CN104222493B - A kind of compound benefit bacterium peptide and its preparation method and application - Google Patents
A kind of compound benefit bacterium peptide and its preparation method and application Download PDFInfo
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Abstract
The invention discloses a kind of compound benefit bacterium peptide and its preparation method and application, it is compounded leavening by bacterium enzyme and adds in dregs of beans fermentation to and form, described bacterium enzyme compounds leavening and is composited by four kinds of probios, four kinds of protease and carbon source, and four kinds of probios are respectively clostridium butyricum, bacillus pumilus S3, Lactobacillus plantarum and saccharomyces boulardii；Four kinds of protease are respectively acid protease, neutral proteinase, alkali protease and polypeptide protein enzyme.And provide the concrete preparation method of compound benefit bacterium peptide and for preparing the application in the feed for piglet additive.Described compound benefit bacterium peptide product, rich in multiple probio, effectively can produce the small-molecule substance such as polypeptide, amino acid by degradation antigen albumen, and create local flavor, the Antagonism materials such as multiple probiotic organic acid, biotin during the fermentation, not only increase palatability, security, and significantly improve dregs of beans utilization rate and feed nutritive value, also can significantly reduce breast grice diarrhoea rate.
Description
Technical field
The invention belongs to biological enzymolysis and fermentation arts, relate to the preparation method of peptide, be specifically related to a kind of compound benefit bacterium peptide
And its preparation method and application.
Background technology
Dregs of beans, as a kind of important plant protein source, has important application and researching value.But the albumen in soybean
The existence of the anti-nutrient substances such as enzyme inhibitor, phytolectin, Soybean antigen protein, urase, phytic acid, soyabean oligosaccharides, not only
Greatly hinder animal to the digestion of nutritional labeling, absorb and utilize, and severely compromise the health of animal and grow.
Soybean antigen protein is that in soybean processing goods, residual quantity is the highest, and a kind of Soybean Anti-nutritional Factors that anti-oxidant action is the strongest can
Cause animal generation enteron aisle and the allergic reaction of other organ such as infant and mouse, piglet, calf, and cause intestinal mucosa to damage
Wound, there is damaging change in small intestine, and then it is even dead to cause diarrhoea, production performance to decline, and is universally acknowledged
One of eight big food allergens.Soybean antigen protein mainly includes 11S glycinin and 7S β-soybean companion's globulin, Liang Zhezhan
About the 70% of soybean protein content.
In the bioremediation of current dregs of beans, microbe fermentation method and ferment treatment method are the most universal, the present invention
Four kinds of bacterium, four kinds of enzymes are combined with carbon source, using dregs of beans as a kind of matrix cultivating microorganism, promotes that four kinds of probios are big
Amount breeding, and in biological treatment process, effectively degrades and utilizes dregs of beans big molecular antigen albumen, by it degradable is almost
The small-molecule substance such as polypeptide, amino acid, it is easy to animal absorbs, and creates the local flavor materials such as multiple probiotic organic acid, raw
The Antagonism materials such as thing element, clostridium butyricum element, while adding palatability, security, also significantly improve the egg of dregs of beans
White utilization rate and nutritive value, and can reduce breast grice diarrhoea rate, possessed certain functional.
Summary of the invention
The invention provides a kind of compound benefit bacterium peptide and its preparation method and application, described compound benefit bacterium peptide can meet breast
The piglet special dietary demand to dregs of beans raw material.
For achieving the above object, the present invention is achieved by the following technical solutions:
A kind of compound benefit bacterium peptide, it compounds leavening by bacterium enzyme and adds fermentation in dregs of beans to and form, and described bacterium enzyme is compounding to be sent out
Ferment agent is composited by four kinds of probios, four kinds of protease and carbon source, and described four kinds of probios are respectively clostridium butyricum, short and small bud
Spore bacillus S3, Lactobacillus plantarum and saccharomyces boulardii；Described four kinds of protease are respectively acid protease, neutral proteinase, alkali
Property protease and polypeptide protein enzyme；The Classification And Nomenclature of described bacillus pumilus S3 is bacillus pumilus (Bacillus
Pumilus), its deposit number is: CGMCC No.9161.
Further improvement to technique scheme: the bacteria containing amount that described four kinds of probios compound in leavening at bacterium enzyme divides
Not: clostridium butyricum: be not less than 1 × 106CFU/g；Bacillus pumilus S3: be not less than 1 × 106CFU/g；Lactobacillus plantarum: no
Less than 1 × 106CFU/g；Saccharomyces boulardii: be not less than 1 × 105CFU/g。
Further improvement to technique scheme: described four kinds of protease compound the content in leavening respectively at bacterium enzyme
It is: acid protease: be not less than 500U/g；Neutral proteinase: be not less than 1000U/g；Alkali protease: be not less than 4000U/g；
Polypeptide protein enzyme: be not less than 4500U/g.
Further improvement to technique scheme: described carbon source is to account for bacterium enzyme to compound the Portugal of leavening quality 9%-10%
Grape sugar.
Present invention also offers the preparation method of described compound benefit bacterium peptide, it comprises the steps:
(1) in dregs of beans, interpolation accounts for the water formation fermentation bed material of Soybean Meal 50-60%, and inoculation accounts for Soybean Meal 10%
Described bacterium enzyme compounds leavening, mixes formation fermentation materials；
(2) described fermentation materials is stored in tinfoil paper seal in bag, in 200 DEG C of sealings, is placed in 37 DEG C of constant incubators,
Fermentation 72h；
(3) after having fermented, low temperature drying, pulverize and sieve, form described compound benefit bacterium peptide.
Further improvement to technique scheme: in described step (3), fermentation materials is in 40 DEG C of low temperature dryings, pulverizes
60 mesh sieves.
Invention further provides described compound benefit bacterium peptide in preparing the feed addictive for feeding breast piglet
Application.
Described compound benefit bacterium peptide adds in feed to account for feeding quality than the ratio as 5-15%, described compound benefit bacterium peptide
Antagonism pig manure coliform.
Compared with prior art, the advantage of the inventive method and having the technical effect that
1, the bacterium enzyme used by the present invention compounds leavening, is in biological enzymolysis fermentation theoretical foundation, in conjunction with putting into practice for many years
Experience, for producing conventional dregs of beans material characteristic, the selectivity leavening prepared voluntarily；Selectivity bacterium enzyme described in employing compounds
Leavening, biological treatment dregs of beans prepares compound benefit bacterium peptide product.
2, the compound benefit bacterium peptide product prepared by the present invention, rich in multiple probio, including clostridium butyricum, short and small gemma bar
Bacterium S3, Lactobacillus plantarum, saccharomyces boulardii.During its enzymatic hydrolysis and fermentation, in dregs of beans of can effectively degrading, antigen protein produces many
The small-molecule substance such as peptide, amino acid, and produce local flavor, the Antagonism materials such as multiple probiotic organic acid, biotin, not only increase
Add palatability, security, and gemma class probio has had good storage stability.
3, the compound benefit bacterium peptide product prepared by the present invention, is remarkably improved dregs of beans utilization rate and feed nutritive value, tool
There is pig manure coliform resistance, also can significantly reduce grice diarrhoea rate.
4, bacterium involved in the present invention, enzyme etc., broadly fall into feed addictive category, wide material sources, and safety is easy to get, low cost
Honest and clean.
5, the method for the invention simple and reasonable steps, swift to operate, there is good industrializing implementation prospect.
After reading in conjunction with the accompanying the detailed description of the invention of the present invention, other advantages of the present invention will become clearer from.
Accompanying drawing explanation
Fig. 1 shows that No. 1 dregs of beans compound benefit bacterium peptide-probiotics bacterial amount composition of preparation after biological treatment is stable with half a year
Property.
Fig. 2 shows that No. 2 dregs of beans compound benefit bacterium peptide-probiotics bacterial amount composition of preparation after biological treatment is stable with half a year
Property.
Fig. 3 shows that dregs of beans raw material is prepared as the antigen protein SDS-PAGE of compound benefit bacterium peptide after biological treatment.
Fig. 4 shows that dregs of beans raw material is prepared as compound benefit bacterium peptide rich in probiotic organic acid after biological treatment.
Fig. 5 shows that dregs of beans raw material is prepared as compound benefit bacterium peptide after biological treatment can antagonism pig manure coliform.
Fig. 6 shows that dregs of beans raw material is prepared as compound benefit bacterium peptide after biological treatment and can significantly reduce breast grice diarrhoea rate.
Fig. 7 is the production technological process of bacillus pumilus S3.
Detailed description of the invention
For being best understood from the function of this compound benefit bacterium peptide product, below in conjunction with the accompanying drawings with specific embodiment to the present invention's
Product is quoted feature and is further described in detail.
Embodiment 1
One, the separation screening of bacillus pumilus S3
In May, 2013, from Jiangzhou, coastal many places gathered the water sample in shrimp aquaculture region, bed mud, gut of shrimp in September, 2013
Ight soil.Bed mud, gut of shrimp fecal specimens be 10 times of dilutions in SPSS, are fully ground, and taking supernatant is sample
Liquid；Water sample is directly as sample liquid；Sample liquid is inoculated in respectively in 2216E fluid nutrient medium and dilutes after 30 DEG C of enrichment culture 48h,
Coat and 2216E flat board carries out the separation of bacterial strain, purifying, filter out the 26 pure bacterium of strain altogether.
Having purified single strain uses flat board dibbling method to screen the bacterium that flys up and down.With vibrio parahaemolytious as indicator bacteria, by pair haemolysis arc
Bacterium is inoculated in 2216E fluid nutrient medium cultivation 18h, and dilution is prepared as 105-106The bacteria suspension of CFU/mL, takes 0.1mL coating
2216E flat board, standby.With sterile toothpick picking bacterial strain to be screened, point is connected to containing in bacterium flat board, and 25 DEG C of incubated 24h see
Examine and a little connect whether periphery of bacterial colonies obvious inhibition zone or the area of coverage occurs.
Use dibbling method to screen 3 strains the 26 strain bacteriums being separated to and vibrio parahaemolytious had the bacterial strain of antagonistic action,
Wherein S3 has stronger antagonistic action to vibrio parahaemolytious, can substantially suppress the growth of pathogen, produces around pathogen
Inhibition zone the most clearly.
Through Bacterial Physiological biochemical method Preliminary Identification, bacterial strain S3 is bacillus, its biological characteristics: Gram's staining
The positive, the colony diameter about 2mm formed on nutrient agar panel, neat in edge is smooth, middle fold, milky.Thalline shape
State is shaft-like, and gemma is oval, middle life.
Two, the molecular genetics taxonomic identification of bacillus pumilus
First extracting the DNA of bacterial strain S3, with this DNA as template, 16S rRNA universal primer is primer, to 16SrRNA sheet
Duan Jinhang expands, and amplified fragments carries out sequencing.Sequencing result belongs to, with Blast software is relevant to GenBank, the 16S planted
RRNA sequence is compared, and result shows in this bacterial strain 16S rRNA sequence and GenBank gene pool in bacillus
The 16s rRNA sequence homology degree of Bacillus pumilus is the highest, and homology reaches 99%.By DNAMAN6.0 pair
In Genbank, the 16S rRNA sequence of existing bacillus carries out phylogenetic analysis, and result shows, bacterial strain S316S
RRNA with Bacillus pumilus homology is the highest, it is determined that this bacterial strain is the bacillus pumilus of bacillus.
The bacterial strain S3 screened is carried out culture presevation, depositary institution: China Committee for Culture Collection of Microorganisms is general
Logical microorganism center (CGMCC)；Address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica；
Preservation date: on May 13rd, 2014；The deposit number of bacillus pumilus Bacillus pumilus is CGMCC No.9161.
Three, the composition of compound benefit bacterium peptide
Compound benefit bacterium peptide of the present invention is compounded leavening by bacterium enzyme to be added in dregs of beans fermentation to and forms, and described bacterium enzyme compounds
Leavening is composited by four kinds of probios, four kinds of protease and carbon source, and described four kinds of probios are respectively clostridium butyricum, short and small
Bacillus S3, Lactobacillus plantarum, saccharomyces boulardii；Described four kinds of protease are respectively acid protease, neutral proteinase, alkali
Property protease, polypeptide protein enzyme；The Classification And Nomenclature of described bacillus pumilus S3 is bacillus pumilus (Bacillus
Pumilus), its deposit number is: CGMCC No.9161.Described carbon source is to account for bacterium enzyme to compound leavening quality 9%-10%
Glucose, the glucose crop carrier of interpolation and carbon source, to meet the growth demand of various bacterium, and promote lactic acid, acetic acid, butyric acid
Generation etc. probiotic organic acid.
Described four kinds of probios compound the bacteria containing amount in leavening respectively at bacterium enzyme: clostridium butyricum: it is not less than 1 ×
106CFU/g；Bacillus pumilus S3: be not less than 1 × 106CFU/g；Lactobacillus plantarum: be not less than 1 × 106CFU/g；Bradley ferment
Female bacterium: be not less than 1 × 105CFU/g.Described four kinds of protease compound the content in leavening respectively at bacterium enzyme: acidic protein
Enzyme: be not less than 500U/g；Neutral proteinase: be not less than 1000U/g；Alkali protease: be not less than 4000U/g；Polypeptide protein enzyme:
It is not less than 4500U/g.
Clostridium butyricum used in the present invention, Lactobacillus plantarum and saccharomyces boulardii are the bacterium powder of commercial goods, institute
Acid protease, neutral proteinase, alkali protease and the polypeptide protein enzyme used is the enzyme preparation of commercial goods, and institute
Stating bacillus pumilus S3 from row filter and to preserve, its technological process of production is as shown in Figure 7.
Four, the preparation method of compound benefit bacterium peptide of the present invention comprises the steps:
(1) in dregs of beans, interpolation accounts for the water formation fermentation bed material of Soybean Meal 50-60%, and inoculation accounts for Soybean Meal 10%
Described bacterium enzyme compounds leavening, mixes formation fermentation materials；
(2) described fermentation materials is stored in tinfoil paper seal in bag, in 200 DEG C of sealings, is placed in 37 DEG C of constant incubators,
Fermentation 72h.
In described step (2), fermentation materials is in and fully seals in environment, and in front 24h, in material, still remaining has oxygen, good
The bacillus pumilus S3 of oxygen mushrooms out with facultative aerobic saccharomyces boulardii；During 24h-48h, along with the continuous consumption of oxygen
With the gradually generation of other gases, amphimicrobian Lactobacillus plantarum gradually occupies advantage, and saccharomycete bacterium also progresses into fermentation
Pattern, can produce around here the multiple probiotic organic acid such as lactic acid, acetic acid；During 48h-72h, after oxygen is completely depleted, detest completely
Oxygen bacterium clostridium butyricum gradually occupies advantage, and secretes butyric acid, is conducive to improving animal intestinal.Under totally sealed environment, 72h
Inside achieve the relay growth of four kinds of characteristic probios, multiple probiotic material can be produced.
In described step (2), fermentation materials is in and fully seals in environment, bacterium dominant growths different in different time sections,
Environment pH is caused also to gradually change, the trend essentially reduced.When early stage pH is higher than 5, alkali protease and polypeptide protein
Enzyme can play certain enzymolysis, after later stage pH is less than 5, is mainly played biodegradable merit by neutral proteinase with acid protease
Energy.The restriction enzyme site of four kinds of enzymes is different, can effective degradation antigen albumen, sour molten albumen, alkaline hydrolysis protein ratio are greatly improved,
Increase free aminoacid content, improve the external protein digestibility of product.
In described step (2), fermentation materials is in and fully seals in environment, and pH is gradually lowered, and four kinds of enzymes alternately work,
And restriction enzyme site is different, can effective degradation antigen albumen, sour molten albumen, alkaline hydrolysis protein ratio are greatly improved, increase free
Amino acid content, improves the external protein digestibility of product.
(3) after having fermented, in 40 DEG C of low temperature dryings, pulverize 60 mesh sieves, form compound benefit bacterium peptide product；
In described step (3), before and after low temperature drying processes, final compound benefit bacterium peptide product compared with fermentation wet feed, fourth
Acid clostridium, bacillus pumilus S3, Lactobacillus plantarum, saccharomyces boulardii etc. have a certain proportion of survival, Bacillus survival rate
Higher than Viable detection under room temperature condition in 89%, and half a year higher than 90%.
Described compound benefit bacterium peptide product is rich in probio, organic acid etc., and has miscellaneous bacteria resistance, joins for the feed for piglet
System, adds with 5-15% ratio, can significantly reduce grice diarrhoea rate.
Described compound benefit bacterium peptide product, possesses certain pig manure coliform resistance.
With the method for the invention, No. 1 is given birth to No. 2 two kinds of dregs of beans raw materials (being commercially available common dregs of beans raw material)
Thing processes, and obtains No. 1 beneficial bacterium peptides compound with No. 2 two kinds.
Embodiment 2
When measuring fermentation 72h respectively, described compound benefit bacterium peptide product, room temperature after fermentation termination wet feed, low temperature drying are deposited
The content of four kinds of bacterium in compound benefit bacterium peptide product after June.Utilize tryptone-sulphite-seromycin culture medium, use interlayer
Cultivation, measures clostridium butyricum bacterium amount；Utilize nutrient agar, use method of dilution butteron on plate, measure bacillus pumilus S3
Bacterium amount；Utilize MRS culture medium, use pour plate method, measure Lactobacillus plantarum bacterium amount；Utilize YPD culture medium, use dilution to be coated with
Cloth method, measures saccharomyces boulardii bacterium amount.In view of four kinds of bacterium bacterium amounts differ greatly, represent bacterium amount with log CFU/g.
It is stable with half a year that Fig. 1 illustrates No. 1 dregs of beans compound benefit bacterium peptide-probiotics bacterial amount composition of preparation after biological treatment
Property, Fig. 2 illustrates compound benefit bacterium peptide-probiotics bacterial amount composition and stability half a year of No. 2 dregs of beans preparation after biological treatment.Low
Before and after temperature drying and processing, final compound benefit bacterium peptide product has a certain proportion of survival, Qi Zhongya with each bacterium in fermentation wet feed
After spore bacterium drying survival rate is higher than depositing half a year under 89%, and room temperature condition, Viable detection is higher than 90%.No. 1 compound benefit bacterium
In the fermentation wet feed of peptide, clostridium butyricum bacterium amount reaches 1.08 × 106CFU/g, after low temperature drying, containing clostridium butyricum 9.7 × 105CFU/
G, bacterium amount after survival rate reaches 89.81%, and half a year of drying still keeps 8.8 × 105CFU/g, survival rate reaches 90.72%, and this is described
In product, clostridium butyricum heat resistance is strong, and good stability which greatly improves the possibility that this bacterium plays a role in animal body；
Bacillus pumilus S3 is similar with clostridium butyricum, and in fermentation wet feed, bacterium amount reaches 4.5 × 107CFU/g, after low temperature drying, bacterium amount reaches
4.2×107CFU/g, after drying, survival rate reaches 93.34%, and after room temperature deposits half a year, bacterium amount still keeps 4 × 107CFU/g, viable bacteria is deposited
Motility rate reaches 95.24%, illustrates that its spore is allowed a choice through drying course, and tolerance is higher, has preferable stability, can increase
The application potential of this product strong；In the fermentation wet feed of No. 1 compound benefit bacterium peptide, Lactobacillus plantarum bacterium amount reaches 3.3 × 107CFU/g,
Declining 1 order of magnitude in drying course, bacterium amount is 3.83 × 106CFU/g, after room temperature deposits half a year, still keep 1.96 ×
106CFU/g, the viable bacteria bacterium amount order of magnitude is unchanged, and survival rate reaches 51.17%, illustrates in compound benefit bacterium peptide product containing more plant
Lactobacillus, though having loss but being resistant to the storage of certain time；In the fermentation wet feed of No. 1 compound benefit bacterium peptide, saccharomyces boulardii amount
Up to 7.5 × 105CFU/g, but its tolerance is poor, and in low temperature drying milling product, bacterium amount reaches 3.5 × 104CFU/g, room temperature is deposited
Put saccharomycete only up to 4.1 × 10 after half a year3CFU/g.In No. 2 compound benefit bacterium peptides, the content of each bacterium is survived before and after ratio, drying
Rate, room temperature deposit before and after half a year the indexs such as Viable detection, similar with No. 1 compound benefit bacterium peptide, but its Lactobacillus plantarum bacterium amount
Slightly higher, other bacterium bacterium amounts are lower slightly.
Embodiment 3
Utilize the qualitative degraded situation determining antigen protein of SDS-PAGE (polyacrylamide gel electrophoresis), utilize ELISA
(enzyme-linked immunosorbent assay) kit measures the clearance of antigen protein.TCA (trichloroacetic acid extraction) is utilized to survey
Fixed sour molten albumen (accounting for crude protein, %)；With reference to GB/T 19541-2004 feed Soybean Meal, 0.2%KOH is utilized to measure KOH molten
Xie Du, measures with reference to GB GBT-8314-2002 free amino acid total amount, measures free aminoacid content；Utilize agriculture section of China
SDS-II Bionic digestive system for monogastric animals of Beijing animal and veterinary research institute of institute research and development, the external protein digestibility of simulated determination.
The SDS-PAGE qualitative determination result of antigen protein is as it is shown on figure 3, ELISA quantified results is as shown in table 1.1
Number from the band depth slightly difference of No. 2 different protein protomers of dregs of beans raw material, glycinin, β-soybean companion globulin, antigen
The concrete measured value of albumen is also slightly different, but dregs of beans raw material and biological treatment after compound benefit bacterium peptide product between contrast become
Gesture is similar: after biological treatment, and the big molecular antigen albumen of compound benefit bacterium peptide is the most degradable for small molecular protein (egg
White molecular weight is less than 14KDa), from No. 1 with No. 2 dregs of beans raw materials to compound benefit bacterium peptide product, the antigen egg that ELISA kit measures
White clearance also respectively reaches 91.98% and 93.21%.
After table 2 shows that two kinds of dregs of beans are treated and is prepared as compound benefit bacterium peptide product, sour molten albumen (accounting for crude protein, %),
The change of the indexs such as KOH solubility, free amino acid, nonruminant external Bionic digestion rate.From No. 1 with No. 2 dregs of beans raw materials to
Compound benefit bacterium peptide product, the amplification of sour molten albumen/cp is respectively 27.64% and 29.15%, and the amplification of KOH solubility is respectively
14.13% and 16.81%, the amplification of free amino acid is respectively 9.34% and 9.26%, and the amplification of external protein digestibility is divided
It is not 13.82% and 13.51%.The change of these indexs, mainly has benefited from selectivity bacterium enzyme and compounds the leavening life to dregs of beans
Thing enzymolysis and fermentation process, also the most degradable with high molecular weight protein before corresponding for small molecular protein result of the test.
This compound benefit bacterium peptide product is for the preparation of breast baby pig feedstuff, advantageously in absorption and the digestion of protide nutrition, and can be effective
Avoid the irritability reaction that big molecular antigen albumen causes.
Table 1 shows that dregs of beans raw material is prepared as the antigen protein ELISA kit mensuration of compound benefit bacterium peptide after biological treatment
Result
Table 2 shows that dregs of beans raw material is prepared as the compound benefit molten albumen of acid of bacterium peptide, KOH solubility after biological treatment, dissociates
The index changes such as amino acid and external protein digestibility
Embodiment 4
With the method for the invention, No. 1 is carried out a biological disposal upon with No. 2 two kinds of dregs of beans raw materials, obtain No. 1 and No. 2 two kinds
Compound benefit bacterium peptide.Use HPLC high performance liquid chromatography, the lactic acid in product, acetic acid, butyric acid content are analyzed.
Fig. 4 shows that dregs of beans raw material is prepared as compound benefit bacterium peptide rich in probiotic organic acid after biological treatment.Lactic acid is one
Plant important antimicrobial compound, the antagonism of pathogenic bacteria is played an important role, also there is promotion growth of animal, extend feed guarantor
The matter phase, prevent the effect of feed mold.The lactic acid contents of No. 2 compound benefit bacterium peptides are slightly above No. 1 compound benefit bacterium peptide, this result and its
Lactobacillus plantarum bacterium amount is slightly higher corresponding.In two kinds of compound benefit bacterium peptide products, certain density acetic acid is had to produce, adjustable ring
Border pH, and form the extraordinary local flavor of product.Butyric acid is specifically produced by clostridium butyricum, it is possible to decrease hindgut pH, simultaneously Competitive assays
The growth of harmful clostridium such as clostridieum welchii.Additionally, butyric acid can also promote large intestine and intestinal epithelial cell breeding, there are reparation intestines
Road mucous membrane, the function of promotion immunity.The compound benefit bacterium peptide relative to No. 2, containing more butyric acid shuttle in No. 1 compound benefit bacterium peptide product
Bacterium, thus also create more butyric acid.These probiotic organic acids, can promote that benefit materials absorbs, and product is had
There are the complex functions such as germ antagonism, intestinal regeneration, Immune-enhancing effect.
Embodiment 5
With the method for the invention, No. 1 is carried out a biological disposal upon with No. 2 two kinds of dregs of beans raw materials, obtain No. 1 and No. 2 two kinds
Compound benefit bacterium peptide.On the methylene blue culture medium of Yihong, with the pig manure coliform of even spread for target bacterium, with dregs of beans raw material with multiple
The monogastric animal bionic digestion simulated solution closing benefit bacterium peptide product is antagonism comparative group, before and after investigating biological treatment, if having anti-
Bacterium property material produces.
The result of Antagonism contrast test is as it is shown in figure 5, show that dregs of beans raw material is prepared as compound benefit bacterium after biological treatment
Peptide can antagonism pig manure coliform.Left figure shows that No. 1 compound benefit bacterium peptide and No. 1 dregs of beans raw material are compared, and inhibition zone, by not having, increases
For 29+0.4mm, amplification is notable；Right figure shows that No. 2 compound benefit bacterium peptides and No. 2 dregs of beans raw materials are compared, and inhibition zone, by not having, increases
For 27+0.3mm, amplification also ratio is more significant.These antibacterial material, mainly include butyric acid and clostridium butyricum that clostridium butyricum secretes
The materials such as the lactic acid that the bacteriocin of element, bacillus pumilus S3 secretion, Lactobacillus plantarum are secreted.Initial guess, this compound benefit bacterium
Peptide product is applied in the feed for piglet, can reach the purpose of the pathogenic microorganism growth and breedings such as suppression coliform.
Embodiment 6
With the method for the invention, No. 1 is carried out a biological disposal upon with No. 2 two kinds of dregs of beans raw materials, obtain No. 1 and No. 2 two kinds
Compound benefit bacterium peptide.On on May 10th, 2014 to June 13, in plant of Hong Lan village, villages and small towns, Pingdu City south, Qingdao, 30 ages in days are broken
Milk piglet, carries out piglet culture efficiency contrast test.Control group uses the piglet dregs of beans corn type that in Weifang, base feed factory provides
Basal diet；Two test group are combined benefit bacterium peptide product Substitution for Soybean Meal corn type basal diets with No. 1 of 10% with No. 2 respectively；
With diarrhea rate as index, before and after investigating biological treatment, if can effectively alleviate grice diarrhoea.
Zooperal result as shown in Figure 6, shows that dregs of beans raw material is prepared as compound benefit bacterium peptide after biological treatment, can show
Write and reduce breast piglet diarrhea of pigs rate.Control group piglet feeds common dregs of beans corn type basal diet, containing more antigen egg
In vain, the ANFs such as trypsin inhibitor, easily cause the stress reactions such as breast grice diarrhoea, in this test, control group is suffered from diarrhoea
Rate reaches 19.06%+0.25%.In two test group, substitute basis day with No. 1 of 10% with No. 2 compound benefit bacterium peptide products respectively
Grain, can significantly reduce the ratio of normal meal, reduces the consumption of the ANFs such as big molecular antigen, and rich in clostridium butyricum,
The probios such as bacillus pumilus, the probiotic organic acid such as butyric acid, lactic acid, can antagonism germ, reduce the stress reaction such as diarrhoea
Occurring, result displays that, two test group the most do not occur grice diarrhoea phenomenon.The method of the invention, can be to dregs of beans raw material
The ANFs such as middle antigen protein carry out the most biodegradable, and rich in multiple prebiotic substance, are prepared as feed product and feed
After piglet, grice diarrhoea rate can be significantly reduced.
Above example is only in order to illustrate technical scheme, rather than is limited；Although with reference to aforementioned reality
Execute example the present invention has been described in detail, for the person of ordinary skill of the art, still can be to aforementioned enforcement
Technical scheme described in example is modified, or wherein portion of techniques feature is carried out equivalent；And these are revised or replace
Change, do not make the essence of appropriate technical solution depart from the spirit and scope of claimed technical solution of the invention.
Claims (7)
1. a compound benefit bacterium peptide, it is characterised in that it is compounded leavening by bacterium enzyme and adds fermentation in dregs of beans to and form, described bacterium
Enzyme compounds leavening and is composited by four kinds of probios, four kinds of protease and carbon source, and described four kinds of probios are respectively butyric acid shuttle
Bacterium, bacillus pumilus S3, Lactobacillus plantarum and saccharomyces boulardii；Described four kinds of protease are respectively acid protease, neutrality
Protease, alkali protease and polypeptide protein enzyme；The Classification And Nomenclature of described bacillus pumilus S3 is bacillus pumilus
(Bacillus pumilus), its deposit number is: CGMCC No. 9161；
Described four kinds of probios compound the bacteria containing amount in leavening respectively at bacterium enzyme: clostridium butyricum: be not less than 1 × 106 CFU/
g；Bacillus pumilus S3: be not less than 1 × 106CFU/g；Lactobacillus plantarum: be not less than 1 × 106CFU/g；Saccharomyces boulardii:
It is not less than 1 × 105CFU/g；
Described four kinds of protease compound the content in leavening respectively at bacterium enzyme: acid protease: be not less than 500U/g；Neutral
Protease: be not less than 1000U/g；Alkali protease: be not less than 4000U/g；Polypeptide protein enzyme: be not less than 4500U/g.
Compound benefit bacterium peptide the most according to claim 1, it is characterised in that: described carbon source compounds leavening quality for accounting for bacterium enzyme
The glucose of 9%-10%.
3. the preparation method of the compound benefit bacterium peptide described in any one of claim 1-2, it is characterised in that it comprises the steps:
(1) in dregs of beans, interpolation accounts for the water formation fermentation bed material of Soybean Meal 50-60%, and inoculation accounts for the described of Soybean Meal 10%
Bacterium enzyme compounds leavening, mixes formation fermentation materials；
(2) described fermentation materials is stored in tinfoil paper seal in bag, in 200 DEG C of sealings, is placed in 37 DEG C of constant incubators, send out
Ferment 72h；
(3) after having fermented, low temperature drying, pulverize and sieve, form described compound benefit bacterium peptide.
The preparation method of compound benefit bacterium peptide the most according to claim 3, it is characterised in that: fermentate in described step (3)
Expect, in 40 DEG C of low temperature dryings, to pulverize 60 mesh sieves.
5. the compound benefit bacterium peptide described in any one of claim 1-2 is preparation answering in the feed addictive feeding breast piglet
With.
Compound benefit bacterium peptide the most according to claim 5 is used for the application feeding in the feed addictive of breast piglet in preparation,
It is characterized in that: described compound benefit bacterium peptide adds in feed to account for feeding quality than the ratio as 5-15%.
Compound benefit bacterium peptide the most according to claim 5 is used for the application feeding in the feed addictive of breast piglet in preparation,
It is characterized in that: described compound benefit bacterium peptide antagonism pig manure coliform.
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| CN104872379B (en) * | 2015-05-18 | 2018-07-24 | 青岛尚德生物技术有限公司 | A kind of fermentate and its fermentation process adding the molten slurry of fish in corn-soybean meal diet |
| CN106922948A (en) * | 2015-12-29 | 2017-07-07 | 丰益(上海)生物技术研发中心有限公司 | The production technology of fermented bean dregs is carried out suitable for edible fat production factory environment |
| CN105876154A (en) * | 2016-04-22 | 2016-08-24 | 宜昌市昌伟饲料科技有限公司 | Antibiotic-free feed for livestock and poultry and production method thereof |
| CN106472932A (en) * | 2016-10-18 | 2017-03-08 | 李德田 | A kind of complex microorganism beverage and preparation method thereof |
| CN106962609A (en) * | 2017-04-01 | 2017-07-21 | 福建龙岩闽雄生物科技股份有限公司 | A kind of direct putting type biological fermentation feed additive and its production and use |
| CN110804564B (en) * | 2019-11-18 | 2020-10-09 | 江苏三仪生物工程有限公司 | Preparation of clostridium butyricum powder and detection culture medium thereof |
| CN114617190A (en) * | 2022-03-15 | 2022-06-14 | 广东海洋大学 | Functional feed protein prepared from euphausia superba and method thereof |
| CN114617191A (en) * | 2022-03-15 | 2022-06-14 | 广东海洋大学 | Functional feed prepared from euphausia superba and method thereof |
| WO2023175153A1 (en) * | 2022-03-18 | 2023-09-21 | Fumi Ingredients B.V. | A microbial cell extract, method for obtaining said microbial cell extract and use of said microbial cell extract |
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