CN107950900A - A kind of preparation method of the compound fermented soya bean of Hyperfibrinolysis - Google Patents
A kind of preparation method of the compound fermented soya bean of Hyperfibrinolysis Download PDFInfo
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- CN107950900A CN107950900A CN201711371278.XA CN201711371278A CN107950900A CN 107950900 A CN107950900 A CN 107950900A CN 201711371278 A CN201711371278 A CN 201711371278A CN 107950900 A CN107950900 A CN 107950900A
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L11/00—Pulses, i.e. fruits of leguminous plants, for production of food; Products from legumes; Preparation or treatment thereof
- A23L11/50—Fermented pulses or legumes; Fermentation of pulses or legumes based on the addition of microorganisms
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/20—Reducing nutritive value; Dietetic products with reduced nutritive value
- A23L33/21—Addition of substantially indigestible substances, e.g. dietary fibres
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Abstract
The invention discloses a kind of preparation method of the compound fermented soya bean of Hyperfibrinolysis, combines using soybean with ginseng and campanulaceae and is formed as fermentation substrate, the fermentation of seeded with pure bacterium bacterium solution.Ginseng active ingredient can lift zymogenic activity, obtain plasmin activity height, the fermented soya bean of soft mouth feel, while the ginsenoside that human body can not directly absorb is changed into absorbable component by zymophyte.Contain platycogenic acid in campanulaceae, campanulaceae diacid, the components such as polygalic acid can neutralize the ammonia produced in fermentation process, yeasting is set to be maintained in suitable pH value range, be conducive to keep the activity of microbial enzyme, promote bioconversion of the microorganism to ginsenoside, improve bioavilability, the fibrinolytic of fermentate can be strengthened, strengthen its thrombolytic effect, coordinate the effect of campanulaceae inducting the medicines into affected channel, tunning active ingredient can be made to be easy to reach lesions position and play the effect of more preferable, since component has neutralized the ammonia produced in fermentation process and achievees the purpose that deodorization in campanulaceae, fermented soya bean taste is set more easily to receive.
Description
Technical field
The invention belongs to technical field of health care food, in particular to a kind of preparation of the compound fermented soya bean of Hyperfibrinolysis
Method, using soybean, ginseng, campanulaceae mixture as matrix inoculation bacterium solution fermentation, obtains a kind of fibrinolysin content height, thrombolytic effect
It is stronger, without ammonia taste and the high new fermented soya bean of ginseng effective component's absorption efficiency.
Background technology
Ginseng is the dry root and root of Araliaceae Panax herbaceous plant ginseng (Panax ginsong C.A.Meyer)
The effect of stem, has and reinforces vital energy, and veins takes off admittedly, reinforces the spleen to benefit the lung, and promotes the production of body fluid to quench thirst, tranquilize the mind and promote the intelligence.Modern pharmacological studies have shown that people
Ginseng can improve immunity of organisms, prevent and treat cardiovascular and cerebrovascular disease, have and resist myocardial ischemia and Antishock function, and can be bright
It is aobvious to reduce blood glucose and blood fat etc..The main component of ginseng is ginsenoside, and oral ginsenoside is hardly absorbed by the body utilization,
The conversion for having to pass through enteric microorganism is just absorbed by the body utilization.Therefore, many scholars realize ginseng soap by experiment in vitro
The microorganism conversion of glycosides, such as changes into rare ginsenoside Rg by ginsenoside3Or Ginsenoside compound K, ginsenoside Rg1Change into
Ginsenoside Rh1Etc..On August 29th, 2012, the Ministry of Public Health issue the 17th command, and approval ginseng (artificial growth) is eaten for new resources
Product, its source are 5 years and the ginseng of less than 5 years artificial growths, and kind is Araliaceae, Panax, and Leaf-feeding insects are root and rhizome,
Daily amount is≤3 grams/day, therefore can be as the raw material of food.
Campanulaceae is the dry root of Campanulaceae campanulaceae (Platycodon grandiflorus (Jacq.) A.Dc.), is belonged to
In integration of drinking and medicinal herbs class Chinese medicine, there is the effect of facilitaing lung, relieving sore-throat, eliminating the phlegm, apocenosis.Campanulaceae is drugs rendering other drugs to act on the upper-jiao, and medicine-carried uplink, has
Promoting the circulation of qi in width, the pent-up of qi-regulating point, the stagnant effect of the stasis of blood of promoting circulation of blood point so that gas row then blood.Campanulaceae principle active component is soap
Glycoside, it has been found that nearly 40 kinds of saponin component, wherein platycogenic acid (platcodic acid) class, campanulaceae diacid
(platcogenic acid) class and polygalic acid (polygalacic acid) class are main Types.Platycodin is lived with surface
Property, the leaching content of refractory components in medicine can be increased by solubilization, and it is collected at gas-liquid, in solid liquid interface, there is drop
The effect of low aqueous solution surface tension, it is not to be uniformly distributed and be collected on some cell surfaces in vivo to make it, so that
Change membrane passage, strengthen the uptake of other drugs.Therefore, the solubilising work that the surface-active of platycodin is determined
With the inherence basis for the influence to permeability of cell membrane being campanulaceae leading action.Platycogenic acid, campanulaceae diacid in campanulaceae, polygala
The components such as acid can be such that yeasting is maintained in constant pH value range, be conducive to maintain the activity of microbial enzyme, promote micro- life
Thing improves bioavilability, can strengthen the fibrinolytic of fermentate, so as to strengthen its thrombolysis to the bioconversion of ginsenoside
Effect.Meanwhile campanulaceae has the function that inducting the medicines into affected channel, tunning active ingredient can be made to be easy to reach lesions position and play more
The effect of good.
Fermented soya bean, ancient times are known as " deep and remote bean ", are Chinese Han nationality's characteristic fermented bean products flavorings.Generally steamed with soya bean or black soya bean
After boiling, formed using Mucor, aspergillus or bacterial fermentation, be chiefly used in seasoning, be divided into bacterial fermented douchi and mould type fermented soya bean, had
There are different physiological roles, Fibrinolytic Enzyme in Douchi contained therein has preferable fibrinolytic.Fermented soya bean contain abundant protein
(20%), fatty (7%) and carbohydrate (25%), and a variety of amino acid containing needed by human body, also contain multi mineral
The nutriment such as matter and vitamin.Fermented soya bean also make one increase appetite with its distinctive fragrance, promote to absorb, can not only season, and
And it can be used as medicine.Traditional Chinese medicine thinks that fermented soya bean are mild-natured, sweet and slightly bitter taste, there is relieving restlessness in inducing sweat and dispelling exogenous evils, clearing heat for promoting eruption, width, Xuan Yu removing toxic substances
Effect, the headache that can cure cold, it is uncomfortable in chest it is tired vomit, the illness such as typhoid fever fever and chills and food poisoning.Fermented soya bean can usually produce during the fermentation
Ammonifying stink, makes people be difficult to receive, and the method that someone is deodorized with ethanol is deodorized, but can so lose osajin isoreactivity into
Divide so as to reduce its effect.
The content of the invention
The object of the present invention is to provide a kind of preparation method of the compound fermented soya bean of Hyperfibrinolysis, by soybean and ginseng and campanulaceae
Combination is used as fermentation substrate, and the fermentation of seeded with pure bacterium bacterium solution forms.
A kind of preparation method of the compound fermented soya bean of Hyperfibrinolysis, it is characterised in that comprise the following steps：
Step 1: prepare fermentation substrate and campanulaceae extracting solution：
By ginseng coarse powder according to solid-to-liquid ratio 1：8 ratio adds water to cook 2 times, every time 1 it is small when, it is dense after merging filtrate after filtering
The 25%-75% for being reduced to original volume obtains Ginseng extract；Soybean is cleaned after draining, be added to above-mentioned ginseng and campanulaceae extraction
When immersion 10-18 is small in liquid, soybean is packed after fully exhausting extracting solution, sterilize 30-40 minutes in 121 DEG C, obtain fermentation base
Matter；
By campanulaceae coarse powder according to solid-to-liquid ratio 1：8 ratio adds water to cook 2 times, every time 1 it is small when, it is dense after merging filtrate after filtering
The 25%-75% for being reduced to original volume obtains campanulaceae extracting solution；
Wherein, the dry weight ratio of soybean, ginseng and campanulaceae is 1：0.05-0.2：0.05-0.1；
Step 2: pure culture ferments：
It is 10 that fermentation substrate prepared by step 1 is inoculated with 2ml concentration according to every 250g soaking soybeans7-109CFU/ml's is withered
The pure culture bacterium of any one in careless bacillus reference culture, the mould reference culture of umbrella branch pears head or Lichtheimia reference culture
Liquid, shakes up, in 25-40 DEG C culture, when pH value reaches higher than 11, add campanulaceae extracting solution be uniformly mixed, continue culture 36 to
72 obtain the compound fermented soya bean of Hyperfibrinolysis when small.
Preferably, the bacillus subtilis reference culture be selected from Bacillus subtilis CICC10446,
Bacillus natto CICC 20132, Bacillus natto CICC 10260 or Bacillus subtilis
Any one in subsp.subtilis CICC 20640；The mould reference culture of the umbrella branch pears head is selected from Absidia
Corymbifera CICC 40500, Absidia corymbifera CICC 3106 or Absidia corymbifera
Any one in CICC 40517；The Lichtheimia reference culture for Lichtheimia ramose CICC 41570 or
Lichtheimia sp.CICC 41600。
A kind of preparation method of the compound fermented soya bean of Hyperfibrinolysis, it is characterised in that comprise the following steps：
Step 1: prepare fermentation substrate：
Soybean is cleaned, with clear water immersion 10-18 it is small when, after draining, by ginseng with campanulaceae co-grinding into fine powder, with leaching
The soybean that soaks mixes, and pack, sterilizes 30 to 40 minutes in 121 DEG C, obtain fermentation substrate, wherein, soybean, ginseng and campanulaceae
Dry weight ratio is 1：0.05-0.2：0.05-0.1；
Step 2: pure culture ferments：
It is 10 that fermentation substrate prepared by step 1 is inoculated with 2ml concentration according to every 250g soaking soybeans7-109CFU/ml's is withered
The pure culture bacterium of any one in careless bacillus reference culture, the mould reference culture of umbrella branch pears head or Lichtheimia reference culture
Liquid, shakes up, and the compound fermented soya bean of Hyperfibrinolysis are obtained when 25-40 DEG C of culture 72-120 is small.
Preferably, the bacillus subtilis reference culture be selected from Bacillus subtilis CICC10446,
Bacillus natto CICC 20132, Bacillus natto CICC 10260 or Bacillus subtilis
Any one in subsp.subtilis CICC 20640；The mould reference culture of the umbrella branch pears head is selected from Absidia
Corymbifera CICC 40500, Absidia corymbifera CICC 3106 or Absidia corymbifera
Any one in CICC 40517；The Lichtheimia reference culture for Lichtheimia ramose CICC 41570 or
Lichtheimia sp.CICC 41600。
A kind of preparation method of the compound fermented soya bean of Hyperfibrinolysis, it is characterised in that comprise the following steps：
Step 1: prepare fermentation substrate：
Ginseng and campanulaceae coarse powder are mixed, according to solid-to-liquid ratio 1：8 ratio adds water to cook 2 times, every time 1 it is small when, after filtering
The 25%-75% that original volume is concentrated into after merging filtrate obtains ginseng and campanulaceae extracting solution；Soybean is cleaned after draining, be added to
When immersion 10-18 is small in above-mentioned ginseng and campanulaceae extracting solution, soybean is set to be packed after fully exhausting extracting solution, sterilize 30- in 121 DEG C
40 minutes, fermentation substrate is obtained, wherein, the dry weight ratio of soybean, ginseng and campanulaceae is 1：0.05-0.2：0.05-0.1；
Step 2: pure culture ferments：
It is 10 that fermentation substrate prepared by step 1 is inoculated with 2ml concentration according to every 250g soaking soybeans7-109CFU/ml's is withered
The pure culture bacterium of any one in careless bacillus reference culture, the mould reference culture of umbrella branch pears head or Lichtheimia reference culture
Liquid, shakes up, and the compound fermented soya bean of Hyperfibrinolysis are obtained when 25-40 DEG C of culture 72-120 is small.
Preferably, the bacillus subtilis reference culture be selected from Bacillus subtilis CICC10446,
Bacillus natto CICC 20132, Bacillus natto CICC 10260 or Bacillus subtilis
Any one in subsp.subtilis CICC 20640；The mould reference culture of the umbrella branch pears head is selected from Absidia
Corymbifera CICC 40500, Absidia corymbifera CICC 3106 or Absidia corymbifera
Any one in CICC 40517；The Lichtheimia reference culture for Lichtheimia ramose CICC 41570 or
Lichtheimia sp.CICC 41600。
A kind of preparation method of the compound fermented soya bean of Hyperfibrinolysis, it is characterised in that comprise the following steps：
Step 1: prepare fermentation substrate：
Ginseng and campanulaceae coarse powder are mixed, according to solid-to-liquid ratio 1：8 ratio adds the ethanol solution refluxing extraction 2 of 50-80%
It is secondary, every time 1 it is small when, after filtering merging filtrate recycling ethanol obtain an extracting solution；Every 100 grams of residues add water 800 milliliters and decoct
Boil 2 times, every time 1 it is small when filtering after merging filtrate obtain secondary raffinate, extracting solution will be uniformly mixed twice and be concentrated into original volume
25%-75% obtain ginseng and campanulaceae extracting solution；Soybean is cleaned after draining, be added in above-mentioned ginseng and campanulaceae extracting solution
When immersion 10-18 is small, soybean is packed after fully exhausting extracting solution, sterilize 30-40 minutes in 121 DEG C, obtain fermentation substrate, its
In, the dry weight ratio of soybean, ginseng and campanulaceae is 1：0.05-0.2：0.05-0.1；
Step 2: pure culture ferments：
It is 10 that fermentation substrate prepared by step 1 is inoculated with 2ml concentration according to every 250g soaking soybeans7-109CFU/ml's is withered
The pure culture bacterium of any one in careless bacillus reference culture, the mould reference culture of umbrella branch pears head or Lichtheimia reference culture
Liquid, shakes up, and the compound fermented soya bean of Hyperfibrinolysis are obtained when 25-40 DEG C of culture 72-120 is small.
Preferably, the bacillus subtilis reference culture be selected from Bacillus subtilis CICC10446,
Bacillus natto CICC 20132, Bacillus natto CICC 10260 or Bacillus subtilis
Any one in subsp.subtilis CICC 20640；The mould reference culture of the umbrella branch pears head is selected from Absidia
Corymbifera CICC 40500, Absidia corymbifera CICC 3106 or Absidia corymbifera
Any one in CICC 40517；The Lichtheimia reference culture for Lichtheimia ramose CICC 41570 or
Lichtheimia sp.CICC 41600。
The present invention has technical advantage：
1) ginseng active ingredient can lift zymogenic activity, obtain plasmin activity height, the fermented soya bean of soft mouth feel, at the same time
The ginsenoside that human body can not directly absorb is changed into absorbable component by zymophyte.
2) ammonia produced in fermentation process can be neutralized, is made containing components such as platycogenic acid, campanulaceae diacid, polygalic acids in campanulaceae
Yeasting is maintained in suitable pH value range, is conducive to keep the activity of microbial enzyme, promotes microorganism to ginsenoside
Bioconversion, improve bioavilability, the fibrinolytic of fermentate can be strengthened, strengthen its thrombolytic effect, coordinate campanulaceae to draw
Medicine enter through effect, can make tunning active ingredient be easy to reach lesions position and play the effect of more preferable.
3) due to having neutralized the ammonia produced in fermentation process containing components such as platycogenic acid, campanulaceae diacid, polygalic acids in campanulaceae,
The purpose of deodorization is can reach, the fermented soya bean taste of preparation is more easily received.
Embodiment
To make those skilled in the art more fully understand technical scheme, below by embodiment to the present invention
A kind of preparation method of the compound fermented soya bean of Hyperfibrinolysis provided is described in detail.The compound fermented soya bean of 1 Hyperfibrinolysis of embodiment
Preparation
Step 1: prepare fermentation substrate：
Prepare soybean 100g, ginseng 10g, campanulaceae 5g, soybean cleaned, with clear water immersion 18 it is small when, after draining, by ginseng
With campanulaceae co-grinding into fine powder, packed after being mixed with soaked soybean, sterilize 30 minutes in 121 DEG C, obtain fermentation substrate.
Step 2: pure culture ferments：
It is 10 that fermentation substrate prepared by step 1 is inoculated with 2ml concentration according to every 250g soaking soybeans7-109CFU/ml's
10446 pure culture bacterium solutions of Bacillus subtilis CICC, shake up, and obtaining Hyperfibrinolysis when 37 DEG C of cultures 72 are small answers
Close fermented soya bean.
According to the proportioning conduct that soybean 100g, soybean 100g and ginseng 10g, soybean 100g and campanulaceae 5g are replaced in step 1
Fermentation substrate prepares fermented soya bean with method respectively, and measures plasmin activity, as control experiment.
It is understood that in step 1, preparing the dry weight ratio of the soybean used in fermentation substrate, ginseng and campanulaceae can be
1：0.05-0.2：0.05-0.1.
It is understood that in step 2, the bacillus subtilis reference culture Bacillus subtilis that are inoculated with
CICC 10446 alternatively into Bacillus natto CICC 20132, Bacillus natto CICC 10260 or
Any one in Bacillus subtilis subsp.subtilis CICC 20640, above-mentioned standard bacterial strain buy from
Chinese industrial Microbiological Culture Collection administrative center.
It is understood that in step 2, the bacillus subtilis reference culture Bacillus subtilis that are inoculated with
CICC 10446 is alternatively into the mould reference culture Absidia corymbifera CICC 40500 of umbrella branch pears head, Absidia
Any one in corymbifera CICC 3106 or Absidia corymbifera CICC40517, above reference culture
Buy from Chinese industrial Microbiological Culture Collection administrative center.
It is understood that in step 2, the bacillus subtilis reference culture Bacillus subtilis that are inoculated with
CICC 10446 is alternatively into Lichtheimia reference culture Lichtheimia ramose CICC41570 or Lichtheimia
Sp.CICC 41600, above reference culture are bought from Chinese industrial Microbiological Culture Collection administrative center.
The preparation of the compound fermented soya bean of 2 Hyperfibrinolysis of embodiment
Step 1: prepare fermentation substrate：
Ginseng 15g and campanulaceae 8g coarse powder are mixed, according to solid-to-liquid ratio 1：8(g：Ml ratio) adds water to cook 2 times, and every time 1
Hour, 120ml is concentrated into after merging filtrate obtain ginseng and campanulaceae extracting solution after filtering；100g soybean is cleaned after draining, add
Enter into above-mentioned ginseng and campanulaceae extracting solution immersion 12 it is small when, soybean is packed after fully exhausting extracting solution, in 121 DEG C sterilize 40
Minute, obtain fermentation substrate.
Step 2: pure culture ferments：
It is 10 that fermentation substrate prepared by step 1 is inoculated with 2ml concentration according to every 250g soaking soybeans7-109CFU/ml's
40500 pure culture bacterium solutions of Absidia corymbifera CICC, shake up, and Hyperfibrinolysis is obtained when 28 DEG C of cultures 96 are small
Compound fermented soya bean.
According to the proportioning conduct that soybean 100g, soybean 100g and ginseng 15g, soybean 100g and campanulaceae 8g are replaced in step 1
Fermentation substrate prepares fermented soya bean with method respectively, and measures plasmin activity, as control experiment.
It is understood that in step 1, preparing the dry weight ratio of the soybean used in fermentation substrate, ginseng and campanulaceae can be
1：0.05-0.2：0.05-0.1.
It is understood that in step 2, the mould reference culture Absidia corymbifera of umbrella branch pears head that are inoculated with
CICC 40500 is alternatively into Absidia corymbifera CICC 3106 or Absidia corymbifera CICC
40517, above reference culture is bought from Chinese industrial Microbiological Culture Collection administrative center.
It is understood that in step 2, the mould reference culture Absidia corymbifera of umbrella branch pears head that are inoculated with
CICC 40500 is alternatively into bacillus subtilis reference culture Bacillus subtilis CICC 10446, Bacillus
Natto CICC 20132, Bacillus natto CICC 10260 or Bacillus subtilis subsp.subtilis
Any one in CICC 20640, above reference culture is bought from Chinese industrial Microbiological Culture Collection administrative center.
It is understood that in step 2, the mould reference culture Absidia corymbifera of umbrella branch pears head that are inoculated with
CICC 40500 is alternatively into Lichtheimia reference culture Lichtheimia ramose CICC 41570 or Lichtheimia
Sp.CICC 41600, above reference culture are bought from Chinese industrial Microbiological Culture Collection administrative center.
The preparation of the compound fermented soya bean of 3 Hyperfibrinolysis of embodiment
Step 1: prepare fermentation substrate：
Ginseng 10g and campanulaceae 10g coarse powder are mixed, according to solid-to-liquid ratio 1：8(g：Ml ratio) adds 80% ethanol solution
Refluxing extraction 2 times, every time 1 it is small when, after filtering merging filtrate recycling ethanol obtain an extracting solution；Every 100 grams of residues add water
800 milliliters decoct 2 times, and merging filtrate obtains secondary raffinate after being filtered when 1 is small every time, will be uniformly mixed concentration by extracting solution twice
Ginseng and campanulaceae extracting solution are obtained to 130ml；100g soybean is cleaned after draining, be added in above-mentioned ginseng and campanulaceae extracting solution
Soak 18 it is small when, soybean is packed after fully exhausting extracting solution, in 121 DEG C sterilize 35 minutes, obtain fermentation substrate.
Step 2: pure culture ferments：
It is 10 that fermentation substrate prepared by step 1 is inoculated with 2ml concentration according to every 250g soaking soybeans7-109CFU/ml's
41570 pure culture bacterium solutions of Lichtheimia ramose CICC, shake up, and Hyperfibrinolysis is obtained when 25 DEG C of cultures 120 are small
Compound fermented soya bean.
It is compared to according to matching somebody with somebody for soybean 100g, soybean 100g and ginseng 10g, soybean 100g and campanulaceae 10g is replaced in step 1
Fermented soya bean are prepared with method respectively for fermentation substrate, and measure plasmin activity, as control experiment.
It is understood that in step 1, preparing the dry weight ratio of the soybean used in fermentation substrate, ginseng and campanulaceae can be
1：0.05-0.2：0.05-0.1.
It is understood that in step 2, the Lichtheimia reference culture Lichtheimia ramose CICC that are inoculated with
41570 alternatively into Lichtheimia sp.CICC 41600, and above reference culture is bought from Chinese industrial microorganism fungus kind
Preservation administrative center.
It is understood that in step 2, the Lichtheimia reference culture Lichtheimia ramose CICC that are inoculated with
41570 alternatively into bacillus subtilis reference culture Bacillus subtilis CICC10446, Bacillus natto
CICC 20132, Bacillus natto CICC 10260 or Bacillus subtilis subsp.subtilis CICC
Any one in 20640, above reference culture is bought from Chinese industrial Microbiological Culture Collection administrative center.
It is understood that in step 2, the Lichtheimia reference culture Lichtheimia ramose CICC that are inoculated with
41570 alternatively into the mould reference culture Absidia corymbifera CICC 40500 of umbrella branch pears head, Absidia
Any one in corymbifera CICC 3106 or Absidia corymbifera CICC40517, above reference culture
Buy from Chinese industrial Microbiological Culture Collection administrative center.
The preparation of the compound fermented soya bean of 4 Hyperfibrinolysis of embodiment
Step 1: prepare fermentation substrate：
By ginseng 10g coarse powder according to solid-to-liquid ratio 1：8(g：Ml ratio) adds water to cook 2 times, every time 1 it is small when, closed after filtering
And it is concentrated into 120ml after filtrate and obtains Ginseng extract；100g soybean is cleaned after draining, be added in above-mentioned Ginseng extract
Soak 18 it is small when, soybean is packed after fully exhausting extracting solution, in 121 DEG C sterilize 30 minutes, obtain fermentation substrate.
Step 2: pure culture ferments：
It is 107-109CFU/ml's that fermentation substrate prepared by step 1 is inoculated with 2ml concentration according to every 250g soaking soybeans
20640 pure culture bacterium solutions of bacillus subtilis reference culture Bacillus subtilis subsp.subtilis CICC, shake
Even, in 35 DEG C of cultures, every the pH value of 12h measure one time fermentation things, as a result pH value reaches up to 11 during fermentation 48h, and has
There is denseer ammonia odor taste.
Step 3: add campanulaceae extracting solution secondary fermentation
By campanulaceae 6g coarse powder according to solid-to-liquid ratio 1：8(g：Ml ratio) adds water to cook 2 times, every time 1 it is small when, merge after filtering
25ml is concentrated into after filtrate and obtains campanulaceae extracting solution, it is degerming with 0.22 μm of membrane filtration, add the one time fermentation that step 2 obtains
In thing, shake up and continue to cultivate 48h in 37 DEG C, up to no ammonia taste (pH value 8) and have a compound fermented soya bean of Hyperfibrinolysis.
According to the one time fermentation thing obtained in step 3 with the hydrochloric acid 5ml replacement campanulaceae extracting solution additions step 2 of 1mol/L
In with method prepare fermented soya bean, and plasmin activity is measured, as control experiment.
According to the proportioning conduct that soybean 100g, soybean 100g and ginseng 10g, soybean 100g and campanulaceae 6g are replaced in step 1
Fermentation substrate prepares fermented soya bean according to step 1 and step 2 with method, and measures plasmin activity, as control experiment.
It is understood that in step 1, soybean, ginseng dry weight ratio used in preparation fermentation substrate can be 1：0.05-
0.2。
It is understood that in step 2, the bacillus subtilis reference culture Bacillus subtilis that are inoculated with
Subsp.subtilis CICC 20640 are alternatively into Bacillus subtilis CICC 10446, Bacillus natto
Any one of CICC 20132, Bacillus natto CICC 10260, above reference culture is bought micro- from Chinese industrial
Biological inoculum preservation administrative center.
It is understood that in step 2, the bacillus subtilis reference culture Bacillus subtilis that are inoculated with
Subsp.subtilis CICC 20640 are alternatively into the mould reference culture Absidia corymbifera CICC of umbrella branch pears head
40500th, it is any one in Absidia corymbifera CICC 3106 or Absidia corymbifera CICC 40517
Kind, above reference culture is bought from Chinese industrial Microbiological Culture Collection administrative center.
It is understood that in step 2, the bacillus subtilis reference culture Bacillus subtilis that are inoculated with
Subsp.subtilis CICC 20640 are alternatively into Lichtheimia reference culture Lichtheimia ramose CICC
41570 or Lichtheimia sp.CICC 41600, above reference culture are bought from Chinese industrial Microbiological Culture Collection pipe
Reason center.
It is understood that in step 3, soybean, campanulaceae dry weight ratio used in preparation fermentation substrate can be 1：0.05-
0.1。
The experimentation that product plasmin activity is measured in above-described embodiment 1-4 is as follows：
Experimental facilities：
Fast several MF3 types all-in-one multifunctional machines (Hangzhou Xunshu It Co., Ltd), DHP9272 constant incubators (Shanghai one
Permanent scientific instrument Co., Ltd), DL-CJ-2N superclean benches (Beijing Dong Lianhaer instrument manufacturings Co., Ltd), BSA224S
Electronic balance (German Sai Duolisi).
Reagent：
Urokinase, fibrin ferment, bovine fibrinogen, agarose, Corning culture dishes φ=90mm, sodium chloride, di(2-ethylhexyl)phosphate
Hydrogen sodium, disodium hydrogen phosphate.
Experimental procedure：
The preparation of urokinase standard solution：
Urokinase 1 (1000IU/ branch) is taken, every ml is made into containing 100,80,60,40,20IU with 0.9% sodium chloride solution
Standard solution.
The preparation of sample solution：
Take compound fermented soya bean 2g to add 0.9% sodium chloride solution 10ml to mix, 4 DEG C it is static 1 it is small when, it is stand-by.
The preparation of working solution：
By phosphate buffer (0.01mol/L) and 0.9% sodium chloride solution according to volume ratio 1:17 mix, to obtain the final product.
The preparation of 1% agarose solution：
Precision weighs agarose 1g, adds working solution 100ml, dissolves by heating, and is kept the temperature in 50 DEG C of water-baths.
The preparation of fibrinogen solution：
Precision weighs fibrinogen 0.0576g, adds working solution 19.2ml, is configured to 0.3% solution, and in 50 DEG C of water
Bath insulation.
The preparation of thrombin solution：
Precision weighs fibrin ferment, adds 0.9% sodium chloride solution to be made into the solution of 1BP concentration.
The preparation of agarose-fibrin plate：
By the agarose solution 19.2ml and fibrinogen solution 19.2ml and thrombin solution of 50 DEG C of water-bath insulations
1.6ml is mixed, and is poured into rapidly in the plate of 2 9cm × 9cm, and room temperature places 30min, and solidification forms.
Plasmin activity measures：
Draw the urokinase standard solution and each 10 μ l of sample solution supernatant of various concentrations respectively with microsyringe, point
Other dibbling is on same agarose-fibrin plate, capping, and 18h are incubated in 37 DEG C, and it is vertical two straight that solusphere is measured after taking-up
Footpath, using two diameter products as ordinate, urokinase standard items units is abscissa, draws standard curve.It is molten according to test sample
The product of vertical two diameter is enclosed, is contrasted with standard curve, calculates plasmin activity.
Embodiment 1, embodiment 2, embodiment 3,4 plasmin activity measurement result of embodiment such as form 1：
Form 1
From form 1 as it can be seen that the fermented soya bean that embodiment 1, embodiment 2, embodiment 3, embodiment 4 obtain under similarity condition, add
Fibrinolytic enzyme activity is improved after entering ginseng, campanulaceae, and the Fibrinolytic Enzyme in Douchi that wherein soybean individually adds ginseng to obtain is lived than independent soybean
The obtained Fibrinolytic Enzyme in Douchi that ferments, which is lived, has been respectively increased 1.48 times, 1.52 times, 1.54 times；Soybean is individually added into the beans that campanulaceae obtains
Fermented soya beans, salted or other wise fibrinolytic enzyme activity lives than the Fibrinolytic Enzyme in Douchi that independent fermented soybean obtains and has been respectively increased 1.12 times, 1.17 times, 1.19 times；It is and big
Beans add the Fibrinolytic Enzyme in Douchi work that ginseng and campanulaceae obtain and significantly improve at the same time, the Fibrinolytic Enzyme in Douchi obtained than independent fermented soybean
Work has been respectively increased 2.06 times, 2.08 times, 2.13 times；The Fibrinolytic Enzyme in Douchi that campanulaceae extracting solution secondary fermentation obtains is added to live than big
Individually ferment obtained Fibrinolytic Enzyme in Douchi of beans lives and improves 2.49 times, it is seen that ginseng and campanulaceae mutual reinforcement between can improve the fibre of fermented soya bean for use
Molten activity, and addition progress secondary fermentation is better after campanulaceae extracting solution, not only improves the fibrinolytic of fermented soya bean, additionally it is possible to protect
The pH value of microbial fermentation is held, reduces ammonia odor taste.Can reduce the pH value of yeasting although adding hydrochloric acid and carrying out secondary fermentation,
But plasmin activity have dropped 56% than the fermented soya bean fibrinolytic for adding campanulaceae extracting solution progress secondary fermentation to obtain, it is seen that ferment
During add campanulaceae and not only natural environmental-protective but also yeasting can have been maintained and improved bioactivity.
It is understood that the principle that embodiment of above is intended to be merely illustrative of the present and the exemplary implementation that uses
Mode, but the present invention is not limited thereto.For those skilled in the art, the essence of the present invention is not being departed from
In the case of refreshing and essence, various changes and modifications can be made therein, these variations and modifications are also considered as protection scope of the present invention.
Claims (8)
1. a kind of preparation method of the compound fermented soya bean of Hyperfibrinolysis, it is characterised in that comprise the following steps：
Step 1: prepare fermentation substrate and campanulaceae extracting solution：
By ginseng coarse powder according to solid-to-liquid ratio 1：8 ratio adds water to cook 2 times, every time 1 it is small when, be concentrated into after filtering after merging filtrate
The 25%-75% of original volume obtains Ginseng extract；Soybean is cleaned after draining, be added in above-mentioned ginseng and campanulaceae extracting solution
When immersion 10-18 is small, soybean is packed after fully exhausting extracting solution, sterilize 30-40 minutes in 121 DEG C, obtain fermentation substrate；
By campanulaceae coarse powder according to solid-to-liquid ratio 1：8 ratio adds water to cook 2 times, every time 1 it is small when, be concentrated into after filtering after merging filtrate
The 25%-75% of original volume obtains campanulaceae extracting solution；
Wherein, the dry weight ratio of soybean, ginseng and campanulaceae is 1：0.05-0.2：0.05-0.1；
Step 2: pure culture ferments：
It is 10 that fermentation substrate prepared by step 1 is inoculated with 2ml concentration according to every 250g soaking soybeans7-109The withered grass bud of CFU/ml
The pure culture bacterium solution of any one in spore bacillus reference culture, the mould reference culture of umbrella branch pears head or Lichtheimia reference culture, shakes
It is even, in 25-40 DEG C culture, when pH value reaches higher than 11, add campanulaceae extracting solution be uniformly mixed, continue culture 36 to 72 it is small when
Obtain the compound fermented soya bean of Hyperfibrinolysis.
2. the preparation method of the compound fermented soya bean of Hyperfibrinolysis according to claim 1, it is characterised in that the withered grass gemma
Bacillus reference culture be selected from Bacillus subtilis CICC 10446, Bacillus natto CICC 20132,
Appointing in Bacillus natto CICC 10260 or Bacillus subtilis subsp.subtilis CICC 20640
Meaning is a kind of；The mould reference culture of the umbrella branch pears head is selected from Absidia corymbifera CICC 40500, Absidia
Any one in corymbifera CICC 3106 or Absidia corymbifera CICC 40517；The horizontal stalk is mould
Category reference culture is Lichtheimia ramose CICC 41570 or Lichtheimia sp.CICC 41600.
3. a kind of preparation method of the compound fermented soya bean of Hyperfibrinolysis, it is characterised in that comprise the following steps：
Step 1: prepare fermentation substrate：
Soybean is cleaned, with clear water immersion 10-18 it is small when, after draining, by ginseng with campanulaceae co-grinding into fine powder, with soaking
Soybean mix, pack, in 121 DEG C sterilize 30 to 40 minutes, obtain fermentation substrate, wherein, the dry weight of soybean, ginseng and campanulaceae
Than for 1：0.05-0.2：0.05-0.1；
Step 2: pure culture ferments：
It is 10 that fermentation substrate prepared by step 1 is inoculated with 2ml concentration according to every 250g soaking soybeans7-109The withered grass bud of CFU/ml
The pure culture bacterium solution of any one in spore bacillus reference culture, the mould reference culture of umbrella branch pears head or Lichtheimia reference culture, shakes
It is even, obtain the compound fermented soya bean of Hyperfibrinolysis when 25-40 DEG C of culture 72-120 is small.
4. the preparation method of the compound fermented soya bean of Hyperfibrinolysis according to claim 3, it is characterised in that the withered grass gemma
Bacillus reference culture be selected from Bacillus subtilis CICC 10446, Bacillus natto CICC 20132,
Appointing in Bacillus natto CICC 10260 or Bacillus subtilis subsp.subtilis CICC 20640
Meaning is a kind of；The mould reference culture of the umbrella branch pears head is selected from Absidia corymbifera CICC 40500, Absidia
Any one in corymbifera CICC 3106 or Absidia corymbifera CICC 40517；The horizontal stalk is mould
Category reference culture is Lichtheimia ramose CICC 41570 or Lichtheimia sp.CICC 41600.
5. a kind of preparation method of the compound fermented soya bean of Hyperfibrinolysis, it is characterised in that comprise the following steps：
Step 1: prepare fermentation substrate：
Ginseng and campanulaceae coarse powder are mixed, according to solid-to-liquid ratio 1：8 ratio adds water to cook 2 times, every time 1 it is small when, merge after filtering
The 25%-75% that original volume is concentrated into after filtrate obtains ginseng and campanulaceae extracting solution；Soybean is cleaned after draining, be added to above-mentioned
When immersion 10-18 is small in ginseng and campanulaceae extracting solution, soybean is packed after fully exhausting extracting solution, sterilize in 121 DEG C 30-40 points
Clock, obtains fermentation substrate, wherein, the dry weight ratio of soybean, ginseng and campanulaceae is 1：0.05-0.2：0.05-0.1；
Step 2: pure culture ferments：
It is 10 that fermentation substrate prepared by step 1 is inoculated with 2ml concentration according to every 250g soaking soybeans7-109The withered grass bud of CFU/ml
The pure culture bacterium solution of any one in spore bacillus reference culture, the mould reference culture of umbrella branch pears head or Lichtheimia reference culture, shakes
It is even, obtain the compound fermented soya bean of Hyperfibrinolysis when 25-40 DEG C of culture 72-120 is small.
6. the preparation method of the compound fermented soya bean of Hyperfibrinolysis according to claim 5, it is characterised in that the withered grass gemma
Bacillus reference culture be selected from Bacillus subtilis CICC 10446, Bacillus natto CICC 20132,
Appointing in Bacillus natto CICC 10260 or Bacillus subtilis subsp.subtilis CICC 20640
Meaning is a kind of；The mould reference culture of the umbrella branch pears head is selected from Absidia corymbifera CICC 40500, Absidia
Any one in corymbifera CICC 3106 or Absidia corymbifera CICC 40517；The horizontal stalk is mould
Category reference culture is Lichtheimia ramose CICC 41570 or Lichtheimia sp.CICC 41600.
7. a kind of preparation method of the compound fermented soya bean of Hyperfibrinolysis, it is characterised in that comprise the following steps：
Step 1: prepare fermentation substrate：
Ginseng and campanulaceae coarse powder are mixed, according to solid-to-liquid ratio 1：8 ratio adds the ethanol solution refluxing extraction 2 times of 50-80%, often
It is secondary 1 it is small when, after filtering merging filtrate recycling ethanol obtain an extracting solution；Every 100 grams of residues add water 800 milliliters and decoct 2 times,
Merging filtrate obtains secondary raffinate after being filtered when 1 is small every time, and extracting solution twice is uniformly mixed to the 25%- for being concentrated into original volume
75% obtains ginseng and campanulaceae extracting solution；Soybean is cleaned after draining, be added in above-mentioned ginseng and campanulaceae extracting solution and soak 10-
18 it is small when, soybean is packed after fully exhausting extracting solution, in 121 DEG C sterilize 30-40 minutes, obtain fermentation substrate, wherein, greatly
The dry weight ratio of beans, ginseng and campanulaceae is 1：0.05-0.2：0.05-0.1；
Step 2: pure culture ferments：
It is 10 that fermentation substrate prepared by step 1 is inoculated with 2ml concentration according to every 250g soaking soybeans7-109The withered grass bud of CFU/ml
The pure culture bacterium solution of any one in spore bacillus reference culture, the mould reference culture of umbrella branch pears head or Lichtheimia reference culture, shakes
It is even, obtain the compound fermented soya bean of Hyperfibrinolysis when 25-40 DEG C of culture 72-120 is small.
8. the preparation method of the compound fermented soya bean of Hyperfibrinolysis according to claim 7, it is characterised in that the withered grass gemma
Bacillus reference culture be selected from Bacillus subtilis CICC 10446, Bacillus natto CICC 20132,
Appointing in Bacillus natto CICC 10260 or Bacillus subtilis subsp.subtilis CICC 20640
Meaning is a kind of；The mould reference culture of the umbrella branch pears head is selected from Absidia corymbifera CICC 40500, Absidia
Any one in corymbifera CICC 3106 or Absidia corymbifera CICC 40517；The horizontal stalk is mould
Category reference culture is Lichtheimia ramose CICC 41570 or Lichtheimia sp.CICC 41600.
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| CN109480195A (en) * | 2018-12-05 | 2019-03-19 | 遵义市刘胡子食品有限公司 | A kind of zi ba pepper fermented soya bean and preparation method thereof |
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