CN106721054A - The method that forage protein is prepared using alcohol effluent and crop material - Google Patents
The method that forage protein is prepared using alcohol effluent and crop material Download PDFInfo
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- CN106721054A CN106721054A CN201611154585.8A CN201611154585A CN106721054A CN 106721054 A CN106721054 A CN 106721054A CN 201611154585 A CN201611154585 A CN 201611154585A CN 106721054 A CN106721054 A CN 106721054A
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- stalk
- solid
- alcohol effluent
- crop material
- fermentation
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- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 title claims abstract description 38
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 25
- 102000004169 proteins and genes Human genes 0.000 title claims abstract description 25
- 239000000463 material Substances 0.000 title claims abstract description 24
- 238000000034 method Methods 0.000 title claims abstract description 20
- 239000004459 forage Substances 0.000 title claims abstract description 18
- 238000004176 ammonification Methods 0.000 claims abstract description 13
- 230000007062 hydrolysis Effects 0.000 claims abstract description 9
- 238000006460 hydrolysis reaction Methods 0.000 claims abstract description 9
- 238000010563 solid-state fermentation Methods 0.000 claims abstract description 7
- 239000007787 solid Substances 0.000 claims description 43
- 238000000855 fermentation Methods 0.000 claims description 33
- 230000004151 fermentation Effects 0.000 claims description 33
- 239000000843 powder Substances 0.000 claims description 25
- 235000015099 wheat brans Nutrition 0.000 claims description 23
- 239000007788 liquid Substances 0.000 claims description 22
- 239000001963 growth medium Substances 0.000 claims description 21
- 241000235646 Cyberlindnera jadinii Species 0.000 claims description 20
- 239000010902 straw Substances 0.000 claims description 18
- 239000000047 product Substances 0.000 claims description 17
- 239000006228 supernatant Substances 0.000 claims description 15
- 240000008042 Zea mays Species 0.000 claims description 14
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 14
- XDOFQFKRPWOURC-UHFFFAOYSA-N 16-methylheptadecanoic acid Chemical compound CC(C)CCCCCCCCCCCCCCC(O)=O XDOFQFKRPWOURC-UHFFFAOYSA-N 0.000 claims description 12
- 241000228245 Aspergillus niger Species 0.000 claims description 12
- 240000006439 Aspergillus oryzae Species 0.000 claims description 12
- 235000002247 Aspergillus oryzae Nutrition 0.000 claims description 12
- 239000000203 mixture Substances 0.000 claims description 11
- 238000000926 separation method Methods 0.000 claims description 11
- 244000063299 Bacillus subtilis Species 0.000 claims description 10
- 235000014469 Bacillus subtilis Nutrition 0.000 claims description 10
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 10
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 10
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 10
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 10
- 235000005822 corn Nutrition 0.000 claims description 10
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims description 10
- 239000002609 medium Substances 0.000 claims description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 10
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 9
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 claims description 9
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims description 9
- 239000004202 carbamide Substances 0.000 claims description 9
- 241000894006 Bacteria Species 0.000 claims description 7
- 235000021355 Stearic acid Nutrition 0.000 claims description 6
- 238000005273 aeration Methods 0.000 claims description 6
- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 claims description 6
- OQCDKBAXFALNLD-UHFFFAOYSA-N octadecanoic acid Natural products CCCCCCCC(C)CCCCCCCCC(O)=O OQCDKBAXFALNLD-UHFFFAOYSA-N 0.000 claims description 6
- 239000008117 stearic acid Substances 0.000 claims description 6
- 241000228212 Aspergillus Species 0.000 claims description 5
- 241000209140 Triticum Species 0.000 claims description 5
- 235000021307 Triticum Nutrition 0.000 claims description 5
- 230000004913 activation Effects 0.000 claims description 5
- 235000011114 ammonium hydroxide Nutrition 0.000 claims description 5
- 239000000306 component Substances 0.000 claims description 5
- 230000003413 degradative effect Effects 0.000 claims description 5
- 235000014655 lactic acid Nutrition 0.000 claims description 5
- 239000004310 lactic acid Substances 0.000 claims description 5
- 239000002689 soil Substances 0.000 claims description 5
- 230000001954 sterilising effect Effects 0.000 claims description 5
- 239000000725 suspension Substances 0.000 claims description 5
- 235000016383 Zea mays subsp huehuetenangensis Nutrition 0.000 claims description 4
- 235000009973 maize Nutrition 0.000 claims description 4
- 238000012549 training Methods 0.000 claims description 4
- 240000007594 Oryza sativa Species 0.000 claims description 3
- 235000007164 Oryza sativa Nutrition 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- 235000009566 rice Nutrition 0.000 claims description 3
- 102000005575 Cellulases Human genes 0.000 claims 1
- 108010084185 Cellulases Proteins 0.000 claims 1
- 239000012535 impurity Substances 0.000 claims 1
- 238000012958 reprocessing Methods 0.000 claims 1
- 210000002700 urine Anatomy 0.000 claims 1
- 238000009423 ventilation Methods 0.000 claims 1
- 108090000790 Enzymes Proteins 0.000 abstract description 9
- 102000004190 Enzymes Human genes 0.000 abstract description 9
- 238000011161 development Methods 0.000 abstract description 6
- 230000008901 benefit Effects 0.000 abstract description 4
- 238000004519 manufacturing process Methods 0.000 abstract description 4
- 244000144972 livestock Species 0.000 abstract description 3
- 201000010099 disease Diseases 0.000 abstract 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract 1
- 230000002708 enhancing effect Effects 0.000 abstract 1
- 230000036039 immunity Effects 0.000 abstract 1
- 230000000968 intestinal effect Effects 0.000 abstract 1
- 238000004064 recycling Methods 0.000 abstract 1
- 229940088598 enzyme Drugs 0.000 description 8
- 108010059892 Cellulase Proteins 0.000 description 4
- 244000068988 Glycine max Species 0.000 description 4
- 235000010469 Glycine max Nutrition 0.000 description 4
- 229940106157 cellulase Drugs 0.000 description 4
- 229910052799 carbon Inorganic materials 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 230000000050 nutritive effect Effects 0.000 description 3
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- 235000013339 cereals Nutrition 0.000 description 2
- 239000007857 degradation product Substances 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 230000002255 enzymatic effect Effects 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 241000235342 Saccharomycetes Species 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 108010089934 carbohydrase Proteins 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 235000019621 digestibility Nutrition 0.000 description 1
- 230000001079 digestive effect Effects 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 235000013601 eggs Nutrition 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 235000021050 feed intake Nutrition 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 235000019629 palatability Nutrition 0.000 description 1
- -1 pectase Proteins 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000007634 remodeling Methods 0.000 description 1
- 239000010865 sewage Substances 0.000 description 1
- 239000002002 slurry Substances 0.000 description 1
- 230000031068 symbiosis, encompassing mutualism through parasitism Effects 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
- 239000002023 wood Substances 0.000 description 1
- 229920001221 xylan Polymers 0.000 description 1
- 150000004823 xylans Chemical class 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
-
- 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
Landscapes
- Fodder In General (AREA)
Abstract
The present invention is not high for crop material fodder availability, the phenomenon of alcohol effluent pollution, it is this method the invention discloses forage protein is prepared using alcohol effluent and crop material, with the objective demand for meeting stalk comprehensive utilization, livestock feed albumen demand, alcohol effluent harmless treatment and utilize.The present invention is made using main using three big steps, respectively ammonification, complex enzyme hydrolysis, the step solid state fermentation of multi-cultur es three, its advantage is have the advantages that enhancing development, improve efficiency of feed utilization, strengthen immunity, improve intestinal microecology, prevent disease, realize the feeding resource high-value-use of crop material and harmless treatment and the recycling of alcohol effluent, be conducive to the health green scientific development of feed manufacturing industry, have broad application prospects.
Description
Technical field
The present invention relates to a kind of method that utilization alcohol effluent and crop material prepare forage protein, belong to feed technology and add
Work field.
Background technology
Now with the improvement of people's living standards and the attention to quality of life, to meat, egg, milk, aquatic products demand
Increasing, grain and Feed Manufacturing face a severe test, and feed protein resource will be increasingly deficient,
Therefore grain is improved to the transformation efficiency of livestock products and the utilization rate of feed, feed resource is expanded, and develops unconventional feeding
Material is extremely urgent, and protein raw material is the basis of feed industrial development, and the in short supply and price of China's protein raw material is surging
Through one of the bottleneck as restriction animal husbandry development, be this strategy for being used aiming at existing protein sources, improve its utilization
Rate, excavate as much as possible feed using nutritional ingredient, fundamentally alleviate feedstuff situation in short supply, according to grinding
Study carefully, by using alcohol effluent and crop material, both are added in feed, the protein content in feed, straw can be strengthened
Stalk is a kind of unconventional property feed resource, and the coarse hard, palatability of its quality is poor, digestive utilization ratio is low, nutritive value is not high,
But after through rationally processing, feed intake, digestibility and nutritive value can be improved, feed plant-eating animal or as preparing complete feed
Base-material, plays the role of good to growth of animal, development and weightening, the raising price of deed and economic benefit.Therefore it is fine using stalk
Dimension element production nutritive value feed higher has bright prospects.Meanwhile, alcohol swallow is evaporated in waste liquid containing a large amount of unemployed
Nutritional ingredient protein, fat, carbohydrate, mineral matter etc. one by one, therefore try to be used to produce protein feed by slops recovery,
Just turn into the striving direction of scientific and technical personnel.Current alcohol effluent is changed into more than feed using separation of solid and liquid, the technique of concentrate drying,
Substantial amounts of sewage is produced, energy consumption is serious, developing energy-saving fodder complete utilization using alcohol effluent turns into alcohol industry development most
Have the utilization ways of application value, it is therefore necessary to which invention is a kind of to prepare forage protein using alcohol effluent and crop material
Method.
The content of the invention
The present invention is for crop material fodder availability is high, the with serious pollution present situation of alcohol effluent, there is provided Yi Zhongli
The method that forage protein is prepared with alcohol effluent and crop material, with meet stalk comprehensive utilization, livestock feed albumen demand,
Alcohol effluent harmless treatment and the objective demand for utilizing.Therefore, above-mentioned purpose of the present invention is to give by the following technical programs
Realize：
The present invention provides a kind of method that utilization alcohol effluent and crop material prepare forage protein, it is characterised in that specifically include
Following steps：
（1）The basic handling of feedstuff：First by crushed stalk：Choose without the stalk for going mouldy, remove the magazines such as soil, crush
Cross 60 mesh sieve.
（2）Separation of solid and liquid：Alcohol effluent natural subsidence separation of solid and liquid, obtains supernatant and fermentation vinasse.
（3）Stalk treatment：
By straw ammoniation：The ammoniacal liquor of stalk 3 ~ 6% is added, alcohol effluent supernatant, adjustment moisture content to 50 ~ 65%, mixing is added
Uniformly, ammonification 2 ~ 6 days under closed environment are placed in.
Stalk is digested：Stalk pH after adjustment ammonification is it is essential to ensure that be between 4.0 ~ 5.0, to add cellulase, wood poly-
Carbohydrase, pectase, enzyme concentration are respectively 2 ~ 3%, 4 ~ 5%, 2 ~ 4%, are well mixed, and digest 10 ~ 15h, hydrolysis temperature control 35 ~
40℃。
（4）Prepare solid medium：Stalk after enzymolysis is mixed with fermentation vinasse, and adds corn pulp, wheat bran, soya-bean cake
Powder, urea, ammonium sulfate etc., adjust culture medium, it is ensured that its water content 100 ~ 120% with supernatant, and it is 7.0 or so to adjust initial pH
（5）Mould treatment is aspergillus activation：Fresh wheat bran is taken, fine powder is weeded out with 40 mesh sieve, by wheat bran：Water is according to 1：2 ratio
Example is carried out, and is fully mixed thoroughly and not only without dry powder but also occur without into the phenomenon of bulk.It is divided in 2000mL triangular flasks, every
The bottled wet wheat bran about 100g of 2000mL triangles, sterilize 45 ~ 65min under 0.1MPa gauge pressures, is cooled to 30 ~ 35 DEG C, and each three
Access inclined-plane aspergillus oryzae cv-8, aspergillus niger sp-1 spores that 2 ~ 3 rings have been activated in the bottle of angle, the control of culture medium product temperature (35 ±
1) DEG C every 10 ~ 15h shaking flasks once, make its sufficient contact, until stopping prepares spore suspension after spore grows.
（6）Solid fermentation：By aspergillus oryzae 3 ~ 4%, aspergillus niger 8 ~ 12%, isostearic acid 0.05-0.08%, different stearic acid
0.02-0.05% is inoculated into mix in proportion, is mixed thoroughly, is cultivated at a temperature of 30 ~ 37 DEG C, aerlbic culture 2 ~ 4 days.
（7）Candida utili is activated：Candida utili is connected in YPD culture mediums, 30 DEG C, under 180r/min rotating speeds
Constant temperature shaking flask 24 hours；Then with 1:100 ratios, seed liquor is connected in YPD culture mediums, obtains saccharomyces cerevisiae and Candida utilis
Yeast liquid seeds；
（8）Bacillus subtilis activates：24h is cultivated under the conditions of 30 ~ 35 DEG C using LB culture mediums, liquid seeds are obtained.
（9）Secondary solid is fermented：Candida utili, bacillus subtilis are seeded to by 4 ~ 6%, 3 ~ 4% inoculative proportions
Culture medium, 30 DEG C or so are cultivated 3 ~ 4 days.
（10）Three solid fermentations：Solid lactic acid bacterium powder is seeded to culture medium by 4 ~ 6% inoculative proportion, 29 ~ 35 DEG C are detested
Oxygen culture 24h.
（11）Dry：Low-temperature aeration is dried after fermentation ends.
（13）Crush：Dried fermented feed is crushed and is got product.
Above-mentioned stalk can be one or more in wheat stalk, maize straw, rice straw.
Above-mentioned steps（4）Middle solid-state fermentation culture medium component is：Straw degradative product 30 ~ 35%, vinasse 20 ~ 25%, corn
Slurry 8 ~ 20%, wheat bran 2.5 ~ 3.5%, beancake powder 9 ~ 11%, urea 4 ~ 6%, ammonium sulfate 7 ~ 11%.
Above-mentioned steps（6）Middle solid medium is without sterilizing.
The present invention provides a kind of method that utilization alcohol effluent and crop material prepare forage protein, the advantage is that：It is logical
The ammoniated treatment and complex enzyme hydrolysis of stalk are crossed, cellulose, xylan that microorganism is difficult to utilize etc. is degraded to reduced sugar, through surveying
The content of reducing sugar in complex enzyme hydrolysis ensuing crop stalk is determined more than 40%, up to 42.56%, than the simple ammonification of current report
Treatment or complex enzyme hydrolysis facture improve more than 20%, and the growth and breeding that content of reducing sugar rises to microorganism provides abundance
Carbon source, is degraded the bacterium in stalk using aspergillus niger and aspergillus oryzae, and it is mutual that the saccharomycete in mould and vinasse is mixed
Favour symbiosis makes stalk protein content improve a lot, using in mixed fermentation with various bacterium, the sugar of enzymatic catalysis generation immediately by
The Institute of Micro-biology of sugar fermentation utilizes, and thus maintains the concentration of degradation product, eliminates the degradation product that enzymatic synthesis effect is subject to
Effect is checked, feedback inhibition of the reacting final product to enzyme is also relieved, fermentation process is shortened, using stepwise fermentation, zymophyte
Strain requires different, different reaction controlling conditions to temperature, pH, logical oxygen etc., improves the growth rate of thalline, shortens fermentation
Time, a small amount of isostearic acid and different stearic acid are added in first time solid fermentation process, make aspergillus oryzae, aspergillus niger enzyme activity produce
Synergy, further increases percent protein, and the present invention uses solid state fermentation, simple production process, non-environmental-pollution.
Specific embodiment
First, specific embodiment
Embodiment 1
A kind of method that utilization alcohol effluent and crop material prepare forage protein, specific preparation process is as follows：
（1）The basic handling of feedstuff：First by crushed stalk：Choose without the stalk for going mouldy, remove the magazines such as soil, crush
Cross 60 mesh sieve.
（2）Separation of solid and liquid：Alcohol effluent natural subsidence separation of solid and liquid, obtains supernatant and fermentation vinasse.
（3）Stalk treatment：
By straw ammoniation：The ammoniacal liquor of stalk 3% is added, alcohol effluent supernatant is added, adjustment moisture content to 50 ~ 65%, mixing is equal
It is even, it is placed in ammonification 2 days under closed environment.
Stalk is digested：Stalk pH after adjustment ammonification is 4.0, adds cellulase, zytase, pectase, enzyme concentration
Respectively %, 4%, 2%, are well mixed, and digest 10h, and hydrolysis temperature is controlled at 35 DEG C.
（4）Prepare solid medium：Stalk after enzymolysis is mixed with fermentation vinasse, and adds corn pulp, wheat bran, soya-bean cake
Powder, urea, ammonium sulfate etc., adjust culture medium, it is ensured that its water content 100% with supernatant, and it is 7.0 to adjust initial pH.
（5）Mould treatment is aspergillus activation：Fresh wheat bran is taken, fine powder is weeded out with 40 mesh sieve, by wheat bran：Water is according to 1：2
Ratio carry out, fully mix thoroughly and not only without dry powder but also occur without into the phenomenon of bulk.It is divided in 2000mL triangular flasks, every
The bottled wet wheat bran about 100g of 2000mL triangles, sterilize 45min under 0.1MPa gauge pressures, is cooled to 30 DEG C, in each triangular flask
Inclined-plane aspergillus oryzae cv-8, the aspergillus niger sp-1 spores for accessing that 2 rings have activated, the control of culture medium product temperature (35 ± 1) DEG C every
10 ~ 15h shaking flasks once, make its sufficient contact, until stopping prepares spore suspension after spore grows.
（6）Solid fermentation：Aspergillus oryzae 3%, aspergillus niger 82%, isostearic acid 0.05%, different stearic acid 0.02% are inoculated with
To in mix, mix thoroughly, cultivated at a temperature of 30 DEG C, aerlbic culture 2 days.
（7）Candida utili is activated：Candida utili is connected in YPD culture mediums, 30 DEG C, under 180r/min rotating speeds
Constant temperature shaking flask 24 hours；Then with 1:100 ratios, seed liquor is connected in YPD culture mediums, obtains saccharomyces cerevisiae and Candida utilis
Yeast liquid seeds；
（8）Bacillus subtilis activates：24h is cultivated under the conditions of 30 DEG C using LB culture mediums, liquid seeds are obtained.
（9）Secondary solid is fermented：Candida utili, bacillus subtilis are seeded to culture by 4%, 3% inoculative proportion
Base, 30 DEG C or so are cultivated 3 days.
（10）Three solid fermentations：Solid lactic acid bacterium powder is seeded to culture medium, 29 DEG C of anaerobism trainings by 4% inoculative proportion
Support 24h.
（11）Dry：Low-temperature aeration is dried after fermentation ends.
（12）Crush：Dried fermented feed is crushed and is got product.
Above-mentioned stalk can be wheat stalk, rice straw.
Above-mentioned steps（4）Middle solid-state fermentation culture medium component is：Straw degradative product 30%, vinasse 20%, corn pulp 8%, bran
Skin 2.5%, beancake powder 9%, urea 4%, ammonium sulfate 7%.
Above-mentioned steps（6）Middle solid medium is without sterilizing.
Embodiment 2
A kind of method that utilization alcohol effluent and crop material prepare forage protein, it is characterised in that specifically include following steps：
（1）The basic handling of feedstuff：First by crushed stalk：Choose without the stalk for going mouldy, remove the magazines such as soil, crush
Cross 60 mesh sieve.
（2）Separation of solid and liquid：Alcohol effluent natural subsidence separation of solid and liquid, obtains supernatant and fermentation vinasse.
（3）Stalk treatment：
By straw ammoniation：The ammoniacal liquor of stalk 4% is added, adds alcohol effluent supernatant, adjustment moisture content to be well mixed to 58%,
It is placed in ammonification 5 days under closed environment.
Stalk is digested：Stalk pH modulation 4.5 after adjustment ammonification, adds cellulase, zytase, pectase, enzyme-added
Amount is respectively 2.5%, 3.5%, 3%, is well mixed, and digests 12h, and hydrolysis temperature is controlled at 38 DEG C.
（4）Prepare solid medium：Stalk after enzymolysis is mixed with fermentation vinasse, and adds corn pulp, wheat bran, soya-bean cake
Powder, urea, ammonium sulfate etc., adjust culture medium, it is ensured that its water content 110% with supernatant, and it is 7.0 to adjust initial pH.
（5）Mould treatment is aspergillus activation：Fresh wheat bran is taken, fine powder is weeded out with 40 mesh sieve, by wheat bran：Water is according to 1：2
Ratio carry out, fully mix thoroughly and not only without dry powder but also occur without into the phenomenon of bulk.It is divided in 2000mL triangular flasks, every
The bottled wet wheat bran about 100g of 2000mL triangles, sterilize 50min under 0.1MPa gauge pressures, is cooled to 32 DEG C, in each triangular flask
Inclined-plane aspergillus oryzae cv-8, the aspergillus niger sp-1 spores for accessing that 3 rings have activated, the control of culture medium product temperature (35 ± 1) DEG C every
12h shaking flasks once, make its sufficient contact, until stopping prepares spore suspension after spore grows.
（6）Solid fermentation：Aspergillus oryzae 4%, aspergillus niger 10%, isostearic acid 0.05%, different stearic acid 0.02% are inoculated with
To in mix, mix thoroughly, cultivated at a temperature of 35 DEG C, aerlbic culture 3 days.
（7）Candida utili is activated：Candida utili is connected in YPD culture mediums, 30 DEG C, under 180r/min rotating speeds
Constant temperature shaking flask 24 hours；Then with 1:100 ratios, seed liquor is connected in YPD culture mediums, obtains saccharomyces cerevisiae and Candida utilis
Yeast liquid seeds；
（8）Bacillus subtilis activates：24h is cultivated under the conditions of 34 DEG C using LB culture mediums, liquid seeds are obtained.
（9）Secondary solid is fermented：Candida utili, bacillus subtilis are seeded to culture by 5%, 4% inoculative proportion
Base, 30 DEG C or so are cultivated 4 days.
（10）Three solid fermentations：Solid lactic acid bacterium powder is seeded to culture medium, 35 DEG C of anaerobism trainings by 6% inoculative proportion
Support 24h.
（11）Dry：Low-temperature aeration is dried after fermentation ends.
（12）Crush：Dried fermented feed is crushed and is got product.
Above-mentioned stalk can be wheat stalk, maize straw.
Above-mentioned steps（4）Middle solid-state fermentation culture medium component is：Straw degradative product 33%, vinasse 24%, corn pulp 15%,
Wheat bran 3.2%, beancake powder 10%, urea 5%, ammonium sulfate 9%.
Above-mentioned steps（6）Middle solid medium is without sterilizing.
Embodiment 3
The present invention provides a kind of method that utilization alcohol effluent and crop material prepare forage protein, it is characterised in that specifically include
Following steps：
（1）The basic handling of feedstuff：First by crushed stalk：Choose without the stalk for going mouldy, remove the magazines such as soil, crush
Cross 60 mesh sieve.
（2）Separation of solid and liquid：Alcohol effluent natural subsidence separation of solid and liquid, obtains supernatant and fermentation vinasse.
（3）Stalk treatment：
By straw ammoniation：The ammoniacal liquor of stalk 6% is added, adds alcohol effluent supernatant, adjustment moisture content to be well mixed to 65%,
It is placed in ammonification 6 days under closed environment.
Stalk is digested：Stalk pH 5.0 after adjustment ammonification, adds cellulase, zytase, pectase, enzyme concentration point
Not Wei 3%, 5%, 4%, be well mixed, digest 15h, hydrolysis temperature control at 40 DEG C.
（4）Prepare solid medium：Stalk after enzymolysis is mixed with fermentation vinasse, and adds corn pulp, wheat bran, soya-bean cake
Powder, urea, ammonium sulfate etc., adjust culture medium, it is ensured that its water content 120% with supernatant, and it is 7.0 or so to adjust initial pH
（5）Mould treatment is aspergillus activation：Fresh wheat bran is taken, fine powder is weeded out with 40 mesh sieve, by wheat bran：Water is according to 1：2 ratio
Example is carried out, and is fully mixed thoroughly and not only without dry powder but also occur without into the phenomenon of bulk.It is divided in 2000mL triangular flasks, every
The bottled wet wheat bran about 100g of 2000mL triangles, sterilize 65min under 0.1MPa gauge pressures, is cooled to 30 ~ 35 DEG C, each triangular flask
Middle to access inclined-plane aspergillus oryzae cv-8, aspergillus niger sp-1 spores that 3 rings have been activated, the control of culture medium product temperature is every in (35 ± 1) DEG C
Every 15h shaking flasks once, its sufficient contact is made, until stopping prepares spore suspension after spore grows.
（6）Solid fermentation：Aspergillus oryzae 4%, aspergillus niger 12%, isostearic acid 0.08%, different stearic acid 0.02% are inoculated with
To in mix, mix thoroughly, cultivated at a temperature of 37 DEG C, aerlbic culture 4 days.
（7）Candida utili is activated：Candida utili is connected in YPD culture mediums, 30 DEG C, under 180r/min rotating speeds
Constant temperature shaking flask 24 hours；Then with 1:100 ratios, seed liquor is connected in YPD culture mediums, obtains saccharomyces cerevisiae and Candida utilis
Yeast liquid seeds；
（8）Bacillus subtilis activates：24h is cultivated under the conditions of 35 DEG C using LB culture mediums, liquid seeds are obtained.
（9）Secondary solid is fermented：Candida utili, bacillus subtilis are seeded to training by 6%, 3 ~ 4% inoculative proportions
Base is supported, 30 DEG C or so are cultivated 4 days.
（10）Three solid fermentations：Solid lactic acid bacterium powder is seeded to culture medium, 35 DEG C of anaerobism by 4 ~ 6% inoculative proportion
Culture 24h.
（11）Dry：Low-temperature aeration is dried after fermentation ends.
（12）Crush：Dried fermented feed is crushed and is got product.
Above-mentioned stalk can be wheat stalk, maize straw.
Above-mentioned steps（4）Middle solid-state fermentation culture medium component is：Straw degradative product 35%, vinasse 25%, corn pulp 20%,
Wheat bran 3.5%, beancake powder 11%, urea 6%, ammonium sulfate 11%.
Above-mentioned steps（6）Middle solid medium is without sterilizing.
Above example is merely to illustrate technical scheme, rather than is limited；Although with reference to foregoing reality
Example is applied to being described in detail by invention, but for the person of ordinary skill of the art, still can be to foregoing reality
Apply the technical scheme described in example to modify, or equivalent is carried out to which part technical characteristic；And to these modifications
Or replace, do not make the spirit and scope of the essence disengaging claimed technical solution of the invention of appropriate technical solution.
Described above not limitation of the present invention, the present invention is also not limited to the example above.The art it is common
In essential scope of the invention, change, remodeling, addition or the replacement made should also belong to protection of the invention to technical staff
Scope.
Claims (7)
1. a kind of method that utilization alcohol effluent and crop material prepare forage protein, it is characterized in that：The method includes following step
Suddenly：
1）The basic handling of feedstuff：First by crushed stalk：Choose without the stalk for going mouldy, remove the impurity such as soil, crush
Cross 60 mesh sieve；
2）Separation of solid and liquid：Alcohol effluent natural subsidence separation of solid and liquid, obtains supernatant and fermentation vinasse；
3）The reprocessing of stalk：
By straw ammoniation：The ammoniacal liquor of stalk 3 ~ 6% is added, alcohol effluent supernatant, adjustment moisture content to 50 ~ 65%, mixing is added
Uniformly, ammonification 2 ~ 6 days under closed environment are placed in；
Stalk is digested：Stalk pH after adjustment ammonification is it is essential to ensure that be between 4.0 ~ 5.0, to add 2 ~ 3% cellulases, 4 ~ 5%
Zytase, 2 ~ 4% pectases, are well mixed, and digest 10 ~ 15h, and hydrolysis temperature is controlled at 35 ~ 40 DEG C；
4）Prepare solid medium：Stalk after enzymolysis is mixed with fermentation vinasse, and adds corn pulp, wheat bran, beancake powder, urine
Element, ammonium sulfate etc., adjust culture medium, it is ensured that its water content 100 ~ 120% with supernatant, and it is 7.0 or so to adjust initial pH；
5）Mould treatment is aspergillus activation：Fresh wheat bran is taken, fine powder is weeded out with 40 mesh sieve, by wheat bran：Water is according to 1：2 ratio
Carry out, fully mix thoroughly and not only without dry powder but also occur without into the phenomenon of bulk.It is divided in 2000mL triangular flasks, every 2000mL
The bottled wet wheat bran about 100g of triangle, sterilize 45 ~ 65min under 0.1MPa gauge pressures, 30 ~ 35 DEG C is cooled to, in each triangular flask
Inclined-plane aspergillus oryzae cv-8, aspergillus niger sp-1 spores that 2 ~ 3 rings have been activated are accessed, the control of culture medium product temperature is every in (35 ± 1) DEG C
Every 10 ~ 15h shaking flasks once, its sufficient contact is made, until stopping prepares spore suspension after spore grows；
6）Solid fermentation：By aspergillus oryzae 3 ~ 4%, aspergillus niger 8 ~ 12%, isostearic acid 0.05-0.08%, different stearic acid 0.02-
0.05% is inoculated into mix in proportion, mixes thoroughly, is cultivated at a temperature of 30 ~ 37 DEG C, aerlbic culture 2 ~ 4 days；
7）Candida utili is activated：Candida utili is connected in YPD culture mediums, 30 DEG C, constant temperature under 180r/min rotating speeds
Shaking flask 24 hours；Then with 1:100 ratios, seed liquor is connected in YPD culture mediums, obtains saccharomyces cerevisiae and candida utili
Liquid seeds；
8）Bacillus subtilis activates：24h is cultivated under the conditions of 30 ~ 35 DEG C using LB culture mediums, liquid seeds are obtained；
9）Secondary solid is fermented：Candida utili, bacillus subtilis are seeded to culture by 4 ~ 6%, 3 ~ 4% inoculative proportions
Base, 30 DEG C or so are cultivated 3 ~ 4 days；
10）Three solid fermentations：Solid lactic acid bacterium powder is seeded to culture medium, 29 ~ 35 DEG C of anaerobism trainings by 4 ~ 6% inoculative proportion
Support 24h；
11）Dry：Low-temperature aeration is dried after fermentation ends；
12）Crush：Dried fermented feed is crushed and is got product.
2. the method that a kind of utilization alcohol effluent according to claim 1 and crop material prepare forage protein, its feature
It is：Stalk can be one or more in wheat stalk, maize straw, rice straw.
3. the method that a kind of utilization alcohol effluent according to claim 1 and crop material prepare forage protein, its feature
It is：Step 4）Middle solid-state fermentation culture medium component is：Straw degradative product 30 ~ 35%, vinasse 20 ~ 25%, corn pulp 8 ~ 20%, bran
Skin 2.5 ~ 3.5%, beancake powder 9 ~ 11%, urea 4 ~ 6%, ammonium sulfate 7 ~ 11%.
4. the method that a kind of utilization alcohol effluent according to claim 1 and crop material prepare forage protein, its feature
It is：Stalk pH after the step adjustment ammonification is it is essential to ensure that between 4.0 ~ 5.0.
5. the method that a kind of utilization alcohol effluent according to claim 1 and crop material prepare forage protein, its feature
It is：Step 6）Solid fermentation is it is essential to ensure that appropriate, in good time ventilation.
6. the method that a kind of utilization alcohol effluent according to claim 1 and crop material prepare forage protein, its feature
It is：Step 6）Middle solid medium is without sterilizing.
7. the method that a kind of utilization alcohol effluent according to claim 1 and crop material prepare forage protein, its feature
It is, step 11）After middle fermentation ends, it is ensured that low-temperature aeration is dried.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108795795A (en) * | 2018-05-04 | 2018-11-13 | 吉林农业大学 | A kind of lactobacteria-containing Thermal degradation stalk composite bacteria agent and its application |
| CN111387340A (en) * | 2020-03-31 | 2020-07-10 | 广西农垦金光乳业有限公司 | Feed for improving milk yield of dairy cows and preparation method thereof |
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| CN104886340A (en) * | 2015-07-08 | 2015-09-09 | 青岛嘉瑞生物技术有限公司 | Process for preparing liquid protein feed by utilizing waste alcoholic fermentation liquor |
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| CN105901282A (en) * | 2016-04-18 | 2016-08-31 | 福建美家园生物科技股份有限公司 | A method of preparing animal protein feed from potato dregs through fermentation and the feed prepared by the method |
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| CN104256057A (en) * | 2014-10-01 | 2015-01-07 | 青岛嘉瑞生物技术有限公司 | Method for preparing feed proteins by utilizing alcohol waste liquor and crop straws |
| CN104886340A (en) * | 2015-07-08 | 2015-09-09 | 青岛嘉瑞生物技术有限公司 | Process for preparing liquid protein feed by utilizing waste alcoholic fermentation liquor |
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| CN111387340A (en) * | 2020-03-31 | 2020-07-10 | 广西农垦金光乳业有限公司 | Feed for improving milk yield of dairy cows and preparation method thereof |
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