CN107455548A - A kind of method that edible mushroom solid state fermentation Soybean Meal prepares feeding level protein additive - Google Patents
A kind of method that edible mushroom solid state fermentation Soybean Meal prepares feeding level protein additive Download PDFInfo
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- CN107455548A CN107455548A CN201710799961.7A CN201710799961A CN107455548A CN 107455548 A CN107455548 A CN 107455548A CN 201710799961 A CN201710799961 A CN 201710799961A CN 107455548 A CN107455548 A CN 107455548A
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
- A23J1/14—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from leguminous or other vegetable seeds; from press-cake or oil-bearing seeds
- A23J1/148—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from leguminous or other vegetable seeds; from press-cake or oil-bearing seeds by treatment involving enzymes or microorganisms
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- 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
- A23K10/12—Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/142—Amino acids; Derivatives thereof
- A23K20/147—Polymeric derivatives, e.g. peptides or proteins
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Abstract
The invention belongs to feed additive field, and in particular to a kind of method that edible mushroom solid state fermentation Soybean Meal prepares feeding level protein additive.Including following steps：A. first class inoculum is prepared by main fermentation strain liquid fermentation of dust cover ghost toadstool；B. first class inoculum prepares second class inoculum by main raw material by solid fermentation of wheat；C. second class inoculum solid state fermentation Soybean Meal prepares feeding level protein additive.Compared with traditional zymotic technique, the preparation method has the characteristics of fermentation period is short, zymotechnique is stable and albumen potency is high, while tunning free from extraneous odour, viscosity is low, is easy to storage and processing.
Description
Technical field
The invention belongs to feed additive field, and in particular to a kind of edible mushroom solid state fermentation Soybean Meal prepares feeding level egg
The method of white additive.
Background technology
Soybean is one of main edible oil source in China as important industrial crops.Soybean Meal is soybean by squeezing
Remaining residue after oily PROCESS FOR TREATMENT, because of its higher nutritive value, the trophic structure of comparatively perfect and without unsaturation
The features such as aliphatic acid, is usually added in feed as protein sources.The Soybean Meal compared with the industrial wood waste of other oil expression plants
Main advantage be embodied in its higher yield, relatively low carbon-nitrogen ratio and complete amino acid composition relatively.However, in feed
In the actual production of industry, the application of Soybean Meal is but by many restrictions.During protein sources as aquatic feeds, Soybean Meal contains
Amount generally must not exceed 20%；During protein sources as animal feeding-stuff, Soybean Meal bel not applied to the raising of growing animal.Limitation is big
Dregs of beans is containing antitrypsin, uremic in undressed soya-bean cake, dregs of beans in the main reason for feed processing industry extensive use
Enzyme, saponin, goitre incitant etc., harmful effect can be produced to the digestibility and utilization of animal and feed.
Dust cover ghost toadstool has short life cycle, metabolism and genomics background clear as one of typical pattern fungi
The features such as clear, its mycelium and the equal edible of childhood fructification.Compared with unicellular probiotics, the mycelia of edible fungi has
Stronger robustness and the ability of transformation medium matrix.
The content of the invention
In order to solve the technical problem of Soybean Meal solid-state fermentation process, the invention provides a kind of edible mushroom solid state fermentation is big
The method that dregs of beans prepares feeding level protein additive, this method fermentation strain based on dust cover ghost toadstool, is mixed by solid-state
Zymotechnique improves the feed coefficient of Soybean Meal, prepares the feed protein additive using Soybean Meal as raw material.
The technical scheme is that：A kind of feeding level protein additive of edible mushroom solid state fermentation Soybean Meal, with tepetate ghost
Agaric is that feeding level protein additive, the addition of described feeding level albumen is prepared in main fermentation strain solid state fermentation Soybean Meal
Trypsin ihhibitor≤0.25 (mgg in agent-1)-1, albumen size is reduced to below 20kD percentage ＞
45%, histidine content accounts for mass percent >=4% of dry, while more rich in vitamin A, B1, the B2 lacked in raw material
Kind vitamin and Thick many candies, wherein, vitamin A content 0.40-0.90mg/100g；Vitamin B1 content is 0.50mg-
0.10/100g；Vitamin B2 content is 0.50mg-0.90/100g；Thick many candies content is 120-180mg/100g.
First class inoculum specifically is prepared by main fermentation strain liquid fermentation of dust cover ghost toadstool, first class inoculum is again with wheat
Second class inoculum is prepared for main raw material by solid fermentation, solid state fermentation Soybean Meal obtains feeding level protein additive to second class inoculum again.
Using dust cover ghost toadstool as main fermentation strain, while for the different aminoacids demand of different raising objects, addition
One or several kinds in following dust cover ghost toadstool symbiosis bacterial strain are as assisted fermentation bacterial strain：Aspergillus oryzae, bafillus natto, production protein
Candida and amphimicrobian lactic acid bacteria.
The method for preparing the described feeding level protein additive of edible mushroom solid state fermentation Soybean Meal, based on dust cover ghost toadstool
Fermentation strain liquid fermentation is wanted to prepare first class inoculum, first class inoculum prepares two level bacterium by main raw material by solid fermentation of wheat again
Kind, solid state fermentation Soybean Meal obtains feeding level protein additive to second class inoculum again.
The method of the described preparation feeding level protein additive of edible mushroom solid state fermentation Soybean Meal, including following steps：
D) preparation of first class inoculum：Dust cover ghost toadstool parent species activate on PDA plate, inoculation fermentation culture medium, 25-35
DEG C, 100-150r/min isothermal vibration culture 72-96 hours；
E) preparation of second class inoculum：After the completion of first class inoculum activation, inoculation wheat culture medium, 25-35 DEG C, quiescent culture,
96-120 hours；
F) Soybean Meal ferments：Soybean Meal medium sterilization is simultaneously cooled to room temperature, by Soybean Meal culture medium quality of material
5%-10% accesses ready second class inoculum, is fermented 48-96 hours in 25-35 DEG C, and feeding level albumen addition is obtained after drying
Agent.
In the step a, fermentation medium is to add magnesium sulfate into potato standard medium by mass percentage
0.1%-0.3% and calcium chloride 1%-3%.
In the step b, wheat culture medium forms by percentage to the quality is：Ripe fresh band wheat bran wheat berry 40-
50%th, glucose 1-3%, yeast extract 0.1%-0.5%, magnesium sulfate 0.1-0.3%, calcium chloride 1-3% remaining be water.
In the step c, Soybean Meal culture medium forms by percentage to the quality is：Soybean Meal 35-45%, corn flour 3-
5%th, wheat bran 1-5%, sucrose 1-2%, calcium carbonate 2-5%, magnesium sulfate 0.1-0.3% remaining be water.
Described Soybean Meal is that particle diameter is less than or equal to 20 purposes.
Beneficial effect
After fermented processing, the ANFs in Soybean Meal significantly reduces, molecular weight of albumen reduces, sensitization reduces,
Nutritive value improves, palatability improves.It is embodied in：Trypsin ihhibitor reduces by more than 90%, and albumen size reduces
To below 20kD, histidine content improves more than 200%, at the same rich in vitamin A, B, D, E, and mineral element with it is thick more
Sugar.
The Antagonistic protein of Coprinus cinereus secretion has the ability for destroying prokaryote wall, in bean pulp fermentation and storage
During safely and steadly suppress harmful microbe growth metabolism.
The first class inoculum culture medium that the present invention uses can be used for the multiple-microorganism of the invention used individually or mixed fermentation；
For half-natural solid medium, the culture medium can effectively keep spawn activity, prevent that strain is early the second class inoculum culture medium used
Decline.
Prepared by the method that a kind of edible mushroom solid state fermentation Soybean Meal provided by the invention prepares feeding level protein additive
Fermentation soybean dregs of rice surface color is pure white, texture is tough, has the distinctive delicate fragrance of ghost agaric mycelium, while a large amount of mycelium are formed
Hydrophobic structure enable its for a long time floating water surface, should not disperse, be splendid fishing protein feed source.
The fermentation soybean dregs of rice that based on Coprinus cinereus prepared by fermentation strain and traditional mould or unicellular probiotics fermention
Dregs of beans is compared and had the advantage that：First, its amino acid pattern is optimized, and relieving must amino acid starvation under specified conditions
The nutrition short -board effect brought；Secondly, the antagonistic substance using trypsin inhibitor as representative significantly reduces, and improves protein
Digestibility；Finally, the technique prepare fermentation soybean dregs of rice color is pure white, smell delicate fragrance, is not easy adhesion, is not easy the moisture absorption.It is comprehensive
Upper described, the zymotechnique disclosure satisfy that feed addictive industry for nutrition, the requirement of flavor, safety, has filled up feeding hair
The blank in ferment Soybean Meal market.
Embodiment
The edible mushroom solid state fermentation Soybean Meal method for preparing feeding level protein additive of the present invention, using dust cover ghost toadstool as
Basal fermentation bacterial strain, aspergillus oryzae, bafillus natto, candida utili are added according to the amino acid requirement that raising object is different
And the one or more in amphimicrobian lactic acid bacteria mix hair with solid-state as assisted fermentation bacterial strain by liquid mixed fermentation
Two steps of ferment improve the nutrition performance of Soybean Meal.
Including following steps：
A) preparation of first class inoculum：Parent species activate on PDA plate, inoculation fermentation culture medium, 25-35 DEG C, 100-150r/
Min isothermal vibration culture 72-96 hours.Fermentation medium is to add sulfuric acid into potato standard medium by mass percentage
Magnesium 0.1%-0.3% and calcium chloride 1%-3%, can influence ferment effect, potato if not adding magnesium sulfate and calcium chloride
Leaching liquor 90%-96%, glucose 1%-2%, yeast extract 0.3%,.
B) preparation of second class inoculum：After the completion of first class inoculum activation, inoculation wheat culture medium, 25-35 DEG C, quiescent culture,
96-120 hours.
C) Soybean Meal ferments：Soybean Meal medium sterilization is simultaneously cooled to room temperature, by Soybean Meal culture medium quality of material
5%-10% accesses ready second class inoculum, is fermented 48-96 hours in 25-35 DEG C.Pancreas in the feeding level protein additive of finished product
Protease inhibitor≤0.25 (mgg-1)-1, albumen size is reduced to below 20kD percentage ＞ 45%, organizes ammonia
Acid content accounts for mass percent >=4% of dry, and 250%-400% is improved compared with raw material dregs of beans；It is rich in raw material and lacks simultaneously
Vitamin A, multivitamin and the Thick many candies such as B1, B2.Wherein, vitamin A is improved to 0.40- by 0mg/100g
0.90mg/100g；Vitamin B1 is improved to 0.50mg-0.10/100g by 0.45mg/100g；Vitamin B2 is by 0.2mg/100g
Improve to 0.50mg-0.90/100g；Thick many candies are improved to 120-180mg/100g by 75mg/100g.
The dust cover ghost toadstool of the present invention is bought from American Type Culture Collecti (American Type Culture
Collection), deposit number is ATCC 18450, and collection address is American Type Culture
Collection(ATCC),P.O.Box 1549,Manassas,VA 20108USA；Aspergillus oryzae is bought from Chinese industrial microorganism
DSMZ, deposit number are CICC 2014, and collection address is Xiaoyun Road, Chaoyang District, Beijing City 32；Natto bud
Spore bacillus is bought from Chinese industrial Culture Collection, and deposit number is CICC 10023, and collection address is north
Jing Shi Chaoyang Districts Xiao Yun roads 32；Candida utili is bought from Chinese industrial Culture Collection, deposit number
CICC 1314, collection address are Xiaoyun Road, Chaoyang District, Beijing City 32；Lactic acid bacteria is bought from Chinese industrial microorganism fungus kind
Collection, deposit number CICC21790, collection address are Xiaoyun Road, Chaoyang District, Beijing City 32.
Embodiment 1
The single culture dust cover ghost toadstool fermentation soybean dregs of rice make feed protein additive
Dust cover ghost toadstool purchase is protected from American Type Culture Collecti (American Type Culture Collection)
It is ATCC 18450 to hide numbering, and collection address is American Type Culture Collection (ATCC),
P.O.Box 1549,Manassas,VA 20108 USA。
Parent species are inoculated in primary-seed medium from dust cover ghost toadstool mycelium after activating on PDA plate, 35 DEG C,
150r/min isothermal vibrations culture 96 hours.First class inoculum activation after the completion of, with 10% inoculum concentration be inoculated with wheat culture medium, 35
DEG C, quiescent culture, 120 hours.Soybean Meal medium sterilization is simultaneously cooled to room temperature, by the 10% of Soybean Meal culture medium quality of material
Ready second class inoculum is accessed, is fermented 96 hours in 35 DEG C.50 DEG C of dryings 6 hours.
The fermentation index of table 1
Embodiment 2
Dust cover ghost toadstool makes feed protein additive with bafillus natto mixed fermentation Soybean Meal
Dust cover ghost toadstool purchase is protected from American Type Culture Collecti (American Type Culture Collection)
It is ATCC 18450 to hide numbering, and collection address is American Type Culture Collection (ATCC),
P.O.Box 1549,Manassas,VA 20108 USA；Aspergillus oryzae purchase is protected from Chinese industrial Culture Collection
It is CICC 2014 to hide numbering, and collection address is Xiaoyun Road, Chaoyang District, Beijing City 32.
Parent species are inoculated in first order seed from dust cover ghost toadstool mycelium and aspergillus oryzae cell after being activated on PDA plate
Culture medium, 32 DEG C, 150r/min isothermal vibrations culture 72 hours.After the completion of first class inoculum activation, wheat is inoculated with 10% inoculum concentration
Grain culture medium, 32 DEG C, quiescent culture, 96 hours.Soybean Meal medium sterilization is simultaneously cooled to room temperature, by Soybean Meal culture medium material
The 10% ready second class inoculum of access of quality, ferments 96 hours in 32 DEG C.50 DEG C of dryings 6 hours.
The fermentation index of table 2
Embodiment 3
Dust cover ghost toadstool, lactic acid bacteria mixed fermentation Soybean Meal make feed protein additive
Dust cover ghost toadstool purchase is protected from American Type Culture Collecti (American Type Culture Collection)
It is ATCC 18450 to hide numbering, and collection address is American Type Culture Collection (ATCC),
P.O.Box 1549,Manassas,VA 20108 USA；Lactic acid bacteria purchase is protected from Chinese industrial Culture Collection
It is CICC 21790 to hide numbering, and collection address is Xiaoyun Road, Chaoyang District, Beijing City 32.
Parent species are from the lactic acid bacteria bacterium activated in dust cover ghost toadstool mycelium after being activated on PDA plate and MRS culture mediums
Body, is inoculated in primary-seed medium, 32 DEG C, 100r/min isothermal vibrations culture 96 hours.After the completion of first class inoculum activation, with
10% inoculum concentration inoculation wheat culture medium, 32 DEG C, quiescent culture, 120 hours.Soybean Meal medium sterilization is simultaneously cooled to room temperature,
By the 10% ready second class inoculum of access of Soybean Meal culture medium quality of material, fermented 72 hours in 32 DEG C.50 DEG C of dryings 6 are small
When.
The fermentation index of table 3
Embodiment 4
Dust cover ghost toadstool makes feed protein additive with candida utili mixed fermentation Soybean Meal
Dust cover ghost toadstool purchase is protected from American Type Culture Collecti (American Type Culture Collection)
It is ATCC 18450 to hide numbering, and collection address is American Type Culture Collection (ATCC),
P.O.Box 1549,Manassas,VA 20108 USA；Candida utili is bought from Chinese industrial Microbiological Culture Collection
The heart, deposit number are CICC 1314, and collection address is Xiaoyun Road, Chaoyang District, Beijing City 32；Lactic acid bacteria purchase is certainly Chinese
Research for Industrial Microbial Germ collection, deposit number are CICC 21790, and collection address is Xiaoyun Road, Chaoyang District, Beijing City
No. 32.
Parent species are from dust cover ghost toadstool mycelium after being activated on PDA plate and the Candida utilis ferment of LB culture mediums activation
Mother, is inoculated in primary-seed medium, 35 DEG C, 150r/min isothermal vibrations culture 96 hours.After the completion of first class inoculum activation, with
10% inoculum concentration inoculation wheat culture medium, 35 DEG C, quiescent culture, 120 hours.Soybean Meal medium sterilization is simultaneously cooled to room temperature,
By the 10% ready second class inoculum of access of Soybean Meal culture medium quality of material, fermented 96 hours in 35 DEG C.50 DEG C of dryings 6 are small
When.
The fermentation index of table 1
Embodiment 5
Dust cover ghost toadstool, lactic acid bacteria, bafillus natto and candida utili common fermentation Soybean Meal make feed protein
Additive
Dust cover ghost toadstool purchase is protected from American Type Culture Collecti (American Type Culture Collection)
It is ATCC 18450 to hide numbering, and collection address is American Type Culture Collection (ATCC),
P.O.Box 1549,Manassas,VA 20108 USA；Aspergillus oryzae purchase is protected from Chinese industrial Culture Collection
It is CICC 2014 to hide numbering, and collection address is Xiaoyun Road, Chaoyang District, Beijing City 32；Bafillus natto purchase is certainly Chinese
Research for Industrial Microbial Germ collection, deposit number are CICC 10023, and collection address is Xiaoyun Road, Chaoyang District, Beijing City
No. 32；Candida utili is bought from Chinese industrial Culture Collection, and deposit number is CICC 1314, in preservation
Heart location is Xiaoyun Road, Chaoyang District, Beijing City 32；Lactic acid bacteria purchase is compiled from Chinese industrial Culture Collection, preservation
Number it is CICC 21790, collection address is Xiaoyun Road, Chaoyang District, Beijing City 32.
Parent species are activated from dust cover ghost toadstool mycelium after being activated on PDA plate and aspergillus oryzae cell, MRS culture mediums
Lactic acid bacteria and the candida utili of LB culture mediums activation, primary-seed medium is inoculated in, 35 DEG C, 150r/min constant temperature shakes
Swing culture 96 hours.First class inoculum activation after the completion of, with 10% inoculum concentration be inoculated with wheat culture medium, 35 DEG C, quiescent culture, 120
Hour.Soybean Meal medium sterilization is simultaneously cooled to room temperature, and ready two are accessed by the 10% of Soybean Meal culture medium quality of material
Level strain, ferments 96 hours in 35 DEG C.50 DEG C of dryings 6 hours.
The fermentation index of table 5
It is good using the present invention fermentation soybean dregs of rice smell delicate fragrance, the palatability that prepare, retain the intrinsic nutrition of the raw soybeans dregs of rice into
Its antagonism factor level is substantially reduced while dividing not destroyed.In addition contained albumen is converted into small peptide mostly in Soybean Meal
Form so that its intrinsic antigen protein inactivation, avoid the immune response easily triggered in mammal cub feeding process and ask
Topic.
Claims (9)
1. a kind of feeding level protein additive of edible mushroom solid state fermentation Soybean Meal, it is characterised in that using dust cover ghost toadstool be main
Fermentation strain solid state fermentation Soybean Meal is prepared feeding level protein additive, tryptose in described feeding level protein additive
Enzyme inhibition factor≤0.25 (mgg-1)-1, albumen size is reduced to below 20kD percentage ＞ 45%, and histidine contains
Amount accounts for mass percent >=4% of dry, while rich in vitamin A, B1, B2 multivitamin and thick lacked in raw material
Polysaccharide, wherein, vitamin A content 0.40-0.90mg/100g；Vitamin B1 content is 0.50mg-0.10/100g；Vitamin
B2 contents are 0.50mg-0.90/100g；Thick many candies content is 120-180mg/100g.
2. the feeding level protein additive of edible mushroom solid state fermentation Soybean Meal according to claim 1, it is characterised in that specific
It is to prepare first class inoculum by main fermentation strain liquid fermentation of dust cover ghost toadstool, first class inoculum is consolidated by primary raw material of wheat again
State fermentation prepares second class inoculum, and solid state fermentation Soybean Meal obtains feeding level protein additive to second class inoculum again.
3. the feeding level protein additive of edible mushroom solid state fermentation Soybean Meal according to claim 1 or 2, it is characterised in that
Using dust cover ghost toadstool as main fermentation strain, while for the different aminoacids demand of different raising objects, add following tepetate
One or several kinds in ghost agaric symbiosis bacterial strain are as assisted fermentation bacterial strain：Aspergillus oryzae, bafillus natto, candida utili
And amphimicrobian lactic acid bacteria.
4. the method for any described feeding level protein additives of edible mushroom solid state fermentation Soybean Meal of claim 1-3 is prepared, its
It is characterised by, prepares first class inoculum by main fermentation strain liquid fermentation of dust cover ghost toadstool, first class inoculum is again based on wheat
Raw material by solid fermentation is wanted to prepare second class inoculum, solid state fermentation Soybean Meal obtains feeding level protein additive to second class inoculum again.
5. the method according to claim 3 for preparing the feeding level protein additive of edible mushroom solid state fermentation Soybean Meal, it is special
Sign is, including following steps：
A) preparation of first class inoculum：Dust cover ghost toadstool parent species activate on PDA plate, inoculation fermentation culture medium, 25-35 DEG C,
100-150r/min isothermal vibration culture 72-96 hours；
B) preparation of second class inoculum：After the completion of first class inoculum activation, inoculation wheat culture medium, 25-35 DEG C, quiescent culture, 96-
120 hours；
C) Soybean Meal ferments：Soybean Meal medium sterilization is simultaneously cooled to room temperature, by the 5%- of Soybean Meal culture medium quality of material
The 10% ready second class inoculum of access, ferments 48-96 hours in 25-35 DEG C, feeding level protein additive is obtained after drying.
6. the method according to claim 5 for preparing the feeding level protein additive of edible mushroom solid state fermentation Soybean Meal, it is special
Sign is：In the step a, fermentation medium is to add magnesium sulfate into potato standard medium by mass percentage
0.1%-0.3% and calcium chloride 1%-3%.
7. the method according to claim 5 for preparing the feeding level protein additive of edible mushroom solid state fermentation Soybean Meal, it is special
Sign is：In the step b, wheat culture medium forms by percentage to the quality is：Ripe fresh band wheat bran wheat berry 40-
50%th, glucose 1-3%, yeast extract 0.1%-0.5%, magnesium sulfate 0.1-0.3%, calcium chloride 1-3% remaining be water.
8. the method according to claim 5 for preparing the feeding level protein additive of edible mushroom solid state fermentation Soybean Meal, it is special
Sign is：In the step c, Soybean Meal culture medium forms by percentage to the quality is：Soybean Meal 35-45%, corn flour 3-
5%th, wheat bran 1-5%, sucrose 1-2%, calcium carbonate 2-5%, magnesium sulfate 0.1-0.3% remaining be water.
9. the method according to claim 8 for preparing the feeding level protein additive of edible mushroom solid state fermentation Soybean Meal, it is special
Sign is：Described Soybean Meal is that particle diameter is less than or equal to 20 purposes.
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Application publication date: 20171212 |