CN113229399A - Method for biologically degrading rapeseed meal toxin and improving nutritive value thereof - Google Patents
Method for biologically degrading rapeseed meal toxin and improving nutritive value thereof Download PDFInfo
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- 235000019779 Rapeseed Meal Nutrition 0.000 title claims abstract description 81
- 239000004456 rapeseed meal Substances 0.000 title claims abstract description 79
- 238000000034 method Methods 0.000 title claims abstract description 34
- 239000003053 toxin Substances 0.000 title claims abstract description 24
- 231100000765 toxin Toxicity 0.000 title claims abstract description 24
- 230000000593 degrading effect Effects 0.000 title claims abstract description 7
- 230000000050 nutritive effect Effects 0.000 title abstract description 6
- 238000000855 fermentation Methods 0.000 claims abstract description 176
- 230000004151 fermentation Effects 0.000 claims abstract description 167
- 239000007787 solid Substances 0.000 claims abstract description 58
- 241000228245 Aspergillus niger Species 0.000 claims abstract description 56
- 240000006439 Aspergillus oryzae Species 0.000 claims abstract description 54
- 235000002247 Aspergillus oryzae Nutrition 0.000 claims abstract description 54
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 51
- 241000193830 Bacillus <bacterium> Species 0.000 claims abstract description 50
- 244000199866 Lactobacillus casei Species 0.000 claims abstract description 46
- 235000013958 Lactobacillus casei Nutrition 0.000 claims abstract description 46
- 229940017800 lactobacillus casei Drugs 0.000 claims abstract description 46
- 240000002791 Brassica napus Species 0.000 claims abstract description 40
- 235000004977 Brassica sinapistrum Nutrition 0.000 claims abstract description 40
- 238000002156 mixing Methods 0.000 claims abstract description 40
- 239000001963 growth medium Substances 0.000 claims abstract description 39
- 235000007164 Oryza sativa Nutrition 0.000 claims abstract description 26
- 235000009566 rice Nutrition 0.000 claims abstract description 26
- 239000002609 medium Substances 0.000 claims abstract description 19
- 238000001035 drying Methods 0.000 claims abstract description 12
- 239000007788 liquid Substances 0.000 claims abstract description 12
- 230000000694 effects Effects 0.000 claims description 39
- 238000004321 preservation Methods 0.000 claims description 30
- 241000209094 Oryza Species 0.000 claims description 25
- 239000002994 raw material Substances 0.000 claims description 22
- 241000194108 Bacillus licheniformis Species 0.000 claims description 21
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 claims description 18
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 18
- 244000005700 microbiome Species 0.000 claims description 16
- 230000001954 sterilising effect Effects 0.000 claims description 16
- 235000019764 Soybean Meal Nutrition 0.000 claims description 15
- 239000004455 soybean meal Substances 0.000 claims description 15
- 108090000145 Bacillolysin Proteins 0.000 claims description 14
- 244000063299 Bacillus subtilis Species 0.000 claims description 14
- 235000014469 Bacillus subtilis Nutrition 0.000 claims description 14
- 108091005507 Neutral proteases Proteins 0.000 claims description 14
- 102000035092 Neutral proteases Human genes 0.000 claims description 14
- 102000004190 Enzymes Human genes 0.000 claims description 13
- 108090000790 Enzymes Proteins 0.000 claims description 13
- 239000005862 Whey Substances 0.000 claims description 13
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- 229940088598 enzyme Drugs 0.000 claims description 13
- 239000000843 powder Substances 0.000 claims description 13
- 238000011081 inoculation Methods 0.000 claims description 12
- 238000011282 treatment Methods 0.000 claims description 12
- 238000012807 shake-flask culturing Methods 0.000 claims description 10
- 229910000019 calcium carbonate Inorganic materials 0.000 claims description 9
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 9
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 9
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 9
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 9
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 9
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 9
- 108010059892 Cellulase Proteins 0.000 claims description 8
- 101710121765 Endo-1,4-beta-xylanase Proteins 0.000 claims description 8
- 229940106157 cellulase Drugs 0.000 claims description 8
- 235000016709 nutrition Nutrition 0.000 claims description 7
- 108091005658 Basic proteases Proteins 0.000 claims description 6
- 239000007858 starting material Substances 0.000 claims description 6
- 238000004659 sterilization and disinfection Methods 0.000 claims description 6
- 244000144972 livestock Species 0.000 claims description 5
- 238000002360 preparation method Methods 0.000 claims 1
- 241001465754 Metazoa Species 0.000 abstract description 12
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- IMQLKJBTEOYOSI-UHFFFAOYSA-N Phytic acid Natural products OP(O)(=O)OC1C(OP(O)(O)=O)C(OP(O)(O)=O)C(OP(O)(O)=O)C(OP(O)(O)=O)C1OP(O)(O)=O IMQLKJBTEOYOSI-UHFFFAOYSA-N 0.000 description 4
- 238000011161 development Methods 0.000 description 4
- 239000011790 ferrous sulphate Substances 0.000 description 4
- 235000003891 ferrous sulphate Nutrition 0.000 description 4
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 description 4
- 229910000359 iron(II) sulfate Inorganic materials 0.000 description 4
- 235000002949 phytic acid Nutrition 0.000 description 4
- 239000000467 phytic acid Substances 0.000 description 4
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- 235000018553 tannin Nutrition 0.000 description 4
- 239000001648 tannin Substances 0.000 description 4
- 229920001864 tannin Polymers 0.000 description 4
- ZMZDMBWJUHKJPS-UHFFFAOYSA-M Thiocyanate anion Chemical compound [S-]C#N ZMZDMBWJUHKJPS-UHFFFAOYSA-M 0.000 description 3
- 238000010564 aerobic fermentation Methods 0.000 description 3
- 238000001784 detoxification Methods 0.000 description 3
- 235000005911 diet Nutrition 0.000 description 3
- 230000037213 diet Effects 0.000 description 3
- 125000004383 glucosinolate group Chemical group 0.000 description 3
- ZMZDMBWJUHKJPS-UHFFFAOYSA-N hydrogen thiocyanate Natural products SC#N ZMZDMBWJUHKJPS-UHFFFAOYSA-N 0.000 description 3
- 150000002540 isothiocyanates Chemical class 0.000 description 3
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- JVTAAEKCZFNVCJ-REOHCLBHSA-N L-lactic acid Chemical compound C[C@H](O)C(O)=O JVTAAEKCZFNVCJ-REOHCLBHSA-N 0.000 description 2
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- 235000019629 palatability Nutrition 0.000 description 2
- 210000002976 pectoralis muscle Anatomy 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
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- 238000010563 solid-state fermentation Methods 0.000 description 2
- 241000228212 Aspergillus Species 0.000 description 1
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- 108010082495 Dietary Plant Proteins Proteins 0.000 description 1
- 206010018498 Goitre Diseases 0.000 description 1
- VCUFZILGIRCDQQ-KRWDZBQOSA-N N-[[(5S)-2-oxo-3-(2-oxo-3H-1,3-benzoxazol-6-yl)-1,3-oxazolidin-5-yl]methyl]-2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidine-5-carboxamide Chemical compound O=C1O[C@H](CN1C1=CC2=C(NC(O2)=O)C=C1)CNC(=O)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F VCUFZILGIRCDQQ-KRWDZBQOSA-N 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000000433 anti-nutritional effect Effects 0.000 description 1
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- PIBWKRNGBLPSSY-UHFFFAOYSA-L palladium(II) chloride Chemical compound Cl[Pd]Cl PIBWKRNGBLPSSY-UHFFFAOYSA-L 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 238000003825 pressing Methods 0.000 description 1
- 238000010025 steaming Methods 0.000 description 1
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- 210000001685 thyroid gland Anatomy 0.000 description 1
- 238000004448 titration Methods 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 238000009423 ventilation Methods 0.000 description 1
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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
- 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
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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
- A23K20/20—Inorganic substances, e.g. oligoelements
- A23K20/26—Compounds containing phosphorus
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- 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
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- 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
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- 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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Abstract
The invention discloses a method for biologically degrading rapeseed meal toxin and improving the nutritive value thereof, which comprises the following steps: firstly, respectively preparing a solid culture medium and a rapeseed meal fermentation culture medium; averagely dividing the rapeseed meal fermentation medium into two parts, inoculating aspergillus oryzae strain into one part, and fermenting to obtain an aspergillus oryzae fermented product; inoculating Aspergillus niger strain to the other part, and fermenting to obtain Aspergillus niger fermented product; inoculating a bacillus strain into the solid culture medium and fermenting to obtain a bacillus fermentation product; and finally, uniformly mixing the rice koji leavening, the black koji leavening and the bacillus leavening, inoculating lactobacillus casei liquid strain, fermenting, and drying the obtained leavening after the fermentation is finished until the water content is 10-12 wt% to obtain the fermented rapeseed dregs. The invention can reduce the toxin content in the rapeseed dregs and improve the nutritive value of the rapeseed dregs, so that the rapeseed dregs can be better applied to animal feed.
Description
Technical Field
The invention belongs to the technical field of microbial fermentation, and particularly relates to a method for biodegrading rapeseed meal toxin and improving the nutritive value of the rapeseed meal toxin.
Background
The protein is an important component in animal feed and is generally provided by soybean meal, but the soybean self-sufficiency rate in China is less than 20%, and the protein mainly depends on import. In recent years, with the rapid development of the livestock breeding industry in China and the further expansion of the breeding volume, the supply of conventional feed raw materials, especially protein raw materials, is in short supply, so that the healthy development of the livestock breeding industry in China is limited. According to statistics, 8851 million tons of domestic imported soybeans are increased by 48 million tons in 2019, the imported quantity accounts for about 85% of domestic consumption, and the imported proportion of Brazil, the United states and Argentina is 65%, 19% and 10% respectively. The imported soybeans in China in 2020 do not decrease and increase reversely under the influence of epidemic situations, and the quantity of imported soybeans in China reaches 9280 million tons to the end of 11 months in 2020, which exceeds the synchronization level in 2019. Under the international background that the competition of the current protein feed raw materials is intensified day by day, the traditional corn-soybean meal type animal feed structure in China will be changed, the development of novel protein feed resources is imminent, and the introduction of diversified feed formulas of miscellaneous meal such as rapeseed meal and the like will become a new normal state of the livestock breeding industry in China.
The rapeseed dregs are byproducts of rapeseed after oil pressing, the crude protein content of the rapeseed dregs is 35% -45%, and the rapeseed dregs are rich in amino acid, so the rapeseed dregs are an important vegetable protein resource. However, the rapeseed dregs contain more toxic and harmful substances, such as isothiocyanate, thiocyanate, sinapine, tannin, phytic acid and the like, which not only affect the palatability of the daily ration and the utilization of other nutrient substances, but also can cause the thyroid enlargement of animals and inhibit the growth of the animals, thereby greatly limiting the application of the rapeseed dregs in the daily ration of the animals. The microbial solid state fermentation can effectively reduce the content of antinutritional factors in the rapeseed dregs, and has higher development value in the aspect of replacing the soybean dregs. However, the existing rapeseed meal fermentation has the problems of different fermentation conditions, larger difference of fermentation effects and the like. Meanwhile, the problems of degrading the crude protein of the rapeseed dregs and detoxifying are difficult to solve by using a single microbial strain. Therefore, a method is needed at present, which can reduce the toxin content in the rapeseed dregs and improve the nutritive value of the rapeseed dregs, so that the rapeseed dregs can be better applied to animal feeds.
Disclosure of Invention
Aiming at the prior art, the invention aims to provide a method for biologically degrading rapeseed meal toxin and improving the nutritional value of the rapeseed meal toxin. The method adopts a first aerobic and then anaerobic two-stage solid state fermentation mode, and can ensure the enzymatic activity of microorganisms by utilizing the environment with relatively low moisture and high ventilation rate in the aerobic fermentation stage, and the high-temperature and high-humidity environment in the anaerobic fermentation stage can promote the enzymolysis. Meanwhile, the lactobacillus casei is utilized for anaerobic fermentation, so that products such as L-lactic acid with biological activity can be generated through metabolism, and the materials can be prevented from growing mixed bacteria under an acidic condition.
In order to achieve the purpose, the invention adopts the following technical scheme:
in a first aspect of the present invention, there is provided a method for biodegrading rapeseed meal toxin and improving the nutritional value thereof, comprising the steps of:
(1) mixing the solid raw material with water until the water content is 45-50 wt%, and sterilizing to obtain a bacillus solid fermentation culture medium; mixing rapeseed meal, whey powder and water until the water content is 42-46 wt%, and sterilizing to obtain a rapeseed meal fermentation culture medium;
(2) averagely dividing the rapeseed meal fermentation medium into two parts, inoculating aspergillus oryzae strains to one part, and fermenting to obtain an aspergillus oryzae fermented product; inoculating Aspergillus niger strain to the other part, and fermenting to obtain Aspergillus niger fermented product; inoculating a bacillus strain into the bacillus solid fermentation culture medium and fermenting to obtain a bacillus fermentation product;
(3) uniformly mixing the rice starter fermentation product, the black starter fermentation product and the bacillus fermentation product, inoculating lactobacillus casei liquid strain, and fermenting to obtain an anaerobic fermentation product after fermentation;
(4) and (4) drying the fermented product obtained in the step (3) until the moisture content is 10-12 wt%, so as to obtain the fermented rapeseed meal.
Preferably, in step (1), the solid raw material comprises the following raw materials in percentage by weight:
80% of bran, 17% of rice hull, 2% of soybean meal, 0.15% of monopotassium phosphate, 0.2% of magnesium sulfate and 0.65% of light calcium carbonate.
Preferably, in the step (1), the addition amount of the whey powder is 0.1% of the total mass of the rapeseed dregs; the sterilization temperature is 105-121 ℃, and the time is 15-30 min. The bacillus solid fermentation culture medium and the rapeseed meal fermentation culture medium are obtained under the same sterilization treatment conditions, and the sterilization temperature and the sterilization time are the same.
Preferably, in the step (2), the Aspergillus oryzae strain is obtained by solid culture until the number of Aspergillus oryzae spores reaches 30 hundred million/g and the activity of neutral protease reaches 2000U/g;
the inoculation amount of the aspergillus oryzae strain is 0.5-1.0% of half of the mass of the rapeseed dreg fermentation culture medium, and the aspergillus oryzae is cultured for 30-32 hours at 28-30 ℃ to obtain a rice koji fermentation product; the aspergillus oryzae is purchased from China industrial microorganism strain preservation management center, and the strain preservation number is as follows: and CICC 2013.
Preferably, in the step (2), the Aspergillus niger strains are obtained by solid culture until the number of Aspergillus niger spores reaches 10 hundred million/g, the activity of xylanase reaches 1000U/g, and the activity of cellulase reaches 100U/g;
the inoculation amount of the aspergillus niger accounts for 1.0-1.5% of the half mass of the rapeseed dreg fermentation culture medium, and the aspergillus niger is cultured for 36-40h at 28-30 ℃ to obtain a aspergillus niger fermented product; the Aspergillus niger is purchased from China industrial microorganism strain preservation management center, and the strain preservation number is as follows: CICC 2041.
Preferably, in step (2), the bacillus strain is obtained by culturing bacillus licheniformis or bacillus subtilis in a shake flask, and the number of the cultured bacillus strain is 1 × 1010cfu/mL; the mass ratio of the inoculation amount of the bacillus strain to the bacillus solid fermentation culture medium is (2-3) mL: 100g, fermenting at 37-39 deg.C for 44-48h to make the number of bacillus reach 2 × 1010cfu/g, the enzyme activity of neutral protease reaches 3000U/g, and the enzyme activity of alkaline protease reaches 5000U/g, so as to obtain a bacillus fermentation product; the bacillus licheniformis is purchased from China industrial microorganism strain preservation management center, and the strain preservation number is as follows: CICC 21886; the bacillus subtilis is purchased from China industrial microorganism strain preservation management center, and the strain preservation number is as follows: CICC 20872.
Preferably, in the step (3), the rice koji fermentation product, the black koji fermentation product and the bacillus fermentation product are respectively according to (55-65): (25-30): (10-15) mass ratio.
Preferably, in the step (3), the lactobacillus casei liquid strain is obtained by culturing lactobacillus casei in a shake flask, and the bacterial count of the lactobacillus casei is cultured to 6 × 109cfu/mL; the ratio of the inoculation amount of the lactobacillus casei fermentation liquid to the total mass of the rice koji fermentation product, the black koji fermentation product and the bacillus fermentation product is (3-5) mL: 100g of the total weight of the mixture; the fermentation temperature is not lower than 30 deg.C and not higher than 40 deg.C, and the fermentation time is 3-4 d; the lactobacillus casei is purchased from China industrial microorganism strain preservation management center, and the strain preservation number is as follows: CICC 6117.
In a second aspect of the present invention, there is provided the fermented rapeseed meal produced by the above-mentioned method.
In a third aspect of the invention, the invention provides an application of fermented rapeseed dregs in preparing livestock feed.
The invention has the beneficial effects that:
(1) the multi-strain stepwise fermentation adopted by the invention can degrade toxic and harmful substances such as glucosinolate, tannin, phytic acid and the like in the rapeseed dregs by utilizing a complex enzyme system of microorganisms in the fermentation process, and can hydrolyze cell wall polysaccharide, crude protein and the like, so that the organic body can absorb more easily, and the nutritional value of the rapeseed dregs is improved.
(2) The content of the crude protein of the fermented rapeseed meal obtained by the invention reaches more than 44 percent, is equivalent to the protein content of the soybean meal, can be used as a protein feed raw material to replace the soybean meal, and reduces the breeding cost. Meanwhile, the fermented rapeseed dregs are added into animal feed, so that the production performance of animals can be improved, the total thorax clearance rate, the pectoral muscle rate and the leg muscle rate are improved, and the economic benefit is increased.
Detailed Description
It should be noted that the following detailed description is exemplary and is intended to provide further explanation of the disclosure. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.
As introduced in the background technology, the rapeseed dregs contain more toxic and harmful substances, such as isothiocyanate, thiocyanate, sinapine, tannin, phytic acid and the like, which not only influence the palatability of the daily ration and the utilization of other nutrient substances, but also cause the goiter of animals and inhibit the growth of the animals, thereby greatly limiting the application of the rapeseed dregs in the daily ration of the animals.
Based on this, the invention aims to provide a method for biologically degrading rapeseed meal toxin and improving the nutritional value of the rapeseed meal toxin. The method of the invention is finally obtained by screening strains and screening fermentation culture modes (see test 1): firstly, respectively preparing a solid culture medium and a rapeseed meal fermentation culture medium; averagely dividing the rapeseed meal fermentation medium into two parts, inoculating aspergillus oryzae strain into one part, and fermenting to obtain an aspergillus oryzae fermented product; inoculating Aspergillus niger strain to the other part, and fermenting to obtain Aspergillus niger fermented product; inoculating a bacillus licheniformis strain or a bacillus subtilis strain to the solid culture medium, and fermenting to obtain a bacillus fermentation product; and finally, uniformly mixing the rice koji leavening, the black koji leavening and the bacillus leavening, inoculating lactobacillus casei leavening liquid, fermenting, and drying the leavening obtained after the fermentation is finished until the moisture content is 10-12 wt% to obtain the fermented rapeseed dregs.
Test 1
The rapeseed meal, the whey powder and the water are uniformly mixed according to the mass percentage of the invention to obtain the rapeseed meal fermentation medium. Firstly, inoculating the strains in the table 1 into a rapeseed meal fermentation culture medium (the inoculation amount is 1wt percent) for aerobic fermentation, fermenting for 34h at 28-30 ℃, and calculating the removal rate of toxin according to the content of the toxin in the rapeseed meal before and after the detection of the fermentation, wherein the removal rate is (the content before the treatment-the content after the treatment)/the content before the treatment is multiplied by 100 percent; the results are shown in Table 1.
TABLE 1
According to the table 1, selecting aspergillus oryzae and aspergillus niger with the optimal rapeseed meal toxin removal rate, and fermenting the rapeseed meal with the same mass respectively to obtain aspergillus oryzae leavening and aspergillus niger leavening; mixing Aspergillus oryzae fermented product and Aspergillus niger fermented product according to the ratio of 1: 1 to obtain a mixture, inoculating the strains in the table 2 to perform anaerobic fermentation, wherein the inoculum size is 3mL of strains inoculated to every 100g of the mixture, controlling the initial fermentation temperature to be 30-32 ℃, and fermenting for 1 d. Calculating the removal rate of the toxin according to the content of the toxin in the rapeseed dregs before and after the fermentation, wherein the removal rate is (the content before the treatment-the content after the treatment)/the content before the treatment is multiplied by 100 percent; the results are shown in Table 2.
TABLE 2
As can be seen from table 2, the ratio of aspergillus oryzae fermented product to aspergillus niger fermented product was 1: 1, the mass ratio of the components is mixed and added with lactobacillus casei for fermentation, so that the effect of removing the toxin from the rapeseed dregs is the best.
Mixing bran, rice hulls, soybean meal, monopotassium phosphate, magnesium sulfate, light calcium carbonate and water according to the mass ratio of the solid culture medium to obtain a solid culture medium, inoculating the strains in the table 3 into the solid culture medium, and performing aerobic fermentation to obtain a solid fermentation product (the inoculation amount is 3% according to mass volume percentage, and the fermentation is carried out for 46 hours at 37-39 ℃). Mixing the solid fermentation product, the aspergillus oryzae fermentation product and the aspergillus niger fermentation product according to the proportion of 1: 1: 1 to obtain a mixture, inoculating the strains in the table 2 to perform anaerobic fermentation, wherein the inoculation amount is that 3mL of strains are inoculated to every 100g of the mixture, the fermentation starting temperature is controlled to be 30-32 ℃, and the fermentation is performed for 1 d. Calculating the removal rate of the toxin according to the content of the toxin in the rapeseed dregs before and after the fermentation, wherein the removal rate is (the content before the treatment-the content after the treatment)/the content before the treatment is multiplied by 100 percent; the results are shown in Table 3.
TABLE 3
As can be seen from table 3, the solid fermentate obtained by inoculating the strains in table 3 with the aspergillus oryzae fermentate and aspergillus niger fermentate was mixed in the ratio of 1: 1: 1, then inoculating the strains in the table 2 for anaerobic fermentation, and the best effect is that the solid fermentation medium is inoculated with bacillus licheniformis or bacillus subtilis to obtain a bacillus fermentation product, and the aspergillus fermentation product and the aspergillus niger fermentation product are mixed and inoculated with lactobacillus casei for anaerobic fermentation to obtain fermented rapeseed meal. The most reduced average toxicity was obtained.
The method comprises the following steps of uniformly mixing rice koji leavening, black koji leavening and bacillus leavening according to different mass ratios, inoculating 4% (volume mass percentage) of lactobacillus casei leavening liquid, controlling the initial fermentation temperature to be 30-32 ℃, culturing for 3d, and obtaining leavening after fermentation is finished; calculating the removal rate of the toxin, and finding that when the mass percentage of the rice koji leavening, the black koji leavening and the bacillus leavening is 65: 27.5: the removal rate of the toxin is highest when the concentration is 12.5. The removal rates of isothiocyanate, thiocyanate, sinapine, tannin and phytic acid are 80.01%, 76.28%, 81.10%, 86.47% and 96.03%, respectively.
In order to make the technical solutions of the present application more clearly understood by those skilled in the art, the technical solutions of the present application will be described in detail below with reference to specific embodiments.
The test materials used in the examples of the present invention are all conventional in the art and commercially available.
Description of the drawings: the aspergillus oryzae is purchased from China industrial microorganism strain preservation management center, and the strain preservation number is as follows: CICC 2013;
the aspergillus niger is purchased from China industrial microorganism strain preservation management center, and the strain preservation number is as follows: CICC 2041;
the bacillus licheniformis is purchased from China industrial microorganism strain preservation management center, and the strain preservation number is as follows: CICC 21886;
the bacillus subtilis is purchased from China industrial microorganism strain preservation management center, and the strain preservation number is as follows: CICC 20872;
the lactobacillus casei is purchased from China industrial microorganism strain preservation management center, and the strain preservation number is as follows: CICC 6117.
In the examples, the inoculation amounts of the fermentation seed liquids of the bacillus licheniformis, the bacillus subtilis and the lactobacillus casei are volume mass percent. For example: 4% of the Lactobacillus casei fermentation broth was inoculated, meaning that 4mL of Lactobacillus casei fermentation broth was inoculated per 100g of substrate.
Example 1:
(1) respectively culturing the fermentation seed solutions of the bacillus licheniformis and the lactobacillus casei by adopting a shake flask culture method, and culturing the bacterial count of the bacillus licheniformis to 1 multiplied by 1010cfu/mL, Lactobacillus casei count up to 6X 109cfu/mL; the solid culture method is adopted to respectively culture aspergillus oryzae and aspergillus niger strains, so that the number of aspergillus oryzae spores is 30 hundred million/g, the activity of neutral protease is 2000U/g, the number of aspergillus niger spores is 10 hundred million/g, the activity of xylanase is 1000U/g, and the activity of cellulase is 100U/g.
(2) Uniformly mixing 80g of bran, 17g of rice hull, 2g of soybean meal, 0.15g of monopotassium phosphate, 0.2g of magnesium sulfate and 0.65g of light calcium carbonate to obtain a solid raw material. Mixing the solid raw material with water until the water content is 47%, mixing 100g of rapeseed meal, 0.1g of whey powder and water until the water content is 44%, and sterilizing at 121 ℃ for 30min to obtain a solid fermentation culture medium and a rapeseed meal fermentation culture medium.
(3) Equally dividing the rapeseed meal fermentation medium into two parts according to the mass, inoculating 0.75 wt% of aspergillus oryzae strain into one part, and culturing at 29 ℃ for 31h to obtain the aspergillus oryzae fermentation product. Inoculating 1.25 wt% Aspergillus niger strain to another part, and culturing at 29 deg.C for 38 hr to obtain Aspergillus niger fermented product. Inoculating 2.5% Bacillus licheniformis strain in solid fermentation medium, and culturing at 38 deg.C for 46 hr to make the number of Bacillus to 2 × 1010cfu/g, the enzyme activity of neutral protease reaches 3000U/g, and the enzyme activity of alkaline protease reaches 5000U/g, so that the bacillus fermentation product is obtained.
(4) Respectively mixing rice koji leavening, black koji leavening and bacillus leavening according to the ratio of 60: 27.5: uniformly mixing the materials according to the mass ratio of 12.5, inoculating 4% lactobacillus casei fermentation liquor, controlling the initial fermentation temperature to be 32-38 ℃, culturing for 3d, and obtaining a fermentation product after the fermentation is finished.
(5) And (4) drying the fermented product obtained in the step (4) at 45-50 ℃ until the water content is 11%, so as to obtain the fermented rapeseed meal.
Example 2
(1) Respectively culturing the seed fermentation solutions of Bacillus subtilis and Lactobacillus casei by shake flask culture method to obtain Bacillus subtilis with count of 1 × 1010cfu/mL, Lactobacillus casei count up to 6X 109cfu/mL; the solid culture method is adopted to respectively culture aspergillus oryzae and aspergillus niger strains, so that the number of aspergillus oryzae spores is 30 hundred million/g, the activity of neutral protease is 2000U/g, the number of aspergillus niger spores is 10 hundred million/g, the activity of xylanase is 1000U/g, and the activity of cellulase is 100U/g.
(2) Uniformly mixing 80g of bran, 17g of rice hull, 2g of soybean meal, 0.15g of monopotassium phosphate, 0.2g of magnesium sulfate and 0.65g of light calcium carbonate to obtain a solid raw material. Mixing the solid raw material with water until the water content is 45%, mixing 100g of rapeseed meal, 0.1g of whey powder and water until the water content is 46%, and sterilizing at 121 ℃ for 30min to obtain a solid fermentation culture medium and a rapeseed meal fermentation culture medium.
(3) Equally dividing the rapeseed meal fermentation medium into two parts according to the mass, inoculating 0.5 wt% aspergillus oryzae strain into one part, and culturing at 30 ℃ for 32h to obtain the aspergillus oryzae fermentation product. Inoculating 1.5 wt% Aspergillus niger strain to the other part, culturing at 28 deg.C for 40 hr,obtaining the black koji leavening. Inoculating 2% Bacillus subtilis strain in solid fermentation medium, and culturing at 39 deg.C for 48 hr to make the number of Bacillus reach 2 × 1010cfu/g, the enzyme activity of neutral protease reaches 3000U/g, and the enzyme activity of alkaline protease reaches 5000U/g, so that the bacillus fermentation product is obtained.
(4) Respectively mixing rice koji fermented product, black koji fermented product and bacillus fermented product according to the ratio of 55: 30: 15, inoculating 5% lactobacillus casei fermentation liquor, controlling the initial fermentation temperature to be 31-34 ℃, culturing for 4 days, and obtaining a fermentation product after the fermentation is finished.
(5) And (4) drying the fermented product obtained in the step (4) at 45-50 ℃ until the water content is 12%, so as to obtain the fermented rapeseed meal.
Example 3
(1) Respectively culturing the fermentation seed solutions of the bacillus licheniformis and the lactobacillus casei by adopting a shake flask culture method, and culturing the bacterial count of the bacillus licheniformis to 1 multiplied by 1010cfu/mL, Lactobacillus casei count up to 6X 109cfu/mL; the solid culture method is adopted to respectively culture aspergillus oryzae and aspergillus niger strains, so that the number of aspergillus oryzae spores is 30 hundred million/g, the activity of neutral protease is 2000U/g, the number of aspergillus niger spores is 10 hundred million/g, the activity of xylanase is 1000U/g, and the activity of cellulase is 100U/g.
(2) Uniformly mixing 80g of bran, 17g of rice hull, 2g of soybean meal, 0.15g of monopotassium phosphate, 0.2g of magnesium sulfate and 0.65g of light calcium carbonate to obtain a solid raw material. Mixing the solid raw material with water until the water content is 50%, mixing 100g of rapeseed meal, 0.1g of whey powder and water until the water content is 42%, and sterilizing at 121 ℃ for 30min to obtain a solid fermentation culture medium and a rapeseed meal fermentation culture medium.
(3) Equally dividing the rapeseed meal fermentation medium into two parts according to the mass, inoculating 1.0 wt% of aspergillus oryzae strain into one part, and culturing for 30h at 28 ℃ to obtain the aspergillus oryzae fermentation product. Inoculating 1.0 wt% Aspergillus niger strain to another part, and culturing at 30 deg.C for 36 hr to obtain Aspergillus niger fermented product. Inoculating 3% Bacillus licheniformis strain in solid fermentation medium, and culturing at 37 deg.C for 48 hr to make the number of Bacillus to 2 × 1010cfu/g, the enzyme activity of neutral protease reaches 3000U/g, and the enzyme activity of alkaline protease reaches 5000U/g, so that the bacillus fermentation product is obtained.
(4) Respectively mixing rice koji leavening, black koji leavening and bacillus leavening according to the ratio of 65: 25: 10, inoculating 3 percent lactobacillus casei fermentation liquor, controlling the initial fermentation temperature to be 32-38 ℃, culturing for 4 days, and obtaining a fermentation product after the fermentation is finished.
(5) And (4) drying the fermented product obtained in the step (4) at 45-50 ℃ until the water content is 10%, so as to obtain the fermented rapeseed meal.
Comparative example 1
Preparing rapeseed meal by using a physical detoxification method: adding rapeseed meal into an autoclave, adding water to moisten until the water content is 30%, introducing steam, treating at 121 deg.C for 2 hr, taking out, and air drying to obtain physical detoxification method rapeseed meal.
Comparative example 2
Preparing rapeseed meal by using a chemical detoxification method: taking rapeseed dregs, weighing ferrous sulfate according to 0.5-1% of the weight of the rapeseed dregs, dissolving the ferrous sulfate in water of 1/2 weight of the rapeseed dregs, pouring the ferrous sulfate into the rapeseed dregs after the ferrous sulfate is fully dissolved, uniformly mixing, and storing for 1h at room temperature. Steaming at 105 deg.C for 30min, taking out, and air drying to obtain chemical detoxication method rapeseed meal.
Comparative example 3
(1) Respectively culturing the fermentation seed solutions of the bacillus licheniformis and the lactobacillus casei by adopting a shake flask culture method, and culturing the bacterial count of the bacillus licheniformis to 1 multiplied by 1010cfu/mL, Lactobacillus casei count up to 6X 109cfu/mL; the solid culture method is adopted to respectively culture aspergillus oryzae and aspergillus niger strains, so that the number of aspergillus oryzae spores is 30 hundred million/g, and the number of aspergillus niger spores is 10 hundred million/g.
(2) Uniformly mixing 80g of bran, 17g of rice hull, 2g of soybean meal, 0.15g of monopotassium phosphate, 0.2g of magnesium sulfate and 0.65g of light calcium carbonate to obtain a solid raw material. Mixing the solid raw material with water until the water content is 47%, mixing 100g of rapeseed meal, 0.1g of whey powder and water until the water content is 44%, and sterilizing at 121 ℃ for 30min to obtain a solid fermentation culture medium and a rapeseed meal fermentation culture medium.
(3) Equally dividing the rapeseed meal fermentation medium into two parts according to the mass, inoculating 0.75 wt% of aspergillus oryzae strain into one part, and culturing at 29 ℃ for 72h to obtain the aspergillus oryzae fermentation product. Inoculating 1.25 wt% Aspergillus niger strain to another part, and culturing at 29 deg.C for 72 hr to obtain Aspergillus niger fermented product. Inoculating 2.5% Bacillus licheniformis strain in the solid fermentation medium, and culturing at 38 deg.C for 72 hr to obtain Bacillus licheniformis fermented product.
(4) Respectively mixing rice koji leavening, black koji leavening and bacillus leavening according to the ratio of 60: 27.5: uniformly mixing the materials according to the mass ratio of 12.5, inoculating 4% lactobacillus casei fermentation liquor, controlling the initial fermentation temperature to be 32-38 ℃, culturing for 6d, and obtaining a fermentation product after the fermentation is finished.
(5) And (4) drying the fermented product obtained in the step (4) at 45-50 ℃ until the water content is 11%, so as to obtain the fermented rapeseed meal.
Comparative example 4
(1) Respectively culturing the seed fermentation solutions of Bacillus subtilis and Lactobacillus casei by shake flask culture method to obtain Bacillus subtilis with count of 1 × 1010cfu/mL, Lactobacillus casei count up to 6X 109cfu/mL; the solid culture method is adopted to respectively culture aspergillus oryzae and aspergillus niger strains, so that the number of aspergillus oryzae spores is 30 hundred million/g, and the number of aspergillus niger spores is 10 hundred million/g.
(2) Uniformly mixing 80g of bran, 17g of rice hull, 2g of soybean meal, 0.15g of monopotassium phosphate, 0.2g of magnesium sulfate and 0.65g of light calcium carbonate to obtain a solid raw material. Mixing the solid raw material with water until the water content is 47%, mixing 100g of rapeseed meal, 0.1g of whey powder and water until the water content is 44%, and sterilizing at 121 ℃ for 30min to obtain a solid fermentation culture medium and a rapeseed meal fermentation culture medium.
(3) Equally dividing the rapeseed meal fermentation medium into two parts according to the mass, inoculating 0.75 wt% of aspergillus oryzae strain into one part, and culturing at 29 ℃ for 12h to obtain the aspergillus oryzae fermentation product. Inoculating 1.25 wt% Aspergillus niger strain to another part, and culturing at 29 deg.C for 12 hr to obtain Aspergillus niger fermented product. Inoculating 2.5% Bacillus subtilis strain in solid fermentation culture medium, and culturing at 38 deg.C for 6 hr to obtain Bacillus fermentation product.
(4) Respectively mixing rice koji leavening, black koji leavening and bacillus leavening according to the ratio of 60: 27.5: uniformly mixing the materials according to the mass ratio of 12.5, inoculating 4% lactobacillus casei fermentation liquor, controlling the initial fermentation temperature to be 32-38 ℃, culturing for 1d, and obtaining a fermentation product after the fermentation is finished.
(5) And (4) drying the fermented product obtained in the step (4) at 45-50 ℃ until the water content is 11%, so as to obtain the fermented rapeseed meal.
Comparative example 5
(1) Respectively culturing lactobacillus casei fermentation seed liquid by adopting a shake flask culture method, and culturing the lactobacillus casei bacterial count to 6 multiplied by 109cfu/mL; the solid culture method is adopted to respectively culture aspergillus oryzae and aspergillus niger strains, so that the number of aspergillus oryzae spores is 30 hundred million/g, the activity of neutral protease is 2000U/g, the number of aspergillus niger spores is 10 hundred million/g, the activity of xylanase is 1000U/g, and the activity of cellulase is 100U/g.
(2) 100g of rapeseed meal, 0.1g of whey powder and water are mixed until the water content is 44%, and the mixture is sterilized for 30min at 121 ℃ to obtain the rapeseed meal fermentation medium. Inoculating 0.75 wt% of Aspergillus oryzae and 1.25 wt% of Aspergillus niger to the rapeseed dreg fermentation culture medium, and culturing at 29 ℃ for 36h to obtain a fermented product.
(3) Inoculating 4% lactobacillus casei fermentation liquor to the fermentation product, controlling the initial fermentation temperature to be 32-38 ℃, culturing for 3d, and obtaining the fermentation product after the fermentation is finished.
(4) And (4) drying the fermented product obtained in the step (3) at 45-50 ℃ until the water content is 11%, so as to obtain the fermented rapeseed meal.
Comparative example 6
(1) Respectively culturing lactobacillus casei fermentation seed liquid by adopting a shake flask culture method, and culturing the lactobacillus casei bacterial count to 6 multiplied by 109cfu/mL; the solid culture method is adopted to respectively culture aspergillus oryzae and aspergillus niger strains, so that the number of aspergillus oryzae spores is 30 hundred million/g, the activity of neutral protease is 2000U/g, the number of aspergillus niger spores is 10 hundred million/g, the activity of xylanase is 1000U/g, and the activity of cellulase is 100U/g.
(2) 100g of rapeseed meal, 0.1g of whey powder and water are mixed until the water content is 44%, and the mixture is sterilized for 30min at 121 ℃ to obtain the rapeseed meal fermentation medium. Inoculating 0.75 wt% of aspergillus oryzae and 1.25 wt% of aspergillus niger to the rapeseed meal fermentation medium, inoculating 4% of lactobacillus casei fermentation liquor, controlling the initial fermentation temperature to be 32-38 ℃, and culturing for 30h to obtain a fermented product.
(3) And (3) drying the fermented product obtained in the step (2) at 45-50 ℃ until the water content is 11% to obtain the fermented rapeseed meal.
Comparative example 7
(1) Respectively culturing the fermentation seed solutions of the bacillus licheniformis and the lactobacillus casei by adopting a shake flask culture method, and culturing the bacterial count of the bacillus licheniformis to 1 multiplied by 1010cfu/mL, Lactobacillus casei count up to 6X 109cfu/mL; the solid culture method is adopted to respectively culture aspergillus oryzae and aspergillus niger strains, so that the number of aspergillus oryzae spores is 30 hundred million/g, the activity of neutral protease is 2000U/g, the number of aspergillus niger spores is 10 hundred million/g, the activity of xylanase is 1000U/g, and the activity of cellulase is 100U/g.
(2) Uniformly mixing 80g of bran, 17g of rice hull, 2g of soybean meal, 0.15g of monopotassium phosphate, 0.2g of magnesium sulfate and 0.65g of light calcium carbonate to obtain a solid raw material. Mixing the solid raw material with water until the water content is 47%, mixing 100g of rapeseed meal, 0.1g of whey powder and water until the water content is 44%, and sterilizing at 121 ℃ for 30min to obtain a solid fermentation culture medium and a rapeseed meal fermentation culture medium.
(3) Inoculating 0.75 wt% of Aspergillus oryzae and 1.25 wt% of Aspergillus niger to the rapeseed dreg fermentation culture medium, and culturing at 29 ℃ for 38h to obtain a mixed fermentation product. Inoculating 2.5% Bacillus licheniformis strain in solid fermentation medium, and culturing at 38 deg.C for 46 hr to make the number of Bacillus to 2 × 1010cfu/g, the enzyme activity of neutral protease reaches 3000U/g, and the enzyme activity of alkaline protease reaches 5000U/g, so that the bacillus fermentation product is obtained.
(4) And (3) mixing the mixed fermentation obtained in the step (3) with a bacillus fermentation product, inoculating 4% of lactobacillus casei fermentation liquor, controlling the initial fermentation temperature to be 32-38 ℃, culturing for 3d, and obtaining the fermentation product after the fermentation is finished.
(5) And (4) drying the fermented product obtained in the step (4) at 45-50 ℃ until the water content is 11% to obtain the fermented rapeseed meal.
Test example 1
Taking the rapeseed dregs obtained in the examples 1-3 and the comparative examples 1-7, and determining the glucosinolate content before and after the fermentation of the rapeseed dregs by adopting a palladium chloride colorimetric method; determining the crude protein content of the sample by using a full-automatic azotometer; determining the content of acid soluble protein by referring to a determination method of GB/T22492-; the total acid content (based on organic acid) before and after fermentation was determined by titration. The results are shown in Table 4.
Table 4 (oven dry foundation)
As can be seen from the data in Table 4, the contents of glucosinolates in the fermented rapeseed meals prepared in examples 1 to 3 were lower than those in the comparative examples, and the contents of crude proteins, acid-soluble proteins and organic acids were higher than those in comparative examples 1 to 7, indicating that the nutritional value of the fermented rapeseed meals was improved.
Test example 2
360 healthy Aijia Yijia (AA) mixed chicks of 1 day old are selected and randomly divided into 3 treatments, each treatment is 6 times repeated, and each treatment is 20 times repeated. The test adopts a single-factor completely random test design, the control group is fed with basic diet which is corn-soybean meal type diet, and the nutrition level is prepared according to NRC (1994) broiler standard. The experimental group recorded 5% example 1, 10% example 1 by replacing the equivalent amount of soybean meal with 5% and 10% of the fermented rapeseed meal prepared in example 1, respectively. The test period is 42d, and the breeding is divided into two stages: the early stage 1-21d and the late stage 22-42 d.
The test results are shown in tables 5-6.
TABLE 5 influence of fermented rapeseed meal on broiler growth Performance
| Item | Control group | 5% example 1 | 10% of example 1 |
| Body weight (g) | 2234.0 | 2236.8 | 2238.0 |
| Average daily gain ADG/(g/d) | 52.41 | 52.46 | 52.47 |
| Average daily food intake ADFI/(g/d) | 94.91 | 92.37 | 92.66 |
| Material weight ratio F/G | 1.81 | 1.76 | 1.77 |
As can be seen from table 5, compared to the control group, the weight of the broiler chicken can be increased and the feed conversion ratio can be reduced by adding fermented rapeseed meal instead of an equal amount of soybean meal to the broiler chicken diet.
TABLE 6 influence of fermented rapeseed meal on slaughter Performance of broiler chickens
As can be seen from table 6, the fermented rapeseed meal can improve the slaughter rate, the total smoldering rate, the pectoral muscle rate and the leg muscle rate of the broiler chickens compared to the control group. Compared with soybean meal, the rapeseed meal has lower market price, can reduce the feed cost and increase the benefit.
The above description is only a preferred embodiment of the present application and is not intended to limit the present application, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, improvement and the like made within the spirit and principle of the present application shall be included in the protection scope of the present application.
Claims (10)
1. A method for biologically degrading rapeseed meal toxin and improving the nutritional value of the rapeseed meal toxin is characterized by comprising the following steps:
(1) mixing the solid raw material with water until the water content is 45-50 wt%, and sterilizing to obtain a bacillus solid fermentation culture medium; mixing rapeseed meal, whey powder and water until the water content is 42-46 wt%, and sterilizing to obtain a rapeseed meal fermentation culture medium;
(2) averagely dividing the rapeseed meal fermentation medium into two parts, inoculating aspergillus oryzae strains to one part, and fermenting to obtain an aspergillus oryzae fermented product; inoculating Aspergillus niger strain to the other part, and fermenting to obtain Aspergillus niger fermented product; inoculating a bacillus strain into the bacillus solid fermentation culture medium and fermenting to obtain a bacillus fermentation product;
(3) uniformly mixing the rice starter fermentation product, the black starter fermentation product and the bacillus fermentation product, inoculating lactobacillus casei liquid strain, and fermenting to obtain an anaerobic fermentation product after fermentation;
(4) and (4) drying the anaerobic fermentation product obtained in the step (3) until the water content is 10-12 wt%, so as to obtain the fermented rapeseed dregs.
2. The method according to claim 1, wherein in step (1), the solid raw materials comprise the following raw materials in percentage by weight:
80% of bran, 17% of rice hull, 2% of soybean meal, 0.15% of monopotassium phosphate, 0.2% of magnesium sulfate and 0.65% of light calcium carbonate.
3. The method according to claim 1, wherein in the step (1), the whey powder is added in an amount of 0.1% of the total mass of the rapeseed meal; the temperature of the sterilization treatment is 105-121 ℃, and the time of the sterilization treatment is 15-30 min.
4. The method according to claim 1, wherein in the step (2), the Aspergillus oryzae strain is obtained by solid culture until the number of Aspergillus oryzae spores reaches 30 hundred million/g and the neutral protease activity reaches 2000U/g;
the inoculation amount of the aspergillus oryzae strain is 0.5-1.0 wt% of half of the mass of the rapeseed dreg fermentation culture medium, and the aspergillus oryzae is cultured for 30-32 hours at 28-30 ℃ to obtain a rice koji fermentation product; the aspergillus oryzae is purchased from China industrial microorganism strain preservation management center, and the strain preservation number is as follows: and CICC 2013.
5. The method according to claim 1, wherein in step (2), the Aspergillus niger strains are obtained by solid culture until the number of Aspergillus niger spores reaches 10 hundred million/g, the xylanase activity reaches 1000U/g, and the cellulase activity reaches 100U/g;
the inoculation amount of the aspergillus niger accounts for 1.0-1.5 wt% of half of the mass of the rapeseed dreg fermentation culture medium, and the aspergillus niger is cultured for 36-40h at 28-30 ℃ to obtain a aspergillus niger fermented product; the Aspergillus niger is purchased from China industrial microorganism strain preservation management center, and the strain preservation number is as follows: CICC 2041.
6. The method according to claim 1, wherein in step (2), the Bacillus species is obtained by shake flask culture of Bacillus licheniformis or Bacillus subtilis, and the number of cultured Bacillus species is 1X 1010cfu/mL; the mass ratio of the inoculation amount of the bacillus strain to the bacillus solid fermentation culture medium is (2-3) mL: 100g, fermenting at 37-39 deg.C for 44-48h to make the number of bacillus reach 2 × 1010cfu/g, the enzyme activity of neutral protease reaches 3000U/g, and the enzyme activity of alkaline protease reaches 5000U/g, so as to obtain a bacillus fermentation product; the bacillus licheniformis is purchased from China industrial microorganism strain preservation management center, and the strain preservation number is as follows: CICC 21886; bacillus subtilis purchased from Chinese industrial microorganismThe strain preservation management center comprises the following strain preservation numbers: CICC 20872.
7. The method according to claim 1, wherein in the step (3), the rice koji fermentation product, the black koji fermentation product and the bacillus fermentation product are respectively prepared according to the following steps (55-65): (25-30): (10-15) mass ratio.
8. The method of claim 1, wherein in step (3), the liquid strain of Lactobacillus casei is obtained by shake flask culture of Lactobacillus casei, and the number of Lactobacillus casei is 6 × 109cfu/mL; the ratio of the inoculation amount of the lactobacillus casei liquid strain to the total mass of the rice starter fermentation product, the black starter fermentation product and the bacillus fermentation product is (3-5) mL: 100g, the initial temperature of fermentation is 30-40 ℃, and the total fermentation time is 3-4 d; the lactobacillus casei is purchased from China industrial microorganism strain preservation management center, and the strain preservation number is as follows: CICC 6117.
9. A fermented rapeseed meal produced by the method according to any one of claims 1 to 8.
10. Use of the fermented rapeseed meal according to claim 8 for the preparation of livestock feed.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114698726A (en) * | 2022-04-01 | 2022-07-05 | 播恩集团股份有限公司 | Low-toxin high-nutrition rapeseed meal, method for degrading toxins in rapeseed meal through segmented fermentation and application |
| CN115316491A (en) * | 2022-08-25 | 2022-11-11 | 山东泰山生力源集团股份有限公司 | Preparation method of feed additive, feed additive and feed |
| CN115736092A (en) * | 2022-12-14 | 2023-03-07 | 山东泰山生力源集团股份有限公司 | Method for improving glutamine content in feed through solid state fermentation |
| CN115851465A (en) * | 2022-12-14 | 2023-03-28 | 湖北省生物农药工程研究中心 | Composite fermentation inoculant, method for preparing fermented rapeseed meal for feed and application of composite fermentation inoculant |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS61166385A (en) * | 1985-01-18 | 1986-07-28 | Ajinomoto Co Inc | Treatment of rapeseed cake |
| CN102652529A (en) * | 2012-04-10 | 2012-09-05 | 湖北省农业科学院农产品加工与核农技术研究所 | Method for obtaining active polypeptide by carrying out multi-strain compound solid state fermentation on common rapeseed meal |
| CN102823726A (en) * | 2012-09-07 | 2012-12-19 | 北京大北农科技集团股份有限公司 | Biological fermentation method capable of improving protein content of cottonseed meal and removing toxicity |
| CN105494890A (en) * | 2014-09-26 | 2016-04-20 | 侯文燕 | Enzymolysis and fermentation combined method for improving feeding nutritional value of rapeseed meal |
| CN106819359A (en) * | 2016-12-28 | 2017-06-13 | 乐山恒峰华邦生物科技有限公司 | One kind fermentation rapeseed meal and its production method |
| CN109123093A (en) * | 2018-09-13 | 2019-01-04 | 圣百奥（山东）生物科技有限公司 | A kind of mixed fermentation miscellaneous dregs of rice and its production technology and application |
-
2021
- 2021-04-28 CN CN202110488825.2A patent/CN113229399B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS61166385A (en) * | 1985-01-18 | 1986-07-28 | Ajinomoto Co Inc | Treatment of rapeseed cake |
| CN102652529A (en) * | 2012-04-10 | 2012-09-05 | 湖北省农业科学院农产品加工与核农技术研究所 | Method for obtaining active polypeptide by carrying out multi-strain compound solid state fermentation on common rapeseed meal |
| CN102823726A (en) * | 2012-09-07 | 2012-12-19 | 北京大北农科技集团股份有限公司 | Biological fermentation method capable of improving protein content of cottonseed meal and removing toxicity |
| CN105494890A (en) * | 2014-09-26 | 2016-04-20 | 侯文燕 | Enzymolysis and fermentation combined method for improving feeding nutritional value of rapeseed meal |
| CN106819359A (en) * | 2016-12-28 | 2017-06-13 | 乐山恒峰华邦生物科技有限公司 | One kind fermentation rapeseed meal and its production method |
| CN109123093A (en) * | 2018-09-13 | 2019-01-04 | 圣百奥（山东）生物科技有限公司 | A kind of mixed fermentation miscellaneous dregs of rice and its production technology and application |
Non-Patent Citations (1)
| Title |
|---|
| 姜锡瑞等: "《生物发酵产业技术》", vol. 1, 北京：中国轻工业出版社, pages: 484 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114698726A (en) * | 2022-04-01 | 2022-07-05 | 播恩集团股份有限公司 | Low-toxin high-nutrition rapeseed meal, method for degrading toxins in rapeseed meal through segmented fermentation and application |
| CN114698726B (en) * | 2022-04-01 | 2022-11-25 | 播恩集团股份有限公司 | Low-toxin high-nutrition rapeseed meal, method for degrading toxins of rapeseed meal through segmented fermentation and application |
| CN115316491A (en) * | 2022-08-25 | 2022-11-11 | 山东泰山生力源集团股份有限公司 | Preparation method of feed additive, feed additive and feed |
| CN115316491B (en) * | 2022-08-25 | 2024-01-16 | 山东泰山生力源集团股份有限公司 | Preparation method of feed additive, feed additive and feed |
| CN115736092A (en) * | 2022-12-14 | 2023-03-07 | 山东泰山生力源集团股份有限公司 | Method for improving glutamine content in feed through solid state fermentation |
| CN115851465A (en) * | 2022-12-14 | 2023-03-28 | 湖北省生物农药工程研究中心 | Composite fermentation inoculant, method for preparing fermented rapeseed meal for feed and application of composite fermentation inoculant |
| CN115851465B (en) * | 2022-12-14 | 2024-01-30 | 湖北省生物农药工程研究中心 | Composite fermentation inoculant, method for preparing fermented rapeseed meal for feeding and application of composite fermentation inoculant |
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