CN107319129A - A kind of preparation of fermented bean dregs rich in astaxanthin and feed - Google Patents
A kind of preparation of fermented bean dregs rich in astaxanthin and feed Download PDFInfo
- Publication number
- CN107319129A CN107319129A CN201710355273.1A CN201710355273A CN107319129A CN 107319129 A CN107319129 A CN 107319129A CN 201710355273 A CN201710355273 A CN 201710355273A CN 107319129 A CN107319129 A CN 107319129A
- Authority
- CN
- China
- Prior art keywords
- astaxanthin
- fermented bean
- bean dregs
- rich
- dregs
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
- A23K10/37—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms from waste material
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K40/00—Shaping or working-up of animal feeding-stuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K40/00—Shaping or working-up of animal feeding-stuffs
- A23K40/10—Shaping or working-up of animal feeding-stuffs by agglomeration; by granulation, e.g. making powders
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/30—Feeding-stuffs specially adapted for particular animals for swines
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/60—Feeding-stuffs specially adapted for particular animals for weanlings
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
Abstract
The invention discloses a kind of preparation and application method of the fermented bean dregs rich in astaxanthin.The present invention is using ocean rhodotorula as principal leavening agent, using dregs of beans as primary raw material, by steps such as the activation of zymotic fluid, low moisture fermentation and after-ripening, mixing, granulations, prepares fermented bean dregs and its feed product rich in astaxanthin.The technological process of the present invention is easy to operate; Feed Manufacturing, processing link can be directly entered; reduce the energy consumption of drying process; reduce processing cost; and the prebiotic factor such as the astaxanthin produced during protected fermentation, fermented bean dregs transit link is reduced, is easy to the application and popularization of feed product; both it is adapted to feed factory large-scale production, also is adapted for scale raising directly production and feeds.The beneficial effects of the invention are as follows：The nutritional need of weanling pig, milking sow is met, the immunity of animal is improved, the generation of grice diarrhoea is reduced, improves the milk water quality of sow, and then improve the production performance of suckling pig.
Description
Technical field
The invention belongs to technical field of microbial fermentation, field of feed is also related to, and in particular to one kind is rich in
The preparation of the fermented bean dregs of astaxanthin and feed.
Background technology
With developing rapidly for aquaculture, the demand to protein raw materials increases increasingly.And high-quality protein raw material such as fish meal etc.
By raw material sources, the influence of many factors such as market supply, supply is increasingly not enough.The new protein sources for substituting fish meal are found, it is special
It is not to utilize the cheap some or all of replacement fish meal of plant protein source.In numerous plant protein sources, dregs of beans is due to egg
White matter content height, affordable, supply are sufficient and turn into best selection, as the base stock for preparing livestock and poultry feed.
Fermented bean dregs overcome weakness of the dregs of beans albumen in nutrition using biofermentation zymolysis technique, by beneficial micro-
Biological agent improves the solubility of dregs of beans albumen, beneficial to digestion；Reduce the molecular weight of albumen in dregs of beans, a part therein
Small peptide level even amino acid levels have been reached, directly can have been absorbed by animal；Fermented bean dregs have certain aromatic odor and
Delicate flavour, there is certain attractant effect, and palatability is preferable；And the non-starch polysaccharide of some in dregs of beans is obtained during the fermentation
Decompose, be conducive to animal digestion.Fermented bean dregs have been widely used in feedstuff industry, as the optimal selection for substituting fish meal.
Strain is developed and zymolysis technique, directly affects the quality of fermented bean dregs.The conventional strain of fermented bean dregs is withered grass bud
Spore bacillus, bacillus licheniformis, Lactobacillus plantarum, saccharomycete etc..Ocean rhodotorula refers to saccharomycete present in nature marine site,
Its thalline is nutritious and comprehensive, rich in protein, glycogen, unrighted acid, animal larval growth hormone and with astaxanthin
Based on carotenoid, be that excellent astaxanthin (astaxanthin) production bacterium, natural pigment source, the animal of great potential are raised
Expect albumen and food additives.Astaxanthin is 3,3 '-dihydroxy -4,4 '-diketo-β, β '-carrotene, is terpenes insatiable hunger
And compound, with very strong scavenging activated oxygen, promote antibody generation, increase host immunity and gorgeous red work
With.The maximum market of astaxanthin is culture fishery and poultry farming, be mainly used as the shellfish such as fish and shrimp crab and
The feed addictive of poultry, can improve the nutrient digestibility and immunity of piglet, reduce the generation of diarrhoea, and then improve piglet
Production performance, and the daily gain of the feed intake of milking sow, milk water quality and suckling pig can be improved.Strengthen research and development
The application of astaxanthin and astaxanthin product in pig field has boundless prospect.
The content of the invention
The present invention discloses a kind of fermented bean dregs rich in astaxanthin, its preparation method and feed, and the preparation method is simple, have
Effect ground improves dregs of beans nutrient composition content, especially improves the content of astaxanthin in fermented bean dregs.
A kind of preparation method technical scheme of fermented bean dregs rich in astaxanthin of the present invention is as follows：
1) the ocean rhodotorula zymotic fluid rich in astaxanthin is prepared；
2) composite zymocyte liquid used in cooperative fermentation is prepared；
3) fermentation substrate is prepared；
4) by step 2) obtained by composite zymocyte liquid be seeded to step 3) in fermentation substrate in, and adjust aqueous
Amount, is well mixed；
5) by step 4) inoculation bacterium solution after fermentation substrate fermented under conditions of temperature control, ferment completion produce fermentation
Dregs of beans.
Wherein, ocean rhodotorula is seeded to ocean rhodotorula fluid nutrient medium, 25-30 DEG C of temperature, rotating speed is 150-
250rpm shaken cultivations are used as ocean rhodotorula zymotic fluid in 44-48 hours；It is preferred that, the ocean rhodotorula is rhodotorula mucilaginosa
CGMCC No.13012。
Rhodotorula mucilaginosa (Rhodotorula mucilaginosa) new strains in the present invention, in September in 2016 19 days
It is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, abbreviation CGMCC, address：Chaoyang District, Beijing City
The institute 3 of North Star West Road 1, Institute of Microorganism, Academia Sinica, deposit number is CGMCC No.13012.
Further, the concentration of each composition is in described ocean rhodotorula culture medium：Molasses 20-30g/L, peptone
1.5-2.5g/L, yeast extract 1.5-2.5g/L, ammonium sulfate 2.5-5.0g/L, potassium dihydrogen phosphate 1.0-1.5g/L, magnesium sulfate monohydrate
0.05-0.15g/L, ferrous sulfate 0.1-0.15g/L, sodium chloride 10-15g/L, pH are adjusted to 5.5-6.0.
Wherein, bacillus bacterium powder, lactic acid bacteria bacterium powder are added in activating solution, activates 8-10h, prepare activating solution；
Ocean rhodotorula fermented bacterium and activating solution are using volume ratio as 2:After 1 is well mixed, composite zymocyte liquid is formed.
Wherein, described activating solution contains following components：1~5g/L of peptone, 4~9g/L of yeast extract, sodium chloride 3.5~
4.5g/L, brown sugar 9~12g/L, pH are adjusted to 6.5-7.0.
Wherein, the bacillus is by bacillus subtilis (Bacillus subtilis) CGMCC No.8148, lichens
Bacillus (Bacillus licheniformis) CGMCC No.2383 compoundings are formed, and the concentration of bacillus subtilis is 200
Hundred million CFU/g, bacillus licheniformis concentration is 10,000,000,000 CFU/g；The lactic acid bacteria is by enterococcus faecalis (Enterococcus
Faecalis) CGMCC No.2385, Lactobacillus plantarum (Lactobacillus plantarum) compounding are formed, enterococcus faecalis
Concentration is 20,000,000,000 CFU/g, and Lactobacillus plantarum is 10,000,000,000 CFU/g.Concentration of the Bacillus bacterium powder in activating solution is：5-10g/L,
The concentration of lactic acid bacteria bacterium powder is：5-10g/L.
Fermentation substrate is configured by following mass percents：Dregs of beans (46%) 70-95%, molasses 1-5%, maize flour 3-7%,
Ammonium sulfate 0.5%, dipotassium hydrogen phosphate 0.2%, bitter salt 0.1%, calcium carbonate 0.1%, acid protease 0.04~
0.05%, cellulase 0.01~0.03%, amylase 0.02~0.04% are mixed to prepare fermentation substrate；
Further, the preparation method of the above-mentioned fermented bean dregs rich in astaxanthin, the enzyme activity of described acid protease
Power is 50,000IU/g；The vigor of cellulase is 6000IU/ grams, and the enzyme activity of amylase is 7000IU/g.
By step 3) obtained by composite zymocyte liquid be seeded to step 4) in fermentation substrate in, inoculum concentration be 3~6%
(mass percent), and it is well mixed to 35~50% (mass percents) to adjust water content,
By step 5) fermentation substrate after bacterium solution is inoculated with by the way of ton bag or fermentation vat, in 25 DEG C to 45 DEG C of bar
Under part, after fermentation 48-96h, fermented bean dregs semi-finished product are formed；
The present invention also provides a kind of fermented bean dregs rich in astaxanthin, and it is obtained by above-mentioned preparation method；Described hair
Ferment dregs of beans has stronger sour fragrance；Moisture 35-50%, pH are in 4.0~5.5, crude protein content 25-40% (quality
Percentage), more former dregs of beans crude protein content improves 5-15%.Ocean rhodotorula mother bacterium number 1.5 × 10 after fermentation7More than cfu/g,
Content astaxanthin is more than 125.43 μ g/kg, lactic acid bacteria number 6.5 × 108More than cfu/g, gemma bacterium number 1.0 × 108Cfu/g with
On；The more former dregs of beans reduction by 70~90% of vomitoxin.
Described fermented bean dregs have stronger sour fragrance；Moisture preferred 35-42%, pH be preferably 4.52~
5.28, crude protein content is preferably 35.36~37.25% (mass percents), and more former dregs of beans crude protein content improves 10.63-
16.54%.Ocean rhodotorula mother bacterium number preferably 1.5 × 10 after fermentation7~2.5 × 108More than cfu/g, content astaxanthin is preferably
125.43~547.26 μ g/kg, lactic acid bacteria number is preferably 6.5 × 108~2.1 × 109Cfu/g, gemma bacterium number is preferably 1.2 ×
108-3.5×108cfu/g；The more former dregs of beans reduction by 75~90% of vomitoxin.
Further, the present invention also provides a kind of feed rich in astaxanthin, and it is prepared from by following methods：
By corn, dregs of beans is handled by primary dcreening operation, and above-mentioned raw materials are crushed into powdering with 2.5mm sieve, into treating dispensing
Storehouse, by the corn after crushing, dregs of beans and wheat bran, and described fermented bean dregs rich in astaxanthin the hair for preparing of preparation method
Ferment dregs of beans is proportionally added into mixer by aniseed burden balance, and fish meal, calcium monohydrogen phosphate, stone flour, salt, L- then are relied into ammonia respectively
Acid, threonine, compound premixed feed, Choline Chloride weigh addition mixer from micro-balance or manually, after mixing 10-15 seconds again
Add after vegetable oil is well mixed and pelletized into storehouse to be pelletized.
Wherein, it is well mixed, is carried out under conditions of vapour pressure 0.2Mpa, 75 DEG C of temperature by the incorporation time of 120 seconds
Granulation, grinding particle size is 2.0-3.0mm.
Compared with prior art, the present invention has following advantage：
1, using the ocean rhodotorula strain with independent intellectual property right, combines microbial fermentation, the use of enzyme preparation,
The content of crude protein and sour molten albumen is improved, mycotoxin contamination is reduced.
In 2 fermentation process, the use of ocean rhodotorula and lactic acid bacteria adds the sour fragrance of fermentation, improves the suitable of feed
Mouth property, improves the beneficial bacteriums such as the feed intake of animal, also, lactic acid bacteria and certain adjustment effect is played to the enteron aisle of livestock and poultry.Bud
The enzyme and antibacterial material of spore bacterium secretion, can improve the digestibility of young animal and sow, and can improve animal intestinal tract, to dynamic
Thing production performance has facilitation.
3 use by ocean rhodotorula during the fermentation, adds the content of astaxanthin in fermented bean dregs, reduces young
Diarrhea of pigs, and the immunity and oxidation resistance of the effect raising animal using astaxanthin, improve weaned piglets.
4 the easily operated fermentation method such as wrap by ton, reduce and take up an area and personnel attrition, to the less demanding of equipment, system
Preparation Method is simple and easy to apply, energy-conserving and environment-protective, reduces the use cost of fermented bean dregs.
5 use low moisture zymotechniques, carry out dispensing with feedstuff, mix, modulate, pelletize, cool down, and fermented bean dregs are partly
Finished product is directly entered feed manufacturing link, reduces the energy consumption for drying link, and produced during cost of transportation, and protected fermentation
The prebiotic factor, be easy to fermented bean dregs in the application and popularization of scale raising and feed factory.
The fermented bean dregs rich in astaxanthin of the present invention are applied on weanling pig, can reduce the diarrhoea of piglet, are strengthened
The content of immunoglobulin, improves the immunity of animal, has facilitation to the production performance of animal.Energy is applied on sow
The reproductive performance of sow is enough improved, the generation of suckling pig diarrhoea is reduced, increases weaning weight, and improve lactoprotein in breast milk, breast
Fat and lactose content, reduce piglet stress.
Brief description of the drawings
Fig. 1 show rhodotorula mucilaginosa Y4-5 (HYH4-5-21) strain passage carotenoid content measurement result.
Embodiment
Following examples are used to illustrate the present invention, but are not limited to the scope of the present invention.
The separation screening High Yield of Carotenoid ocean rhodotorula bacterial strain of embodiment 1
Take and come from Fujian, the water sample in Jiangsu fish pond and the shrimp pool, mud sample and animal intestinal tract sample, park mud sample and mangrove leaf
Deng, take 1mL fluid samples or 1g solid samples in the 9mL YPD fluid nutrient mediums of 18ml test tubes, add Benzylpenicillin sodium salt (800,000
Every bottle of sterile purified water of unit is diluted to 1mL, addition 1uL/ test tubes), after 85 DEG C of water bath processing 10min.Test tube is put into temperature
28 DEG C, rotating speed is to obtain enrichment culture liquid after shaken cultivation 16h under the conditions of 180rpm.
Enrichment culture liquid is carried out by gradient dilution using coubling dilution, is coated on YPD solid mediums, is positioned over 28
Pink, the red or orange single bacterium colony grown on 48h in DEG C incubator, picking flat board is rule, and is purified twice, is obtained
The single bacterium colony that must be lived, totally 47 plants of bacterium, strain glycerol stocks.
Embodiment 2 determines biological dry weight and carotenoid output, screens outstanding bacterial strain
In super-clean bench, 47 plants of strain to be tested single bacterium colonies are accessed in screening and culturing medium, repeat number is 3.Cultural method is：
250mL shaking flasks fill liquid 50mL, 28 DEG C of cultivation temperature, rotating speed 180rpm, incubation time 36h.
Wherein, fluid nutrient medium P3 formulas are：Yeast extract 15g, epsom salt 1g, sucrose 40g, seawater 1000mL (sea
Salt 20g).Solid medium agar is 20g/L.
Dry cell weight assay method：Zymotic fluid 10ml, 8000r/min centrifugation 5min is taken, supernatant is removed, adds distilled water
Washing three times, then 8000r/min centrifugation 5min, remove supernatant, dry mycelium (unit g/L) are produced in 50 degree of drying to constant weights.
The extracting method of carotenoid：1) wall-breaking method -- ultrasonic assistant acid heat method：Dry mycelium is taken to be put into band cap test tube
In, 3mol/L hydrochloric acid (needing to prepare) is added according to the amount of 15ml/g dry myceliums, 40min is soaked.In immersion process by test tube extremely
In under Ultrasonic Conditions.After broken wall is completed, acetone is added into cell fragment.2) 2 times of hydrochloric acid shell-broken liquid volumes are used as leaching
Extract, soaks 30min, 4000r/min centrifugation 15min, gained supernatant is the carotenoid leaching liquor of acetone.Will extraction
Liquid is determined with 484nm.
Carotenoid content determines (μ g/g)=OD × D × V/ (0.16 × W)
(D is extension rate；V is organic solvent volume；W is extraction dry cell weight；0.16 is organic solvent delustring system
Number.) determine absorbance, unit：μg/g.
9 plants of preferred strain measurement results such as table 1, from table 1, bacterial strain HYH4-5 dry cell weights and total carotinoid production
Amount is all highest.
9 bacterial strain before the biological dry weight of table 1 and carotenoid output
The ocean rhodotorula bacterial strain HYH4-5 of embodiment 3 ITS rRNA gene sequencings
The bacterial strain HYH4-5 of acquisition is subjected to ITS rRNA gene sequencings, sequence is as shown in SEQ ID No.1.By base
Because of the defeated GenBank of sequence, analysis is compared to all sequences in database by Blast programs.As a result find, the red ferment in ocean
Mother strains HYH4-5 ITS rRNA gene orders belong to type strain with rhodotorula mucilaginosa higher similitude.
The ocean rhodotorula bacterial strain HYH4-5 of embodiment 4 Analysis of The Physiological And Biochemical Properties
Ocean rhodotorula new strains HYH4-5 in the present invention, its physio-biochemical characteristics are as follows：Azymic glucose, malt
Any one of sugar, galactolipin, sucrose, lactose, gossypose and melibiose.Can to glucose, maltose, sucrose, gossypose,
Melibiose, cellobiose, trehalose, xylose, D-R and L-arabinose are assimilated, to galactolipin, lactose and can
Soluble starch can not be assimilated.
In summary, it is rhodotorula mucilaginosa (Rhodotorula to determine ocean rhodotorula bacterial strain HYH4-5
mucilaginosa)。
Embodiment 5 obtains rhodotorula mucilaginosa mutagenic strain Y4-5
Mutagenesis is carried out to ocean rhodotorula HYH4-5 using nitrosoguanidine, operating method is as follows：
1) bacteria suspension is prepared.Take shake flask culture 10mL 4000rpm to centrifuge 10min and collect yeast.Cultural method：250mL
Shaking flask fills liquid 50mL, 28 DEG C of cultivation temperature, rotating speed 180rpm, incubation time 24h.With 0.1mol/L pH5.5 buffered sodium citrates
Liquid is washed 2 times, and cell is suspended with citric acid solution, and it is 10 that cell concentration, which is made,8CFU/g bacteria suspension.
2) nitrosoguanidine stoste is prepared.2-3 drops acetone soln is added dropwise into 10mg nitrosoguanidines, 10mL is then added
0.1mol/L pH5.5 sodium citrate buffer solutions are allowed to be completely dissolved.Experiment determines that nitroso guanidine solution mutagenesis concentration is 42 μ g/
During mL, cell lethality is 95% or so, now mutation rate highest.
3) mutagenesis is carried out.Take above-mentioned bacteria suspension 10mL, add nitroso guanidine solution 0.44mL, make its in bacteria suspension most
Final concentration reaches 42 μ g/mL, is incubated 30min in 28 DEG C, abandoning supernatant is centrifuged after taking-up immediately, with 0.1mol/L pH7.0 phosphorus
Sour potassium buffer solution is washed 2 times makes thalline be suspended in 24 DEG C of overnight incubations with 10mL P3 fluid nutrient mediums again.
4) mutant strain is screened：By the appropriate dilution spread of cultured bacterium solution in (after sterilizing solid on culture medium P3 solid plates
Body culture medium P3, is added in diphenylamine solution, final concentration of 25 μ g/mL when being cooled to 45 DEG C or so：Suppress yeast synthesis class recklessly
The ability of radish element, the concentration can kill 90% cell).After 24 DEG C of culture 7d, bacterium colony to be formed, pick out red deeper
Bacterium colony, line preserves culture, determines mutant strain total carotinoid content.As shown in Table 2, bacterial strain HYH4-5-14 carotenoid
Yield highest, bacterial strain HYH4-5-21 carotenoid total amount is highest, therefore elects HYH4-5-21 as product candidate strains,
It is renumbered as Y4-5.
The rhodotorula mucilaginosa HYH4-5 bacterial strain mutant strain carotenoid content measurement results of table 2
Mutant strain stability test and preservation.By Y4-5 inoculations in preserving bacterial strain, continuous passage on P3 solid slopes
10 times, passage every time determines its carotenoid content.As a result Fig. 1 is seen.As a result show that Y4-5 has mitotic stability.The present invention
In rhodotorula mucilaginosa (Rhodotorula mucilaginosa) new strains Y4-5, be preserved in China within 19th in September in 2016
Microbiological Culture Collection administration committee common micro-organisms center, abbreviation CGMCC, address：BeiChen West Road, Chaoyang District, BeiJing City 1
Institute 3, Institute of Microorganism, Academia Sinica, deposit number is CGMCC No.13012.
The rhodotorula mucilaginosa Y4-5 tolerances of embodiment 6 are determined
(1) low temperature tolerant is tested:Respectively at 12 DEG C, 15 DEG C, 18 DEG C, 21 DEG C, 24 DEG C, 28 DEG C, in P3 solid mediums
On flat board, gradient (10 is carried out-1、10-2、10-3、10-4) the red zymotic fluid dibbling growth experiment in ocean (and with liquid-transfering gun draw fermentation it is dilute
Liquid 2uL is released, is seeded on culture medium flat plate), incubation time is 36h, and observation upgrowth situation (bacterium colony size is during which carried out per 4h
And color), characterize its cold tolerance.
Observe result：At different temperatures, zymotic fluid bacterial strain can normal growth, but temperature be 12-18 DEG C in the range of
Than the 21-28 DEG C speed of growth is relatively slow (being presented as same dilution gradient zymotic fluid, same time bacterium colony is smaller), but temperature it is more low more
Be conducive to the generation (colony colour is deeper) of pigment, illustrate the bacterial strain in low temperature, the amount that carotenoid is produced is unaffected.
(2) high-salt tolerance is tested：It is respectively 0 ‰, 10 ‰, 15 ‰, 20 ‰, 25 ‰, 30 ‰, 35 ‰ P3 solids in salinity
On culture medium flat plate, gradient (10 is carried out-1、10-2、10-3、10-4) nutrient solution dibbling growth experiment (and with liquid-transfering gun draw culture it is dilute
Liquid 2uL is released, is seeded on culture medium flat plate), incubation time 36h, 28 DEG C of cultivation temperature.Period carries out observation upgrowth situation per 4h
(bacterium colony size and color), characterizes its high-salt tolerance.
Observe result：Under different salinity, zymotic fluid bacterial strain can normal growth, but salinity be 20-30 ‰ in the range of
Growth result is optimal (being presented as same dilution gradient zymotic fluid, same time bacterium colony is big, and colony colour is deep).
The ocean rhodotorula bacterial strain Y4-5 of embodiment 7 pilot scale fermentation culture
(1) prepared by one-level shake-flask seed liquid：In super-clean bench, the ocean rhodotorula bacterial strain single bacterium colony for separating acquisition is inoculated with
Into fluid nutrient medium P3,50mL, 180rpm, 28 DEG C of culture 30h of 250mL shaking flasks dress liquid.
(2) prepared by second-level shake flask seed liquor：The bacterial strain seed liquor prepared in step (1) is accessed according to 10% inoculum concentration
Fermented with second-level shake flask, 1000mL shaking flasks dress liquid 300mL, 28 DEG C of fermentation temperature, rotating speed 200rpm, fermentation time 24h；
(3) ferment tank：The bacterial strain seed liquor prepared in step (2) is fermented according to 5% inoculum concentration access 100L
Fermented in tank (liquid amount is 60L), 28 DEG C of fermentation temperature, rotating speed 300rpm fills pressure 0.05Mpa, ventilating ratio is 1:0.5,
Initial pH value is 6.0 or so, fermentation time 24h.
Under this pilot plant conditions, the yield for measuring carotenoid is 10.45mg/L, and protein content is 41%.
A kind of preparation of the fermented bean dregs rich in astaxanthin of embodiment 8
Ocean rhodotorula CGMCC No.13012 are seeded to ocean rhodotorula fluid nutrient medium, 28 DEG C of temperature, rotating speed is
180rpm shaken cultivations are used as ocean rhodotorula zymotic fluid in 45 hours；The concentration of each composition in described ocean rhodotorula culture medium
For：Molasses 20g/L, peptone 2.5g/L, yeast extract 1.5g/L, ammonium sulfate 5.0g/L, potassium dihydrogen phosphate 1.0g/L, sulfuric acid monohydrate
Magnesium 0.15g/L, ferrous sulfate 0.1g/L, sodium chloride 10g/L, pH are adjusted to 6.0.
Wherein, bacillus bacterium powder, lactic acid bacteria bacterium powder are added in activating solution, activates 8-10h, prepare activating solution；It is described
Activating solution contain following components：Peptone 5g/L, yeast extract 5g/L, sodium chloride 3.5g/L, brown sugar 10g/L, pH adjust to
6.5.The bacillus is by bacillus subtilis (Bacillus subtilis) CGMCC No.8148, bacillus licheniformis
(Bacillus licheniformis) CGMCC No.2383 compoundings are formed, and the concentration of bacillus subtilis is 20,000,000,000 CFU/g,
Bacillus licheniformis concentration is 10,000,000,000 CFU/g；The lactic acid bacteria is by enterococcus faecalis (Enterococcus faecalis) CGMCC
No.2385, Lactobacillus plantarum (Lactobacillus plantarum) compounding are formed, and the concentration of enterococcus faecalis is 20,000,000,000 CFU/
G, Lactobacillus plantarum is 10,000,000,000 CFU/g.Concentration of the Bacillus bacterium powder in activating solution is：5g/L, the concentration of lactic acid bacteria bacterium powder
For：10g/L.
Ocean rhodotorula fermented bacterium, bacillus and the lactic acid bacteria activating solution of preparation are using volume ratio as 2:1 is well mixed
Afterwards, composite zymocyte liquid is formed.
Fermentation substrate is configured by following mass percents：Dregs of beans (46%) 93%, molasses 1%, maize flour 5%, ammonium sulfate
0.5%, dipotassium hydrogen phosphate 0.2%, bitter salt 0.1%, calcium carbonate 0.1%, acid protease 0.04%, cellulase
0.04%th, amylase 0.02%, is mixed to prepare fermentation substrate.The enzyme activity of described acid protease is 50,000IU/g；It is fine
The vigor of the plain enzyme of dimension is 6000IU/ grams, and the enzyme activity of amylase is 7000IU/g.
Composite zymocyte liquid is seeded in fermentation substrate, inoculum concentration be 3% (mass percent), and adjust water content to
40% (mass percent) is well mixed, by the way of ton bag or fermentation vat, under conditions of 30 DEG C, after fermentation 72h, shape
Into fermented bean dregs semi-finished product.
A kind of preparation of the fermented bean dregs rich in astaxanthin of embodiment 9
Ocean rhodotorula CGMCC No.13012 are seeded to ocean rhodotorula fluid nutrient medium, 30 DEG C of temperature, rotating speed is
150rpm shaken cultivations are used as ocean rhodotorula zymotic fluid in 44 hours；The concentration of each composition in described ocean rhodotorula culture medium
For：Molasses 30g/L, peptone 1.5g/L, yeast extract 2.5g/L, ammonium sulfate 5.0g/L, potassium dihydrogen phosphate 1.0g/L, sulfuric acid monohydrate
Magnesium 0.10g/L, ferrous sulfate 0.1g/L, sodium chloride 10g/L, pH are adjusted to 5.5.
Wherein, bacillus bacterium powder, lactic acid bacteria bacterium powder are added in activating solution, activates 8h, prepare activating solution；Described work
Change liquid and contain following components：Peptone 5g/L, yeast extract 6g/L, sodium chloride 4.5g/L, brown sugar 10g/L, pH are adjusted to 6.5.Institute
Bacillus is stated by bacillus subtilis (Bacillus subtilis) CGMCC No.8148, bacillus licheniformis
(Bacillus licheniformis) CGMCC No.2383 compoundings are formed, and the concentration of bacillus subtilis is 20,000,000,000 CFU/g,
Bacillus licheniformis concentration is 10,000,000,000 CFU/g；The lactic acid bacteria is by enterococcus faecalis (Enterococcus faecalis) CGMCC
No.2385, Lactobacillus plantarum (Lactobacillus plantarum) compounding are formed, and the concentration of enterococcus faecalis is 20,000,000,000 CFU/
G, Lactobacillus plantarum is 10,000,000,000 CFU/g.Concentration of the Bacillus bacterium powder in activating solution is：10g/L, the concentration of lactic acid bacteria bacterium powder
For：5g/L.
Fermentation substrate is configured by following mass percents：Dregs of beans (46%) 92%, molasses 2%, maize flour 5%, ammonium sulfate
0.5%, dipotassium hydrogen phosphate 0.2%, bitter salt 0.1%, calcium carbonate 0.1%, acid protease 0.05%, cellulase
0.03%th, amylase 0.02%, is mixed to prepare fermentation substrate.The enzyme activity of described acid protease is 50,000IU/g；It is fine
The vigor of the plain enzyme of dimension is 6000IU/ grams, and the enzyme activity of amylase is 7000IU/g.
Composite zymocyte liquid is seeded in fermentation substrate, inoculum concentration be 5% (mass percent), and adjust water content to
35% (mass percent) is well mixed, by the way of ton bag or fermentation vat, under conditions of 25 DEG C, after fermentation 96h, shape
Into fermented bean dregs semi-finished product.
A kind of preparation of the fermented bean dregs rich in astaxanthin of embodiment 10
Ocean rhodotorula CGMCC No.13012 are seeded to ocean rhodotorula fluid nutrient medium, 25 DEG C of temperature, rotating speed is
250rpm shaken cultivations are used as ocean rhodotorula zymotic fluid in 48 hours；The concentration of each composition in described ocean rhodotorula culture medium
For：Molasses 30g/L, peptone 2.5g/L, yeast extract 2.5g/L, ammonium sulfate 2.5g/L, potassium dihydrogen phosphate 1.2g/L, sulfuric acid monohydrate
Magnesium 0.05g/L, ferrous sulfate 0.15g/L, sodium chloride 15g/L, pH are adjusted to 6.0.
Wherein, bacillus bacterium powder, lactic acid bacteria bacterium powder are added in activating solution, activates 10h, prepare activating solution；Described
Activating solution contains following components：Peptone 2.5g/L, yeast extract 9g/L, sodium chloride 4.0g/L, brown sugar 12g/L, pH adjust to
7.0.The bacillus is by bacillus subtilis (Bacillus subtilis) CGMCC No.8148, bacillus licheniformis
(Bacillus licheniformis) CGMCC No.2383 compoundings are formed, and the concentration of bacillus subtilis is 20,000,000,000 CFU/g,
Bacillus licheniformis concentration is 10,000,000,000 CFU/g；The lactic acid bacteria is by enterococcus faecalis (Enterococcus faecalis) CGMCC
No.2385, Lactobacillus plantarum (Lactobacillus plantarum) compounding are formed, and the concentration of enterococcus faecalis is 20,000,000,000 CFU/
G, Lactobacillus plantarum is 10,000,000,000 CFU/g.Concentration of the Bacillus bacterium powder in activating solution is：10g/L, the concentration of lactic acid bacteria bacterium powder
For 10g/L.
Ocean rhodotorula fermented bacterium, bacillus and the lactic acid bacteria activating solution of preparation are using volume ratio as 2:1 is well mixed
Afterwards, composite zymocyte liquid is formed.
Fermentation substrate is configured by following mass percents：Dregs of beans (46%) 87%, molasses 5%, maize flour 7%, ammonium sulfate
0.5%, dipotassium hydrogen phosphate 0.2%, bitter salt 0.1%, calcium carbonate 0.1%, acid protease 0.05%, cellulase
0.03%th, amylase 0.02%, is mixed to prepare fermentation substrate.The enzyme activity of described acid protease is 50,000IU/g；It is fine
The vigor of the plain enzyme of dimension is 6000IU/ grams, and the enzyme activity of amylase is 7000IU/g.
Composite zymocyte liquid is seeded in fermentation substrate, inoculum concentration be 5% (mass percent), and adjust water content to
50% (mass percent) is well mixed, by the way of ton bag or fermentation vat, under conditions of 40 DEG C, after fermentation 48h, shape
Into fermented bean dregs semi-finished product.
The measure of the fermented bean dregs index of embodiment 11
The Indexs measure of the obtained fermented bean dregs semi-finished product rich in astaxanthin the results are shown in Table 2.Wherein, crude protein content is
Determined by GB/T 6432-1994；PH is measured by pH meter；Marine yeast bacterium number content is according to GB4789.15-2010 methods
Measure, lactic acid bacteria bacterium number content is measured according to GB4789.35-2010 methods, Bacillus bacterium number content is according to NY/T 2131-
2012nd, NY/T 1461-2007 methods are measured.Content astaxanthin is measured according to DB23/T 1275-2008 methods.Vomitoxin
Determine and carried out with reference to vomitoxin detection kit operational procedure.
The situation of change of index before and after the bean pulp fermentation of table 2
Application of fermented bean dregs of the embodiment 12 rich in astaxanthin in weanling pig feed
Feed formulation：Feed formulation is constituted and nutritive water with reference to NRC (2012) and swine rearing standard (2004), experiment daily ration
It is flat to be shown in Table 3.By corn, dregs of beans is handled by primary dcreening operation, and above-mentioned raw materials are crushed into powdering with 2.5mm sieve, into treating dispensing
Storehouse, by the corn after crushing, dregs of beans, and the described fermented bean dregs semi-finished product rich in astaxanthin are pressed by aniseed burden balance
Ratio adds mixer, then respectively by fish meal, whey powder, calcium monohydrogen phosphate, stone flour, salt, 1B, threonine, color ammonia
Acid, 0.5% compound premixed feed, Choline Chloride, chrome green, zinc oxide are weighed to add from micro-balance or manually and mixed
Machine, mixing is added after soybean oil is well mixed after 10-15 seconds and pelletized into storehouse to be pelletized.Commercially available fermented bean dregs are by Shanghai
Certain company provides.
Wherein, incorporation time is 120 seconds, vapour pressure 0.2Mpa, is pelletized, pelletized under conditions of 75 DEG C of modulator temperature
Ultramicro grinding is carried out after cooling again, grinding particle size is (400-500 μm).
The experiment daily ration composition of table 3 and trophic level
Note：0.5% compound premixing closes batch mixing to provide Cu >=40g, Fe >=1040mg, Mn >=14.4g per kg complete feeds,
Se >=120g, I >=160mg, cobalt >=200mg, iron >=36.8g, moisture≤10%.Vitamin A (KIU) >=2000, vitamin D3
(KIU) >=375, vitamin E (IU) >=5300, vitamin K >=400mg, vitamin B1>=450mg, vitamin B2>=600mg, dimension
Raw element B6>=530mg, vitamin B12>=6.4mg, folic acid >=216mg, biotin >=8.8mg, nicotinic acid >=7800mg, calcium pantothenate >=
6000mg。
Experimental animal and feeding management：Health (Du × length × big) 21 ages in days of this experiment from 144 (male and female half and half)
Weanling pig, experimental period 10d.Experiment is carried out on Tangshan, Hebei Province Yutian County pig farm.Using totally enclosed type pig house, She Neiwen
Degree, humidity, ventilation, carbon dioxide and ammonia density Automated condtrol, give up temperature and are maintained at 24-26 DEG C during experiment.Test piglet point
Column is raised, slatted floor, the adjustable hopper of stainless steel, nipple-shaped drinker.Using free choice feeding and the feed mode of drinking-water.
Experimental design：144 healthy weanling pigs are grouped into 4 processing by body weight and sex completely random, each place
6 repetitions are managed, 6 pigs (male and female half and half) are each repeated.Daily ration treatment group is respectively：(1) control group：Corn-soybean-commercially available hair
Ferment dregs of beans basal diet；(2) the fermented bean dregs group of 12% embodiment 8 is added；(3) the fermented bean dregs group of 12% embodiment 9 is added；(4)
Add the fermented bean dregs group of 12% embodiment 10.Ensure that the crude protein level of each test group is consistent.
Testing index and assay method：Average daily gain (ADG), average daily gain (ADFI), feed-weight ratio.Experimental period
Between, in order to observe the order of severity of diarrhea of weaned piglets, record excrement index daily, such as table 4, when index be 4,5 items regard as
Diarrhoea.
1 pig, every the blood sampling of pig vena cava anterior 10mL, 3000r/ are randomly selected on the day of off-test from each repetition
Min centrifugation 10min prepare blood serum sample, after packing freezing deposit in -80 DEG C it is to be measured.Immunoglobulin A, immunoglobulin G and
The measure of immunoglobulin M carries out EUSA according to the explanation of quantitative determination reagent kit (R＆D, China).
The excrement index of table 4
| Excrement index | Diarrhoea degree | Excrement state |
| 1 | Constipation | Sheep manure shape (spherical) |
| 2 | Normally | Strip is molded |
| 3 | Laxativeness | Excrement is in viscous atherosclerotic |
| 4 | Mild diarrhea | Excrement is in sposh pulpous state |
| 5 | Severe diarrhea | Excrement is in watery |
Statistical method：Test data statistics designs ANOVA processes progress side using the balance test in SAS9.0 software kits
Difference is analysed, and the Multiple range test of average is carried out using Duncan methods.
Result of the test：
Physiochemical indice：As shown in table 5, control group, the fermented bean dregs group of embodiment 8, the fermented bean dregs group of embodiment 9 and embodiment
The equal no significant differences of IgA and IgM in 10 fermented bean dregs group piglet bloods, each test group of IgG indexs is above control group, shows this
The fermented bean dregs of invention can improve the immunoglobulin level of weanling pig, strengthen animal immunizing power.
The fermented bean dregs of table 5 substitute shadow noon of the dregs of beans to weanling pig physiochemical indice in daily ration
Production performance：Commercially available fermented bean dregs are to Growth Performance of Weaning Piglets in the fermented bean dregs replacement daily ration of astaxanthin-containing
Influence the results are shown in Table 6.Each group piglet original body mass does not have difference.As a result show, each test group ratio of fermented bean dregs of astaxanthin-containing
Control group daily gain and daily ingestion amount increase significantly (P<0.05), difference is not notable between each test group.Each test group with
Control group has downward trend compared to feedstuff-meat ratio, the not notable (P of difference between embodiment 8 and the fermented bean dregs test group of embodiment 9>
0.05), but with control group significant difference (P<0.05).According to upper table data display, the fermented bean dregs of astaxanthin-containing substitute daily ration
Middle common fermentation dregs of beans can improve the growth performance of weanling pig, and reduce the grice diarrhoea rate after wean.
Commercially available fermented bean dregs are to Growth Performance of Weaning Piglets and diarrhea rate in the fermented bean dregs replacement daily ration of the astaxanthin-containing of table 6
Influence
Application of fermented bean dregs of the embodiment 13 rich in astaxanthin in the sow feed of scale pig farm
Feed formulation：Feed formulation is with reference to NRC (2012) and Chinese lean meat species milking sow feeding standard (2004), experiment
Daily ration is constituted and trophic level is shown in Table 7.Feed be scale pig farm from dispensing, by corn, dregs of beans, wheat bran, fish meal 3mm sieve
After crushing, batch bin is added, fermented bean dregs semi-finished product, premix and soybean oil rich in astaxanthin is then proportionally added into, mixed
It can be fed after closing uniformly.
Table 7 tests daily ration and trophic level
Note：4% compound premixing closes batch mixing and provides Cu >=5g for often kg complete feeds, Fe >=130mg, Zn >=5g, and Mn >=
1.8g, Se >=15g, I >=20mg, cobalt >=25mg, calcium >=160-270g, iron >=4.6g, phosphorus >=38g, moisture≤10%.Vitamin
A (KIU) >=253, vitamin D3(KIU) >=47, vitamin E (IU) >=662, vitamin K >=53mg, vitamin B1>=58mg, dimension
Raw element B2>=75mg, vitamin B6>=67mg, vitamin B12>=0.8mg, folic acid >=27mg, biotin >=1.1mg, nicotinic acid >=
975mg, calcium pantothenate >=750mg, Choline Chloride >=5g.
Experimental animal and feeding management：Test site is carried out in boar plant of In Shunyi District of Beijing, chooses kind phase
Same, age, body condition and the expected date of childbirth, close to the sow 48 of 110 days or so the gestational period in seven days, are randomly divided into 4 groups, every group 12
Head sow, every sow is 1 repetition.Respectively control group (basal diet), the fermented bean dregs test group of embodiment 8, embodiment 9
Fermented bean dregs test group, the fermented bean dregs test group of embodiment 10.Test and opened from the antenatal childbirth house for going to thorough disinfection for 7 days of sow
Begin until weaned piglet, raises the individual pen in the position limiting fence of full crack formula and raise, cleaning is dried in house, air permeability is good, temperature
It is suitable 25-28 DEG C.Feeding management and immune programme for children are performed by large-scale pig farm uniform requirement during experiment.From turn childbirth house to
Childbirth, sow limits feeding according to body condition in 2.0-2.5kg daily.Not feeding on the day of Farrowing, the 1st day and the 2nd day often after childbirth
Day feeding 1-2kg, later increase 1.0kg daily, the 6th day free choice feeding (slightly remaining material in hopper after having enough to be defined), daily
Spirit, the appetite situation of viewing test pig, record sow whether there is constipation phenomenon, it is found that sick pig isolates treatment in time.Sow is in son
Breeding house is gone to after pig wean (21 days)
Testing index and assay method：Record before childbirth and lactation period sow feed intake, calculate average daily gain, record
Litter size, young number living, mummy number, stillborn foetus number, childbirth duration (stages of labor), weigth at birth, calculate newborn piglet survival rate
(the young number/nest litter size of just life), nascent individual counterpoise.According to being adjusted in sow number born alive and galactopoiesis situation group, sow band is young
Number records the young number of band, weaned piglet head number, the weight of weaning litter of every sow, calculates suckling pig and averagely increase day by day at 10 or so
The individual counterpoise of heavy, wean, number of weaned (during nest is tested, in order to observe the order of severity of diarrhea of weaned piglets, are recorded daily
Excrement index, index 4,5 regard as diarrhoea.Coefficients statistics of suffering from diarrhoea standard is with reference to table 4.
20th day (peak of lactation in theory) after Farrowing, often processing randomly selects four-head pig respectively before, during and after
The nipple collection milk sample at 3 positions, every pig gathers 20m1 or so, -20 DEG C of preservations.With quick analysis of milk composition instrument
(MILKYWAY-CP2) testing index：Lactoprotein, butter oil, lactose.
Statistical method：Test data statistics designs ANOVA processes progress side using the balance test in SAS9.0 software kits
Difference is analysed, and the Multiple range test of average is carried out using Duncan methods.
Result of the test：
Fermented bean dregs substitute influence of the dregs of beans to sow and suckling pig growth performance in lactation daily ration and are shown in Table 8.Each group is female
Pigsty produces strong young number, nascent individual counterpoise without significant difference.Significantly risen using the average daily feed intake of sow after fermented bean dregs
Height (P<0.05) without significant difference, but between each embodiment.After addition fermented bean dregs sow it is disconnected match somebody with somebody between be separated with the trend of shortening, but
With control group without significant difference.Using the average daily gain of suckling pig after fermented bean dregs (P is significantly improved compared with control group<
0.05), diarrhea rate is remarkably decreased (P<0.05).Result of the test shows that fermented bean dregs of the addition rich in astaxanthin are substituted in daily ration
The feed intake of sow can be improved, and improves the average daily gain of suckling pig, the generation of suckling pig diarrhoea is reduced.
The fermented bean dregs of table 8 substitute influence of the dregs of beans to sow and suckling pig production performance and diarrhea rate in daily ration
There is dregs of beans to milking sow milk composition in fermented bean dregs replacement lactation daily ration rich in astaxanthin carries in various degree
It is high.The content of wherein lactoprotein and butter oil significantly improves (P compared with control group<0.05).The lactose level of fermented bean dregs group is more right
According to group in the trend risen.Fermented bean dregs rich in astaxanthin can improve lactoprotein and butter oil in lactation period sow milk
Level.
The measure of table 9 lactation period sow breast milk biochemical indicator
Described above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, without departing from the technical principles of the invention, some improvements and modifications can also be made, these improvements and modifications
Also it should be regarded as protection scope of the present invention.
Sequence table
<110>Beijing Wei Nong bio tech ltd
<120>A kind of preparation of fermented bean dregs rich in astaxanthin and feed
<130>
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 620
<212> DNA
<213>Rhodotorula mucilaginosa（Rhodotorula mucilaginosa）
<400> 1
cttccgtagg gtgaacctgc ggaaggatca ttagtgaata taggacgtcc aacttaactt 60
ggagtccgaa ctctcacttt ctaaccctgt gcatttgttt gggatagtaa ctctcgcaag 120
agggcgaact cctattcact tataaacaca aagtctatga atgtattaaa ttttataaca 180
aaataaaact ttcaacaacg gatctcttgg ctctcgcatc gatgaagaac gcagcgaaat 240
gcgataagta atgtgaattg cagaattcag tgaatcatcg aatctttgaa cgcaccttgc 300
gctccatggt attccgtgga gcatgcctgt ttgagtgtca tgaatacttc aaccctcctc 360
tttcttaatg attgaagagg tgtttggttt ctgagcgctg ctggccttta gggtctagct 420
cgttcgtaat gcattagcat ccgcaatcga acttcggatt gacttggcgt aatagactat 480
tcgctgagga attctagtct tcggactaga gccgggttgg gttaaaggaa gcttctaatc 540
agaatgtcta cattttaaga ttagatctca aatcaggtag gactacccgc tgaacttaag 600
catatcaata agcggaggaa 620
Claims (10)
1. a kind of preparation method of the fermented bean dregs rich in astaxanthin, it is characterised in that comprise the steps：
1) the ocean rhodotorula zymotic fluid rich in astaxanthin is prepared；
2) composite zymocyte liquid used in cooperative fermentation is prepared；
3) fermentation substrate is prepared；
4) by step 2) obtained by composite zymocyte liquid be seeded to step 3) in fermentation substrate in, and adjust water content, mix
Close uniform；
5) by step 4) inoculation bacterium solution after fermentation substrate fermented under conditions of temperature control, ferment completion produce fermentation beans
The dregs of rice.
2. the preparation method of the fermented bean dregs according to claim 1 rich in astaxanthin, it is characterised in that by the red ferment in ocean
Mother is seeded to ocean rhodotorula fluid nutrient medium, 25-30 DEG C of temperature, and rotating speed is 150-250rpm, and shaken cultivation is made for 44-48 hours
The standby ocean rhodotorula zymotic fluid, the ocean rhodotorula is rhodotorula mucilaginosa CGMCC No.13012.
3. the preparation method of the fermented bean dregs according to claim 1 rich in astaxanthin, it is characterised in that prepare collaboration hair
Composite zymocyte liquid specifically includes following steps used in ferment：
Bacillus bacterium powder, lactic acid bacteria bacterium powder are added in activating solution, 8-10h is activated, prepares activating solution；
By ocean rhodotorula zymotic fluid and activating solution using volume ratio as 2:After 1 is well mixed, composite zymocyte liquid is formed.
4. the preparation method of the fermented bean dregs according to claim 3 rich in astaxanthin, it is characterised in that：Described activation
Liquid contains following components：1~5g/L of peptone, 4~9g/L of yeast extract, 3.5~4.5g/L of sodium chloride, brown sugar 9~12g/L, pH
Adjust to 6.5-7.0.
5. the preparation method of the fermented bean dregs according to claim 3 rich in astaxanthin, it is characterised in that the gemma bar
Bacterium is by bacillus subtilis (Bacillus subtilis) CGMCC No.8148, bacillus licheniformis (Bacillus
Licheniformis) CGMCC No.2383 are constituted, and the concentration of bacillus subtilis is 200,000,000,000 CFU/g, bacillus licheniformis
Concentration is 200,000,000,000 CFU/g；The lactic acid bacteria is by enterococcus faecalis (Enterococcus faecalis) CGMCC No.2385, plant
Thing lactobacillus (Lactobacillus plantarum) is constituted, and the concentration of enterococcus faecalis is 20,000,000,000 CFU/g, and Lactobacillus plantarum is
10000000000 CFU/g, concentration of the Bacillus bacterium powder in activating solution is：5-10g/L, the concentration of lactic acid bacteria bacterium powder is：5-10g/L.
6. the preparation method of the fermented bean dregs according to claim 1 rich in astaxanthin, it is characterised in that：Step 3) it is described
Fermentation substrate, by following mass percents configure：Dregs of beans 70-95%, molasses 1-5%, maize flour 3-7%, ammonium sulfate
0.5%, dipotassium hydrogen phosphate 0.2%, bitter salt 0.1%, calcium carbonate 0.1%, acid protease 0.04~0.05% are fine
The plain enzyme 0.01~0.03% of dimension, amylase 0.02~0.04%, the enzyme activity of described acid protease is 50,000IU/g；It is fine
The vigor of the plain enzyme of dimension is 6000IU/ grams, and the enzyme activity of amylase is 7000IU/g.
7. the preparation method of the fermented bean dregs according to claim 1 rich in astaxanthin, it is characterised in that：Composite zymocyte
Liquid is seeded in fermentation substrate, and inoculum concentration is 3~6%, and adjusts water content to 35~50%, is well mixed, using ton bag or
The mode of person's fermentation vat, under conditions of 25 DEG C to 45 DEG C, ferment 48-96h.
8. a kind of fermented bean dregs rich in astaxanthin, it is characterised in that as the preparation method system described in claim any one of 1-7
It is standby to form, wherein, moisture 35-50%, pH are in 4.0~5.5, crude protein content 25-40%, ocean rhodotorula bacterium number 1.5
×107More than cfu/g, content astaxanthin is more than 125.43 μ g/kg, lactic acid bacteria number 6.5 × 108More than cfu/g, gemma bacterium number
1.0×108More than cfu/g.
9. a kind of feed rich in astaxanthin, it is characterised in that：It is prepared from by following methods：
By corn, dregs of beans is handled by primary dcreening operation, and above-mentioned raw materials are crushed into powdering with 2.5mm sieve, will into batch bin is treated
Corn after crushing, dregs of beans, and the described fermented bean dregs semi-finished product rich in astaxanthin, are added in proportion by aniseed burden balance
Enter mixer, then respectively by fish meal, whey powder, calcium monohydrogen phosphate, stone flour, salt, 1B, threonine, tryptophan,
0.5% compound premixed feed, Choline Chloride, chrome green, zinc oxide are weighed from micro-balance or manually adds mixer, mixed
Add after soybean oil is well mixed and pelletized into storehouse to be pelletized after closing 10-15 seconds, wherein, the content of fermented bean dregs is 5-
20%.
10. the feed according to claim 9 rich in astaxanthin, it is characterised in that in vapour pressure 0.2Mpa, modulator temperature
Pelletized under conditions of 75 DEG C of degree, carry out ultramicro grinding after granulation cooling again, grinding particle size is 400-500 μm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710355273.1A CN107319129A (en) | 2017-05-19 | 2017-05-19 | A kind of preparation of fermented bean dregs rich in astaxanthin and feed |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710355273.1A CN107319129A (en) | 2017-05-19 | 2017-05-19 | A kind of preparation of fermented bean dregs rich in astaxanthin and feed |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107319129A true CN107319129A (en) | 2017-11-07 |
Family
ID=60193591
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710355273.1A Pending CN107319129A (en) | 2017-05-19 | 2017-05-19 | A kind of preparation of fermented bean dregs rich in astaxanthin and feed |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107319129A (en) |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107699527A (en) * | 2017-11-21 | 2018-02-16 | 广东海洋大学 | A kind of probiotics micro-ecological formulation and its preparation method and application |
| CN108441452A (en) * | 2018-04-13 | 2018-08-24 | 天津博菲德科技有限公司 | A kind of store pig compound microbial culture starter and its application |
| CN109997955A (en) * | 2019-04-15 | 2019-07-12 | 广东大广生物科技有限公司 | A kind of farrowing sow fermented feed and preparation method thereof |
| CN111165652A (en) * | 2020-01-19 | 2020-05-19 | 博益德（北京）生物科技有限公司 | Fermented feed for river crabs as well as preparation method and application of fermented feed |
| TWI715312B (en) * | 2019-11-26 | 2021-01-01 | 國立屏東科技大學 | Method for manufacturing raw material of feed |
| CN113249425A (en) * | 2021-05-18 | 2021-08-13 | 青岛润达生物科技有限公司 | Method for improving high yield of astaxanthin by rhodotorula benthica strain |
| CN115558694A (en) * | 2022-07-18 | 2023-01-03 | 好氧探索(武汉)生物科技有限公司 | Astaxanthin-rich vegetable protein and solid fermentation production method thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2008042338A2 (en) * | 2006-09-28 | 2008-04-10 | Microbia, Inc. | Production of carotenoids in oleaginous yeast and fungi |
| CN101433275A (en) * | 2008-12-16 | 2009-05-20 | 中国海洋大学 | Method for preparing Rhodotorula benthica fermentation feed |
| CN104845893A (en) * | 2015-05-13 | 2015-08-19 | 威海利达生物科技有限公司 | Rhodotorula mucilaginosa and application thereof to fermentation production of malus micromalus astaxanthin |
-
2017
- 2017-05-19 CN CN201710355273.1A patent/CN107319129A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2008042338A2 (en) * | 2006-09-28 | 2008-04-10 | Microbia, Inc. | Production of carotenoids in oleaginous yeast and fungi |
| CN101433275A (en) * | 2008-12-16 | 2009-05-20 | 中国海洋大学 | Method for preparing Rhodotorula benthica fermentation feed |
| CN104845893A (en) * | 2015-05-13 | 2015-08-19 | 威海利达生物科技有限公司 | Rhodotorula mucilaginosa and application thereof to fermentation production of malus micromalus astaxanthin |
Non-Patent Citations (1)
| Title |
|---|
| 李爱科: "《中国蛋白质饲料资源》", 31 January 2013, 中国农业大学出版社 * |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107699527A (en) * | 2017-11-21 | 2018-02-16 | 广东海洋大学 | A kind of probiotics micro-ecological formulation and its preparation method and application |
| CN107699527B (en) * | 2017-11-21 | 2021-01-08 | 广东海洋大学 | Probiotic micro-ecological preparation and preparation method and application thereof |
| CN108441452A (en) * | 2018-04-13 | 2018-08-24 | 天津博菲德科技有限公司 | A kind of store pig compound microbial culture starter and its application |
| CN109997955A (en) * | 2019-04-15 | 2019-07-12 | 广东大广生物科技有限公司 | A kind of farrowing sow fermented feed and preparation method thereof |
| TWI715312B (en) * | 2019-11-26 | 2021-01-01 | 國立屏東科技大學 | Method for manufacturing raw material of feed |
| CN111165652A (en) * | 2020-01-19 | 2020-05-19 | 博益德（北京）生物科技有限公司 | Fermented feed for river crabs as well as preparation method and application of fermented feed |
| CN113249425A (en) * | 2021-05-18 | 2021-08-13 | 青岛润达生物科技有限公司 | Method for improving high yield of astaxanthin by rhodotorula benthica strain |
| CN115558694A (en) * | 2022-07-18 | 2023-01-03 | 好氧探索(武汉)生物科技有限公司 | Astaxanthin-rich vegetable protein and solid fermentation production method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107319129A (en) | A kind of preparation of fermented bean dregs rich in astaxanthin and feed | |
| CN1281147C (en) | Microbial fodder additive and its preparing method and use | |
| CN103981118B (en) | A kind of bacillus subtilis feed addictive and its preparation method and application | |
| CN103275907B (en) | Bacillus amyloliquefacien and preparation method and application thereof | |
| CN101638627B (en) | Bacillus subtilis and application thereof in biological feed additives | |
| CN102488084B (en) | Method for preparing protein feed by fermentation on straw and cake using aerobic mixed bacteria | |
| CN106010990B (en) | A kind of rhodotorula mucilaginosa and its fermentation culture medium and application | |
| CN106260540B (en) | Biological feed for creep feed and creep feed | |
| CN103798553B (en) | Environment-friendly pig feed additive and preparation method thereof | |
| CN101671638B (en) | New strain of bifidobacterium and fermentative preparation method and application thereof | |
| CN104012758B (en) | biological fermentation feed and preparation method thereof | |
| CN106260504B (en) | Method for producing microbial fermentation wet feed by using beer yeast paste | |
| CN102907563B (en) | Method for preparing high-activity probiotic preparation for livestock breeding | |
| CN104293696A (en) | Enterococcus faecalis HEW-A131 and application thereof | |
| CN104286400A (en) | Method for preparing protein feed from straw and inedible vegetables through mixed bacteria aerobic fermentation | |
| CN107683972B (en) | Improve the fermented feed and its preparation method and application of milking sow production performance | |
| CN103907749A (en) | Biological fermentation active carrier type composite premix feed as well as preparation method and application thereof | |
| CN106721261A (en) | One kind is used for swine rearing mixed fermentation fiber feedstuff and preparation method thereof | |
| CN107455336A (en) | A kind of pig concentration of suitable the Northwest efficiently cultivates the swine rearing system of feeding pattern and suitable the Northwest | |
| CN102925374B (en) | Saccharomyces cerevisiae new strains, and the feed addictive containing it, batch | |
| CN110317753A (en) | A kind of bacillus megaterium microbial inoculum, liquid fermentation process and its application | |
| CN105861615A (en) | Production method of amino acid bioactive peptide for aquatic products | |
| CN104686797B (en) | The preparation method of feed probiotic microcapsule | |
| CN109170149A (en) | A method of microecological fermented forage is produced using antibiotic milk | |
| CN114190486A (en) | Microbial fermented feed for preventing and treating constipation of sows and sow breeding method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20171107 |