CN100337557C - Method for producing soy bean peptide liquid food and biosis bacteria tablet using soy bean fermentation - Google Patents
Method for producing soy bean peptide liquid food and biosis bacteria tablet using soy bean fermentation Download PDFInfo
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- CN100337557C CN100337557C CNB2004100197586A CN200410019758A CN100337557C CN 100337557 C CN100337557 C CN 100337557C CN B2004100197586 A CNB2004100197586 A CN B2004100197586A CN 200410019758 A CN200410019758 A CN 200410019758A CN 100337557 C CN100337557 C CN 100337557C
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Abstract
The present invention relates to a method for producing soybean peptide liquid food and biosis bacterium tablets by soybean fermentation, which belongs to edible natto bacillus fermented soybean products. In the present invention, the method for producing soybean peptide liquid food and biosis bacterium tablets solves the problems that the security of bacteria is poor, the fermentation capacity of biosis bacteria at present is low, the cost of culture media is high, etc. The natto bacilli capable of producing natto kinase are used as bacteria. Soybean flour or soybean cakes or soybean paste or soybean milk is used as a culture medium. After being fermented and separated, the materials are made into liquid food, namely the mixture of natto kinase-contained oybean peptides, oligopeptides and short peptides, and beneficial bacterium tablets which contain rich natto bacilli and soybean fibrin. The present invention provides the healthcare food which has rich nutrition and peculiar flavors. The present invention can be widely used for a plurality of the technical fields of medicine, food, feed supplement, etc., has the advantages of simple preparation processes, short period, low cost and is suitable for commercial production. In addition, the present invention develops an efficient path for the industrial development of protein peptides and has great economical and social benefits.
Description
Technical field
The invention belongs to the method for utilizing bean product fermenting and producing Soybean Peptide liquid food and biosis bacteria tablet, being particularly related to a kind of employing bacillus natto is bacterial classification, with soybean bean powder, soybean soya-bean cake, soybean meal or soybean milk is culture medium, the preparation method of fermenting and producing Soybean Peptide liquid food and biosis bacteria tablet.
Technical background:
Soybean Peptide liquid food and Bacillus Natto Beneficial Fungus Tablet are emerging high-quality health foods.The protein content of Soybean Peptide is about 85%, and its amino acid is formed almost completely identical with soybean protein.But compare with soybean protein, Soybean Peptide have better physicochemical property as easily digest and assimilate, low antigenicity, enhancing human immunological competence etc., and contain some physiological activator different physiological roles arranged in body.Active peptide of soybean protein equals that the part human body is finished difficult digestion process and takes external carrying out, and is emerging nourishing healthy source, and the research to Soybean Peptide starts from the seventies in the world, and wherein the U.S. and Japan all are in the prostatitis of development Soybean Peptide production and utilization.China studied progressively in recent years and enlivened, and Soybean Peptide production obtains fast development.At present, the method for Soybean Peptide enzymolysis commonly used prepares.But the enzymolysis selectivity is stronger, and percent hydrolysis is on the low side, and there is bitter taste in hydrolysate, is difficult for being accepted by the people, and does not have fibrinolytic.
Probiotics tablets is to use beneficial bacterium, and its cultivation propagation, concentrate drying are made microbial inoculum.Qualified probio should have following two characteristics, promptly contains microorganism alive and can improve host health by playing a role in oral cavity, the intestines and stomach.Basic demand is a high security to bacterial classification, and nontoxic, nothing disables, and do not have to cause a disease, and has no drug resistance, side effect such as no drug residue; Must be the beneficial bacterium of living, in cultivation and in the organism, easily breed that processing still has high viability after handling; Acid, alkali, bile there are tolerance, can avoid the influence of hydrochloric acid in gastric juice, bile in mould inhibitor, antioxidant and feed process and the animal intestinal; The field planting ability is good in enteron aisle, fast growth; Can produce lactic acid or other antibacterial substance.Biosis bacteria tablet often exists and produces pathology and the toxicological test that bacterial classification does not pass through strictness at present, and long-term the use has toxic and side effects such as drug resistance, " three cause "; Some beneficial bacterium is difficult for propagation in cultivation and in the organism, it is low that the back survival rate is handled in processing, and quality can not get guaranteeing.
At present, the production method of Soybean Peptide mainly contains: three kinds of microbe fermentation methods, enzymatic isolation method, chemical method, wherein advanced with microbe fermentation method, it can be when producing peptide the bitter taste factor of modified peptides, simultaneously, microorganism is effectively eliminated the composition of these two kinds influence digestion and taste with the ANFs in the raw material (as KTI and BBI) degraded.But also have some problems, it is few and enzyme activity is low to produce the enzyme class as the bacterial classification that adopted, and fermentability is poor, low productivity can only adopt meticulousr raw material to ferment, and tunning does not obtain comprehensive utilization, only utilize a part in the zymotic fluid, cause the product price height.In sum, the low and production cost height of fermentability is to influence fermentation method production Soybean Peptide and apply key.For this reason, researching and developing a kind of economical and practical production method is the task of top priority.
Summary of the invention
At the problems referred to above, the invention solves bacterial classification safe differential, the low high problem of raw materials used cost cost that reaches of fermentability, provide and utilized bacillus natto to be bacterial classification, with the bean product is culture medium production Soybean Peptide liquid food and Bacillus Natto Beneficial Fungus Tablet, and making that resulting biosis bacteria tablet security is good, viable count is high, peptide liquid food has fibrinolytic.
Technical scheme:
The method of fermenting and producing Soybean Peptide liquid food and biosis bacteria tablet, it is characterized in that adopting bacillus natto (Bacillus natto, Chinese industrial microorganism fungus kind preservation center AS 1.107) for producing bacterial strain, fermentation medium is a raw material with soybean bean powder, soybean meal, soybean-cake flour or soybean milk mainly, through seed culture, and fermented and cultured, centrifugal or filter, drying process obtains having Soybean Peptide liquid food and solid Bacillus Natto Beneficial Fungus Tablet, and concrete steps are as follows:
(1) preparation zymotic fluid
The preparation of bacterial classification
After 18-24 hour, preserve stand-by by 4 ℃ of refrigerators 35-37 on the agar slant ℃ of cultivation for bacillus natto.110-125 ℃ of slant medium sterilising conditions, 15-20 minute;
Wherein slant medium is: beef extract 5.0-10.0g
Peptone 10.0-20.0g
Sodium chloride 5.0-10.0g
Agar 18.0-20.0g
Water 1000ml
pH 7.0-7.2
Seed culture
To connect a ring at the cultured bacterial classification of slant medium and join in the seed culture medium 110-125 ℃ of seed culture medium sterilising conditions, 15-20 minute; Under 200rpm rotating speed on the shaking table, 35-37 ℃ after shaken cultivation 20-28 hour, can obtain being fit to the inoculum of inoculation.
Wherein seed culture medium is: peptone 10.0-15.0g
Glucose 20.0-30.0g
K
2HPO
4 1.0-1.5g
MgSO
4 0.5-0.75g
Water 1000ml
pH 7.0-7.2
Fermented and cultured
(volume ratio, inoculum concentration ml/ml) are inoculated in the fresh fermentation medium through sterilization treatment, cultivate for 30-40 ℃ and carry out deep fermentation in 60-72 hour with 2%-5% with above-mentioned inoculum.During the fermentation, should control the pH value at 6.0-8.0; The adjusting of pH between yeast phase, the concrete condition of looking zymotic fluid pH, and, add acid or aqueous slkali realization by stream according to the fermentation needs.Dissolved oxygen is regulated by the control ventilation, and dissolved oxygen should be controlled at 20-40%.
Check the gemma production rate during fermentation ends, require the gemma production rate to reach more than 70%.
Need to prove:
Adjusting pH value method is that the adjusting of initial pH can be that the 1.0-2.5% phosphate buffer is realized by adding concentration.The adjusting of pH between yeast phase, used acid solution such as phosphoric acid solution, used alkali lye such as ammoniacal liquor or urea.When can transferring to pH 6.0-6.3 or 7.7-8.0 to 48 hours, fermentation stimulate gemma to produce.
After fermentation was to 48 hours, can reduce dissolved oxygen, the control dissolved oxygen at 20-30% so that stimulate the generation of gemma.
After fermentation 48 hours, can reduce the thermal stimulus gemma and produce, temperature is controlled at 25-32 ℃, not only can promote gemma to produce, also help the stability that keeps Nattokinase, it is not degraded.
When fermentation medium adopted soybean-cake flour to be raw material, its prescription was: soybean-cake flour 20-80g, glucose 20-40g, MgSO
40.5-1.0g, Na
2HPO
46.0-12.0g, NaH
2PO
41.0-2.0g, CaCl
20.2-0.4g, 1000ml water pH7.0-7.2.
When fermentation medium adopted the soybean bean powder to be raw material, its prescription was: soybean bean powder 20-80g, glucose 20-40g, MgSO
40.5-1.0g, Na
2HPO
46.0-12.0g, NaH
2PO
41.0-2.0g, CaCl
20.2-0.4g, 1000ml water, pH7.0-7.2.
When fermentation medium adopted soybean meal to be raw material, its prescription was: soybean meal 20-80g, glucose 20-40g, MgSO
40.5-1.0g, Na
2HPO
46.0-12.0g, NaH
2PO
41.0-2.0g, CaCl
20.2-0.4g, 1000ml water, pH7.0-7.2.
When fermentation medium adopted soybean milk to be raw material, its prescription was: soybean milk 125-250ml, glucose 20-40g, MgSO
40.5-1.0g, Na
2HPO
46.0-12.0g, NaH
2PO
41.0-2.0g, CaCl
20.2-0.4g, water supplies 1000ml, pH7.0-7.2.
Above-mentioned soybean-cake flour, soybean bean powder, soybean meal must be levigate, and it is stand-by to cross the 80-120 mesh sieve, and fermentation medium is taked 115-125 ℃, sterilization in 15-25 minute.
(2) preparation Soybean Peptide liquid food and biosis bacteria tablet
Separation of Solid and Liquid
After the fermentation ends, after sterility test is determined pollution-free assorted bacterium, adopted in refrigerated centrifugation, membrane filter method or the centrifugal filtration process one or both to unite to use and carry out Separation of Solid and Liquid.When adopting membrane filter method to separate solid-liquid, reduce as far as possible that suspension sees through film in the zymotic fluid.When adopting the gravity centrifugal treating, detect the centrifuged supernatant viable count, select the rotating speed should be, make that suspension is centrifugal in the zymotic fluid gets off, when centrifugal filtration process is handled, can centrifugally combine, in centrifugal process, finish filtration simultaneously with filtration greater than 5000r/min.
Preparation Soybean Peptide liquid food
Clarified supernatant is measured Nattokinase enzyme activity in the liquid with the fibrin plate method, requires Nattokinase content to reach 100-500U/ml, and soybean peptide ammino acid content is 20-120g/L.After the assay was approved, add a small amount of food additives and be deployed into Soybean Peptide liquid food;
Annotate: the Nattokinase enzyme activity unit is defined as: decomposition 1nmol is dissolved in the concentration 5 * 10 in the 0.1mol/L phosphate buffer solution (pH7.4) in 37 ℃ of 1min
-4The Nattokinase amount of the H-D-Val-Leu-Lys-Pna of mol/L is 1U.Perhaps the active unit with urokinase or fibrinolysin represents.
The preparation biosis bacteria tablet
Separation is obtained bacterium mud carry out count plate, adopt vacuum freezedrying, fluidized bed drying, spray-drying or boulton process to carry out drying, carry out count plate after the drying once more, require the viable bacteria loss to be not more than 80-85%.Add excipient such as starch, lightweight sodium carbonate etc., get final product after fully stirring Bacillus Natto Beneficial Fungus Tablet.
Beneficial effect:
The present invention utilizes microbial fermentation technology, rely on the bacillus natto breeding fast, advantages such as gemma is produced in easily cultivation, and metabolic capability is strong, make culture medium with cheap raw material, by the control of fermenting of specifically fermentation technology, zymotic fluid is after Separation of Solid and Liquid, and centrifugate is made Soybean Peptide liquid food, bacterium mud is made the bacillus natto probiotics tablets, and fermented product obtains comprehensive utilization.Obtain that the biosis bacteria tablet security is good, viable count is high, and peptide liquid food tool fibrinolytic.
The bacterial classification that this production method adopted is a bacillus natto, is mutation of bacillus subtilis, so bafillus natto has gemma, thereby can be acidproof, alkaline-resisting, high temperature resistant and anti-extruding all can keep stability highly in pelletization and acid gastric environment; It has pathogenic bacteria effects such as the salmonella of inhibition, typhoid fever bacterium, dysentery bacillus and O157:H7 Escherichia coli; And because bacillus natto is long-term as producing the natto bacterial classification, it has tight security.Because beans can induce bacillus natto to produce Nattokinase, protease, lipase, amylase, phytases etc. are enzyme more than 100 kinds, so bacillus natto utilizes soya-bean cake, dregs of beans, bean powder, cheap raw materials such as soya-bean milk have higher fermentability when fermenting, not only solve the high problem of the existing cost of fermentation method production Soybean Peptide, and be also to be rich in a large amount of Nattokinases in the Soybean Peptide liquid food, to cause Soybean Peptide liquid food to have anti-cholesterol, antithrombotic forms, has the strong external and interior fibrinolytic double effects of body simultaneously, make the invention provides a kind of nutritious, the easy health food that absorbs, can be widely used in medicine, food, numerous areas such as feed addictive have tempting economic outlook; And this preparation method's technology is simple, and the cycle is short, and cost is low, is fit to suitability for industrialized production.Thereby, have than large economy benefit and social benefit for the protein peptides industrial development provides effective way.
The specific embodiment
Embodiment 1:
(1) preparation zymotic fluid
The preparation of bacterial classification
The production bacterial classification is bacillus natto (Bacillus natto, AS 1.107).With bacillus natto on the agar slant 37 ℃ cultivate 18 hours after, microscopy, thalli growth is good, 4 ℃ of refrigerators are preserved stand-by.110 ℃ of slant medium sterilising conditions, 20 minutes.
Wherein slant medium is: beef extract 5.0g
Peptone 10.0g
Sodium chloride 5.0g
Agar 18.0g
Water 1000ml
pH 7.0-7.2
Seed culture
To connect a ring at the cultured bacterial classification of slant medium and join in the seed culture medium 110 ℃ of seed culture medium sterilising conditions, 20 minutes; Under 200rpm rotating speed on the shaking table, 37 ℃ of shaken cultivation obtained being fit to the inoculum of inoculation after 20 hours.
Wherein seed culture medium is: peptone 10.0g
Glucose 20.0g
K
2HPO
4 1.0g
MgSO
4 0.5g
Water 1000ml
pH 7.0-7.2
Fermented and cultured
Inoculum is inoculated in fresh fermentation medium with 2% inoculum concentration, cultivates for 40 ℃ and carried out deep fermentation in 60 hours.During the fermentation, should control the pH value at 6.0-8.0.Regulating pH value way is: the adjusting of initial pH can be that 1.0% phosphate buffer is realized by adding concentration; The adjusting of pH between yeast phase, the concrete condition of looking zymotic fluid pH, and, add acid or aqueous slkali realization by stream according to the fermentation needs.Used acid is phosphoric acid solution, and used alkali is ammoniacal liquor.Dissolved oxygen is regulated by the control ventilation, and dissolved oxygen should be controlled at 20-40%, and the gemma production rate reaches 70%.
Wherein fermentation medium is: soybean-cake flour 20g
Glucose 20g
MgSO
4 0.5g
Na
2HPO
4 6.0g
NaH
2PO
4 1.0g
CaCl
2 0.2g
Water 1000ml
pH 7.0-7.2
Soybean-cake flour must be levigate, and it is stand-by to cross 100 mesh sieves.Fermentation medium is taked 115 ℃, sterilization in 25 minutes.
(2) Soybean Peptide liquid food and biosis bacteria tablet
Separation of Solid and Liquid
After the fermentation ends, after sterility test is determined pollution-free assorted bacterium, adopted refrigerated centrifugation to carry out Separation of Solid and Liquid, centrifugal 15 minutes of 4 ℃ of following 5000r/min.
Preparation Soybean Peptide liquid food
Clarified supernatant is measured Nattokinase enzyme activity in the liquid with the fibrin plate method.Measure soybean peptide ammino acid content in the solution with amino acid determining instrument.Nattokinase content is 250U/ml, and soybean peptide ammino acid content is 6g/L.After the assay was approved, add a small amount of albumen sugar and citric acid and be deployed into Soybean Peptide liquid food.
The preparation biosis bacteria tablet
Separation is obtained bacterium mud carry out count plate, carry out count plate once more after the employing vacuum freezedrying, the viable bacteria loss is not more than 80%.It adds excipient starch, get final product after fully stirring Bacillus Natto Beneficial Fungus Tablet.This biosis bacteria tablet viable count reaches 1.1 hundred million/g.
Embodiment 2:
During the preparation of bacterial classification, with bacillus natto on agar slant 36 ℃ cultivated 20 hours, the slant medium sterilising conditions is 125 ℃, 15 minutes.
Wherein slant medium is: beef extract 10.0g
Peptone 20.0g
Sodium chloride 10.0g
Agar 20.0g
Water 1000ml
pH 7.0-7.2
During seed culture, 125 ℃ of seed culture medium sterilising conditions, 15 minutes; Cultivated 24 hours for 36 ℃.
Wherein seed culture medium is: peptone 15.0g
Glucose 30.0g
K
2HPO
41.5g
MgSO
4 0.75g
Water 1000ml
pH 7.0-7.2
Inoculum is inoculated in through 120 ℃ with 4% inoculum concentration, and the fresh fermentation medium of 20 minutes sterilization treatment is cultivated for 35 ℃ and was carried out deep fermentation in 66 hours.The adjusting of initial pH can be that 1.8% phosphate buffer is realized by adding concentration.Fermentation to 48 hour is elevated to 7.7-8.0 with pH.
Fermentation medium wherein: soybean-cake flour 50g
Glucose 30g
MgSO
4 0.7g
Na
2HPO
4 9.0g
NaH
2PO
4 1.5g
CaCl
2 0.3g
Water 1000ml
pH 7.0-7.2
Fermentation medium is taked 120 ℃, sterilization in 20 minutes.Excipient adopts the lightweight sodium carbonate.Other are with embodiment 1.
The Nattokinase enzyme activity is 200U/ml in the mensuration liquid, and soybean peptide ammino acid content is 7g/L weight, and the Bacillus Natto Beneficial Fungus Tablet viable count reaches 1.5 hundred million/g.
Embodiment 3:
During the preparation of bacterial classification, with bacillus natto on agar slant 35 ℃ cultivated 24 hours.The slant medium sterilising conditions is 115 ℃, 18 minutes.
Slant medium wherein: beef extract 7.5g
Peptone 15g
Sodium chloride 7.5g
Agar 19.0g
Water 1000ml
pH 7.0-7.2
During seed culture, 125 ℃ of seed culture medium sterilising conditions, 15 minutes; Cultivated 20 hours for 35 ℃.
Seed culture medium wherein: peptone 12.5g
Glucose 25.0g
K
2HPO
4 1.3g
MgSO
4 0.6g
Water 1000ml
pH 7.0-7.2
Inoculum is inoculated in fresh fermentation medium through sterilization treatment with 5% inoculum concentration, cultivates for 30 ℃ and carried out deep fermentation in 72 hours.The adjusting of initial pH can be that 2.5% phosphate buffer is realized by adding concentration.Ferment and can reduce dissolved oxygen in 48 hours, the control dissolved oxygen at 20-30% so that stimulate the generation of gemma.
Fermentation medium wherein: soybean-cake flour 80g
Glucose 40g
MgSO
4 1.0g
Na
2HPO
4 12.0g
NaH
2PO
4 2.0g
CaCl
2 0.4g
Water 1000ml
pH 7.0-7.2
Fermentation medium is taked 125 ℃, sterilization in 15 minutes.After the fermentation ends, after sterility test is determined pollution-free assorted bacterium, adopted membrane filter method to carry out Separation of Solid and Liquid, other are with embodiment 1.
The Nattokinase enzyme activity is 300U/ml in the mensuration liquid, and soybean peptide ammino acid content is 10g/L weight, and the biosis bacteria tablet viable count reaches 1.2 hundred million/g.
Embodiment 4:
Fermentation medium: soybean meal 20g
Glucose 20g
MgSO
4 0.5g
Na
2HPO
4 6.0g
NaH
2PO
4 1.0g
CaCl
2 0.2g
Water 1000ml
pH 7.0-7.2
Soybean meal must be levigate, and it is stand-by to cross 100 mesh sieves.
After the fermentation to 48 hour, pH is transferred to 7.7-8.0; Reduce dissolved oxygen, the control dissolved oxygen so that stimulate the generation of gemma, reduces temperature to 25 ℃ at 20-30%.Other are with embodiment 1.
The Nattokinase enzyme activity is 430U/ml in the mensuration liquid, and soybean peptide ammino acid content is 7g/L weight, and the biosis bacteria tablet viable count reaches 1.3 hundred million/g.
Embodiment 5:
Fermentation medium: soybean meal 50g
Glucose 30g
MgSO
4 0.7g
Na
2HPO
4?9.0g
NaH
2PO
4?1.5g
CaCl
2 0.3g
Water 1000ml
pH 7.0-7.2
Soybean meal must be levigate, and it is stand-by to cross 100 mesh sieves.
Fermentation to 48 hour can transfer to pH 6.0-6.3 stimulates gemma to produce.Other are with embodiment 1.
The Nattokinase enzyme activity is 360U/ml in the mensuration liquid, and soybean peptide ammino acid content is 9g/L weight, and the biosis bacteria tablet viable count reaches 1.1 hundred million/g.
Embodiment 6:
Fermentation medium: soybean meal 80g
Glucose 40g
MgSO
4 1.0g
Na
2HPO
4 12.0g
NaH
2PO
4 2.0g
CaCl
2 0.4g
Water 1000ml
pH 7.0-7.2
Soybean meal must be levigate, and it is stand-by to cross 100 mesh sieves.
Fermentation to 48 hour can reduce dissolved oxygen, and the control dissolved oxygen is reduced to 6.0-6.3 with pH simultaneously at 20-30%, so that stimulate the generation of gemma.Other are with embodiment 1.
The Nattokinase enzyme activity is 350U/ml in the mensuration liquid, and soybean peptide ammino acid content is 6.5g/L weight, and the biosis bacteria tablet viable count reaches 1.3 hundred million/g.
Embodiment 7:
Fermentation medium: soybean bean powder 20g
Glucose 20g
MgSO
4 0.5g
Na
2HPO
4?6.0g
NaH
2PO
4?1.0g
CaCl
2 0.2g
Water 1000ml
pH 7.0-7.2
The soybean bean powder must be levigate, and it is stand-by to cross 100 mesh sieves.
Ferment and to reduce the generation of thermal stimulus gemma after 48 hours, temperature is controlled at 28 ℃.Other are with embodiment 1.
The Nattokinase enzyme activity is 450U/ml in the mensuration liquid, and soybean peptide ammino acid content is 7.8g/L weight, and the biosis bacteria tablet viable count reaches 1.1 hundred million/g.
Embodiment 8:
Fermentation medium: soybean bean powder 50g
Glucose 30g
MgSO
4 0.7g
Na
2HPO
4 9.0g
NaH
2PO
4 1.5g
CaCl
2 0.3g
Water 1000ml
pH 7.0-7.2
The soybean bean powder must be levigate, and it is stand-by to cross 100 mesh sieves.
Fermentation to 48 hour can transfer to 6.0-6.3 with pH, and the control dissolved oxygen is controlled at 28 ℃ with temperature about 20-30%, and other are with embodiment 1.
The Nattokinase enzyme activity is 500U/ml in the mensuration liquid, and soybean peptide ammino acid content is 12g/L weight, and the biosis bacteria tablet viable count reaches 1.5 hundred million/g.
Embodiment 9:
Fermentation medium: soybean bean powder 80g
Glucose 40g
MgSO
4 1.0g
Na
2HPO
4 12.0g
NaH
2PO
4 2.0g
CaCl
2 0.4g
Water 1000ml
pH 7.0-7.2
The soybean bean powder must be levigate, and it is stand-by to cross 100 mesh sieves.
PH can be transferred to 7.7-8.0 after the fermentation to 48 hour; Reduce dissolved oxygen, the control dissolved oxygen so that stimulate the generation of gemma, reduces temperature to 32 ℃ at 20-30%.Other are with embodiment 1.
The Nattokinase enzyme activity is 490U/ml in the mensuration liquid, and soybean peptide ammino acid content is 8g/L weight, and the biosis bacteria tablet viable count reaches 1.3 hundred million/g.
Embodiment 10:
Fermentation medium: soybean milk 125ml
Glucose 20g
MgSO
4 0.5g
Na
2HPO
4 6.0g
NaH
2PO
4 1.0g
CaCl
2 0.2g
Water is supplied 1000ml
pH 7.0-7.2
Reduce dissolved oxygen after the fermentation to 48 hour, the control dissolved oxygen so that stimulate the generation of gemma, reduces temperature to 25 ℃ at 20-30%.Other are with embodiment 1.
The Nattokinase enzyme activity is 250U/ml in the mensuration liquid, and soybean peptide ammino acid content is 7g/L weight, and the biosis bacteria tablet viable count reaches 1.2 hundred million/g.
Embodiment 11:
Fermentation medium: soybean milk 180ml
Glucose 30g
MgSO
4 0.7g
Na
2HPO
4 9.0g
NaH
2PO
4 1.5g
CaCl
2 0.3g
Water is supplied 1000ml
pH 7.0-7.2
PH can be transferred to 7.7-8.0 after the fermentation to 48 hour, reduce temperature to 28 ℃.Other are with embodiment 1.
The Nattokinase enzyme activity is 500U/ml in the mensuration liquid, and soybean peptide ammino acid content is 5g/L weight, and the biosis bacteria tablet viable count reaches 1.4 hundred million/g.
Embodiment 12:
Fermentation medium: soybean milk 250ml
Glucose 40g
MgSO
4 1.0g
Na
2HPO
4 12.0g
NaH
2PO
4 2.0g
CacL
2 0.4g
Water is supplied 1000ml
pH 7.0-7.2
PH can be transferred to 6.0-6.3 after the fermentation to 48 hour, reduce temperature to 32 ℃.Other are with embodiment 1.
The Nattokinase enzyme activity is 480U/ml in the mensuration liquid, and soybean peptide ammino acid content is 10g/L weight, and the biosis bacteria tablet viable count reaches 1.4 hundred million/g.
Claims (5)
1. utilize the method for bean product fermenting and producing Soybean Peptide liquid food and biosis bacteria tablet, it is characterized in that adopting bacillus natto (Bacillus natto) AS 1.107 for producing bacterial strain, fermentation medium is a raw material with soybean bean powder, soybean meal, soybean-cake flour or soybean milk mainly, through seed culture, fermented and cultured, centrifugal or filtration, drying process, obtain having Soybean Peptide liquid food and solid biosis bacteria tablet, concrete steps are as follows:
(1) preparation zymotic fluid
The preparation of bacterial classification
With bacillus natto 35-37 on the agar slant ℃ cultivate 18-24 hour after, it is stand-by that 4 ℃ of refrigerators are preserved, 110-125 ℃ of slant medium sterilising conditions, 15-20 minute;
Seed culture
To connect a ring at the cultured bacterial classification of slant medium and join in the seed culture medium 110-125 ℃ of seed culture medium sterilising conditions, 15-20 minute; Under 200rpm rotating speed on the shaking table, 35-37 ℃ after shaken cultivation 20-28 hour, can obtain being fit to the inoculum of inoculation;
Fermented and cultured
With above-mentioned inoculum with 2-5% (volume ratio; Ml/ml) inoculum concentration is inoculated in the fresh fermentation medium through sterilization treatment, cultivates for 30-40 ℃ and carries out deep fermentation in 60-72 hour; During the fermentation, should control the pH value at 6.0-8.0; The adjusting of pH between yeast phase, the concrete condition of looking zymotic fluid pH, and, add acid or aqueous slkali realization by stream according to the fermentation needs; Dissolved oxygen is regulated by the control ventilation, and dissolved oxygen should be controlled at 20-40%;
Check the gemma production rate during fermentation ends, require the gemma production rate to reach more than 70%;
(2) preparation Soybean Peptide liquid food and biosis bacteria tablet
Separation of Solid and Liquid
After the fermentation ends, after sterility test is determined pollution-free assorted bacterium, adopted in refrigerated centrifugation, membrane filter method or the centrifugal filtration process one or both to unite to use and carry out Separation of Solid and Liquid, when adopting membrane filter method to separate solid-liquid, reduce in the zymotic fluid suspension as far as possible and see through film, when adopting the gravity centrifugal treating, detect the centrifuged supernatant viable count, the selection rotating speed should be greater than 5000r/min, make that suspension is centrifugal in the zymotic fluid gets off, when centrifugal filtration process is handled, centrifugal and filtration can be combined, in centrifugal process, finish filtration simultaneously;
Preparation Soybean Peptide liquid food
Clarified supernatant, measure Nattokinase enzyme activity in the liquid with the fibrin plate method, require Nattokinase content to reach 100-500U/ml, soybean peptide ammino acid content is 20-120g/L weight, after the assay was approved, add a small amount of albumen sugar and citric acid food additives and be deployed into Soybean Peptide liquid food;
(3) preparation biosis bacteria tablet
Separation is obtained bacterium mud carry out count plate, adopt vacuum freezedrying, fluidized bed drying, spray-drying or vacuum drying to carry out drying, carry out count plate after the drying once more, require the viable bacteria loss less than 80-85%, add excipient, get final product after fully stirring Bacillus Natto Beneficial Fungus Tablet.
2. according to the described method of utilizing bean product fermenting and producing Soybean Peptide liquid food and biosis bacteria tablet of claim 1, it is characterized in that: when fermentation medium adopted soybean-cake flour to be raw material, its prescription was: soybean-cake flour 20-80g, glucose 20-40g, MgSO
40.5-1.0g, Na
2HPO
46.0-12.0g, NaH
2PO
41.0-2.0g, CaCl
20.2-0.4g, 1000ml water, pH7.0-7.2;
When fermentation medium adopted the soybean bean powder to be raw material, its prescription was: soybean bean powder 20-80g, glucose 20-40g, MgSO
40.5-1.0g, Na
2HPO
46.0-12.0g, NaH
2PO
41.0-2.0g, CaCl
20.2-0.4g, 1000ml water, pH7.0-7.2;
When fermentation medium adopted soybean meal to be raw material, its prescription was: soybean meal 20-80g, glucose 20-40g, MgSO
40.5-1.0g, Na
2HPO
46.0-12.0g, NaH
2PO
41.0-2.0g, CaCl
20.2-0.4g, 1000ml water, pH7.0-7.2;
When fermentation medium adopted soybean milk to be raw material, its prescription was: soybean milk 125-250ml, glucose 20-40g, MgSO
40.5-1.0g, Na
2HPO
46.0-12.0g, NaH
2PO
41.0-2.0g, CaCl
20.2-0.4g, water supplies 1000ml, pH7.0-7.2;
Above-mentioned soybean-cake flour, soybean bean powder, soybean meal must be levigate, and it is stand-by to cross the 80-120 mesh sieve, and fermentation medium is taked 115-125 ℃, sterilization in 15-25 minute.
3. according to the described method of utilizing bean product fermenting and producing Soybean Peptide liquid food and biosis bacteria tablet of claim 1, it is characterized in that controlling the pH value, adjusting pH value method is: initial pH regulates can be by adding the phosphate buffer realization that concentration be 1.0-2.5%; The adjusting of pH between yeast phase adds acid or aqueous slkali realization by stream, and used acid solution is a phosphoric acid solution, and used alkali lye is ammoniacal liquor or urea, when fermenting to 48 hours, pH can be transferred to 6.0-6.3 or 7.7-8.0.
4. according to the described method of utilizing bean product fermenting and producing Soybean Peptide liquid food and biosis bacteria tablet of claim 1, it is characterized in that dissolved oxygen by controlling the ventilation adjusting, after fermentation was to 48 hours, reduces dissolved oxygen, the control dissolved oxygen is at 20-30%.
5. according to the described method of utilizing bean product fermenting and producing Soybean Peptide liquid food and biosis bacteria tablet of claim 1, it is characterized in that after fermentation 48 hours, can reduce the thermal stimulus gemma and produce, temperature is controlled at 25-32 ℃.
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| CN101214263B (en) * | 2007-12-26 | 2010-06-02 | 浙江大学宁波理工学院 | Method for preparing freeze-dried powder containing bacillus natto and nattokinase |
| CN104170982A (en) * | 2014-07-14 | 2014-12-03 | 山西省医药与生命科学研究院 | Manufacture process of natto milk tablets |
| CN108208309A (en) * | 2017-12-28 | 2018-06-29 | 保龄宝生物股份有限公司 | The production method of low ash content soybean peptide containing profitable probliotics |
| CN108410847B (en) * | 2018-03-13 | 2021-01-29 | 西安交通大学 | Method for preparing nattokinase dry powder and natto bacillus feed based on liquid fermentation method and application |
| CN111480713A (en) * | 2019-01-29 | 2020-08-04 | 金振库 | Natto black tea fungus liquid beverage |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH05336917A (en) * | 1991-07-01 | 1993-12-21 | Nippon Opereetaa Kk | Enzyme-activated odorless 'natto' and its preparation |
| JP2001352929A (en) * | 2000-06-14 | 2001-12-25 | Yamato Yakuhin Kk | Processed food and method for food processing |
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2004
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH05336917A (en) * | 1991-07-01 | 1993-12-21 | Nippon Opereetaa Kk | Enzyme-activated odorless 'natto' and its preparation |
| JP2001352929A (en) * | 2000-06-14 | 2001-12-25 | Yamato Yakuhin Kk | Processed food and method for food processing |
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