CN106174040B - A kind of production method of the rich lactobacillus type fermented soya bean for producing γ-aminobutyric acid - Google Patents
A kind of production method of the rich lactobacillus type fermented soya bean for producing γ-aminobutyric acid Download PDFInfo
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- CN106174040B CN106174040B CN201610600840.0A CN201610600840A CN106174040B CN 106174040 B CN106174040 B CN 106174040B CN 201610600840 A CN201610600840 A CN 201610600840A CN 106174040 B CN106174040 B CN 106174040B
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- 244000068988 Glycine max Species 0.000 title claims abstract description 103
- 235000010469 Glycine max Nutrition 0.000 title claims abstract description 103
- BTCSSZJGUNDROE-UHFFFAOYSA-N gamma-aminobutyric acid Chemical compound NCCCC(O)=O BTCSSZJGUNDROE-UHFFFAOYSA-N 0.000 title claims abstract description 34
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 25
- 229960003692 gamma aminobutyric acid Drugs 0.000 title claims abstract description 18
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- 241000186660 Lactobacillus Species 0.000 title claims abstract description 8
- 229940039696 lactobacillus Drugs 0.000 title claims abstract description 8
- 238000000855 fermentation Methods 0.000 claims abstract description 34
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- 239000000843 powder Substances 0.000 claims abstract description 32
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- 240000006024 Lactobacillus plantarum Species 0.000 claims abstract description 20
- 238000011081 inoculation Methods 0.000 claims abstract description 13
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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
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
Abstract
The invention discloses a kind of production methods of rich lactobacillus type fermented soya bean for producing γ-aminobutyric acid, this method comprises: soya bean is successively cleaned, is impregnated, is drained, after boiling, inoculation bacterium powder carries out koji-making and fermentation, obtain fermented soya bean；Before koji-making, the bacterium powder of inoculation is the Mixed Microbes powder of aspergillus oryzae and aspergillus niger, and the mass ratio of the two is 1.5~4:1；After koji-making, inoculation contains lactobacillus plantarum bacterium solution, then ferments.The method of the present invention carries out koji-making using the Mixed Microbes powder of aspergillus oryzae and aspergillus niger, lactobacillus plantarum bacterium solution is fermented, and cooperate suitable koji-making and zymotechnique, it significantly improves into bent prolease activity, the amino-acid nitrogen content and alpha-aminobutyric acid content that fermentation generates, obtains quality and taste is better and healthier fermented soya bean finished product；Preparation process is simple, substantially reduces fermentation period, reduces production cost.
Description
Technical field
The present invention relates to technical field of fungal fermentation more particularly to a kind of rich lactobacillus type fermented soya bean for producing γ-aminobutyric acid
Production method.
Background technique
Fermented soya bean are the traditional fermented bean products in China, since its nutrition abundant and unique flavor are deep by the majority of consumers
Like；Especially in south, dish can be not only done, but also can be used as flavouring and eaten extensively.Due to fermented soya bean preparation process
In microbial fermentation effect, not only make fermented soya bean protein rich in, fat, carbohydrate, vitamin and mineral
The nutritional ingredients such as matter, also form polypeptide abundant in fermented soya bean, amino acid, oligosaccharides, and the substances such as monosaccharide make fermented soya bean be easier quilt
Absorption of human body promotes digestion, is suitable for the crowd of stomach malabsorption, the old man and dyspeptic children of digestion power decline
It is edible.
Due to areal variation, makes the microorganism that fermented soya bean are utilized and be also not quite similar, mainly there is aspergillus type fermented soya bean, such as Hunan
Liuyang fermented brown bean；Bacterial fermented douchi such as utilizes the Japanese Natto of bafillus natto production；Mucor type fermented soya bean, such as the Yongchuan in Chongqing
Fermented soya bean；Head mold type fermented soya bean, as the day of Indonesia is trained.
Traditional fermented soya bean manufacture craft is: soya bean → immersion → boiling → drains cooling → inoculation → room temperature koji-making → and washes
Bent, addition auxiliary material → rear ferment → fermented soya bean → drying → finished product.Above-mentioned technique is mainly fermented under field conditions (factors), so that fermented soya bean system
The bent time is longer, and the production of fermented soya bean is also limited by season, and most of fermented soya bean all make in temperature higher summer, and
It cannot produce throughout the year, therefore seriously affect the utilization of working shop of enterprise and the market supply and demand and production-scale expansion of fermented soya bean, increase
Production cost is added.
To improve the above problem, it is conducive to production and promotes, researcher optimizes process aspect and set about from the improvement of strain.But
It is that the technique generally takes single bacterium kind koji-making to produce fermented soya bean, the microorganism for participating in fermentation is less, and the enzyme class of secretion is not complete,
So that fermented soya bean of the flavor of fermented soya bean finished product not as good as traditional zymotic.Such as:
Application publication number discloses one kind for the Chinese invention patent application document of CN103621913A and rapidly and efficiently ferments
The production technology of health care fermented soya bean, the production technology is the following steps are included: by wheat bran is mixed after soybeans soaking, in 121 DEG C of sterilizings 30
Minute；For clinker cooling to 40 DEG C or so inoculation Triangle-botde kojis, 30 DEG C of cultures to clinker surface row are bent up to fermented soya bean at white；It will
10% salt water is added in fermented soya bean song, and the water content of fermented soya bean unstrained spirits is made to reach 80%~100%, in 50 DEG C after progress hot fermentation seven days
Fermented soya bean unstrained spirits is i.e. mature；By fermented soya bean unstrained spirits be added 90 DEG C or more filter after hot water thermal insulating 2~4 hours thick soy sauce, remaining solid are
Drench oily fermented soya bean；The Bacillus nattoSawamura liquid (2000IU/mL or more) fermented is added in 0.1-1% ratio and is drenched in oily fermented soya bean,
Obtain basic fermented soya bean；The different spices such as rice wine, capsicum, Chinese prickly ash are added in basic fermented soya bean, can must obtain style fermented soya bean.
Compared with traditional handicraft, above-mentioned technique living contaminants are few, fermentation period is short, product salt content is low, contain Nattokinase
Stablize；And one time fermentation, produces soy sauce and fermented soya bean are not only full of nutrition, and of all shapes and colors, is suitble to large scale fermentation production；
But single flavor is not as good as the fermented soya bean of traditional zymotic.
In addition, the fermented soya bean that industrial production is promoted at present are only able to satisfy the conventional mouthfeel requirement of consumer mostly, for health care
The production of type fermented soya bean is promoted less；Wherein, γ-aminobutyric acid (GABA) has drop blood as the healthy ingredient in food now
The effects of pressure, anti-heart rhythm disorders, adjusting hormone secretion；As living standard is continuously improved, people are more next for health food
More pay attention to, it is even more well received especially to some food that can be adapted to " three high " crowd and eat.
Therefore, it is necessary to further improve existing production technology, a kind of fermented soya bean are provided taste is better the abundant, production cycle
It is shorter, it is more suitable for large-scale promotion, and be able to produce the manufacture craft of the fermented soya bean with certain health care functions.
Summary of the invention
The present invention provides a kind of production methods of rich lactobacillus type fermented soya bean for producing γ-aminobutyric acid, which can not only
Enough shorten fermentation period, reduce production cost, additionally it is possible to significantly improve into bent prolease activity and fermentation during the preparation process
The amino-acid nitrogen content of generation, and a large amount of γ-aminobutyric acid healthy ingredients are generated, so as to improve the flavor of fermented soya bean, improve fermented soya bean
Trophism.
The index that prolease activity is completed as koji-making is decomposed high molecular weight protein substance and is provided predominantly in fermentation process
Preparing, amino-acid nitrogen content is the index that fermentation is completed, and fermentation process is a series of biochemical reaction, such as Maillard reaction,
With the flavor of fermented soya bean and nutritionally relevant.
A kind of production method of the rich lactobacillus type fermented soya bean for producing γ-aminobutyric acid, comprising: soya bean is successively cleaned, is impregnated,
It drains, after boiling, inoculation bacterium powder carries out koji-making and fermentation, obtains fermented soya bean；Before koji-making, the bacterium powder of inoculation is aspergillus oryzae and black song
Mould Mixed Microbes powder, the mass ratio of the two are 1.5~4:1；After koji-making, inoculation contains lactobacillus plantarum (Lactobacillus
Plantarum) the bacterium solution of CGMCC NO.3782, then ferment.
Lactobacillus plantarum (Lactobacillus plantarum) the CGMCC NO.3782 is in 2010 by Liu Peicong
It is isolated in conventional Kimchi, it is accredited as lactobacillus plantarum, and be named as (Lactobacillus plantarum) lp15-2-
1, and it is preserved in Chinese microorganism strain administration committee common micro-organisms center (CGMCC), the preservation time is April 27 in 2010
Day, deposit number is CGMCC NO.3782.Separation, purifying and the identification method of the bacterial strain are in Chinese patent literature
It is disclosed in 201010251108 .X.The present invention is not related to the preservation of the strain.
Further, the aspergillus oryzae is that Shanghai makes 3.042, and the aspergillus niger is As 3.350.
For aspergillus oryzae as brewing power microorganism, producing enzyme is abundant, can generate such as protease, peptase, amylase, fiber
Plain enzyme etc., these enzyme systems play different role to fermented soya bean production, and wherein protease is key enzyme.Aspergillus oryzae produces neutral proteinase
Ability it is stronger, the ability for producing acid protease is relatively weak；And the fermentation process of fermented soya bean is the environment in neutral and slant acidity
Middle completion, need acid protease.
The shortcomings that experiment discovery, the addition of aspergillus niger not only will not influence the fermentation of fermented soya bean, can also make up aspergillus oryzae, it is
Entire fermented soya bean song provides acid protease.Wherein, when the proportion of aspergillus oryzae and aspergillus niger is 4:1, the produced protease activity of koji-making
Power highest.
Experiment also found that the addition of lactobacillus plantarum (Lactobacillus plantarum) CGMCC NO.3782 can also
Enough amino-acid nitrogen contents for further promoting fermented soya bean fermentation to generate, so that the content of γ-aminobutyric acid in fermented soya bean be made further to mention
It is high.
Preferably, the inoculum concentration of the bacterium powder is 1~1.5% in terms of the quality of soya bean.
Preferably, the inoculum concentration of the bacterium solution is 1~2mL/kg, institute in bacterium solution in terms of the quality of fermented soya bean song after koji-making
The concentration for stating lactobacillus plantarum is 107cfu/mL。
Preferably, the temperature of the immersion is 25~35 DEG C, the time is 3~5h.
Preferably, the temperature of the boiling is 120~125 DEG C, the time is 15~25min.
The growth temperature of mould is unsuitable too low, is unfavorable for the growth and metabolism of microorganism；Temperature is unsuitable excessively high, is easy to produce
" burning bent " phenomenon, is unfavorable for microorganism growth, and easily inactivate enzyme.
Fungus growth needs a certain amount of moisture, and suitable humidity can make raw material keep higher moisture, and make microorganism
In equilibrium state, be conducive to the growth of mould in this way, increase yield of enzyme；But humidity is excessive, easily makes moisture mistake in incubator
Height may interfere with the mass exchange of mould and external environment, is unfavorable for the growth of microorganism, declines enzyme activity.
Therefore preferably, the condition of the koji-making are as follows: temperature is 30~35 DEG C, and relative humidity is 75~80%, and the time is
36~60h.It is further preferred that the temperature is 30 DEG C, and relative humidity 75%, time 48h.
In fermented soya bean fermentation process, needing to add certain salt, this not only contributes to the flavor quality of final finished, and
Also there are much relations with microorganism growth.The growth with high salt for being able to suppress microorganism and prolease activity, hinder protein
It decomposes, so that amino-acid nitrogen content declines, so control salt content is most important for fermented soya bean quality of finished.
Preferably, adding salt and water, then ferment after koji-making；Wherein, the additive amount of salt is 40~60g/
kg.It is further preferred that the additive amount of salt is 50g/kg.
Fermented soya bean earlier fermentation is the hydrolysis that the protein in soybean occurs under the catalysis of the enzyme of microorganism secretion, institute
Must have water participation.When water content is too low, it is unfavorable for protein and is hydrolyzed into amino nitrogen, higher moisture is kept to be conducive to egg
The decomposition of white matter；But moisture is excessively high, sauce shape or paste is presented in fermented soya bean, can not keep the original solid granulates of fermented soya bean.Cause
This, control water content is conducive to fermented soya bean fermentation in a certain range.
Preferably, the additive amount of the water is 400~600mL/kg.It is further preferred that the additive amount of water is 500mL/kg.
In fermented soya bean fermentation process, macromolecular substances such as protein generate the amino nitrogen of small molecule under the catalysis of enzyme,
The too low catalysis reaction for being unfavorable for enzyme of temperature, it is excessively high easily to inactivate enzyme.
Relative humidity is related to the water content in fermented soya bean, and only under suitable humidity, fermented soya bean and microorganism are just able to maintain
Appropriate moisture, to be conducive to fermented soya bean fermentation；Humidity is too low, and the moisture in fermented soya bean can be evaporated, and is unfavorable for so micro-
The growth and protein of biology are hydrolyzed into amino nitrogen；Humidity is excessively high, and fermented soya bean water content is high, and fermented soya bean surface is by large quantity of moisture packet
It encloses, is unfavorable for the exchange of interior external substance.In addition, fermentation time will affect the quality of fermented soya bean finished product, such as flavor and taste.
Preferably, the condition of the fermentation are as follows: temperature is 35~40 DEG C, and relative humidity is 60~70%, the time 10
~15d.It is further preferred that temperature is 37 DEG C, and relative humidity 65%, time 14d.
Compared with prior art, the invention has the following advantages:
(1) the method for the present invention carries out koji-making using the Mixed Microbes powder of aspergillus oryzae and aspergillus niger, and lactobacillus plantarum is added
(Lactobacillus plantarum) CGMCC NO.3782 bacterium solution is fermented, and cooperates suitably koji-making and fermentation work
Skill significantly improves into bent prolease activity, the amino-acid nitrogen content and alpha-aminobutyric acid content that fermentation generates, obtains product
Matter and taste is better and healthier fermented soya bean finished product.
(2) the method for the present invention preparation process is simple, substantially reduces fermentation period, reduces production cost.
Detailed description of the invention
Fig. 1 is that different aspergillus oryzaes and black-koji mould powder match the influence for comparing holding fermented blank bean koji-making prolease activity in embodiment 1.
Fig. 2 is influence of the different bacterium powder inoculum concentrations to holding fermented blank bean koji-making prolease activity in embodiment 1.
Fig. 3 is influence of the different starter-making temperatures to holding fermented blank bean koji-making prolease activity in embodiment 1.
Fig. 4 is influence of the different koji-making humidity to holding fermented blank bean koji-making prolease activity in embodiment 1.
Fig. 5 is influence of the different koji-making times to holding fermented blank bean koji-making prolease activity in embodiment 1.
Fig. 6 is to compare the case where two kinds of fermented soya bean fermentations produce amino nitrogen in embodiment 2.
Fig. 7 is γ-aminobutyric acid standard items liquid chromatogram in embodiment 2.
Fig. 8 is γ-aminobutyric acid canonical plotting in embodiment 2.
Fig. 9 is γ-aminobutyric acid liquid chromatogram in lactobacillus type fermented soya bean in embodiment 2.
Specific embodiment
Each index determining method involved in the following example is as follows:
1, the measurement of prolease activity: GB/T 23527-2009；
2, the measurement of amino-acid nitrogen content:
(1) preparation of sample: weighing 5g or so sample, is put into 200mL beaker, and 50mL distilled water is added, uses refiner
It smashes uniformly, is transferred in centrifuge tube, is centrifuged 15min under conditions of 3500r/min, supernatant is moved into 100mL volumetric flask,
Again plus 10mL distilled water is in centrifuge tube precipitating, is centrifuged again, and supernatant pours into volumetric flask, this operation 3 times repeatedly is finally used
Distilled water is settled to 100mL, mixes.
(2) measurement of sample: 10mL sample liquid is drawn in 200mL beaker, 60mL distilled water is added, starts magnetic agitation
Device is added 10mL formalin and is mixed with the NaOH solution titratable acidity meter pH to 8.2 of 0.05mol/L, then uses 0.05mol/L
NaOH be titrated to 9.2, write down the NaOH volume V that pH is consumed from 8.2 to 9.21；Blank assay only needs to change sample into 10mL
Distilled water writes down the NaOH volume V of consumption2.。
Amino-acid nitrogen content:
3, the measurement of γ-aminobutyric acid (GABA)
(1) reagent
0.2%KH2PO4:0.2g is dissolved in 100mL ultrapure water, is filtered with 0.45 μm, and filtrate is collected.
The preparation of sample solution: it accurately weighs 0.05-0.25g and crushes sample, 100mL is settled to water, through ultrasonic dissolution
Later, with 0.22 μm of membrane filtration, filtrate is collected as sample solution.
0.4mol/L borate buffer: accurately weighing 2.47g boric acid, about 80mL water is added, extremely with sodium hydroxide tune pH
10.2, it is settled to 100mL with water, is filtered with 0.45 μm, filtrate is collected.
The preparation of derivative reagent: weighing 0.01g o-phthalaldehyde (OPA), is dissolved with 2.5mL acetonitrile, adds 20 μ L β-sulfydryl
Ethyl alcohol, ultrasonic dissolution.
(2) pre-column derivatization
Precision draws 50uL sample solution and 50 μ L derivative reagents, after 400uL borate buffer hybrid reaction about 2min, adds
Enter 500 μ L 0.2%KH2PO4Reaction is terminated, sample introduction, sample volume are 20 μ L immediately after 30 seconds.
(3) chromatographic condition
Mobile phase
A phase: weighing 8.0g crystallization sodium acetate, is settled to 1000mL with water dissolution；Then 220 μ L triethylamines are added, stir
And the acetic acid tune pH to 7.20+0.02 of dropwise addition 5%；It is eventually adding 5mL tetrahydrofuran, is filtered after mixing spare.
B phase: weighing 8.0g crystallization sodium acetate, is settled to 1000mL with water dissolution；Then be added dropwise 2% acetic acid tune pH extremely
7.20±0.02；Sodium acetate solution: acetonitrile: spare after methanol=1:2:2 (volume ratio) hybrid filtering is pressed again.
Gradient elution table
Column temperature: 40 DEG C；Flow velocity: 1.0mL/min；Detection wavelength: 338nm
(4) standard curve
0.1g γ-aminobutyric acid is accurately weighed, 100mL, as 1.0mg/mL are settled to after being dissolved with water, with 0.45 μm of mistake
Filter is collected filtrate and is diluted, accurate respectively to draw 0.1mg/mL, 0.2mg/mL, 0.4mg/mL, 0.6mg/mL, 0.9mg/mL
Concentration standard liquid carries out chromatography according to the above method, with peak area-concentration of standard solution mapping, draws standard curve and recurrence
Equation, linearly dependent coefficient R are 0.99 or more.
(5) Specimen Determination
10g fermented soya bean are weighed, are put into 200mL beaker, 50mL distilled water is added, is smashed uniformly with refiner, is transferred to centrifugation
Guan Zhong is centrifuged 15min under conditions of 3500r/min, by supernatant move into 100mL volumetric flask in, then plus 10mL distilled water in
It in centrifuge tube precipitating, is centrifuged again, supernatant pours into volumetric flask, this operation 3 times repeatedly is finally settled to distilled water
100mL is mixed；5mL sample solution is drawn, 5mL 0.1mol/L trichloroacetic acid, vortex 2min, then in 40 DEG C of water-baths is added
1h is extracted, 1mL is taken；It is centrifugated (12000r/min, 5min), takes 50 μ L of supernatant, draw 400 μ L of borate buffer, OPA spreads out
500 μ L 0.2%KH are added after raw 50 μ L of agent, hybrid reaction 1min2PO4Reaction is terminated, with 0.22 μm of membrane filtration, is stood after 30 seconds
That is sample introduction, sample volume are 20 μ L, record the retention time and peak area of chromatographic peak, and the derivatization treatment of sample and standard solution is extremely
Sample injection time should be consistent.According to the peak area of chromatographic peak, the concentration of corresponding γ-aminobutyric acid is calculated with external standard method.
4, the aspergillus oryzae of the following example is that Shanghai makes 3.042, and the aspergillus niger is As 3.350, and lactobacillus plantarum is
(Lactobacillus plantarum)CGMCC NO.3782。
Embodiment 1
Using prolease activity as index, to bacterium powder proportion, inoculum concentration, starter-making temperature, relative humidity and koji-making time etc. because
Element is assessed.
1, bacterium powder matches: selecting full raw soybeans 100g, washes with clean water, 300mL, 30 DEG C of water is then added
Impregnate 4h, after draining, access 1g bacterium powder (inoculum concentration 1.0%, w/w), wherein aspergillus oryzae and bacterium powder proportion be set as 5:0,4:1,
3:2,2:3 after koji-making 60h, measure prolease activity under conditions of temperature is 30 DEG C, relative humidity 75%.
As shown in Figure 1, when the proportion of aspergillus oryzae and black-koji mould powder is 4:1, the produced prolease activity highest of koji-making.
2, inoculum concentration: selecting full raw soybeans 100g, wash with clean water, and 300mL, 30 DEG C of water logging is then added
Steep 4h, after draining, access bacterium powder, wherein aspergillus oryzae and black-koji mould powder proportion be 4:1, inoculum concentration be arranged (w/w) be 2.0%,
1.5%, 1.0%, 0.5%, under conditions of temperature is 30 DEG C, relative humidity 75%, after koji-making 60h, measure prolease activity.
As shown in Figure 2, with the increase of inoculum concentration, prolease activity is gradually increased, and is reached most in inoculum concentration for 1.5%
Greatly, but when inoculum concentration further increases, prolease activity but declines instead, it may be possible to which, because inoculum concentration is too big, microorganism is raw
Long excessive, the temperature of generation is higher than its optimum growth temperature, and is limited by the nutriment that can be utilized.So
The inoculum concentration for determining 1.5% is optimum inoculation amount.
3, starter-making temperature: selecting full raw soybeans 100g, wash with clean water, and 300mL, 30 DEG C of water is then added
4h is impregnated, after draining, is accessed bacterium powder (inoculum concentration 1.5%, w/w), wherein aspergillus oryzae and black-koji mould powder proportion are 4:1, In
Temperature setting is 25 DEG C, 30 DEG C, 35 DEG C, 37 DEG C, 40 DEG C, under conditions of relative humidity 75%, after koji-making 60h, measures protease
Vigor.
From the figure 3, it may be seen that fermented soya bean song prolease activity gradually rises as fermentation temperature gradually rises, the enzyme activity at 35 DEG C
Reach maximum, but as temperature further increases, enzyme activity sharply declines.
4, koji-making relative humidity: selecting full raw soybeans 100g, wash with clean water, and is then added 300mL, and 30 DEG C
Water impregnate 4h, after draining, access bacterium powder (inoculum concentration 1.5%, w/w), wherein aspergillus oryzae and black-koji mould powder proportion be 4:
1, under conditions of temperature is 30 DEG C, relative humidity is set as 65%, 70%, 75%, 80% and 85%, after koji-making 60h, measurement
Prolease activity.
From fig. 4, it can be seen that prolease activity declines after first increasing, in relative humidity with the increase of relative humidity again
Reach maximum for 75% enzyme activity.
5, the koji-making time: selecting full raw soybeans 100g, wash with clean water, and 300mL, 30 DEG C of water is then added
4h is impregnated, after draining, is accessed bacterium powder (inoculum concentration 1.5%, w/w), wherein aspergillus oryzae and black-koji mould powder proportion are 4:1, In
Temperature be 30 DEG C, relative humidity be set as under conditions of 75%, respectively koji-making 12h, for 24 hours, after 36h, 48h, 60h and 72h, sampling
Measure prolease activity.
As shown in Figure 5, with the extension of koji-making time, downward trend after first increase is presented in prolease activity, in 48h
Reach maximum.Before 48h, microorganism is growing, so that enzyme activity also gradually increases；In 48h, fungus growth obtains vigorous, production
Enzyme ability also reaches maximum, and enzyme activity also correspondinglys increase, and after 48h, may be limited by available nutriment and
The reason of strain enzymatic productivity decline, prolease activity are gradually reduced.
Embodiment 2
By the research of embodiment 1, optimal holding fermented blank bean koji-making technique is obtained.
Specific step is as follows:
(1) the soya bean particle that 1kg is full is taken, is cleaned with clear water until completely, after 30 DEG C of warm water immersion 4h of 3L is added, dripping
Solid carbon dioxide point, is put into boiling 20min in 121 DEG C of autoclaving pots, is cooled to room temperature, obtain cooked soya bean particle；
(2) 15g Mixed Microbes powder is accessed into cooked soya bean particle, wherein the mass ratio of aspergillus oryzae and black-koji mould powder is
4:1 is uniformly mixed, and is put into koji-making 48h in incubator, and temperature is 30 DEG C, relative humidity 75%；
(3) after koji-making is completed, salt and sterile water are added, in mass, the fermented soya bean song inoculation 1mL of every 1kg contains
107The bacterium solution of cfu/mL lactobacillus plantarum (Lactobacillus plantarum) CGMCC NO.3782, and 50g salt is added
With the sterile water of 500mL, temperature is transferred to 37 DEG C, relative humidity is adjusted to 65%, ferments 12 days, obtains finished product fermented soya bean.
After the completion of koji-making, prolease activity highest can achieve 1158u/g；After fermentation, amino-acid nitrogen content reaches
2.33g/100g, alpha-aminobutyric acid content reach 38mg/kg；Finished product fermented soya bean darkly brown particles shape, there is dense beany flavour,
Soft taste has certain saline taste.
Comparative example 1
A kind of production method of fermented soya bean, the specific steps are as follows:
(1) the soya bean particle that 1kg is full is taken, is cleaned with clear water until completely, after 30 DEG C of warm water immersion 4h of 3L is added, dripping
Solid carbon dioxide point, is put into boiling 20min in 121 DEG C of autoclaving pots, is cooled to room temperature, obtain cooked soya bean particle；
(2) 15g Mixed Microbes powder is accessed into cooked soya bean particle, wherein the mass ratio of aspergillus oryzae and black-koji mould powder is
4:1 is uniformly mixed, and is put into koji-making 48h in incubator, and temperature is 30 DEG C, relative humidity 75%；
(3) after koji-making is completed, salt and sterile water are added, in mass, the fermented soya bean song of every 1kg be added 50g salt and
Temperature is transferred to 37 DEG C by the sterile water of 500mL, and relative humidity is adjusted to 65%, ferments 12 days, obtains finished product fermented soya bean.
The fermented soya bean that embodiment 2 and comparative example 1 are obtained carry out the comparison of amino-acid nitrogen content, as a result as shown in Figure 6.
Claims (1)
1. a kind of production method of the rich lactobacillus type fermented soya bean for producing γ-aminobutyric acid, comprising: successively clean soya bean, impregnate, drip
After dry, boiling, inoculation bacterium powder carries out koji-making and fermentation, obtains fermented soya bean；It is characterized in that, the bacterium powder of inoculation is rice before koji-making
The Mixed Microbes powder of aspergillus and aspergillus niger, the mass ratio of the two are 1.5~4:1；After koji-making, inoculation contains lactobacillus plantarum
The bacterium solution of (Lactobacillus plantarum) CGMCC NO.3782, then ferment；
The aspergillus oryzae is that Shanghai makes 3.042, and the aspergillus niger is As 3.350；
The temperature of the immersion is 25~35 DEG C, and the time is 3~5h；The temperature of the boiling is 120~125 DEG C, the time 15
~25min；
In terms of the quality of soya bean, the inoculum concentration of the bacterium powder is 1~1.5%；In terms of the quality of fermented soya bean song after koji-making, the bacterium solution
Inoculum concentration be 1~2mL/kg, the concentration of lactobacillus plantarum described in bacterium solution is 107cfu/mL；
The condition of the koji-making are as follows: temperature is 30~35 DEG C, and relative humidity is 75~80%, and the time is 36~60h；
After koji-making, salt and water are also added, then ferment；Wherein, the additive amount of salt is 40~60g/kg, the additive amount of water
For 400~600mL/kg；The condition of the fermentation are as follows: temperature be 37~45 DEG C, relative humidity be 60~70%, the time be 10~
15d。
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