CN116195679A - Method for improving quality of rapeseed meal through synergistic fermentation of bacterial enzymes - Google Patents
Method for improving quality of rapeseed meal through synergistic fermentation of bacterial enzymes Download PDFInfo
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- CN116195679A CN116195679A CN202310343072.5A CN202310343072A CN116195679A CN 116195679 A CN116195679 A CN 116195679A CN 202310343072 A CN202310343072 A CN 202310343072A CN 116195679 A CN116195679 A CN 116195679A
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- 238000000855 fermentation Methods 0.000 title claims abstract description 139
- 230000004151 fermentation Effects 0.000 title claims abstract description 139
- 235000019779 Rapeseed Meal Nutrition 0.000 title claims abstract description 118
- 239000004456 rapeseed meal Substances 0.000 title claims abstract description 118
- 238000000034 method Methods 0.000 title claims abstract description 59
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- 238000002791 soaking Methods 0.000 claims description 21
- 239000001963 growth medium Substances 0.000 claims description 20
- 238000010563 solid-state fermentation Methods 0.000 claims description 14
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- LRBQNJMCXXYXIU-QWKBTXIPSA-N gallotannic acid Chemical compound OC1=C(O)C(O)=CC(C(=O)OC=2C(=C(O)C=C(C=2)C(=O)OC[C@H]2[C@@H]([C@@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)O2)OC(=O)C=2C=C(OC(=O)C=3C=C(O)C(O)=C(O)C=3)C(O)=C(O)C=2)O)=C1 LRBQNJMCXXYXIU-QWKBTXIPSA-N 0.000 description 2
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- 239000003440 toxic substance Substances 0.000 description 2
- UMURLIQHQSKULR-UHFFFAOYSA-N 1,3-oxazolidine-2-thione Chemical compound S=C1NCCO1 UMURLIQHQSKULR-UHFFFAOYSA-N 0.000 description 1
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- LEVWYRKDKASIDU-QWWZWVQMSA-N D-cystine Chemical compound OC(=O)[C@H](N)CSSC[C@@H](N)C(O)=O LEVWYRKDKASIDU-QWWZWVQMSA-N 0.000 description 1
- 108010082495 Dietary Plant Proteins Proteins 0.000 description 1
- URXZXNYJPAJJOQ-UHFFFAOYSA-N Erucic acid Natural products CCCCCCC=CCCCCCCCCCCCC(O)=O URXZXNYJPAJJOQ-UHFFFAOYSA-N 0.000 description 1
- 235000019733 Fish meal Nutrition 0.000 description 1
- 238000007696 Kjeldahl method Methods 0.000 description 1
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 108010073771 Soybean Proteins Proteins 0.000 description 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 1
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 1
- 230000009471 action Effects 0.000 description 1
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- 229910052802 copper Inorganic materials 0.000 description 1
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- 238000000354 decomposition reaction Methods 0.000 description 1
- 230000000593 degrading effect Effects 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- DPUOLQHDNGRHBS-KTKRTIGZSA-N erucic acid Chemical compound CCCCCCCC\C=C/CCCCCCCCCCCC(O)=O DPUOLQHDNGRHBS-KTKRTIGZSA-N 0.000 description 1
- 239000004467 fishmeal Substances 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 230000002496 gastric effect Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 description 1
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- 210000004185 liver Anatomy 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
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- 230000004060 metabolic process Effects 0.000 description 1
- 229930182817 methionine Natural products 0.000 description 1
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- 150000002825 nitriles Chemical class 0.000 description 1
- PIBWKRNGBLPSSY-UHFFFAOYSA-L palladium(II) chloride Chemical compound Cl[Pd]Cl PIBWKRNGBLPSSY-UHFFFAOYSA-L 0.000 description 1
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- 238000011160 research Methods 0.000 description 1
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- 235000019710 soybean protein Nutrition 0.000 description 1
- 239000011593 sulfur Substances 0.000 description 1
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- 210000001685 thyroid gland Anatomy 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
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- 229910052725 zinc Inorganic materials 0.000 description 1
- 239000011701 zinc Substances 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
- 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
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/14—Pretreatment of feeding-stuffs with enzymes
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/20—Animal feeding-stuffs from material of animal origin
- A23K10/22—Animal feeding-stuffs from material of animal origin from fish
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/20—Animal feeding-stuffs from material of animal origin
- A23K10/26—Animal feeding-stuffs from material of animal origin from waste material, e.g. feathers, bones or skin
-
- 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
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
- A23L5/20—Removal of unwanted matter, e.g. deodorisation or detoxification
- A23L5/21—Removal of unwanted matter, e.g. deodorisation or detoxification by heating without chemical treatment, e.g. steam treatment, cooking
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
- A23L5/20—Removal of unwanted matter, e.g. deodorisation or detoxification
- A23L5/25—Removal of unwanted matter, e.g. deodorisation or detoxification using enzymes
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
- A23L5/20—Removal of unwanted matter, e.g. deodorisation or detoxification
- A23L5/28—Removal of unwanted matter, e.g. deodorisation or detoxification using microorganisms
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
- A23L5/30—Physical treatment, e.g. electrical or magnetic means, wave energy or irradiation
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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 relates to the technical field of microbial fermentation, and discloses a method for improving the quality of rapeseed meal by synergistic fermentation of bacterial enzymes. Pretreating rapeseed meal, carrying out solid fermentation by using an inert adsorption carrier, adding sterile water and an enzyme preparation into a fermentation product after fermentation, carrying out liquid enzymolysis, and drying the enzymolysis product. The method can reduce the anti-nutritional factors in the rapeseed meal and improve the content of small peptides in the rapeseed meal so as to improve the quality of the rapeseed meal.
Description
Technical Field
The invention relates to the technical field of microbial fermentation, in particular to a method for improving the quality of rapeseed meal by synergistic fermentation of bacterial enzymes.
Background
Rapeseed meal is a byproduct remaining after the oil is made from rapeseed. The rapeseed meal is a high-quality vegetable protein feed resource, the protein content of the rapeseed meal is 35% -42%, the amino acid composition is reasonable, and the rapeseed protein is full-value protein and almost has no limiting amino acid. Compared with soybean meal, the rapeseed cake (meal) has higher content of methionine, cystine and other sulfur-containing amino acids, the lysine content is only slightly lower than that of soybean protein, and meanwhile, the rapeseed meal contains rich mineral substances, wherein the content is the highest in common vegetable feed. Therefore, the rapeseed meal is a very valuable potential protein raw material of livestock and poultry feed.
However, the rapeseed meal contains various anti-nutritional factors such as thioglycoside, phytic acid, erucic acid, tannic acid and the like, so that the application of the rapeseed meal in the food and feed industries is limited. Wherein, thioglycoside is a main anti-nutritional factor, and after decomposition, oxazolidinethione, isothiocyanate, nitrile and other toxic substances are generated, which have toxic effects on animal thyroid, liver and kidney; the phytic acid can be combined with nutrient substances such as protein, copper, zinc, calcium and the like to form complex which is difficult to digest, so that the nutritive value of the rapeseed meal is reduced; on the other hand, because of the low small peptide content in the rapeseed meal protein, compared with the soybean meal and fish meal proteins, the rapeseed meal protein is more difficult to digest.
In order to improve the quality of the rapeseed meal, a great deal of researches have been carried out on the rapeseed meal by a plurality of scholars at home and abroad for reducing the content of anti-nutritional factors of thioglycoside and phytic acid in the rapeseed meal and improving the content of small peptide, and the early physical method and the chemical method can effectively reduce toxic substances, but have the problems of reagent residue, nutrition loss and the like. Although the enzymolysis method can improve the content of peptide substances in the rapeseed meal, partial bitter peptide exists in the product, so that the taste of the feed is affected, and the treatment cost is increased.
Disclosure of Invention
In view of the above, aiming at the technical problems of low nutritive value and high content of anti-nutritional factors of the existing rapeseed meal feed, a method for preparing the rapeseed meal feed by synergistic fermentation of bacterial enzymes is provided, wherein the content of anti-nutritional factors of thioglycoside and phytic acid in the rapeseed meal can be reduced, the content of small peptides in the rapeseed meal can be improved, and the quality of the rapeseed meal feed can be improved.
In order to achieve the above purpose, the invention adopts the following technical scheme:
a method for improving the quality of rapeseed dregs by the synergistic fermentation of bacterial enzymes comprises the steps of preprocessing the rapeseed dregs, carrying out solid state fermentation by using an inert adsorption carrier, adding sterile water and an enzyme preparation into a fermentation product after the fermentation is finished for liquid enzymolysis, and drying the enzymolysis product.
In a further technical scheme of the invention, the method specifically comprises the following steps:
(1) pretreatment of rapeseed meal: soaking rapeseed meal in water, homogenizing at high temperature and high pressure for 30min before soaking, standing and cooling to room temperature for use;
(2) solid state fermentation: mixing pretreated rapeseed meal and inert adsorption carrier, placing the mixture in a fermentation container, and fermenting, wherein low-intensity alternating magnetic field is adopted for assisting fermentation during fermentation;
(3) liquid enzymolysis: mixing the fermentation product with sterile water, adding neutral protease, performing constant temperature enzymolysis, and drying the enzymolysis product.
In the further technical scheme of the invention, in the rapeseed dreg pretreatment step, the rapeseed dreg is soaked in water for 4-6 hours, high-temperature and high-pressure homogenization treatment is carried out for 30 minutes before the soaking is finished, the pressure is controlled to be 30MPa-50MPa, the temperature is 121 ℃, the treatment time is 30 minutes, and after the treatment is finished, the rapeseed dreg is stood and cooled to room temperature for standby.
In a further technical scheme of the invention, the inert adsorption carrier is oyster shell powder with 200 meshes, and the preparation steps of the inert adsorption carrier are as follows: mixing 200 mesh oyster shell powder and microorganism strain fermentation medium, sterilizing at 121deg.C for 20-30min, and cooling the mixed solution.
In a further technical scheme of the invention, the microorganism strain is selected from single strain or combination of single strain in aspergillus niger, bacillus coagulans, bacillus subtilis, enterococcus faecalis and saccharomycetes.
In a further technical scheme of the invention, the weight ratio of the inert adsorption carrier to the culture medium (culture solution) containing the strain is 1:40.
In a further technical scheme of the invention, the microorganism strain is preferably a combination of a plurality of strains, and the weight of a culture medium (culture solution) containing the strains accounts for 1-40% of the weight (dry weight) of the rapeseed meal raw material, and the combination is respectively 1-10% of Aspergillus niger, 0.5-10% of Bacillus coagulans, 0.1-10% of enterococcus faecalis and 1-10% of microzyme.
In a further technical scheme of the invention, the low-intensity alternating magnetic field assists fermentation, and the specific operation is as follows: and (3) placing the fermentation tank in a magnetic field intensity of 140Gs for fermentation for 4 hours every 12 hours in the fermentation process.
In a further technical scheme of the invention, the addition proportion of the neutral protease is 0.1-0.5% of the mixed raw materials (dry weight).
In a further technical scheme of the invention, the fermentation temperature is controlled to be 45-50 ℃ and the fermentation time is 30-36h.
In a further technical scheme of the invention, the weight ratio of the rapeseed meal to the inert adsorption carrier is 50:1.
In a further technical scheme of the invention, the weight ratio of the fermentation product to the sterile water is 1:1.2-1.5.
In the further technical scheme of the invention, the constant-temperature enzymolysis temperature is controlled at 45 ℃, and the enzymolysis time is controlled at 30-36h.
According to the technical scheme, the invention discloses a method for improving the quality of rapeseed meal by synergistic fermentation of bacterial enzymes, and compared with the prior art, the technical scheme disclosed by the invention has the following technical effects:
the invention reduces the content of anti-nutritional factors in rapeseed meal and improves the content of small peptides in the rapeseed meal so as to improve the quality of the rapeseed meal for feeding.
The method does not affect other nutritional ingredients of the feed, does not reduce the nutritional value of the feed, is easier to be absorbed by animals, is rich in various probiotics, and is more beneficial to improving the gastrointestinal health of cultured animals.
According to the invention, the rapeseed meal is fermented by adopting a method of solid state fermentation and liquid state enzymolysis, and the rapeseed meal is fermented after treatment, so that the fermentation efficiency is higher, various enzyme active substances such as protease, cellulase and phytase are generated in the fermentation process by utilizing microorganism strains after the fermentation is finished, and the enzymolysis reaction is performed without adding excessive enzymes when the next enzymolysis is performed, so that the enzymolysis reaction efficiency is higher and the cost is lower. Therefore, the invention only adds neutral protease to carry out enzymolysis when carrying out enzymolysis reaction, and further can improve the content of small peptide in the rapeseed meal.
Compared with the existing technique for fermenting rapeseed meal by synergistic action of bacterial enzymes, the invention has the advantages that the pretreatment is carried out before the rapeseed meal is fermented, the pretreatment of the rapeseed meal by adopting a treatment mode of soaking and high temperature and high pressure can remove a part of anti-nutritional factors in the rapeseed meal before the fermentation, such as: phytic acid, tannic acid and the like can eliminate various anti-nutritional factors, and the cell walls of the rapeseed meal can be destroyed thoroughly in a pretreatment mode, so that the nutrients in the rapeseed meal are released more easily, and a foundation is laid for the next fermentation.
Compared with direct fermentation in the prior art of fermenting rapeseed meal by the synergistic action of bacterial enzymes, the method adopts low-intensity alternating magnetic field to assist fermentation when the rapeseed meal is fermented, thereby effectively improving the content of small peptides in the rapeseed meal and improving the quality of the rapeseed meal.
The oyster shell powder has high calcium content, is rich in various trace elements, can enrich the nutrition of the feed, and can play a role in promoting metabolism of livestock and poultry, the oyster shell powder has a large number of 2-10 mu m micropores, has strong adsorption capacity, can fully hold and load microorganism strains, provides a stable fermentation environment for microorganisms, shortens fermentation time, improves fermentation efficiency, and meanwhile, the fermented product can be directly subjected to enzymolysis, and the steps of removing inert adsorption carriers are performed differently, so that time and cost are saved.
Detailed Description
The embodiment of the invention discloses a method for improving the quality of rapeseed meal by bacterial-enzyme synergistic fermentation, which comprises the steps of firstly pretreating the rapeseed meal, then carrying out solid state fermentation by using an inert adsorption carrier, adding sterile water and an enzyme preparation into a fermentation product obtained after fermentation, carrying out liquid enzymolysis, and then drying the enzymolysis product, wherein the method specifically comprises the following steps:
(1) pretreatment of rapeseed meal: selecting raw materials meeting the standard requirements of GB/T23736-2009 rapeseed meal for feed, soaking the raw materials in water for 4-6 hours, carrying out high-temperature high-pressure homogenization treatment for 30 minutes before soaking is completed, controlling the pressure to be 30MPa-50MPa, the temperature to be 121 ℃ and the treatment time to be 30 minutes, and standing and cooling to room temperature for standby after the treatment is completed;
(2) solid state fermentation: mixing pretreated rapeseed meal and inert adsorption carrier according to a weight ratio of 50:1, and placing the mixture into a fermentation container for fermentation, wherein the fermentation container can adopt a false bottom ventilated fermentation tank or other fermentation equipment, such as a solid fermentation tank, the loading thickness is preferably 25-50 cm, low-intensity alternating magnetic field is adopted for auxiliary fermentation during fermentation, and the auxiliary fermentation specific operation is as follows: fermenting the fermentation tank in 140Gs magnetic field intensity for 4h at intervals of 12h at 45-50deg.C for 30-36h; under the action of high pressure and high temperature, the pretreated rapeseed meal contains a certain amount of water, so that the rapeseed meal does not need to be added with water for fermentation; the inert adsorption carrier is oyster shell powder, and the preparation steps of the inert adsorption carrier are as follows: mixing 200 mesh oyster shell powder and microorganism strain fermentation medium, sterilizing at 121deg.C for 20-30min, and cooling the mixed solution; the microorganism strain is selected from single strain or combination of Aspergillus niger, bacillus coagulans, bacillus subtilis, enterococcus faecalis and yeast; the strains are inoculated in respective culture mediums according to instructions in advance for culture, and when the strain and the culture mediums are used, the strains and the culture mediums are added into fermentation equipment together, so that the strain sources can be widely used; when the mixed strain is used, the weight of the culture medium (culture solution) containing the strain accounts for 1-40% of the weight (dry weight) of the rapeseed meal raw material, and the weight is respectively 1-10% of aspergillus niger, 0.5-10% of bacillus coagulans, 0.1-10% of enterococcus faecalis and 1-10% of saccharomycetes;
(3) liquid enzymolysis: fermenting to obtain fermentation product, mixing the fermentation product with sterile water at a weight ratio of 1:1.2-1.5, adding 0.1-0.5% neutral protease of mixed raw materials (dry weight) for constant temperature enzymolysis at 45 ℃, controlling the enzymolysis time to 30-36h, and drying the enzymolysis product.
The following description of the technical solutions in the embodiments of the present invention will be clear and complete, and it is obvious that the described embodiments are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention. The methods used in the examples are conventional methods unless otherwise specified.
Example 1
The method for improving the quality of the rapeseed meal by the synergistic fermentation of the bacterial enzymes of the invention is used for preparing the nontoxic high-nutrition rapeseed meal animal feeding protein, and specifically comprises the following processes and parameters by referring to the method:
(1) pretreatment of rapeseed meal: soaking rapeseed meal in water for 4 hours, carrying out high-temperature high-pressure homogenization treatment for 30 minutes before soaking, controlling the pressure at 50MPa and the temperature at 121 ℃ for 30 minutes, standing and cooling to room temperature for standby after the treatment is completed;
(2) solid state fermentation: uniformly mixing oyster shell powder of 200 meshes with a microbial strain fermentation culture medium according to a weight ratio of 1:40, sterilizing at 121 ℃ for 20min, mixing the cooled mixed solution and pretreated rapeseed meal according to a weight ratio of 1:50, placing the mixed solution into a fermentation container, fermenting, placing a fermentation tank into a magnetic field intensity of 140Gs for fermenting for 4h at intervals of 12h, controlling the fermentation temperature at 45 ℃ for 30h, and obtaining a fermentation product after fermentation, wherein the microbial strain comprises 1% of aspergillus niger, 0.5% of bacillus coagulans, 0.1% of enterococcus faecalis and 1% of saccharomycetes according to the weight of the culture medium (culture solution) containing the strain accounting for the weight (dry weight) of the rapeseed meal raw material;
(3) liquid enzymolysis: mixing the fermentation product with sterile water according to a weight ratio of 1:1.2, adding neutral protease accounting for 0.1% of the dry weight of the mixed raw material, performing constant temperature enzymolysis at 45 ℃ for 30 hours, and drying the enzymolysis product.
Example 2
The method for improving the quality of the rapeseed meal by the synergistic fermentation of the bacterial enzymes of the invention is used for preparing the nontoxic high-nutrition rapeseed meal animal feeding protein, and specifically comprises the following processes and parameters by referring to the method:
(1) pretreatment of rapeseed meal: soaking rapeseed meal in water for 6 hours, carrying out high-temperature high-pressure homogenization treatment for 30 minutes before soaking, controlling the pressure at 30MPa and the temperature at 121 ℃ for 30 minutes, standing and cooling to room temperature for standby after the treatment is completed;
(2) solid state fermentation: uniformly mixing oyster shell powder of 200 meshes with a microbial strain fermentation culture medium according to a weight ratio of 1:40, sterilizing at 121 ℃ for 30min, mixing the cooled mixed solution and pretreated rapeseed meal according to a weight ratio of 1:50, placing the mixed solution into a fermentation container, fermenting, placing a fermentation tank into a magnetic field intensity of 140Gs for fermenting for 4h at intervals of 12h, controlling the fermentation temperature at 50 ℃ and the fermentation time at 36h, and obtaining a fermentation product after fermentation, wherein the microbial strain accounts for 40% of Aspergillus niger, 10% of bacillus coagulans, 10% of enterococcus faecalis and 10% of saccharomycetes according to the weight (dry weight) of the culture medium (culture solution) containing the strain;
(3) liquid enzymolysis: mixing the fermentation product with sterile water according to a weight ratio of 1:1.5, adding neutral protease accounting for 0.5% of the dry weight of the mixed raw material, performing constant temperature enzymolysis at 45 ℃ for 36h, and drying the enzymolysis product.
Example 3
The method for improving the quality of the rapeseed meal by the synergistic fermentation of the bacterial enzymes of the invention is used for preparing the nontoxic high-nutrition rapeseed meal animal feeding protein, and specifically comprises the following processes and parameters by referring to the method:
(1) pretreatment of rapeseed meal: soaking rapeseed meal in water for 5 hours, carrying out high-temperature high-pressure homogenization treatment for 30 minutes before soaking, controlling the pressure at 40MPa and the temperature at 121 ℃ for 30 minutes, standing and cooling to room temperature for standby after the treatment is completed;
(2) solid state fermentation: uniformly mixing oyster shell powder of 200 meshes with a microbial strain fermentation culture medium according to a weight ratio of 1:40, sterilizing at 121 ℃ for 25min, mixing the cooled mixed solution and pretreated rapeseed meal according to a weight ratio of 1:50, placing the mixture into a fermentation container, fermenting, placing a fermentation tank into a magnetic field intensity of 140Gs for fermenting for 4h at intervals of 12h, controlling the fermentation temperature at 50 ℃ and the fermentation time at 32h, and obtaining a fermentation product after fermentation, wherein the microbial strain comprises 24.6 percent of Aspergillus niger, 5.5 percent of bacillus coagulans, 4.5 percent of enterococcus faecalis and 8.4 percent of saccharomycetes according to the weight of the culture medium (culture solution) containing the microbial strain accounting for the weight (dry weight) of the rapeseed meal raw material;
(3) liquid enzymolysis: mixing the fermentation product with sterile water according to a weight ratio of 1:1.3, adding neutral protease accounting for 0.3% of the dry weight of the mixed raw material, performing constant-temperature enzymolysis at 45 ℃ for 32 hours, and drying the enzymolysis product.
Comparative example 1
The method for improving the quality of the rapeseed meal by the synergistic fermentation of the bacterial enzymes of the invention is used for preparing the nontoxic high-nutrition rapeseed meal animal feeding protein, and the specific process and parameters are the same as those of the embodiment 3 by referring to the method, and the difference is that: the method only comprises the following steps:
(1) solid state fermentation: uniformly mixing oyster shell powder of 200 meshes with a microbial strain fermentation culture medium according to a weight ratio of 1:40, sterilizing at 121 ℃ for 25min, mixing the mixed liquor with rapeseed meal according to a weight ratio of 1:50, placing the mixed liquor in a fermentation container, fermenting, placing a fermentation tank in a magnetic field intensity of 140Gs for fermenting for 4h every 12h, controlling the fermentation temperature at 45 ℃ and the fermentation time at 32h, and obtaining a fermentation product after fermentation, wherein the microbial strain comprises 24.6% of Aspergillus niger, 5.5% of bacillus coagulans, 4.5% of enterococcus faecalis and 8.4% of saccharomycetes according to the weight of the culture medium (culture solution) containing the strain accounting for the weight (dry weight) of the rapeseed meal raw material;
(2) liquid enzymolysis: mixing the fermentation product with sterile water according to a weight ratio of 1:1.3, adding neutral protease accounting for 0.3% of the dry weight of the mixed raw material, performing constant-temperature enzymolysis at 45 ℃ for 32 hours, and drying the enzymolysis product.
Comparative example 2
The method for improving the quality of the rapeseed meal by the synergistic fermentation of the bacterial enzymes of the invention is used for preparing the nontoxic high-nutrition rapeseed meal animal feeding protein, and the specific process and parameters are the same as those of the embodiment 3 by referring to the method, and the difference is that: the method only comprises the following steps:
(1) pretreatment of rapeseed meal: soaking rapeseed meal in water for 5 hours, carrying out high-temperature high-pressure homogenization treatment for 30 minutes before soaking, controlling the pressure at 40MPa and the temperature at 121 ℃ for 30 minutes, standing and cooling to room temperature for standby after the treatment is completed;
(2) solid state fermentation: fully and uniformly mixing 200-mesh oyster shell powder and a microbial strain fermentation culture medium according to a weight ratio of 1:40, sterilizing at 121 ℃ for 25min, mixing the cooled mixed solution and pretreated rapeseed meal according to a weight ratio of 1:50, placing the mixed solution into a fermentation container, fermenting at a temperature of 45 ℃ for 32h, and obtaining a fermentation product after fermentation, wherein the microbial strain comprises 24.6% of aspergillus niger, 5.5% of bacillus coagulans, 4.5% of enterococcus faecalis and 8.4% of saccharomycetes according to the weight percentage of the culture medium (culture medium) containing the strain in the rapeseed meal raw material (dry weight);
(3) liquid enzymolysis: mixing the fermentation product with sterile water according to a weight ratio of 1:1.3, adding neutral protease accounting for 0.3% of the dry weight of the mixed raw material, performing constant-temperature enzymolysis at 45 ℃ for 32 hours, and drying the enzymolysis product.
Comparative example 3
The method for improving the quality of the rapeseed meal by the synergistic fermentation of the bacterial enzymes of the invention is used for preparing the nontoxic high-nutrition rapeseed meal animal feeding protein, and the specific process and parameters are the same as those of the embodiment 3 by referring to the method, and the difference is that: the method only comprises the following steps:
(1) pretreatment of rapeseed meal: soaking rapeseed meal in water for 5 hours, carrying out high-temperature high-pressure homogenization treatment for 30 minutes before soaking, controlling the pressure at 40MPa and the temperature at 121 ℃ for 30 minutes, standing and cooling to room temperature for standby after the treatment is completed;
(2) solid state fermentation: mixing pretreated rapeseed meal with a microbial strain fermentation culture medium according to a weight ratio of 1:50, placing the mixture into a fermentation container for fermentation, placing a fermentation tank into a magnetic field intensity of 140Gs for fermentation for 4 hours at intervals of 12 hours, controlling the fermentation temperature at 50 ℃ and the fermentation time at 32 hours, and obtaining a fermentation product after fermentation, wherein the microbial strain comprises 24.6% of Aspergillus niger, 5.5% of bacillus coagulans, 4.5% of enterococcus faecalis and 8.4% of saccharomycetes according to the weight of a culture medium (culture solution) containing the microbial strain accounting for the weight (dry weight) of the rapeseed meal raw material;
(3) liquid enzymolysis: mixing the fermentation product with sterile water according to a weight ratio of 1:1.3, adding neutral protease accounting for 0.3% of the dry weight of the mixed raw material, performing constant-temperature enzymolysis at 45 ℃ for 32 hours, and drying the enzymolysis product.
Comparative example 4
The method for improving the quality of the rapeseed meal by the synergistic fermentation of the bacterial enzymes of the invention is used for preparing the nontoxic high-nutrition rapeseed meal animal feeding protein, and the specific process and parameters are the same as those of the embodiment 3 by referring to the method, and the difference is that: the method only comprises the following steps:
(1) pretreatment of rapeseed meal: soaking rapeseed meal in water for 5 hours, carrying out high-temperature high-pressure homogenization treatment for 30 minutes before soaking, controlling the pressure at 40MPa and the temperature at 121 ℃ for 30 minutes, standing and cooling to room temperature for standby after the treatment is completed;
(1) solid state fermentation: fully and uniformly mixing 200-mesh oyster shell powder and a microbial strain fermentation culture medium according to a weight ratio of 1:40, sterilizing at 121 ℃ for 25min, mixing the cooled mixed solution and pretreated rapeseed meal according to a weight ratio of 1:50, placing the mixed solution into a fermentation container, fermenting, placing the fermentation tank into a magnetic field intensity of 140Gs for fermentation for 4h at intervals of 12h, controlling the fermentation temperature at 45 ℃ and the fermentation time at 32h, and obtaining a fermentation product after fermentation; the microbial strains respectively comprise 24.6% of Aspergillus niger, 5.5% of Bacillus coagulans, 4.5% of enterococcus faecalis and 8.4% of saccharomycetes according to the weight of a culture medium (culture solution) containing the strains accounting for the weight (dry weight) of the rapeseed meal raw material.
Product inspection
Physicochemical property measurements were performed on the rapeseed meal prepared in examples 1 to 3 and comparative examples 1 to 4 and the rapeseed meal without any treatment (blank).
The method comprises the following steps of measuring thioglycoside according to a palladium chloride method, and specifically referring to the following documents:
wu Zhengke, liu Guohua, cai Huiyi, etc. screening of thioglycoside detoxified strains and the effect of fermenting rapeseed meal [ J ]. Animal nutrition report, 2018, 30 (1): 313-320.
The method for measuring the phytic acid adopts a ferric trichloride colorimetric method.
The small peptides were determined by reference to the following:
SHI C，ZHANG Y，LU Z，et al.Solid-state fermentationofcorn-soybean meal mixed feed with Bacillus subtilis andEnterococcus faecium for degrading antinutritional factorsand enhancing nutritional value[J].Journal ofAnimal Science and Biotechnology，2017，8(4):50.
the protein content is measured according to the Kjeldahl method.
The measurement results are shown in Table 1
TABLE 1
In the present specification, each embodiment is described in a progressive manner, and each embodiment is mainly described in a different point from other embodiments, and identical and similar parts between the embodiments are all enough to refer to each other. The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention.
Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (10)
1. A method for improving the quality of rapeseed meal by bacterial enzyme synergistic fermentation comprises the steps of firstly, pretreating the rapeseed meal, then carrying out solid state fermentation by using an inert adsorption carrier, obtaining a fermentation product after fermentation, adding sterile water and an enzyme preparation for liquid enzymolysis, and then drying the enzymolysis product; the method is characterized in that: specifically, the method comprises the following steps:
(1) pretreatment of rapeseed meal: soaking rapeseed meal in water, carrying out high-temperature high-pressure homogenization treatment before soaking, and standing and cooling to room temperature for standby after the treatment is finished;
(2) solid state fermentation: mixing the pretreated rapeseed meal and an inert adsorption carrier, and placing the mixture into a fermentation container for fermentation, wherein low-intensity alternating magnetic field is adopted for auxiliary fermentation during fermentation;
(3) liquid enzymolysis: adding sterile water and enzyme preparation into the fermentation product, performing constant-temperature enzymolysis, and drying the enzymolysis product.
2. The method for improving the quality of rapeseed meal by the collaborative fermentation of bacterial enzymes according to claim 1, which is characterized in that: in the pretreatment step of the rapeseed meal, the rapeseed meal is soaked in water for 4-6 hours, high-temperature and high-pressure homogenization treatment is carried out for 30min before the soaking is finished, the pressure is controlled at 30-50 MPa, the temperature is 121 ℃, the treatment time is 30min, and the rapeseed meal is kept stand and cooled to room temperature for standby after the treatment is finished.
3. The method for improving the quality of rapeseed meal by the collaborative fermentation of bacterial enzymes according to claim 1, which is characterized in that: the preparation method of the specific inert adsorption carrier comprises the following steps: mixing 200 mesh oyster shell powder and microorganism strain fermentation medium, sterilizing at 121deg.C for 20-30min, and cooling the mixed solution; the microorganism strain is selected from single strain or combination of Aspergillus niger, bacillus coagulans, bacillus subtilis, enterococcus faecalis and saccharomycetes; the enzyme preparation is neutral protease.
4. The method for improving the quality of rapeseed meal by synergistic fermentation of bacterial enzymes according to claim 3, wherein the method comprises the following steps: the weight ratio of the inert adsorption carrier to the culture medium (culture solution) containing the strain is 1:40.
5. The method for improving the quality of rapeseed meal by the collaborative fermentation of bacterial enzymes according to claim 1, which is characterized in that: the microbial strain is preferably a combination of a plurality of strains, and the weight of a culture medium (culture solution) containing the strains accounts for 1-40% of the weight (dry weight) of the rapeseed meal raw material, and the microbial strain comprises 0.5-10% of aspergillus niger, 0.1-10% of bacillus coagulans, 0.1-10% of enterococcus faecalis and 1-10% of saccharomycetes.
6. The method for improving the quality of rapeseed meal by the collaborative fermentation of bacterial enzymes according to claim 1, which is characterized in that: the low-intensity alternating magnetic field assists fermentation, and the specific operation is as follows: and (3) fermenting the fermentation tank in a magnetic field intensity of 140Gs for 4 hours every 12 hours in the fermentation process, wherein the fermentation temperature is controlled at 45-50 ℃ and the fermentation time is 30-36 hours.
7. The method for improving the quality of rapeseed meal by the collaborative fermentation of bacterial enzymes according to claim 1, which is characterized in that: the addition proportion of the enzyme preparation is 0.1-0.5% of the mixed raw materials (dry weight).
8. The method for improving the quality of rapeseed meal by the collaborative fermentation of bacterial enzymes according to claim 1, which is characterized in that: the weight ratio of the rapeseed meal to the inert adsorption carrier is 50:1.
9. The method for improving the quality of rapeseed meal by the collaborative fermentation of bacterial enzymes according to claim 1, which is characterized in that: the weight ratio of the fermentation product to the sterile water is 1:1.2-1.5.
10. The method for improving the quality of rapeseed meal by the collaborative fermentation of bacterial enzymes according to claim 1, which is characterized in that: the constant temperature enzymolysis temperature is controlled at 45 ℃, and the enzymolysis time is controlled at 30-36h.
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