CN112806486A - Ruminant fermented feed and preparation method thereof - Google Patents
Ruminant fermented feed and preparation method thereof Download PDFInfo
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- 241000282849 Ruminantia Species 0.000 title claims abstract description 31
- 238000002360 preparation method Methods 0.000 title claims abstract description 12
- 240000008042 Zea mays Species 0.000 claims abstract description 69
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims abstract description 69
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims abstract description 69
- 235000005822 corn Nutrition 0.000 claims abstract description 69
- 238000000855 fermentation Methods 0.000 claims abstract description 63
- 230000004151 fermentation Effects 0.000 claims abstract description 59
- 239000007788 liquid Substances 0.000 claims abstract description 41
- 235000012054 meals Nutrition 0.000 claims abstract description 26
- 239000000203 mixture Substances 0.000 claims abstract description 24
- 241000894006 Bacteria Species 0.000 claims abstract description 19
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- 239000002253 acid Substances 0.000 claims abstract description 14
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- 239000001963 growth medium Substances 0.000 claims abstract description 13
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- 108091005508 Acid proteases Proteins 0.000 claims abstract description 11
- 241000194108 Bacillus licheniformis Species 0.000 claims abstract description 11
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- 235000014469 Bacillus subtilis Nutrition 0.000 claims abstract description 11
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- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims abstract description 10
- 229940072205 lactobacillus plantarum Drugs 0.000 claims abstract description 10
- WQZGKKKJIJFFOK-DVKNGEFBSA-N alpha-D-glucose Chemical group OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-DVKNGEFBSA-N 0.000 claims abstract description 7
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- 235000013922 glutamic acid Nutrition 0.000 claims description 17
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- 238000000034 method Methods 0.000 claims description 17
- 239000002351 wastewater Substances 0.000 claims description 16
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 claims description 14
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- QIJRTFXNRTXDIP-UHFFFAOYSA-N (1-carboxy-2-sulfanylethyl)azanium;chloride;hydrate Chemical compound O.Cl.SCC(N)C(O)=O QIJRTFXNRTXDIP-UHFFFAOYSA-N 0.000 claims description 7
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 7
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- 229910052799 carbon Inorganic materials 0.000 claims description 7
- 238000012258 culturing Methods 0.000 claims description 7
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- 235000003891 ferrous sulphate Nutrition 0.000 claims description 7
- 238000011081 inoculation Methods 0.000 claims description 7
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 claims description 7
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims description 7
- 229940099596 manganese sulfate Drugs 0.000 claims description 7
- 239000011702 manganese sulphate Substances 0.000 claims description 7
- 235000007079 manganese sulphate Nutrition 0.000 claims description 7
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 7
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 claims description 7
- 229960001763 zinc sulfate Drugs 0.000 claims description 7
- 229910000368 zinc sulfate Inorganic materials 0.000 claims description 7
- 150000003839 salts Chemical class 0.000 claims description 6
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 claims description 4
- NGWKGSCSHDHHAJ-YPFQVHCOSA-N Liquoric acid Chemical compound C1C[C@H](O)C(C)(C)C2CC[C@@]3(C)[C@]4(C)C[C@H]5O[C@@H]([C@](C6)(C)C(O)=O)C[C@@]5(C)[C@@H]6C4=CC(=O)C3[C@]21C NGWKGSCSHDHHAJ-YPFQVHCOSA-N 0.000 claims description 4
- NGWKGSCSHDHHAJ-UHFFFAOYSA-N Liquoric acid Natural products C1CC(O)C(C)(C)C2CCC3(C)C4(C)CC5OC(C(C6)(C)C(O)=O)CC5(C)C6C4=CC(=O)C3C21C NGWKGSCSHDHHAJ-UHFFFAOYSA-N 0.000 claims description 4
- 239000002609 medium Substances 0.000 claims description 4
- 229960001781 ferrous sulfate Drugs 0.000 claims description 2
- 239000000843 powder Substances 0.000 claims description 2
- 230000003068 static effect Effects 0.000 claims description 2
- 239000006227 byproduct Substances 0.000 abstract description 17
- 239000002994 raw material Substances 0.000 abstract description 8
- 229940088598 enzyme Drugs 0.000 description 16
- 102000004190 Enzymes Human genes 0.000 description 12
- 108090000790 Enzymes Proteins 0.000 description 12
- 241000192031 Ruminococcus Species 0.000 description 12
- 239000000047 product Substances 0.000 description 10
- 238000004519 manufacturing process Methods 0.000 description 9
- 230000000813 microbial effect Effects 0.000 description 7
- 229920002261 Corn starch Polymers 0.000 description 5
- 229920002678 cellulose Polymers 0.000 description 5
- 239000001913 cellulose Substances 0.000 description 5
- 239000008120 corn starch Substances 0.000 description 5
- 239000010903 husk Substances 0.000 description 5
- 244000005700 microbiome Species 0.000 description 5
- 230000008569 process Effects 0.000 description 5
- 230000035755 proliferation Effects 0.000 description 5
- 229920002472 Starch Polymers 0.000 description 4
- 239000003674 animal food additive Substances 0.000 description 4
- 235000013339 cereals Nutrition 0.000 description 4
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- 102000004169 proteins and genes Human genes 0.000 description 4
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- 239000007787 solid Substances 0.000 description 4
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- 229920001221 xylan Polymers 0.000 description 4
- 150000004823 xylans Chemical class 0.000 description 4
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- 235000019750 Crude protein Nutrition 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 108010084185 Cellulases Proteins 0.000 description 2
- 102000005575 Cellulases Human genes 0.000 description 2
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 2
- 108010001817 Endo-1,4-beta Xylanases Proteins 0.000 description 2
- 241000233866 Fungi Species 0.000 description 2
- 102000004139 alpha-Amylases Human genes 0.000 description 2
- 108090000637 alpha-Amylases Proteins 0.000 description 2
- 229940024171 alpha-amylase Drugs 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- HEBKCHPVOIAQTA-NGQZWQHPSA-N d-xylitol Chemical compound OC[C@H](O)C(O)[C@H](O)CO HEBKCHPVOIAQTA-NGQZWQHPSA-N 0.000 description 2
- 238000003912 environmental pollution Methods 0.000 description 2
- 108010038658 exo-1,4-beta-D-xylosidase Proteins 0.000 description 2
- 230000003301 hydrolyzing effect Effects 0.000 description 2
- 239000004310 lactic acid Substances 0.000 description 2
- 235000014655 lactic acid Nutrition 0.000 description 2
- 235000013406 prebiotics Nutrition 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- 108010013043 Acetylesterase Proteins 0.000 description 1
- 108091005658 Basic proteases Proteins 0.000 description 1
- 241000221198 Basidiomycota Species 0.000 description 1
- 108010008885 Cellulose 1,4-beta-Cellobiosidase Proteins 0.000 description 1
- 101710089042 Demethyl-4-deoxygadusol synthase Proteins 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 101710112457 Exoglucanase Proteins 0.000 description 1
- 108010031186 Glycoside Hydrolases Proteins 0.000 description 1
- 102000005744 Glycoside Hydrolases Human genes 0.000 description 1
- 229920002488 Hemicellulose Polymers 0.000 description 1
- 108090001060 Lipase Proteins 0.000 description 1
- 102000004882 Lipase Human genes 0.000 description 1
- 239000004367 Lipase Substances 0.000 description 1
- 108010093941 acetylxylan esterase Proteins 0.000 description 1
- 108010084650 alpha-N-arabinofuranosidase Proteins 0.000 description 1
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 1
- CKLJMWTZIZZHCS-REOHCLBHSA-N aspartic acid group Chemical group N[C@@H](CC(=O)O)C(=O)O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 1
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 1
- 108010047754 beta-Glucosidase Proteins 0.000 description 1
- 102000006995 beta-Glucosidase Human genes 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 235000010980 cellulose Nutrition 0.000 description 1
- 108010080434 cephalosporin-C deacetylase Proteins 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 235000005687 corn oil Nutrition 0.000 description 1
- 239000002285 corn oil Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 229940079919 digestives enzyme preparation Drugs 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 235000019197 fats Nutrition 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- -1 i.e. Proteins 0.000 description 1
- 229920005610 lignin Polymers 0.000 description 1
- 235000019421 lipase Nutrition 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- 239000000618 nitrogen fertilizer Substances 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 229920001542 oligosaccharide Polymers 0.000 description 1
- 150000007965 phenolic acids Chemical class 0.000 description 1
- 239000006041 probiotic Substances 0.000 description 1
- 235000018291 probiotics Nutrition 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
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- 230000002062 proliferating effect Effects 0.000 description 1
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- 239000011573 trace mineral Substances 0.000 description 1
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- 125000000969 xylosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)CO1)* 0.000 description 1
Classifications
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- 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
- A23K50/10—Feeding-stuffs specially adapted for particular animals for ruminants
-
- 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
-
- 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
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/189—Enzymes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- 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
- A23V2400/169—Plantarum
-
- 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
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Health & Medical Sciences (AREA)
- Polymers & Plastics (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
- Organic Chemistry (AREA)
- Wood Science & Technology (AREA)
- Animal Husbandry (AREA)
- Food Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- Mycology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Virology (AREA)
- Botany (AREA)
- Physiology (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
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Abstract
The invention provides a preparation method of a fermented feed for ruminants, which comprises the following steps: 1) mixing two to six of corn bran, corn germ meal, corn sugar residue, sea buckthorn seed meal, palm kernel meal and shinyleaf yellowhorn meal according to a certain proportion to obtain a first mixture; 2) preparing a liquid fermentation culture medium, and sterilizing at high temperature and high pressure; 3) respectively inoculating rumen coccus, eubacteria and rumen coccus amylovorus to three liquid fermentation culture media to prepare corresponding seed solutions; 4) adding bacillus licheniformis, bacillus subtilis, lactobacillus plantarum, saccharomyces cerevisiae, mannase, xylanase, acid protease and acid cellulase into any seed liquid prepared in the step 3), uniformly mixing, adding into the first mixture for fermentation, and obtaining the fermented feed after the fermentation is finished. The invention takes the corn byproducts and the miscellaneous meal as raw materials to carry out biological pretreatment to prepare the ruminant feed, thereby improving the added value of the corn byproducts.
Description
Technical Field
The invention belongs to the technical field of biological feed, and particularly relates to a fermented feed for ruminants and a preparation method thereof.
Background
The corn is one of three main grain crops in the world and is also one of three main grain varieties in China, and the corn serving as the second grain crop in China has the yield accounting for about 35 percent of the total yield of grains in China. Corn in China is mainly used for producing feed for a long time, but with the change of social requirements and the continuous development and progress of corn deep processing industry, the consumption structure of corn gradually changes.
Although the corn deep processing industry can create economic growth, the corn deep processing industry in China has many insufficient places. The main problems are that the industrial chain is not long enough, the added value is not enough, the production efficiency is low and the like in the deep processing production process of the corn. In particular, the corn deep processing byproducts are high in yield, but the economic value of the corn deep processing byproducts is low, wherein byproducts generated in the corn starch production process are a prominent problem. The production of starch by corn is the major mode of the corn processing industry, since the production of starch is around 2000 million tons per year, and the corn starch accounts for about 80-85%. Wherein the byproducts of the wet-process production of the corn starch mainly comprise corn oil, corn germ meal, corn steep liquor, corn husks, corn starch residues, corn DDGS and the like. The produced byproducts can be applied to production and life, and meanwhile, some byproducts which are difficult to directly utilize are also produced: corn bran, corn steep liquor, corn germ meal, corn starch residue and the like, wherein the byproducts are not subjected to deep processing and are generally sold separately and are low in price.
The glutamic acid fermentation wastewater is obtained by extracting glutamic acid and mycoprotein and then evaporating and concentrating in the glutamic acid fermentation production process, and has high COD and NH3The characteristics of N, high salt and low pH, namely three highs and one lowness, are once the symbols of the high and heavy pollution industry in China, the waste water is usually used for preparing nitrogen fertilizer after being developed, the additional value generated by the utilization mode is low, a utilization mode with higher economic benefit needs to be searched, and the biological feed technology is an effective way.
The biological feed is a general name of a novel feed resource and a feed additive which are safe, efficient, environment-friendly and free of residue and are developed by using microbial engineering fermentation by taking feed and a feed additive as objects and taking modern biotechnology such as genetic engineering, protein engineering, fermentation engineering and the like as means. The categories mainly include: feed enzyme preparations, microbial feed additives, functional protein peptides, functional amino acids, organic trace elements, plant and microbial extracts, prebiotics, fermented and enzymolyzed feeds and other biotechnology related products can be classified as biological feed additives and biological fermented feeds. Part of the corn by-products, the miscellaneous meal and the glutamic acid fermentation wastewater are subjected to bacterium-enzyme synergistic fermentation treatment to prepare fermented feed for ruminants, so that the additional value of the raw materials can be effectively increased, the problem of environmental pollution caused by the glutamic acid fermentation wastewater is relieved, and certain economic benefit is brought to enterprises.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides a ruminant fermented feed and a preparation method thereof.
The invention is realized by the following steps:
the invention provides a preparation method of a fermented feed for ruminants, which comprises the following steps:
1) mixing two to six of corn bran, corn germ meal, corn sugar residue, sea buckthorn seed meal, palm kernel meal and shinyleaf yellowhorn meal according to a certain proportion to obtain a first mixture;
2) preparing a liquid fermentation culture medium, and sterilizing at high temperature and high pressure;
3) respectively inoculating rumen coccus, eubacteria and rumen coccus amylovorus into three liquid fermentation culture media to respectively prepare corresponding seed solutions;
4) adding bacillus licheniformis, bacillus subtilis, lactobacillus plantarum, saccharomyces cerevisiae, mannase, xylanase, acid protease and acid cellulase into any seed liquid prepared in the step 3), uniformly mixing, adding into the first mixture for fermentation, and obtaining the fermented feed after the fermentation is finished.
The preparation method comprises the following steps of preparing a ruminant fermented feed by adopting a two-step fermentation method, wherein in the first step, liquid fermentation is carried out, rumen coccus, eubacteria and rumen coccus amylovorus are prepared into seed liquid, and inorganic nitrogen in a nitrogen source is converted into microbial mycoprotein by microorganisms in the microbial proliferation process; and secondly, performing bacterium-enzyme synergistic fermentation, namely adding the bacterium-enzyme into the seed liquid, adding the bacterium-enzyme into the solid raw material, fermenting for a certain time at a certain temperature, and obtaining the fermented feed for the ruminants after the fermentation is finished.
Bacillus licheniformis can produce alpha-amylase, alkaline protease for hydrolyzing starch and protein in feed, and enzyme for hydrolyzing cellulose. The bacillus subtilis itself synthesizes enzymes such as alpha-amylase, protease, lipase, cellulase, etc., and can decompose and utilize residual starch, protein, fat and cellulose in the first mixture. The proliferation of yeast can metabolize to produce acetic acid and ethanol, improve the smell of materials and prevent the growth and reproduction of mold. The proliferation of lactic acid bacteria can be metabolized to produce lactic acid, and the growth and reproduction of mixed bacteria can be inhibited. Monosaccharides generated by enzymolysis of xylan and cellulose by xylanase and acidic cellulase can be used as a carbon source by microorganisms, and the effect of promoting proliferation of the microorganisms is achieved.
The xylanase is a complex enzyme system for degrading xylan molecules, has complex composition and comprises beta-xylanase, beta-D-xylosidase, alpha-L-furan type arabinosidase, acetyl xylan esterase, phenolic acid esterase and the like, wherein the beta-xylanase is a main enzyme for degrading hemicellulose, and the enzyme acts on beta-1, 4-xyloside bonds inside a xylan main chain in an internal mode to degrade xylan into short-chain xylo-oligosaccharides, and a small amount of xylose is generated; and the beta-D-xylosidase acts on short-chain xylooligosaccharide and catalyzes the tail end of the xylooligosaccharide to release xylose residues. Cellulases belong to glycoside hydrolases and are traditionally divided into three groups of components: 1. endoglucanases, commonly known as CX enzymes, and EG enzymes from fungi; 2. exoglucanases, i.e., cellobiohydrolases, commonly known as the C1 enzyme, known as CBH from fungi; 3. beta-glucosidase, abbreviated as BG. Cellulases degrade cellulose as a result of a synergistic interaction between the components of the enzyme.
The acidic protease is widely present in mould, yeast and basidiomycetes, and is rarely found in bacteria, mainly carboxyl protease, the active center of which contains two aspartic acid residues, the content of the acidic amino acid is high, the content of the basic amino acid is low, the optimum pH value is 2.0-4.0, and the optimum temperature is about 40 ℃.
Further, the corn steep liquor and glutamic acid fermentation wastewater are used as nitrogen sources, corn flour and corn bran are used as carbon sources, ferrous sulfate, manganese sulfate, zinc sulfate, cysteine hydrochloride and light calcium carbonate are used as inorganic salts, and tap water is mixed to prepare the liquid fermentation medium.
Further, the liquid fermentation medium comprises, by mass, 5-15 parts of corn steep liquor, 5-15 parts of glutamic acid fermentation wastewater, 5-15 parts of corn flour, 5-15 parts of corn bran, 0.1-0.5 part of ferrous sulfate, 0.1-0.5 part of manganese sulfate, 0.1-0.3 part of zinc sulfate, 0.5-1.5 parts of cysteine hydrochloride, 0.5-1 part of light calcium carbonate and 100 parts of tap water.
Further, in the step 3), the seed liquid needs to be subjected to static culture, the culture temperature is 20-50 ℃, the culture time is 4-72h, and the initial pH is 3-8.
Furthermore, the bacillus licheniformis, the bacillus subtilis, the lactobacillus plantarum, the saccharomyces cerevisiae, the mannase, the xylanase, the acid protease and the acid cellulase are all powder, and the addition amount of the components is 0.01-0.03 part of the bacillus licheniformis, 0.01-0.05 part of the bacillus subtilis, 0.01-0.15 part of the lactobacillus plantarum, 0.01-0.15 part of the saccharomyces cerevisiae, 0.01-0.08 part of the mannase, 0.01-0.08 part of the xylanase, 0.01-0.08 part of the acid protease and 0.01-0.08 part of the acid cellulase according to the parts by mass.
Further, step 4) adopts anaerobic fermentation. Ruminococcus, eubacteria and ruminococcus amylovorus are strictly anaerobic ruminal bacteria, so the medium needs to be maintained in an anaerobic state during fermentation.
Further, the step 3) also comprises the step of mixing 1-20 parts of the seed liquid with 20-30 parts of tap water to prepare an initial seed liquid.
Further, the prepared initial seed solution is added into the first mixture with an inoculation amount of 30-50%, and is placed in an anaerobic fermentation device for culturing for 48-120h at a temperature of 20-60 ℃ after inoculation.
The invention also provides a ruminant fermented feed prepared by the method.
The invention has the following beneficial effects:
1. the invention adopts two-step fermentation of liquid fermentation and solid fermentation, saves production cost, and improves the added value of the corn by-products by using the corn by-products and the miscellaneous meal as raw materials to carry out biological pretreatment to prepare the ruminant feed.
2. The liquid fermentation part respectively takes corn steep liquor and glutamic acid fermentation wastewater as nitrogen sources and corn flour and corn bran as carbon sources, so that the four raw materials are fully treated and utilized by the ruminococcus, the eubacteria and the ruminococcus amylovorus, and a large amount of bacteria are proliferated; the solid fermentation part carries out deep processing on corn husks, corn germ meal, corn steep liquor, corn sugar residues and other miscellaneous meals in the corn byproducts, and the addition of mannase, xylanase, acid protease and acid cellulase carries out predigestion on the corn byproducts and miscellaneous meals, thereby improving the added value of the corn byproducts.
3. The glutamic acid fermentation wastewater is recycled, and the pollution of the glutamic acid fermentation wastewater to the environment is relieved.
4. The prepared ruminant fermented feed is rich in rumen microorganisms, and is beneficial to the health of rumen of the ruminant.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
The embodiment provides a ruminant fermented feed product and a preparation method thereof, and the process comprises the following steps:
1) mixing 40 parts of corn husks, 40 parts of corn germ meal, 10 parts of corn sugar residues and 10 parts of sea buckthorn seed meal which are crushed and sieved by a 30-mesh sieve to obtain a first mixture;
2) taking 5 parts of corn steep liquor and 5 parts of glutamic acid fermentation wastewater as nitrogen sources, taking 5 parts of corn flour and 5 parts of corn bran which are crushed and sieved by a 30-mesh sieve as carbon sources, taking 0.1 part of ferrous sulfate, 0.3 part of manganese sulfate, 0.2 part of zinc sulfate, 0.5 part of cysteine hydrochloride, 1 part of light calcium carbonate as inorganic salts and 100 parts of tap water, uniformly mixing, and performing high-temperature and high-pressure sterilization to obtain a liquid fermentation culture medium;
3) respectively inoculating the ruminococcus, the eubacteria and the ruminococcus amylovorus into the culture medium in the step 2), and culturing at the culture temperature of 25 ℃, the culture time of 16h and the initial pH of 3.5 to prepare seed liquid;
4) respectively mixing 1 part of seed liquid and 30 parts of tap water by weight to obtain initial seed liquid;
5) 0.01 part of bacillus licheniformis, 0.01 part of bacillus subtilis, 0.01 part of lactobacillus plantarum, 0.01 part of saccharomyces cerevisiae, 0.01 part of mannase, 0.01 part of xylanase, 0.01 part of acid protease and 0.01 part of acid cellulase are added into an initial seed solution of ruminococcus as a seed mixture solution, and the seed mixture is added into the first mixture according to the inoculation amount of 30% for fermentation.
6) Carrying out anaerobic culture on the inoculated first mixture for 48 hours at 25 ℃ to obtain a finished product of the fermented feed for the ruminants;
detecting the finished product, wherein the fermentation water content is 40%, the crude protein content is 30%, the acid soluble protein content is 15%, NDF 50%, ADF 18%, the organic acid content is 6%, and the total number of micro-ecological bacteria is 1.2 multiplied by 108cfu/g。
Example 2
The embodiment provides a ruminant fermented feed product and a preparation method thereof, and the process comprises the following steps:
1) mixing 40 parts of corn husks, 38 parts of corn germ meal, 12 parts of corn sugar residues and 10 parts of palm meal which are crushed and sieved by a 30-mesh sieve to obtain a first mixture;
2) taking 10 parts of corn steep liquor and 10 parts of glutamic acid fermentation wastewater as nitrogen sources, taking 10 parts of corn flour and 10 parts of corn bran which are crushed and sieved by a 30-mesh sieve as carbon sources, taking 0.2 part of ferrous sulfate, 0.1 part of manganese sulfate, 0.3 part of zinc sulfate, 1 part of cysteine hydrochloride, 0.5 part of light calcium carbonate as inorganic salts and 100 parts of tap water, uniformly mixing, and performing high-temperature and high-pressure sterilization to obtain a liquid fermentation culture medium;
3) respectively inoculating the ruminococcus, the eubacteria and the ruminococcus amylovorus into the culture medium in the step 2), and culturing at the culture temperature of 30 ℃ for 20h and under the condition of initial pH 4 to prepare seed liquid;
4) respectively taking 5 parts of seed liquid and 20 parts of tap water to mix as initial seed liquid according to parts by weight;
5) 0.02 part of bacillus licheniformis, 0.02 part of bacillus subtilis, 0.05 part of lactobacillus plantarum, 0.05 part of saccharomyces cerevisiae, 0.05 part of mannanase, 0.05 part of xylanase, 0.05 part of acid protease and 0.05 part of acid cellulase are added into an initial seed solution of eubacteria to be used as a seed mixing solution, and the seed mixing solution is added into the first mixture according to the inoculation amount of 40% for fermentation.
6) Anaerobic culturing the inoculated first mixture at 30 ℃ for 72h to obtain a finished product of the fermented feed for the ruminants;
detecting the finished product, wherein the fermentation water content is 45%, the crude protein content is 35%, the acid soluble protein content is 18%, NDF 55%, ADF 20%, the organic acid content is 6.7%, and the total number of micro-ecological bacteria is 1.2 multiplied by 108cfu/g。
Example 3
The embodiment provides a ruminant fermented feed product and a preparation method thereof, and the process comprises the following steps:
1) mixing 45 parts of corn husks, 30 parts of corn germ meal, 15 parts of corn sugar residues and 10 parts of shinyleaf yellowhorn meal which are crushed and sieved by a 30-mesh sieve to obtain a first mixture;
2) taking 15 parts of corn steep liquor and 15 parts of glutamic acid fermentation wastewater as nitrogen sources, taking 15 parts of corn flour and 15 parts of corn bran which are crushed and sieved by a 30-mesh sieve as carbon sources, taking 0.5 part of ferrous sulfate, 0.5 part of manganese sulfate, 0.1 part of zinc sulfate, 1.5 parts of cysteine hydrochloride, 0.75 part of light calcium carbonate as inorganic salts and 100 parts of tap water, uniformly mixing, sterilizing at high temperature and high pressure, and taking the mixture as a liquid fermentation culture medium;
3) respectively inoculating the ruminococcus, the eubacteria and the ruminococcus amylovorus into the culture medium in the step 2), and culturing at the culture temperature of 35 ℃, for 36 hours and under the condition of initial pH 5 to prepare seed liquid;
4) respectively taking 20 parts of seed liquid and 20 parts of tap water to mix as initial seed liquid according to parts by weight;
5) 0.03 part of bacillus licheniformis, 0.05 part of bacillus subtilis, 0.15 part of lactobacillus plantarum, 0.15 part of saccharomyces cerevisiae, 0.08 part of mannase, 0.08 part of xylanase, 0.08 part of acid protease and 0.08 part of acid cellulase are added into an initial seed solution of ruminococcus amylovorus to serve as a seed mixture solution, and the seed mixture is added into the first mixture according to the inoculation amount of 50% for fermentation.
6) Anaerobic culturing the inoculated first mixture at 35 ℃ for 96h to obtain a finished product of the fermented feed for the ruminants;
detecting the finished product, wherein the fermentation water content is 50%, the crude protein content is 38%, the acid soluble protein content is 20%, NDF 60%, ADF 23%, the organic acid content is 7.8%, and the total number of micro-ecological bacteria is 1.0 multiplied by 108cfu/g。
The preparation method comprises the following steps of preparing a ruminant fermented feed by adopting a two-step fermentation method, wherein in the first step, liquid fermentation is carried out, rumen coccus, eubacteria and rumen coccus amylovorus are prepared into seed liquid, and inorganic nitrogen in nitrogen source corn steep liquor and glutamic acid fermentation wastewater is converted into microbial mycoprotein by microorganisms in the microbial proliferation process; and secondly, performing bacterial enzyme synergistic fermentation, namely adding the bacterial enzyme into the seed liquid, adding the bacterial enzyme into the solid raw material, fermenting for a certain time at a certain temperature, degrading cellulose and lignin in the raw material of the feed in the fermentation process, proliferating the rumen probiotics in a large amount, producing the prebiotics in a large amount, and obtaining the fermented feed for the ruminants after the fermentation is finished. The invention obviously improves the nutritive value of the feed raw materials such as the corn by-products, improves the added value of the corn by-products, relieves the problem of environmental pollution caused by glutamic acid fermentation wastewater, and brings certain economic benefit for enterprises.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (9)
1. A preparation method of a fermented feed for ruminants is characterized by comprising the following steps:
1) mixing two to six of corn bran, corn germ meal, corn sugar residue, sea buckthorn seed meal, palm kernel meal and shinyleaf yellowhorn meal according to a certain proportion to obtain a first mixture;
2) preparing a liquid fermentation culture medium, and sterilizing at high temperature and high pressure;
3) respectively inoculating rumen coccus, eubacteria and rumen coccus amylovorus into three liquid fermentation culture media to respectively prepare corresponding seed solutions;
4) adding bacillus licheniformis, bacillus subtilis, lactobacillus plantarum, saccharomyces cerevisiae, mannase, xylanase, acid protease and acid cellulase into any seed liquid prepared in the step 3), uniformly mixing, adding into the first mixture for fermentation, and obtaining the fermented feed after the fermentation is finished.
2. The method of preparing a fermented feed for ruminants according to claim 1, wherein: the liquid fermentation culture medium is prepared by taking corn steep liquor and glutamic acid fermentation wastewater as nitrogen sources, taking corn flour and corn bran as carbon sources, taking ferrous sulfate, manganese sulfate, zinc sulfate, cysteine hydrochloride and light calcium carbonate as inorganic salts and mixing with tap water.
3. A method of preparing a fermented feed for ruminants according to claim 2, wherein: the liquid fermentation medium comprises, by mass, 5-15 parts of corn steep liquor, 5-15 parts of glutamic acid fermentation wastewater, 5-15 parts of corn flour, 5-15 parts of corn bran, 0.1-0.5 part of ferrous sulfate, 0.1-0.5 part of manganese sulfate, 0.1-0.3 part of zinc sulfate, 0.5-1.5 parts of cysteine hydrochloride, 0.5-1 part of light calcium carbonate and 100 parts of tap water.
4. The method of preparing a fermented feed for ruminants according to claim 1, wherein: in the step 3), the seed liquid needs to be subjected to static culture, the culture temperature is 20-50 ℃, the culture time is 4-72h, and the initial pH is 3-8.
5. The method of preparing a fermented feed for ruminants according to claim 1, wherein: the bacillus licheniformis, the bacillus subtilis, the lactobacillus plantarum, the saccharomyces cerevisiae, the mannase, the xylanase, the acid protease and the acid cellulase are all powder, and the addition amount of the components is 0.01-0.03 part of the bacillus licheniformis, 0.01-0.05 part of the bacillus subtilis, 0.01-0.15 part of the lactobacillus plantarum, 0.01-0.15 part of the saccharomyces cerevisiae, 0.01-0.08 part of the mannase, 0.01-0.08 part of the xylanase, 0.01-0.08 part of the acid protease and 0.01-0.08 part of the acid cellulase by mass.
6. The method of preparing a fermented feed for ruminants according to claim 1, wherein: and 4) adopting anaerobic fermentation.
7. A method of preparing a fermented feed for ruminants according to any one of claims 1 to 6, wherein: and step 3) mixing 1-20 parts of seed liquid and 20-30 parts of tap water to prepare initial seed liquid.
8. The method of preparing a fermented feed for ruminants according to claim 7, wherein: the inoculation amount of the prepared initial seed liquid added into the first mixture is 30-50%, and the initial seed liquid is placed in an anaerobic fermentation device for culturing for 48-120h at the temperature of 20-60 ℃ after inoculation.
9. A fermented feed for ruminants prepared by the method of any one of claims 1 to 8.
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