, the production of Hokkaido, Japan keg city and Beijing production
" Japanese flavor Beijing natto "), isolated and purified with General nutrition agar plate and 4 kinds of natto bacterial strains are obtained, use fibrin plate
Method determines the ability that these four natto bacterial strains produce Nattokinase, selects and produces the most strong individuality of Nattokinase ability, -70 DEG C of refrigerators
Bacterial classification is preserved, as production bacterial classification;Production bacterial classification is inoculated in the LB liquid nutrient media of commercialization, 35 DEG C~40 DEG C, per point
150~200 turns of clock shaking table culture 16~24 hours overnight, obtains bafillus natto kind nutrient solution;
2nd, soya bean be soaked in water overnight (soak 24 hours according to soya bean quality is different, the weight after soya bean water suction be which from
2.3~2.7 times of body weight), 121 DEG C of autoclavings 20~30 minutes (Small Grain Soybean can high pressure 20 minutes, big grain soya bean needs
30 minutes, glutinous it is defined so that soya bean after high pressure is soft), it is cooled to 40 DEG C;
3rd, the mass ratio after high pressure on soft glutinous soya bean by 5% accesses Liquid Culture bafillus natto overnight, stirs
After mixing uniformly, 24 hours (soya bean superficial growth has canescence mycoderm) is cultivated, cultivation temperature is 37 DEG C;
4th, by rape pollen Steam by water bath 30 minutes, room temperature is cooled to, then by the rape pollen after cooling and superficial growth
There is the soya bean of canescence mycoderm by 1:5 mass ratio is mixed and stirred, and continues fermentation 36 hours;
5th, the soya bean of fermentation and the mixture of rape pollen are collected, and vacuum is freezed dried 48 hours, vacuum requirement≤10Pa,
Vacuum freezes dried initial temperature for -50 DEG C, after vacuum freezes dried 12 hours, heats up 5 DEG C per hour, after temperature rises to 25 DEG C
Maintain to parching completely, obtain and dry thalline;
6th, thalline will be dry be ground into the fine powder of 100 mesh with SAPMAC method pulverizer, packaging, product is obtained.
1. soya bean of table is pressed different proportion with rape pollen and is mixed, different temperatures culture, the time required to completing fermentation (with
It is standard to produce the distinctive glutinous silk of Bacillus natto, and in table, the 2nd row word is temperature, and the 3rd to 6 row word is to produce the time for sticking silk,
Unit is hour)
2. vacuum of table is freezed dried from -50 DEG C, is gradually increased under different equilibrium temperatures, the time required to drying (hour)
As it can be seen from table 1 different proportion soya bean and rape pollen mixing, all can grow very well at 20~45 DEG C, its
In 37~45 DEG C when, the speed of growth is faster.As can be seen from Table 2, the equilibrium temperature that vacuum is freezed dried is higher, and drying time gets over
Short, but consideration retains the demand of natto kinase activity, and equilibrium temperature control is preferred within 30 DEG C.
Embodiment 2
A kind of production method of natto fermentation pollen, using solid fermentation method, its production stage is:
1st, the Japanese Natto of different brands is selected in the preparation of bafillus natto nutrient solution (natto bacterium solution) respectively, with general
Logical nutrient agar panel is isolated and purified and obtains different natto bacterial strains, is determined the strain of different bacillus nattos with fibrin plate method and is produced
The ability of Nattokinase, selects and produces the most strong individuality of Nattokinase ability, and -70 DEG C of Refrigerator store bacterial classifications, used as production bacterial classification.Raw
Bacterial classification inoculation liquid nutrient media is produced, 35 DEG C, 150 turns of shaking table cultures per minute 16 hours, acquisition bafillus natto kind are trained
Nutrient solution;
2nd, soya bean is soaked in water overnight, and (according to the different immersions of soya bean quality 12~24 hours, the weight after soya bean water suction was
2.3~2.7 times of its own weight), 121 DEG C of autoclavings 20~30 minutes (Small Grain Soybean can high pressure 20 minutes, big grain soya bean
30 minutes are needed, glutinous are defined so that soya bean after high pressure is soft), it is cooled to 40 DEG C;
3rd, the mass ratio after high pressure on soft glutinous soya bean by 8% accesses Liquid Culture bafillus natto overnight, stirs
After mixing uniformly, 16 hours (soya bean superficial growth has canescence mycoderm) is cultivated, cultivation temperature is 37 DEG C;
4th, by pollen pini Steam by water bath 30 minutes, room temperature is cooled to, then the pollen pini after cooling and superficial growth is had ash
The soya bean of white mycoderm presses 1:5 mass ratio is mixed and stirred, and continues fermentation 36 hours;
5th, the soya bean of fermentation and the mixture of rape pollen are collected, and vacuum is freezed dried 48 hours, vacuum requirement≤10Pa,
Vacuum freezes dried initial temperature for -50 DEG C, after vacuum freezes dried 12 hours, heats up 5 DEG C per hour, after temperature rises to 25 DEG C
Maintain to parching completely, obtain and dry thalline;
6th, thalline will be dry be ground into the fine powder of 50 mesh with SAPMAC method pulverizer, packaging, product is obtained.
3. soya bean of table presses different proportion mixed culture with pollen pini, and different temperatures culture, (to produce the time required to completing fermentation
The distinctive glutinous silk of raw Bacillus natto is standard, and in table, the 2nd row word is temperature, and the 3rd to 6 row word is to produce the time for sticking silk, single
Position is hour)
4. vacuum of table is freezed dried from -50 DEG C, is gradually increased under different equilibrium temperatures, the time required to drying (hour)
From table 3 it can be seen that different proportion soya bean and pollen pini mixing, all can grow very well at 20~45 DEG C, wherein
When 37~45 DEG C, the speed of growth is faster.As can be seen from Table 4, the equilibrium temperature that vacuum is freezed dried is higher, and drying time is shorter,
But consideration retains the demand of natto kinase activity, equilibrium temperature control is preferred within 30 DEG C.
Embodiment 3
A kind of production method of natto fermentation pollen, using solution fermentation, its production stage is:
1st, the Japanese Natto of different brands is selected in the preparation of bafillus natto nutrient solution (natto bacterium solution) respectively, with general
Logical nutrient agar panel is isolated and purified and obtains different natto bacterial strains, is determined the strain of different bacillus nattos with fibrin plate method and is produced
The ability of Nattokinase, selects and produces the most strong individuality of Nattokinase ability, and -70 DEG C of Refrigerator store bacterial classifications, used as production bacterial classification.Raw
Bacterial classification inoculation liquid nutrient media is produced, 40 DEG C, 200 turns of shaking table cultures per minute 24 hours, acquisition bafillus natto kind are trained
Nutrient solution;
2nd, soya bean and water are pressed 1:3 mass ratio soaks 24 hours, and soak is homogenized or defibrination, with the 3% of gross mass
Add glycerine, 121 DEG C of autoclavings 30 minutes, it is cooled to 40 DEG C;
3rd, the mass ratio after high pressure in soya bean slurries by 5% accesses Liquid Culture bafillus natto overnight, leads to
Enter filtrated air, oxygen dissolving value is more than 10%, with 150 revs/min of agitated liquid, cultivate 10 hours, cultivation temperature is 37 DEG C;
4th, rape pollen is placed in water and is hotted plate 20 minutes with 100 DEG C, rape pollen boil liquid is added to and above-mentioned has cultivated 10
In the soya bean slurries of hour, the 20%~60% of total amount of liquid is accounted for rape pollen boil liquid, continue fermentation 24 hours;
5th, zymotic fluid is collected, and vacuum is freezed dried 48 hours, and vacuum requirement≤10Pa, vacuum freeze dried initial temperature
For -30 DEG C, after vacuum freezes dried 12 hours, heat up 5 DEG C per hour, temperature is maintained to parching completely after rising to 25 DEG C,
Acquisition dries thalline;
6th, thalline will be dry be ground into the fine powder of 50-100 mesh with SAPMAC method pulverizer, packaging, product is obtained.
5. soya bean of table is pressed different proportion with rape pollen and is mixed, different temperatures culture, the time required to completing fermentation (with
It is standard to produce the distinctive glutinous silk of Bacillus natto, and in table, the 2nd row word is temperature, and the 3rd to 6 row word is to produce the time for sticking silk,
Unit is hour)
Liquid Culture result is similar with solid process, but easily mass and automation, but water content is big, freezes dried
Pressure is also big.
Claims (5)
1. a kind of production method of natto fermentation pollen, it is characterised in that bafillus natto is carried out as nutrient raw material with soya bean
Amplification cultivation, and based on this culture, add pollen to carry out pollen fermentation, specifically include solid fermentation method and liquid fermentation
Two kinds of method:
1) solid fermentation method, comprises the steps:
1a, the preparation of bafillus natto nutrient solution choose the natto raw material of multiple brands respectively, former to various brands natto
Material is isolated and purified with General nutrition agar plate respectively and obtains multiple natto bacterial strains accordingly, is surveyed with fibrin plate method respectively
Fixed each natto bacterial strain produces the ability of Nattokinase, selects and produces the most strong individuality of Nattokinase ability, -70 DEG C of Refrigerator store bacterium
Kind, as production bacterial classification;Production bacterial classification is inoculated in LB liquid nutrient media, 35 DEG C~40 DEG C, 150~200 turns per minute
Shaking table culture overnight obtains bafillus natto kind nutrient solution in 16~24 hours;
1b, soya bean is soaked in water 12~24 hours overnight, 115 DEG C~130 DEG C autoclavings 20~30 minutes, with yellow after high pressure
Beans soft glutinous be defined, be cooled to 40 DEG C;
1c, the mass ratio after high pressure on soft glutinous soya bean by 2%~10% access Liquid Culture bafillus natto overnight,
After stirring, cultivate 8~24 hours, cultivation temperature is 20 DEG C~40 DEG C, has canescence mycoderm to soya bean superficial growth;
1d, by pollen Steam by water bath 30 minutes, be cooled to room temperature, then the pollen after cooling and superficial growth had canescence mycoderm
Soya bean press 1:1~10 mass ratio is mixed and stirred, and continues fermentation 24~48 hours;
The mixture of 1e, the soya bean of collection fermentation and pollen, vacuum are freezed dried 36~48 hours, vacuum requirement≤10Pa, very
Sky freezes dried initial temperature for -30~-50 DEG C, after vacuum freezes dried 12 hours, heats up 5 DEG C per hour, and temperature rises to 25
Maintain after DEG C to parching completely, obtain and dry thalline;
1f, the fine powder of 50~100 mesh thalline will be dry will be ground into SAPMAC method pulverizer, packaging, product is obtained;
2) solution fermentation, comprises the steps:
2a, the preparation of bafillus natto nutrient solution choose the natto raw material of multiple brands respectively, former to various brands natto
Material is isolated and purified with General nutrition agar plate respectively and obtains multiple natto bacterial strains accordingly, is surveyed with fibrin plate method respectively
Fixed each natto bacterial strain produces the ability of Nattokinase, selects and produces the most strong individuality of Nattokinase ability, -70 DEG C of Refrigerator store bacterium
Kind, as production bacterial classification;Production bacterial classification is inoculated in LB liquid nutrient media, 35 DEG C~40 DEG C, 150~200 turns per minute
Shaking table culture overnight obtains bafillus natto kind nutrient solution in 16~24 hours;
2b, soya bean and water are pressed 1:2~5 mass ratio soaks 16~24 hours, and soak is homogenized or defibrination, with gross mass
1%~5% adds glycerine or glucose or wheat bran, 115 DEG C~130 DEG C autoclavings 20~30 minutes, is cooled to 40 DEG C;
2c, the mass ratio after high pressure in soya bean slurries by 2%~5% access Liquid Culture bafillus natto overnight,
Filtrated air is passed through, dissolved oxygen amount is more than 10%, with 100~200 revs/min of agitated liquid to after uniform, cultivates 6~12 hours, training
Foster temperature is 30 DEG C~42 DEG C;
2d, pollen is placed in water and is boiled 15~25 minutes with 100 DEG C, pollen boil liquid is added to and above-mentioned has cultivated 6~12 hours
Soya bean slurries in, account for the 10%~60% of total amount of liquid to pollen boil liquid, continue fermentation 24 hours;
2e, collection zymotic fluid, vacuum are freezed dried 36~48 hours, and vacuum requirement≤10Pa, vacuum freeze dried initial temperature
For -30~-50 DEG C, after vacuum freezes dried 12 hours, heat up 5 DEG C per hour, temperature is maintained after rising to 25 DEG C to parching completely
Till, obtain and dry thalline;
2f, the fine powder of 50~100 mesh thalline will be dry will be ground into SAPMAC method pulverizer, packaging, product is obtained.
2. the production method of natto fermentation pollen according to claim 1, it is characterised in that institute in step 1a and step 2a
The natto raw material of selection is " natto life " board natto, the natto of " the big delicious food in mountain " board and minimum grain natto, the China that Japan produces
" Japanese flavor Beijing " board natto that is produced from Beijing.
3. the production method of natto fermentation pollen according to claim 1, it is characterised in that described pollen is rape flower
Powder, pollen pini, camellia pollen, Chinese date pollen, pollen of Semen Fagopyri Esculenti, zasiokaurin, lotus pollen, Pollen Codonopsis, motherwort pollen, corn poppy
Pollen, sesame pollen, dandelion pollen, pollen powder of chrysanthemun flower, Chinese chestnut pollen, fruit of Chinese magnoliavine pollen, sunflower Bee Pollen, peach blossom pollen, Kazakhstan
One or more combination in close melon pollen, acacia flower powder, May bloom pollen, wild rose pollen.
4. the production method of natto fermentation pollen according to claim 1, it is characterised in that the vacuum frost of tunning
Drying freezes dried machine using vacuum to be carried out, and the initial temperature that vacuum is freezed dried is -50 DEG C, vacuumizes after reaching vacuum,
Maintain to parching after being gradually heating to 20 DEG C~35 DEG C.
5. the production method of natto fermentation pollen according to claim 1, it is characterised in that institute in step 1b and step 2b
Autoclave temperature is 121 DEG C.
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CN109221926A (en) * | 2018-11-08 | 2019-01-18 | 浙江大学 | A kind of highland barley natto processing method |
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CN107523599A (en) * | 2017-09-01 | 2017-12-29 | 兰溪市沉默生物科技有限公司 | The method that fermentation method prepares peony pollen polypeptide |
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