CN109757603A - A kind of method that fermentation soybean grouts prepare pannage - Google Patents
A kind of method that fermentation soybean grouts prepare pannage Download PDFInfo
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- CN109757603A CN109757603A CN201910063107.3A CN201910063107A CN109757603A CN 109757603 A CN109757603 A CN 109757603A CN 201910063107 A CN201910063107 A CN 201910063107A CN 109757603 A CN109757603 A CN 109757603A
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Abstract
The present invention relates to the preparation fields of feed, more particularly to one kind using soybean cake dregs as raw material, method that auxotype pannage is prepared by compound strain solid state fermentation.This method step are as follows: obtain pannage after compound bacteria fermentation being added in dregs of beans, the compound bacteria includes prokaryotic micro-organisms and eukaryotic microorganisms.The prokaryotic micro-organisms includes bafillus natto and Bacillus acidi lactici；And the eukaryotic microorganisms include S. cervisiae and aspergillus niger.Bafillus natto, Bacillus acidi lactici, S. cervisiae and the effective bacterium number ratio of aspergillus niger are 1~3:1~3:1~3:1~3 in the compound bacteria.The compound bacteria is 1 × 10 by concentration7~1 × 108Bafillus natto, Bacillus acidi lactici, S. cervisiae and the aspergillus niger of CFU/mL mixes.The pannage that the present invention is prepared can remove the natural anti-nutritional factors in soybean cake dregs, and have additional nutrients element, improve bioavailability.
Description
Technical field
The present invention relates to the preparation fields of feed, more particularly to one kind using dregs of beans as raw material, pass through compound strain solid-state
The method that fermentation prepares auxotype pannage.
Background technique
China is one of maximum livestock and poultry cultivation producing country in the world, while being also feedstuff especially albumen in the world
The demand big country of matter raw material.Protein feed resource wretched insufficiency in the production of China's feed industry, the high-quality animal egg such as fish meal
White raw material relies primarily on international import, and the reduction increasingly of the global stock of fish at present directly result in fish meal prices it is direct on
Rise.Although fish meal is considered top quality protein raw materials by row insider, by its price, fish meal makes in feed
Still tend to decline with ratio, animal protein price is surging so that sight has been increasingly turned to vegetable protein by people.
A kind of soybean cake dregs plant protein material fixed as abundance, quality is always to constitute animal fodder egg
The main body of Bai Laiyuan, the animal protein such as fish meal resource is increasingly in short supply in addition and holds at high price, and keeps its usage amount limited, promotees
Expand the market demand of soybean cake dregs constantly.But it is main to wrap due to containing in soybean cake dregs there are many natural anti-nutritional factors
Include: urase, trypsin ihhibitor, soybean antigen, hemagglutinin, Flatulent factors etc., some anti-nutritional factors are to heat
It is unstable, soybean cake dregs are carried out most of anti-nutritional factors can be made to inactivate after sufficiently heating.Currently, at the heat of soybean cake dregs
It is general lower (60~80 DEG C) to manage temperature, the anti-nutritional factors that effectively can not be passivated or remove in soybean cake dregs, thus
Limit direct application of the soybean cake dregs in animal feed.
Summary of the invention
(1) technical problems to be solved
In order to solve the above problem of the prior art, the present invention provides one kind using soybean cake dregs as substrate, and compound strain is sent out
The method that ferment prepares the high pannage of nutrient content.
(2) technical solution
In order to achieve the above object, the main technical schemes that the present invention uses include:
A kind of method that fermentation soybean grouts prepare pannage comprising following steps: compound by being added in soybean cake dregs
Pannage is obtained after bacterium fermentation, the compound bacteria includes prokaryotic micro-organisms and eukaryotic microorganisms.
Contain 43.5%~47% thick protein and 5%~7% crude fibre in this programme soybean cake dregs.It is sent out through microorganism
Ferment, protein are decomposed into micromolecule polypeptide, oligopeptides and amino acid, more conducively under the hydrolysis of thallus extracellular protease
It absorbs and utilizes, also, a large amount of mycoprotein can be improved the protein content in soybean cake dregs.Portion of cellulose can be fine
Plain enzyme hydrolysis is tieed up at monosaccharide, to improve the utility value of soybean cake dregs.Soybean cake dregs are during the fermentation, additionally it is possible to generate cream
The organic acids such as acid, butyric acid, acetic acid, succinic acid, the lifes such as having enhances raising animal appetite, and inhibition harmful bacteria is proliferated in the gastrointestinal tract
Manage function.Also, since portion carbohydrate is degraded, soybean cake dregs compact texture becomes loose, and palatability significantly improves.
This programme utilizes the difference of prokaryotic micro-organisms and eukaryotic microorganisms fermenting speed, wherein the fermenting speed of prokaryotic micro-organisms is than true
Core microorganism is fast.Stop fermentation before prokaryotic micro-organisms fermentation starts to generate nuisance, at this point, the fermentation of eukaryotic microorganisms is not
Completely, so that the anti-nutritional factors in soybean cake dregs is degraded to lower content, generate the nutrients such as the protein of high usage,
And soybean cake dregs compact texture becomes loose without excessive fermentation, palatability significantly improves.
The present invention is prepared in the preferred technical solution of method of pannage, and the prokaryotic micro-organisms includes bafillus natto
And Bacillus acidi lactici；And the eukaryotic microorganisms include S. cervisiae and aspergillus niger.
Above-mentioned bafillus natto (Bacillus natto) abbreviation Bacillus natto belongs to bacterium section, bacillus, is withered grass
A subspecies for bacillus.It can produce a variety of high-quality active antibacterial substances during growth and breeding, such as polyglutamic acid (γ-
PGA) beta -glycosidase, bacitracin, bacteriocin, interferon and microorganism etc..Polyglutamic acid (γ-PGA), also known as Bacillus natto glue,
Polyglutamic acid, it is a kind of water solubility, and biodegrade is free of toxicity, uses boiomacromolecule made from microbe fermentation method.
Aspergillus niger: Deuteromycotina, Hyphomycetes, hyphomycetales, Moniliaceae, a Common Species in aspergillus fungi.Extensively
In the general grain being distributed in all over the world, plant product and soil.
In above-mentioned compound bacteria: bafillus natto (Bacillus natto), Chinese industrial Microbiological Culture Collection management
Center (CICC), bacterium numbering CICC23413；Bacillus acidi lactici (Lactobacillus sp.), Chinese industrial microorganism fungus kind protect
It hides administrative center (CICC), bacterium numbering CICC21007；Saccharomyces cerevisiae (Saccharomyces cerevisiae), Chinese work
Industry Microbiological Culture Collection administrative center (CICC), bacterium numbering CICC1882；Aspergillus niger (Aspergillus) China is common micro-
Biological inoculum preservation administrative center (CGMCC), bacterium numbering CGMCC3.3928.
This programme is fermented by the compound bacteria that bafillus natto, Bacillus acidi lactici, S. cervisiae and aspergillus niger form
The soybean cake dregs of low value, using these four microorganisms growth course own metabolism process, and secretion extracellular protein
Enzyme, amylase, cellulase etc. generate natto rich in a variety of nutrition being easily absorbed by organisms such as amino acid, organic acid, oligosaccharides
Ingredient.Also contain several physiological active substances simultaneously, as Nattokinase, superoxide dismutase, isoflavones, saponin(e element, γ-are more
Polyglutamic acid, tocopherol, vitamin etc..The probiotics such as bacillus and lactic acid bacteria can search for food with feed enters animal gastrointestinal tract,
The microecological balance of bacterium colony in gastrointestinal tract can be effectively adjusted, inhibits harmful bacteria proliferation, improves gastrointestinal function, body is improved and exempts from
Epidemic disease ability and disease resistance.In addition the bioavailability of soybean cake dregs can be improved, can directly be answered effectively to remove anti-nutritional factors
For in pannage, and the protein and amino acid, vitamin and other substances generated after fermenting, it enriches in soybean cake dregs
Nutritional ingredient, the final nutritional quality for improving soybean cake dregs improve soybean cake dregs and are applied to pannage in the intracorporal biology benefit of pig
With rate.Therefore, using protokaryon bacterium and eukaryon fungi to carry out compound strain production pannage can be anti-for improvement dregs of beans quality, reduction
Trophic factors content provides effectively feasible approach.
The present invention is prepared in the preferred technical solution of method of pannage, bafillus natto, lactic acid in the compound bacteria
Bacillus, S. cervisiae and aspergillus niger efficient bacterium number ratio are 1~3:1~3:1~3:1~3.
In the range of aforementioned proportion, preparation-obtained pannage, in the feelings that the inhibiting factors content such as tryptose is minimum
Under condition, free aminoacid content and urease activity are best.
The present invention is prepared in the preferred technical solution of method of pannage, and the compound bacteria is 1 × 10 by concentration7~1 ×
108Bafillus natto, Bacillus acidi lactici, S. cervisiae and the aspergillus niger concentration of CFU/mL mixes.
Wherein, the preparation of each strain in compound bacteria specifically:
Bafillus natto is inoculated in MNB agar solid medium culture by plate streaking mode to activate, from
In choose 3 bafillus natto selected to choose single colonie and be inoculated in MNB fluid nutrient medium, be placed in 180r/min shaking table, 37 DEG C of cultures
12h, and further expansion culture to living bacteria count is 1 × 107~1 × 108CFU/mL；S. cervisiae is lined YPD to consolidate
Culture is therefrom selected S. cervisiae single colonie and is inoculated in YPD fluid nutrient medium, be placed in 180r/ in body culture medium to activate
Min shaking table, 25 DEG C are cultivated for 24 hours, and further expansion culture to living bacteria count is 1 × 107~1 × 108CFU/mL；By lactic acid
Bacterium lines culture on MRS solid medium and therefrom selects lactic acid bacteria single colonie to activate and be inoculated in MRS fluid nutrient medium,
42 DEG C of temperature control, Anaerobic culturel 48h is stood, and further expansion culture to living bacteria count is 1 × 107~1 × 108CFU/mL；It will
Aspergillus niger lines culture on PDA solid medium and therefrom selects aspergillus niger to activate and be inoculated in PDA liquid medium, set
In 180r/min shaking table, 25 DEG C of culture 48h, and further expansion culture to aspergillus niger spore number are 1 × 107~1 × 108CFU/
mL。
The present invention is prepared in the preferred technical solution of method of pannage, and the compound bacteria of the addition is the 8 of dregs of beans weight
~12%.
The present invention is prepared in the preferred technical solution of method of pannage, and the soybean cake dregs are successively crushed, by feed liquid
Than adding compound bacteria fermentation after being diluted with water, sterilizing for 1~3:1.
The present invention is prepared in the preferred technical solution of method of pannage, after soybean cake dregs are crushed through high pressure sterilization 15~
After 25min, compound bacteria fermentation is added, the high pressure is 103.4kPa.
The present invention is prepared in the preferred technical solution of method of pannage, and the fermentation is solid state fermentation, the temperature of fermentation
It is 30~35 DEG C, the time of fermentation is 40~80h.
The present invention is prepared in the preferred technical solution of method of pannage, the tunning obtained after the dregs of beans is fermented
It is crushed after the dry drying of 40~80 DEG C of hot-air flows and obtains pannage.
(3) beneficial effect
The beneficial effects of the present invention are:
The prokaryotic micro-organisms bafillus natto and lactic acid bacteria and the speed of growth that the present invention selects the speed of growth fast are slower
Eukaryotic microorganisms saccharomyces cerevisiae and aspergillus niger, combination ferments to soybean cake dregs as compound strain, to remove soybean cake
Natural anti-nutritional factors in the dregs of rice；Protein, cellulose, lignin, the starch etc. in soybean cake dregs are decomposed into small point simultaneously
Biological polypeptide, amino acid, the monosaccharide etc. of son are conducive to raising animal and soybean cake dregs after fermentation are absorbed and utilized, also, ferments
Remaining Yeast protein in soybean cake dregs provides a kind of good protein source.
Detailed description of the invention
Fig. 1: the resulting pannage sensory evaluation scores of compound bacteria different vaccination ratio in embodiment 1；
Fig. 2: different feed liquid is than resulting pannage sensory evaluation scores in embodiment 2；
Fig. 3: the resulting pannage sensory evaluation scores of different fermentations temperature in embodiment 3；
Fig. 4: different fermentations time resulting pannage sensory evaluation scores in embodiment 4；
Fig. 5: free amino acid total amount in different strains fermentation pannage obtained in embodiment 5；
Fig. 6: urease activity in different strains fermentation pannage obtained in embodiment 5；
Fig. 7: trypsin ihhibitor content in different strains fermentation pannage obtained in embodiment 5.
Specific embodiment
In order to preferably explain the present invention, in order to understand, with reference to the accompanying drawing, by specific embodiment, to this hair
It is bright to be described in detail.
Embodiment 1
The method that fermentation soybean grouts prepare pannage, specifically:
S1: the pretreatment of soybean cake dregs, soybean cake dregs add sterile water first through being ground into powder, according still further to the solid-liquid ratio of 3:1
Afterwards, under 103.4kPa high pressure, temperature is 121 DEG C, and sterilizing dregs of beans is obtained after the 25min that sterilizes.
S2: the preparation of compound bacteria: bafillus natto, Bacillus acidi lactici, S. cervisiae and aspergillus niger are activated respectively and
Obtaining concentration after expansion culture is 1 × 107The bacterium solution of CFU/mL；By bafillus natto liquid, Bacillus acidi lactici liquid, saccharomyces cerevisiae
It is 3:2:1:3 compound bacteria that bacterium solution and aspergillus niger liquid obtain efficient bacterium number ratio after mixing according to a certain volume.
Wherein, the activation and expansion culture of each strain in compound bacteria specifically:
Bafillus natto is inoculated in MNB agar solid medium culture by plate streaking mode to activate, from
In select bafillus natto and choose single colonie and be inoculated in MNB fluid nutrient medium, be placed in 180r/min shaking table, 37 DEG C of culture 12h,
And further expansion culture to living bacteria count is 1 × 107CFU/mL；S. cervisiae is lined in YPD solid medium
Culture is therefrom selected S. cervisiae single colonie and is inoculated in YPD fluid nutrient medium to activate, and is placed in 180r/min shaking table, and 25
DEG C culture for 24 hours, and further expansion culture to living bacteria count be 1 × 107CFU/mLL；Lactic acid bacteria is lined into the training of MRS solid
It supports culture on base therefrom to select lactic acid bacteria single colonie to activate and be inoculated in MRS fluid nutrient medium, 42 DEG C of temperature control, stands anaerobism
48h is cultivated, and further expansion culture to living bacteria count is 1 × 107CFU/mL；Aspergillus niger is lined into PDA solid culture
Culture is therefrom selected aspergillus niger and is inoculated in PDA liquid medium on base to activate, and is placed in 180r/min shaking table, 25 DEG C of cultures
48h, and further expansion culture to aspergillus niger spore number is 1 × 107CFU/mL。
S3: fermentation: the composite bacteria liquid that the S2 for the corresponding percentage that its weight is added in the sterilizing dregs of beans that S1 is obtained is obtained into
Row solid state fermentation, the temperature of fermentation are 32 DEG C, and the time of fermentation is 50h, obtain fermentation liquid after fermentation；
S4: the fermentation liquid that S3 is obtained is crushed after the dry drying of 60 DEG C of hot-air flows and obtains pannage.
Difference to verify compound bacteria inoculum concentration in the present embodiment does following reality to the prepared influence for obtaining pannage
It tests:
The weight percent that the composite bacteria liquid being added in the present embodiment step S3 is dregs of beans is set to 3%, 5%,
7%, 10%, 12%, 15%, six groups of corresponding pannages are prepared according to method described in the present embodiment in other conditions, and
According to " GB/T 14698-2017 feed microscopy " and combine the actual conditions of pannage from color, smell, tactile
Three aspects carry out sensory evaluation scores to pannage referring to table 1.
1 sensory evaluation criteria table of table
As a result as shown in Figure 1, the organoleptic quality of pannage also shows the trend gradually improved with the increase for connecing bacterium ratio,
When inoculum concentration is 10%, the organoleptic quality of pannage is best.With connecing further increasing for bacterium ratio, the sense organ product of pannage
Matter then shows downward trend, therefore selecting the inoculum concentration of compound strain is 10%.
Embodiment 2
The method that fermentation soybean grouts prepare pannage, specifically:
S1: the pretreatment of soybean cake dregs, soybean cake dregs are added sterile first through being ground into powder according still further to certain solid-liquid ratio
After water dilution, under 103.4kPa high pressure, temperature is 121 DEG C, and sterilizing dregs of beans is obtained after the 20min that sterilizes.
S2: the preparation of compound bacteria: bafillus natto, Bacillus acidi lactici, S. cervisiae and aspergillus niger are activated respectively and
Obtaining concentration after expansion culture is 1 × 108The bacterium solution of CFU/mL；By bafillus natto liquid, Bacillus acidi lactici liquid, saccharomyces cerevisiae
Bacterium solution and aspergillus niger liquid obtain efficient bacterium number than the compound bacteria for 2:1:3:2 after mixing according to a certain volume.
Wherein, the activation and expansion culture of each strain in compound bacteria specifically:
Bafillus natto is inoculated in MNB agar solid medium culture by plate streaking mode to activate, from
In select bafillus natto and choose single colonie and be inoculated in MNB fluid nutrient medium, be placed in 180r/min shaking table, 37 DEG C of culture 12h,
And further expansion culture to living bacteria count is 1 × 108CFU/mL；S. cervisiae is lined in YPD solid medium
Culture is therefrom selected S. cervisiae single colonie and is inoculated in YPD fluid nutrient medium to activate, and is placed in 180r/min shaking table, and 25
DEG C culture for 24 hours, and further expansion culture to living bacteria count be 1 × 108CFU/mL；Lactic acid bacteria is lined into the training of MRS solid
It supports culture on base therefrom to select lactic acid bacteria single colonie to activate and be inoculated in MRS fluid nutrient medium, 42 DEG C of temperature control, stands anaerobism
48h is cultivated, and further expansion culture to living bacteria count is 1 × 108CFU/mL；Aspergillus niger is lined into PDA solid culture
Culture is therefrom selected aspergillus niger and is inoculated in PDA liquid medium on base to activate, and is placed in 180r/min shaking table, 25 DEG C of cultures
48h, and further expansion culture to aspergillus niger spore number is 1 × 108CFU/mL。
S3: fermentation: the composite bacteria liquid for 10% S2 of its weight will be added in sterilizing dregs of beans that S1 is obtained obtaining, consolidate
State fermentation, the temperature of fermentation are 30 DEG C, and the time of fermentation is 80h, obtain fermentation liquid after fermentation；
S4: the fermentation liquid that S3 is obtained is crushed after the dry drying of 50 DEG C of hot-air flows and obtains pannage.
By soybean cake dregs powder in the present embodiment step S1 according to solid-liquid ratio be 1:0.5,1:0.8,1:1,1:1.2,1:1.5
Sterile water is added, five groups of corresponding pannages are prepared according to method described in the present embodiment in other conditions, and according to " GB/T
14698-2017 feed microscopy " and the actual conditions of pannage is combined to join in terms of color, smell, tactile three
Sensory evaluation scores are carried out to pannage according to table 1.
As shown in Figure 2, when the ratio of dregs of beans and water is 1:1, the organoleptic quality of pannage is optimal for sensory score result.Add
Water shortage or the organoleptic quality that excessively will affect final products.It may be due to Soluble Monosaccharide, albumen in dregs of beans
The nutrients such as matter, amino acid need to be efficiently used by microorganism in aqueous solution state, if water shortage, nutrient
The amount of dissolution is few, insufficient to provide enough nutriments for the rapid growth of microorganism early period.Amount of water excessively will lead to nutrition
Element is diluted with Fungal biodiversity, and quality is dilute soft after will lead to bean pulp fermentation, is that subsequent drying process brings burden.Therefore,
Select optimal solid-liquid ratio for 1:1.
Embodiment 3
The method that fermentation soybean grouts prepare pannage, specifically:
S1: the pretreatment of soybean cake dregs, soybean cake dregs add sterile water first through being ground into powder, according still further to the solid-liquid ratio of 2:1
After dilution, under 103.4kPa high pressure, temperature is 121 DEG C, and sterilizing dregs of beans is obtained after the 15min that sterilizes.
S2: the preparation of compound bacteria: bafillus natto, Bacillus acidi lactici, S. cervisiae and aspergillus niger are activated respectively and
Obtaining concentration after expansion culture is 5 × 107The bacterium solution of CFU/mL；By bafillus natto liquid, Bacillus acidi lactici liquid, saccharomyces cerevisiae
Bacterium solution and aspergillus niger liquid obtain efficient bacterium number than the compound bacteria for 1:3:2:1 after mixing according to a certain volume.
Wherein, the activation and expansion culture of each strain in compound bacteria specifically:
Bafillus natto is inoculated in MNB agar solid medium culture by plate streaking mode to activate, from
In select bafillus natto and choose single colonie and be inoculated in MNB fluid nutrient medium, be placed in 180r/min shaking table, 37 DEG C of culture 12h,
And further expansion culture to living bacteria count is 5 × 107CFU/mL；S. cervisiae is lined in YPD solid medium
Culture is therefrom selected S. cervisiae single colonie and is inoculated in YPD fluid nutrient medium to activate, and is placed in 180r/min shaking table, and 25
DEG C culture for 24 hours, and further expansion culture to living bacteria count be 5 × 107CFU/mL；Lactic acid bacteria is lined into the training of MRS solid
It supports culture on base therefrom to select lactic acid bacteria single colonie to activate and be inoculated in MRS fluid nutrient medium, 42 DEG C of temperature control, stands anaerobism
48h is cultivated, and further expansion culture to living bacteria count is 5 × 107CFU/mL；Aspergillus niger is lined into PDA solid culture
Culture is therefrom selected aspergillus niger and is inoculated in PDA liquid medium on base to activate, and is placed in 180r/min shaking table, 25 DEG C of cultures
48h, and further expansion culture to aspergillus niger spore number is 5 × 107CFU/mL。
S3: fermentation: the composite bacteria liquid progress solid-state that 8% S2 that its weight is added in sterilizing dregs of beans that S1 is obtained is obtained
Fermentation, the temperature of fermentation are 30 DEG C, and the time of fermentation is 40h, obtain fermentation liquid after fermentation；
S4: the fermentation liquid that S3 is obtained is crushed after the dry drying of 40 DEG C of hot-air flows and obtains pannage.
The temperature of fermentation in the present embodiment step S3 is respectively set in 28 DEG C, 30 DEG C, 33 DEG C, 35 DEG C, 37 DEG C,
Five groups of corresponding pannages are prepared according to method and condition described in the present embodiment in his method and condition, and according to " GB/T
14698-2017 feed microscopy " and the actual conditions of pannage is combined to join in terms of color, smell, tactile three
Sensory evaluation scores are carried out to pannage according to table 1.
As a result as shown in Figure 3, when fermentation temperature is relatively low, the sensory evaluation scores of pannage increase presentation increasing trend with temperature,
And it peaks at 33 DEG C.Hereafter significant downward trend is presented in the organoleptic quality of pannage.Its reason is analyzed, it may
When being relatively low due to fermentation temperature, microorganism slow growth, the particularly disadvantageous growth and breeding in bafillus natto and lactic acid bacteria,
Cause the biomass of both bacterial strains very few, influences the organoleptic quality of pannage.In temperature drift, aspergillus niger and saccharomyces cerevisiae
Growth be suppressed, can equally be unfavorable for the fermentation process of dregs of beans.Therefore, the most suitable fermentation temperature of dregs of beans is 33 DEG C.
Embodiment 4
The method that fermentation soybean grouts prepare pannage, specifically:
S1: the pretreatment of soybean cake dregs, soybean cake dregs add sterile water first through being ground into powder, according still further to the solid-liquid ratio of 1:1
After dilution, under 103.4kPa high pressure, temperature is 121 DEG C, and sterilizing dregs of beans is obtained after the 15~25min that sterilizes.
S2: the preparation of compound bacteria: bafillus natto, Bacillus acidi lactici, S. cervisiae and aspergillus niger are activated respectively and
Obtaining concentration after expansion culture is 3 × 107The bacterium solution of CFU/mL；By bafillus natto liquid, Bacillus acidi lactici liquid, saccharomyces cerevisiae
Bacterium solution and aspergillus niger liquid obtain efficient bacterium number than the compound bacteria for 1:3:1:3 after mixing according to a certain volume.
Wherein, the activation and expansion culture of each strain in compound bacteria specifically:
Bafillus natto is inoculated in MNB agar solid medium culture by plate streaking mode to activate, from
In select bafillus natto and choose single colonie and be inoculated in MNB fluid nutrient medium, be placed in 180r/min shaking table, 37 DEG C of culture 12h,
And further expansion culture to living bacteria count is 3 × 107CFU/mL；S. cervisiae is lined in YPD solid medium
Culture is therefrom selected S. cervisiae single colonie and is inoculated in YPD fluid nutrient medium to activate, and is placed in 180r/min shaking table, and 25
DEG C culture for 24 hours, and further expansion culture to living bacteria count be 3 × 107CFU/mL；Lactic acid bacteria is lined into the training of MRS solid
It supports culture on base therefrom to select lactic acid bacteria single colonie to activate and be inoculated in MRS fluid nutrient medium, 42 DEG C of temperature control, stands anaerobism
48h is cultivated, and further expansion culture to living bacteria count is 3 × 107CFU/mL；Aspergillus niger is lined into PDA solid culture
Culture is therefrom selected aspergillus niger and is inoculated in PDA liquid medium on base to activate, and is placed in 180r/min shaking table, 25 DEG C of cultures
48h, and further expansion culture to aspergillus niger spore number is 3 × 107CFU/mL。
S3: fermentation: the obtained composite bacteria liquid of 10% S2 that its weight is added in sterilizing dregs of beans that S1 is obtained is consolidated
State fermentation, the temperature of fermentation are 33 DEG C, obtain fermentation liquid after fermentation to regular hour；
S4: the fermentation liquid that S3 is obtained is crushed after the dry drying of 80 DEG C of hot-air flows and obtains pannage.
By the time of the fermentation in the present embodiment step S3 be respectively set in 40h, 45h, 50h, 55h, 60h, 65h,
Seven groups of corresponding pannages, and foundation is prepared according to method and condition described in the present embodiment in 70h, other methods and condition
" GB/T14698-2017 feed microscopy " and combine the actual conditions of pannage from color, smell, tactile three
Aspect carries out sensory evaluation scores to pannage referring to table 1.
As a result as shown in Figure 4, since fermentation time is 40h, with the extension of fermentation time, the organoleptic quality of pannage
Also gradually improve therewith.When being 60h between when fermenting, the performance optimal of pannage.But with the further extension of fermentation time,
The organoleptic quality of pannage then gradually declines.During the fermentation, bafillus natto and lactic acid bacteria are prokaryotic micro-organisms, life
Long speed is faster than eukaryotic microorganisms saccharomyces cerevisiae and aspergillus niger.Fermentation time is too short, and saccharomyces cerevisiae can not effectively be sent out with aspergillus niger
Wave its biological function；Fermentation time is too long, and bafillus natto and lactic acid bacteria are gradually changed into disadvantage bacterium, and biomass gradually subtracts
It is few, and the microbial fermentations metabolic waste such as some low molecule aldehyde, ketone, organic acid, amine starts to accumulate in the product, leads to pig
The decline of feed organoleptic quality.Therefore, best fermentation time is 60h.
Embodiment 5
The method that fermentation soybean grouts prepare pannage, specifically:
S1: the pretreatment of soybean cake dregs, soybean cake dregs add sterile water first through being ground into powder, according still further to the solid-liquid ratio of 3:1
After dilution, under 103.4kPa high pressure, temperature is 121 DEG C, and sterilizing dregs of beans is obtained after the 20min that sterilizes.
S2: the preparation of compound bacteria: bafillus natto, Bacillus acidi lactici, S. cervisiae and aspergillus niger are activated respectively and
Obtaining concentration after expansion culture is 1 × 107The bacterium solution of CFU/mL；By bafillus natto liquid, Bacillus acidi lactici liquid, saccharomyces cerevisiae
Bacterium solution and aspergillus niger liquid obtain efficient bacterium number than the compound bacteria for 1:1:1:1 after mixing according to a certain volume.
Wherein, the activation and expansion culture of each strain in compound bacteria specifically:
Bafillus natto is inoculated in MNB agar solid medium culture by plate streaking mode to activate, from
In select bafillus natto and choose single colonie and be inoculated in MNB fluid nutrient medium, be placed in 180r/min shaking table, 37 DEG C of culture 12h,
And further expansion culture to living bacteria count is 1 × 107CFU/mL；S. cervisiae is lined in YPD solid medium
Culture is therefrom selected S. cervisiae single colonie and is inoculated in YPD fluid nutrient medium to activate, and is placed in 180r/min shaking table, and 25
DEG C culture for 24 hours, and further expansion culture to living bacteria count be 1 × 107CFU/mL；Lactic acid bacteria is lined into the training of MRS solid
It supports culture on base therefrom to select lactic acid bacteria single colonie to activate and be inoculated in MRS fluid nutrient medium, 42 DEG C of temperature control, stands anaerobism
48h is cultivated, and further expansion culture to living bacteria count is 1 × 107CFU/mL；Aspergillus niger is lined into PDA solid culture
Culture is therefrom selected aspergillus niger and is inoculated in PDA liquid medium on base to activate, and is placed in 180r/min shaking table, 25 DEG C of cultures
48h, and further expansion culture to aspergillus niger spore number is 1 × 107CFU/mL。
S3: fermentation: the obtained composite bacteria liquid of 10% S2 that its weight is added in sterilizing dregs of beans that S1 is obtained is consolidated
State fermentation, the temperature of fermentation are 33 DEG C, and the time of fermentation is 60h, obtain fermentation liquid after fermentation；
S4: the fermentation liquid that S3 is obtained is crushed after the dry drying of 60 DEG C of hot-air flows and obtains pannage.
The pannage that will be prepared according to present implementation replaces with same concentrations with by the compound bacteria in the present embodiment S3
Corresponding pannage is prepared in bafillus natto, Bacillus acidi lactici, S. cervisiae or aspergillus niger respectively, with the present embodiment
The pannage that method is prepared compares, and composite bacteria liquid is replaced with to the sterile distilled water of identical weight, is prepared
Pannage does blank control.By above-mentioned six groups of pork feed, sealing preservation is in -20 DEG C of refrigerator-freezers for detecting after crushing respectively
The measurement of the measurement of index, each index is specific as follows:
(1) it the measurement of amino acid content: refers to " measurement GB/T18246-2000 of amino acid in feed ", using amino
Sour automatic analyzer measurement.Dregs of beans after fermenting in right amount is weighed, 10mL ultrapure water is dissolved in, through ultrasonic cleaner ultrasonic vibration
After 30min, it is centrifuged 10min under the conditions of 12000rpm, takes supernatant after 0.22 μm of filter membrane filters, is surveyed by amino-acid analyzer
Determine free amino acid composition and total content.
(2) phenylamino (BAPA) the active measurement of trypsin ihhibitor: is replaced using benzoyl-DL-arginine-B- nitro
Detection method measures the trypsin ihhibitor activity in pannage.Trypsin ihhibitor can be combined with trypsase,
By measurement substrate B APA under trypsin digestion effect product-p-nitrophenyl amine aqueous solution absorbance for generating, then with
The difference of the standard solution absorbance of not enzyme inhibiting factor indicates repressed tryptic activity, to press down to trypsase
The activity of the factor processed is evaluated.Solution absorbance value is measured at 385nm (or 410nm), it is specified that it is 1 that light absorption value, which reduces 0.01,
A trypsin ihhibitor unit (TIU).
Trypsin ihhibitor content (mg/g)=[(sample OD385nm- sample blank OD385nm)/0.19×1000]
× extension rate × 50/ sample quality (g)；
(3) measurement of urease activity: referring to the survey of urease activity in GB/T 8622-2006 feed bean product
Determine method.
Measurement result is as shown in Fig. 5,6,7, and compound strain shows bigger advantage of fermenting than single bacterial strain, through compound bacteria
Free aminoacid content after strain fermentation in obtained pannage and control group increase significantly, increase to 8.3% by 5.4%；
And urease activity is also remarkably decreased with trypsin ihhibitor content, urease activity drops to 0.075U/g by 0.64U/g,
Trypsin ihhibitor drops to 0.59mg/g by 3.64mg/g.
Claims (9)
1. a kind of method that fermentation soybean grouts prepare pannage, which is characterized in that it is the following steps are included: by soybean cake dregs
Pannage is obtained after compound bacteria fermentation is added, the compound bacteria includes prokaryotic micro-organisms and eukaryotic microorganisms.
2. the method that fermentation soybean grouts as described in claim 1 prepare pannage, it is characterised in that: the prokaryotic micro-organisms
Including bafillus natto and Bacillus acidi lactici；And the eukaryotic microorganisms include S. cervisiae and aspergillus niger.
3. the method that fermentation soybean grouts as claimed in claim 2 prepare pannage, it is characterised in that: received in the compound bacteria
Beans bacillus, Bacillus acidi lactici, S. cervisiae and the effective bacterium number ratio of aspergillus niger are 1~3:1~3:1~3:1~3.
4. the method that fermentation soybean grouts as described in claim 1 prepare pannage, it is characterised in that: the compound bacteria is by dense
Degree is 1 × 107~1 × 108Bafillus natto, Bacillus acidi lactici, S. cervisiae and the aspergillus niger of CFU/mL mixes.
5. the method that fermentation soybean grouts as described in claim 1 prepare pannage, it is characterised in that: the addition it is compound
Bacterium is the 8~12% of dregs of beans weight.
6. the method that fermentation soybean grouts as described in claim 1 prepare pannage, which is characterized in that the soybean cake dregs according to
It is secondary crushed, be by solid-liquid ratio 1~3:1 be diluted with water, sterilize after add compound bacteria and ferment.
7. the method that fermentation soybean grouts as described in claim 1 prepare pannage, it is characterised in that: crush soybean cake dregs
After 15~25min of high pressure sterilization, compound bacteria fermentation is added, the high pressure is 103.4kPa.
8. the method that fermentation soybean grouts as described in claim 1 prepare pannage, it is characterised in that: the fermentation is solid-state
Fermentation, the temperature of fermentation are 30~35 DEG C, and the time of fermentation is 40~80h.
9. the method that fermentation soybean grouts as described in claim 1 prepare pannage, it is characterised in that: the soybean cake dregs warp
The tunning obtained after fermentation crushes after the dry drying of 40~80 DEG C of hot-air flows and obtains pannage.
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| CN110195020A (en) * | 2019-05-28 | 2019-09-03 | 四川天王牧业有限公司 | A kind of preparation method of feed fermentation agent |
| CN112980900A (en) * | 2021-02-24 | 2021-06-18 | 高邮市龙伟食品科技有限公司 | Method for preparing gamma-polyglutamic acid by using waste bean dregs |
| CN113943767A (en) * | 2021-10-12 | 2022-01-18 | 淮阴工学院 | Method for improving yield of surface active element homolog of bacillus natto by using aspergillus niger fermentation liquid |
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