CN106962594B - Selenium-rich fermented soybean meal, preparation method and application - Google Patents
Selenium-rich fermented soybean meal, preparation method and application Download PDFInfo
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- 229910052711 selenium Inorganic materials 0.000 title claims abstract description 82
- 239000011669 selenium Substances 0.000 title claims abstract description 82
- BUGBHKTXTAQXES-UHFFFAOYSA-N Selenium Chemical compound [Se] BUGBHKTXTAQXES-UHFFFAOYSA-N 0.000 title claims abstract description 80
- 235000019764 Soybean Meal Nutrition 0.000 title claims abstract description 57
- 239000004455 soybean meal Substances 0.000 title claims abstract description 57
- 238000002360 preparation method Methods 0.000 title claims abstract description 34
- 238000000855 fermentation Methods 0.000 claims abstract description 47
- 230000004151 fermentation Effects 0.000 claims abstract description 47
- 239000001963 growth medium Substances 0.000 claims abstract description 44
- 244000046052 Phaseolus vulgaris Species 0.000 claims abstract description 34
- 235000010627 Phaseolus vulgaris Nutrition 0.000 claims abstract description 34
- 238000011218 seed culture Methods 0.000 claims abstract description 30
- 239000006041 probiotic Substances 0.000 claims abstract description 28
- 235000018291 probiotics Nutrition 0.000 claims abstract description 28
- 230000000529 probiotic effect Effects 0.000 claims abstract description 25
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims abstract description 21
- 230000000694 effects Effects 0.000 claims abstract description 17
- 241001465754 Metazoa Species 0.000 claims abstract description 14
- 238000000034 method Methods 0.000 claims abstract description 13
- 150000001875 compounds Chemical class 0.000 claims abstract description 9
- 238000009395 breeding Methods 0.000 claims abstract description 7
- 230000001488 breeding effect Effects 0.000 claims abstract description 7
- 229940091258 selenium supplement Drugs 0.000 claims description 75
- 230000001580 bacterial effect Effects 0.000 claims description 42
- 239000007788 liquid Substances 0.000 claims description 36
- BVTBRVFYZUCAKH-UHFFFAOYSA-L disodium selenite Chemical compound [Na+].[Na+].[O-][Se]([O-])=O BVTBRVFYZUCAKH-UHFFFAOYSA-L 0.000 claims description 32
- 239000011781 sodium selenite Substances 0.000 claims description 32
- 229960001471 sodium selenite Drugs 0.000 claims description 32
- 235000015921 sodium selenite Nutrition 0.000 claims description 32
- 241000235646 Cyberlindnera jadinii Species 0.000 claims description 26
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 23
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 22
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 22
- 239000001888 Peptone Substances 0.000 claims description 16
- 108010080698 Peptones Proteins 0.000 claims description 16
- 229940041514 candida albicans extract Drugs 0.000 claims description 16
- 235000019319 peptone Nutrition 0.000 claims description 16
- 239000012138 yeast extract Substances 0.000 claims description 16
- 239000002131 composite material Substances 0.000 claims description 15
- 241000194108 Bacillus licheniformis Species 0.000 claims description 14
- 241000191996 Pediococcus pentosaceus Species 0.000 claims description 14
- 244000057717 Streptococcus lactis Species 0.000 claims description 14
- 101100273269 Thermus thermophilus (strain ATCC 27634 / DSM 579 / HB8) cse3 gene Proteins 0.000 claims description 13
- 101150106467 cas6 gene Proteins 0.000 claims description 13
- 238000011081 inoculation Methods 0.000 claims description 11
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 11
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 11
- 239000002609 medium Substances 0.000 claims description 11
- 238000002156 mixing Methods 0.000 claims description 11
- 239000011780 sodium chloride Substances 0.000 claims description 11
- 238000002703 mutagenesis Methods 0.000 claims description 10
- 231100000350 mutagenesis Toxicity 0.000 claims description 10
- 239000000725 suspension Substances 0.000 claims description 9
- 235000019750 Crude protein Nutrition 0.000 claims description 8
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 8
- 239000008103 glucose Substances 0.000 claims description 8
- 229940099596 manganese sulfate Drugs 0.000 claims description 8
- 239000011702 manganese sulphate Substances 0.000 claims description 8
- 235000007079 manganese sulphate Nutrition 0.000 claims description 8
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 8
- 238000012216 screening Methods 0.000 claims description 8
- 238000003756 stirring Methods 0.000 claims description 8
- 230000001502 supplementing effect Effects 0.000 claims description 7
- 235000014897 Streptococcus lactis Nutrition 0.000 claims description 6
- 238000012258 culturing Methods 0.000 claims description 6
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 6
- PWKSKIMOESPYIA-UHFFFAOYSA-N 2-acetamido-3-sulfanylpropanoic acid Chemical compound CC(=O)NC(CS)C(O)=O PWKSKIMOESPYIA-UHFFFAOYSA-N 0.000 claims description 5
- 229920001817 Agar Polymers 0.000 claims description 5
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 5
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 claims description 5
- 239000008272 agar Substances 0.000 claims description 5
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 5
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 5
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 5
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 5
- 239000001110 calcium chloride Substances 0.000 claims description 5
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 5
- 229910000365 copper sulfate Inorganic materials 0.000 claims description 5
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 claims description 5
- 239000012153 distilled water Substances 0.000 claims description 5
- 238000001035 drying Methods 0.000 claims description 5
- 210000003746 feather Anatomy 0.000 claims description 5
- 239000011790 ferrous sulphate Substances 0.000 claims description 5
- 235000003891 ferrous sulphate Nutrition 0.000 claims description 5
- 239000012737 fresh medium Substances 0.000 claims description 5
- 235000013922 glutamic acid Nutrition 0.000 claims description 5
- 239000004220 glutamic acid Substances 0.000 claims description 5
- 238000005468 ion implantation Methods 0.000 claims description 5
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 claims description 5
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims description 5
- 235000012054 meals Nutrition 0.000 claims description 5
- 235000013379 molasses Nutrition 0.000 claims description 5
- 239000001301 oxygen Substances 0.000 claims description 5
- 229910052760 oxygen Inorganic materials 0.000 claims description 5
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 claims description 5
- 229910000368 zinc sulfate Inorganic materials 0.000 claims description 5
- 229960001763 zinc sulfate Drugs 0.000 claims description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 4
- 238000009630 liquid culture Methods 0.000 claims description 4
- 239000002028 Biomass Substances 0.000 claims description 3
- 102000010911 Enzyme Precursors Human genes 0.000 claims description 3
- 108010062466 Enzyme Precursors Proteins 0.000 claims description 3
- 239000006285 cell suspension Substances 0.000 claims description 3
- 239000011248 coating agent Substances 0.000 claims description 3
- 238000000576 coating method Methods 0.000 claims description 3
- 238000007865 diluting Methods 0.000 claims description 3
- 238000002513 implantation Methods 0.000 claims description 3
- 239000012528 membrane Substances 0.000 claims description 3
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 3
- 239000008223 sterile water Substances 0.000 claims description 3
- 238000005406 washing Methods 0.000 claims description 3
- 241001052560 Thallis Species 0.000 claims description 2
- 238000007605 air drying Methods 0.000 claims description 2
- 229910052757 nitrogen Inorganic materials 0.000 claims description 2
- 235000015097 nutrients Nutrition 0.000 abstract description 8
- 235000016709 nutrition Nutrition 0.000 abstract description 6
- 239000000126 substance Substances 0.000 abstract description 5
- 150000001413 amino acids Chemical class 0.000 abstract description 3
- 230000036737 immune function Effects 0.000 abstract description 3
- 244000005700 microbiome Species 0.000 abstract description 3
- 241000894006 Bacteria Species 0.000 abstract description 2
- 238000010521 absorption reaction Methods 0.000 abstract description 2
- 230000009286 beneficial effect Effects 0.000 abstract description 2
- 230000006378 damage Effects 0.000 abstract description 2
- 230000029087 digestion Effects 0.000 abstract description 2
- 239000002207 metabolite Substances 0.000 abstract description 2
- 230000035764 nutrition Effects 0.000 abstract description 2
- 229940082569 selenite Drugs 0.000 abstract description 2
- MCAHWIHFGHIESP-UHFFFAOYSA-L selenite(2-) Chemical compound [O-][Se]([O-])=O MCAHWIHFGHIESP-UHFFFAOYSA-L 0.000 abstract description 2
- 238000001514 detection method Methods 0.000 description 9
- 239000002054 inoculum Substances 0.000 description 9
- 239000000843 powder Substances 0.000 description 6
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- 239000000047 product Substances 0.000 description 5
- 241000194036 Lactococcus Species 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 3
- 229930006000 Sucrose Natural products 0.000 description 3
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 3
- 235000015278 beef Nutrition 0.000 description 3
- 230000003115 biocidal effect Effects 0.000 description 3
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 3
- 244000144972 livestock Species 0.000 description 3
- 238000011068 loading method Methods 0.000 description 3
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 3
- 235000019796 monopotassium phosphate Nutrition 0.000 description 3
- 239000006872 mrs medium Substances 0.000 description 3
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 3
- 229920000136 polysorbate Polymers 0.000 description 3
- 235000018102 proteins Nutrition 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 239000008213 purified water Substances 0.000 description 3
- 239000001632 sodium acetate Substances 0.000 description 3
- 235000017281 sodium acetate Nutrition 0.000 description 3
- 239000005720 sucrose Substances 0.000 description 3
- 239000008399 tap water Substances 0.000 description 3
- 235000020679 tap water Nutrition 0.000 description 3
- YWYZEGXAUVWDED-UHFFFAOYSA-N triammonium citrate Chemical compound [NH4+].[NH4+].[NH4+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O YWYZEGXAUVWDED-UHFFFAOYSA-N 0.000 description 3
- 235000011046 triammonium citrate Nutrition 0.000 description 3
- 239000001393 triammonium citrate Substances 0.000 description 3
- 239000012137 tryptone Substances 0.000 description 3
- 206010012735 Diarrhoea Diseases 0.000 description 2
- 241000282887 Suidae Species 0.000 description 2
- 235000001014 amino acid Nutrition 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 235000019621 digestibility Nutrition 0.000 description 2
- 238000003912 environmental pollution Methods 0.000 description 2
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- 208000037817 intestinal injury Diseases 0.000 description 2
- 244000144977 poultry Species 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- PMYDPQQPEAYXKD-UHFFFAOYSA-N 3-hydroxy-n-naphthalen-2-ylnaphthalene-2-carboxamide Chemical compound C1=CC=CC2=CC(NC(=O)C3=CC4=CC=CC=C4C=C3O)=CC=C21 PMYDPQQPEAYXKD-UHFFFAOYSA-N 0.000 description 1
- KJDSORYAHBAGPP-UHFFFAOYSA-N 4-(3,4-diaminophenyl)benzene-1,2-diamine;hydron;tetrachloride Chemical compound Cl.Cl.Cl.Cl.C1=C(N)C(N)=CC=C1C1=CC=C(N)C(N)=C1 KJDSORYAHBAGPP-UHFFFAOYSA-N 0.000 description 1
- 208000035240 Disease Resistance Diseases 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 108010064851 Plant Proteins Proteins 0.000 description 1
- 206010039921 Selenium deficiency Diseases 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 238000013329 compounding Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 231100000086 high toxicity Toxicity 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
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- 235000012424 soybean oil Nutrition 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/21—Streptococcus, lactococcus
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/41—Pediococcus
- A23V2400/427—Pentosaceus
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
Landscapes
- Fodder In General (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses selenium-rich fermented soybean meal, a preparation method and application, and the method comprises the following steps: the method comprises the steps of selenium-enriched strain breeding, seed culture solution preparation, fermentation culture, probiotic single strain fermentation, probiotic compound bacteria solution preparation, fresh culture medium addition and high-activity selenium-enriched bean pulp preparation. The invention adopts selenium-enriched yeast and probiotics to ferment the soybean meal together to obtain the fermented soybean meal with high organic selenium content and rich amino acid content. On one hand, the pollution and the damage to the environment and animal bodies caused by directly adding selenite substances into the feed are overcome, on the other hand, the selenium content in the bean pulp can be greatly improved, the bean pulp is rich in biological organic selenium, contains a large amount of beneficial microorganisms and fermentation metabolites, is richer in nutrition, the quality of the bean pulp is obviously improved, the digestion and absorption of the animal to the nutrient substances can be promoted, the immune function of the organism is enhanced, the high-selenium-rich multifunctional bean pulp is manufactured, and the nutritional value and the efficacy of the bean pulp are improved.
Description
Technical Field
The invention relates to fermented soybean meal, in particular to selenium-rich fermented soybean meal, a preparation method and application.
Background
Selenium is a multifunctional nutrient element necessary for maintaining normal life activities of organisms, and the selenium deficiency of animals mainly shows white muscle disease, slow growth and the like. The selenium-rich feed can obviously improve the growth performance of animals, enhance the reproductive performance of the animals, improve the immune function of the animals and improve the survival rate and disease resistance of young animals. At present, the main selenium supplement form in animal feed is sodium selenite, and the sodium selenite has low biological effectiveness, high toxicity and easy environmental pollution, so the use amount of the sodium selenite is strictly limited, for example, Sweden limits the use of the sodium selenite in milk pig feed, and Japan prohibits the addition of the sodium selenite in animal feed. Compared with sodium selenite, the organic selenium produced by using microorganisms has higher biological safety and compatibility and high biological utilization efficiency, and becomes the mainstream development trend of the current selenium supplement industry for livestock.
The soybean meal is a byproduct obtained by extracting soybean oil from soybeans, wherein the content of crude protein is high, the composition of amino acid is balanced, and the soybean meal is the most important plant protein raw material for livestock and poultry, and the dosage of the soybean meal accounts for more than 60% of the dosage of protein feed for livestock and poultry. After the soybean meal is fermented, the protein quality is remarkably improved, the fermented soybean meal is used for replacing the soybean meal in daily ration, the growth performance and nutrient substance digestibility of weaned piglets can be improved, intestinal injury is reduced, and diarrhea is relieved. At present, most of researches related to fermented soybean meal stay on improving the protein quality of the soybean meal, and few reports about the research of selenium-rich fermented soybean meal are reported, so that the further improvement of the nutritional value of the soybean meal is limited. Therefore, the selenium-rich fermented soybean meal is urgently needed to be developed, so that the nutritional value of the soybean meal is further improved, the application of selenium additives such as sodium selenite and the like in cultivation is reduced, the industrial level of feed is improved, and the healthy development of the animal husbandry is promoted.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides selenium-enriched fermented soybean meal, a preparation method and application thereof, wherein the soybean meal is converted into multifunctional soybean meal rich in small peptides and organic selenium nutrient elements.
The invention is realized by the following technical scheme: the invention relates to a preparation method of selenium-rich fermented soybean meal, which comprises the following steps:
(1) selenium-enriched strain breeding
Performing nitrogen ion implantation mutagenesis on Candida utilis, and performing multiple mutagenesis screening to obtain a yeast mutant strain tolerant to high-concentration sodium selenite, namely C.utilis cas 6;
(2) preparation of seed culture solution
Inoculating a candida utilis mutant strain C.utilis cas6 into a slant culture medium to obtain a slant strain, and then inoculating the slant strain into a seed culture medium to obtain a seed culture solution;
(3) fermentation culture
Inoculating the seed culture solution into a fermentation culture medium according to the inoculation amount of 8-12% of the volume percentage, supplementing sodium selenite with the final concentration of 150-200 mg/L into a fermentation product, and continuing fermentation culture for 20-24 hours to obtain a zymophyte solution;
(4) probiotic single strain fermentation
Separately fermenting three strains of Bacillus licheniformis Lincheniforms cas58, pediococcus pentosaceus cas138 and Streptococcus lactis cas168 to obtain respective probiotic zymocyte liquid;
(5) preparation of probiotic composite bacterial liquid
Fermenting bacterial liquids of four strains of selenium-enriched yeast C.utilis cas6, Bacillus licheniformis B.Lincheniforms cas58, Pediococcus pentosaceus P.penttosaceus cas138 and Streptococcus lactis S.lactis cas168 are mixed according to the ratio of 3-5: 2-3: 2-3: 1-2, and obtaining the probiotic composite bacterial liquid, wherein the effective viable count is 8.0 multiplied by 109～1.3×1010cfu/ml；
(6) Fresh media addition
Adding a fresh culture medium-cane molasses into the composite bacterial liquid according to the mass concentration of 2-3%;
(7) preparation of high-activity selenium-rich bean pulp
Mixing bean pulp, feather meal, bran and water according to a weight ratio of 10: 2-3: 1.5-2: and 4-5, adding the probiotic compound bacterial liquid prepared in the step (6) according to the weight ratio of 2-3 per mill after mixing, uniformly mixing and stirring, fermenting for 36-48 hours at the constant temperature of 36-42 ℃, and drying to obtain the selenium-enriched soybean meal.
The step (1) comprises the following steps:
(11) culturing Candida utilis on a YEPD culture plate for 48-72h, picking a single colony to a culture bottle containing a YEPD liquid culture medium, culturing at 180-200 rpm for 48-72h by shaking, centrifuging and washing a bacterial liquid to prepare a single-cell suspension, and ensuring the cell concentration to be 108～109Per mL;
(12) uniformly coating the bacterial suspension in a sterile culture dish, and air-drying to prepare a bacterial membrane;
(13) in N+Performing ion implantation in an implanter with energy of 14-18 KeV;
(14) 6 gradient doses per strain, two plates per dose, N+The implantation doses were 4.0X 10 respectively14、6.0×1014、8.0×1014、1.0×1015、1.2×1015、1.4×1015ion/cm2And making a vacuum control flat plate;
(15) after mutagenesis is finished, adding sterile water to wash the thalli to prepare bacterial suspension;
(16) diluting the bacterial suspension 104～108The yeast mutant strain which is tolerant to the high-concentration sodium selenite is obtained by multiple mutagenesis and screening after being coated on a screening culture medium containing the sodium selenite in times, and the fermentation biomass of the yeast mutant strain is improved by more than 2 times compared with the original strain under the same condition, so that the yeast mutant strain is named as C.utilis cas 6.
In the step (2), the Candida utilis mutant strain C.utilis cas6 is inoculated in a slant culture medium at 25-30 ℃ and cultured for 24-32 h to obtain a slant strain, then the slant strain is inoculated in a seed culture medium at 25-30 ℃ and 180-200 r/min, and shaking culture is carried out for 24-36 h to obtain a seed culture solution.
The slant culture medium comprises the following components: 15-20 g/L of glucose, 15-20 g/L of peptone, 5-10 g/L of yeast extract, 20-30 mg/L of sodium selenite and 12-18 g/L of agar.
The seed culture medium comprises the following components: 10-20 g/L of malt extract, 10-15 g/L of peptone, 5-10 g/L of yeast extract and 30-60 mg/L of sodium selenite.
In the step (3), the fermentation medium comprises the following components: 30-40 g/L of fermentable reducing sugar, 0.035-0.05 g/L of copper sulfate, 0.03-0.04 g/L of manganese sulfate, 0.35-0.45 g/L of ferrous sulfate, 5.5-7.0 g/L of magnesium sulfate, 2.5-3.5 g/L of zinc sulfate, 3.5-5.5 g/L of calcium chloride, 5-7 g/L of ammonium sulfate, 8.5-11 g/L of sodium chloride, 7-11 g/L of glutamic acid, 11-13 g/L of L-cysteine, and the balance of the total amount of the components is 1L of distilled water.
In the step (3), the fermentation process is as follows: and (3) fermenting for 12-14 h to a logarithmic phase at 25-30 ℃ with dissolved oxygen of 50-70% and pH of 5.5-6.0, supplementing sodium selenite with final concentration of 150-200 mg/L into the fermentation product, and continuing fermentation culture for 20-24 h to obtain zymogen liquid.
In the step (4), the Bacillus licheniformis Lincheniforms cas58 is cultured for 24-36 h by adopting an LB culture medium, the inoculation amount is 2-3%, and the rotating speed of a shaking table is 180-200 rpm; the Pediococcus pentosaceus cas138 is cultured for 24-36 h by adopting an MRS culture medium, the inoculation amount is 2-3%, and the rotating speed of a shaking table is 180-200 rpm; the Streptococcus lactis cas168 is cultured for 24-36 hours by adopting a culture medium, the inoculation amount is 2-3%, and the rotating speed of a shaking table is 180-200 rpm.
The selenium-rich fermented soybean meal prepared by the preparation method of the selenium-rich fermented soybean meal.
An application of selenium-rich fermented soybean meal in animal feed.
The invention adopts high selenium-enriched yeast Candida utilis cas6 as a production strain, and uses a liquid culture medium containing hydrolysis sugar and nutrient salt as a fermentation culture medium to convert sodium selenite into organic selenium with safety, no toxicity and high biological effectiveness. And then, compounding the selenium-enriched yeast with probiotic strains such as bacillus licheniformis, pediococcus pentosaceus, streptococcus lactis and the like, fermenting the bean pulp by using the compounded mixed bacterial liquid, and converting the bean pulp into multifunctional bean pulp rich in small peptides and organic selenium nutrient elements through the synergistic action between probiotics.
Compared with the prior art, the invention has the following advantages: the invention adopts selenium-enriched yeast and probiotics to ferment the soybean meal together to obtain the fermented soybean meal with high organic selenium content and rich amino acid content. On one hand, the pollution and the damage to the environment and animal bodies caused by directly adding selenite substances into the feed are overcome, on the other hand, the selenium content in the bean pulp can be greatly improved, the bean pulp is rich in biological organic selenium, contains a large amount of beneficial microorganisms and fermentation metabolites, is richer in nutrition, the quality of the bean pulp is obviously improved, the digestion and absorption of the animal to the nutrient substances can be promoted, the immune function of the organism is enhanced, the high-selenium-rich multifunctional bean pulp is manufactured, and the nutritional value and the efficacy of the bean pulp are improved. Meanwhile, the preparation method has the advantages of simple process, convenient material taking, simple equipment, low cost and higher popularization value.
Detailed Description
The following examples are given for the detailed implementation and specific operation of the present invention, but the scope of the present invention is not limited to the following examples.
Example 1
The bacterial strains of the fermented soybean meal composite probiotic bacterial liquid in the embodiment comprise the following components:
the preparation method of the selenium-rich fermented soybean meal comprises the following steps:
(1) selenium-enriched strain breeding
Culturing Candida utilis C.utilis on YEPD culture plate for 48-72h, picking single colony to 250ml triangular flask containing 100ml YEPD liquid culture medium, culturing at 180-200 rpm for 48-72h with shaking, centrifuging and washing bacterial liquid for 3-4 times to prepare single cell suspension, and making cell concentration be 108～109one/mL. 0.1mL of the bacterial suspension is evenly coated in a sterile culture dish and is placed in an ultra-clean working table to be dried by air to prepare a bacterial membrane. In N+Ion implantation is carried out in the implanter with the energy of 14-18 KeV. 6 gradient doses were made per strain, two plates per dose. N is a radical of+The implantation doses were 4.0X 10 respectively14、6.0×1014、8.0×1014、1.0×1015、1.2×1015、1.4×1015ion/cm2And vacuum control plates were made. After mutagenesis, 1ml of sterile water was added to wash the cells to prepare a bacterial suspension. Diluting the bacterial suspension 105And coating the strain with a screening culture medium containing sodium selenite, and determining the selenium content of the yeast strain by using a 3,3' -diaminobenzidine hydrochloride method. Multiple times of mutagenesis screening are carried out to obtain the tolerant high-concentration subunitThe yeast mutant strain of sodium selenate (200mg/L) has fermentation biomass increased by 2.5 times compared with the original strain under the same condition, and is named as C.utilis cas 6.
(2) Preparation of seed culture solution
Candida utilis C.utilis cas6 is inoculated in a slant culture medium and cultured for 24h at the temperature of 28 ℃ to obtain a slant strain, and the slant culture medium comprises the following components: 15g/L of glucose, 15g/L of peptone, 5g/L of yeast extract, 20mg/L of sodium selenite and 12g/L of agar.
Inoculating the slant strain into 80mL seed culture medium, and performing shake culture at 28 deg.C and 180r/min for 24h to obtain seed culture solution. The seed culture medium comprises the following components: 10g/L of malt extract, 10g/L of peptone, 5g/L of yeast extract and 30mg/L of sodium selenite.
(3) Fermentation culture
Inoculating the seed culture solution into a 50L fermentation tank containing a fermentation culture medium according to the inoculation amount of 8 percent (volume percentage content) (the liquid loading amount is 40 percent), controlling the dissolved oxygen at 28 ℃, maintaining the pH value at 5.5, fermenting for 12 hours to logarithmic phase, supplementing sodium selenite with the final concentration of 150mg/L into the fermentation product, and continuing to ferment and culture for 20 hours to obtain a zymogen solution.
The fermentation medium comprises the following components: 30g/L of fermentable reducing sugar, 0.035g/L of copper sulfate, 0.03g/L of manganese sulfate, 0.35g/L of ferrous sulfate, 5.5g/L of magnesium sulfate, 2.5g/L of zinc sulfate, 3.5g/L of calcium chloride, 5g/L of ammonium sulfate, 8.5g/L of sodium chloride, 7g/L of glutamic acid and 11g/L of L-cysteine, and the balance of the components is made up to 1L by using distilled water.
(4) Fermenting with single strain
Bacillus licheniformis B.Linchenoforms cas58 was cultured in LB medium (tryptone 10g, yeast extract 5g, sodium chloride 10g, purified water 1000ml) for 24h with an inoculum size of 2% and a shaker speed of 180 rpm.
Pediococcus pentosaceus P.penttosaceus cas138 was cultured in MRS medium (peptone 10.0g, beef powder 5.0g, yeast powder 4.0g, glucose 20.0g, Tween 801.0 mL, dipotassium hydrogen phosphate 2.0g, sodium acetate 5.0g, triammonium citrate 2.0g, magnesium sulfate 0.2g, manganese sulfate 0.05g, pure water 1000mL) for 24h with an inoculum size of 2% and a shaker rotation speed of 180 rpm.
Lactic streptococci S.lactis cas168 was cultured in a medium (10 g sucrose, 20g peptone, 10g yeast extract, 30g potassium dihydrogen phosphate, 2g sodium chloride, 0.2g magnesium sulfate, 1000ml pure water) for 24h with an inoculum size of 2% and a shaker rotation speed of 180 rpm.
(5) Preparation of probiotic composite bacterial liquid
After the fermentation of the single strains is finished, four strains of fresh fermentation liquor of microzyme C.utilis cas6, Bacillus licheniformis B.Lincheniforms cas58, Pediococcus pentosaceus P.pentosaceus cas138 and Streptococcus lactis S.lactis 168 are mixed according to the volume ratio of 3:2:2:1 to prepare the probiotic compound bacterial liquid, wherein the effective viable count is 8.0 multiplied by 109cfu/ml。
(6) Fresh media addition
Adding fresh culture medium-cane molasses (containing more than 50% of sugar and more than 85% of brix) into the composite bacterial liquid according to the mass concentration of 2%.
(7) Preparation of high-activity selenium-rich bean pulp
100 kg (dry weight) of soybean meal is weighed, 20 kg of feather meal and 15 kg of bran are added, and 40 kg of tap water and materials are placed overnight for mixing. And (3) after uniformly stirring, adding 0.35 kg of the probiotic compound bacterial liquid prepared in the step (6) according to the proportion of 2 per mill, uniformly mixing and stirring, fermenting for 36 hours at the constant temperature of 36 ℃, and drying by adopting a roller type process (the airflow temperature does not exceed 80 ℃) to prepare the selenium-enriched bean pulp rich in high activity.
The content detection of the high-activity selenium-enriched bean pulp prepared by the embodiment: detecting the content of crude protein in the prepared bean pulp by reference to GB/T6432-1994; and (3) determining the content of the organic selenium in the prepared bean pulp by adopting a conventional detection method. The detection result shows that the selenium-enriched bean pulp contains 45.3% of crude protein, 6.1% of small peptide, 7.2% of water and 8.5mg/kg of organic selenium.
Example 2
The bacterial strains of the fermented soybean meal composite probiotic bacterial liquid in the embodiment comprise the following components:
the preparation method of the selenium-rich fermented soybean meal comprises the following steps:
(1) selenium-enriched strain breeding
The detailed description is the same as example 1.
(2) Preparation of seed culture solution
Candida utilis C.utilis cas6 is inoculated in a slant culture medium and cultured for 30h at the temperature of 28 ℃ to obtain a slant strain, and the slant culture medium comprises the following components: 18g/L of glucose, 18g/L of peptone, 8g/L of yeast extract, 25mg/L of sodium selenite and 15g/L of agar.
Inoculating the slant strain into 80mL seed culture medium, and performing shake culture at 28 deg.C and 190r/min for 30h to obtain seed culture solution. The seed culture medium comprises the following components: 15g/L of malt extract, 12g/L of peptone, 8g/L of yeast extract and 45mg/L of sodium selenite.
(3) Fermentation culture
Inoculating the seed culture solution into a 50L fermentation tank containing a fermentation culture medium according to the inoculation amount of 10% (volume percentage content) (the liquid loading amount is 45%), controlling the dissolved oxygen at 28 ℃, maintaining the pH value at 5.5, fermenting for 13h to logarithmic phase, supplementing sodium selenite with the final concentration of 180mg/L into the fermentation product, and continuing to ferment and culture for 22h to obtain a zymophyte solution.
The fermentation medium comprises the following components: 35g/L of fermentable reducing sugar, 0.04g/L of copper sulfate, 0.035g/L of manganese sulfate, 0.40g/L of ferrous sulfate, 6.5g/L of magnesium sulfate, 2.8g/L of zinc sulfate, 4.5g/L of calcium chloride, 6.0g/L of ammonium sulfate, 9.5g/L of sodium chloride, 8.5g/L of glutamic acid and 11.5g/L of L-cysteine, and the balance of the components are made up to 1L by distilled water.
(4) Fermenting with single strain
Bacillus licheniformis B.Linchenoforms cas58 was cultured in LB medium (tryptone 10g, yeast extract 5g, sodium chloride 10g, purified water 1000ml) for 30h with an inoculum size of 2.5% and a shaker speed of 190 rpm.
Pediococcus pentosaceus P.penttosaceus cas138 was cultured in MRS medium (peptone 10.0g, beef powder 5.0g, yeast powder 4.0g, glucose 20.0g, Tween 801.0 mL, dipotassium hydrogen phosphate 2.0g, sodium acetate 5.0g, triammonium citrate 2.0g, magnesium sulfate 0.2g, manganese sulfate 0.05g, pure water 1000mL) for 30h with an inoculum size of 2.5% and a shaker rotation speed of 190 rpm.
Lactic streptococci S.lactis cas168 was cultured in a medium (10 g sucrose, 20g peptone, 10g yeast extract, 30g potassium dihydrogen phosphate, 2g sodium chloride, 0.2g magnesium sulfate, 1000ml pure water) for 30h with an inoculum size of 2.5% and a shaker rotation speed of 190 rpm.
(5) Preparation of probiotic composite bacterial liquid
After the fermentation of the single strains is finished, four strains of fresh fermentation liquor of microzyme C.utilis cas6, Bacillus licheniformis B.Lincheniforms cas58, Pediococcus pentosaceus P.pentosaceus cas138 and Streptococcus lactis S.lactis 168 are mixed according to the volume ratio of 4:2.5:2.5:1.5 to prepare the probiotic compound bacterial liquid, wherein the effective viable count is 1.05 multiplied by 1010cfu/ml。
(6) Fresh media addition
Adding fresh culture medium-cane molasses (containing more than 50% of sugar and more than 85% of brix) into the composite bacterial liquid according to the mass concentration of 2.5%.
(7) Preparation of high-activity selenium-rich bean pulp
100 kg (dry weight) of soybean meal is weighed, 25 kg of feather meal and 18 kg of bran are added, and 45 kg of tap water and materials are placed overnight for mixing. And (3) after uniformly stirring, adding 0.47 kg of the probiotic compound bacterial liquid prepared in the step (6) according to the proportion of 2.5 per mill, uniformly mixing and stirring, fermenting for 36 hours at the constant temperature of 40 ℃, and drying by adopting a roller type process (the airflow temperature does not exceed 80 ℃) to prepare the selenium-enriched bean pulp rich in high activity.
The content detection of the high-activity selenium-enriched bean pulp prepared by the embodiment: detecting the content of crude protein in the prepared bean pulp by reference to GB/T6432-1994; and (3) determining the content of the organic selenium in the prepared bean pulp by adopting a conventional detection method. The detection result shows that the selenium-enriched soybean meal contains 48% of crude protein, 8% of small peptide, 7.2% of water and 10mg/kg of organic selenium.
Example 3
The fermented soybean meal composite probiotic bacteria of the embodimentThe liquid strain was composed as follows: yeast C.utilis cas 65.0X 109cfu/ml
Bacillus licheniformis b. lincheniforms cas 583.0 × 109cfu/ml
Pediococcus pentosaceus P.penttosaceus 1383.0X 109cfu/ml
Lactic streptococci S.lactis cas 1682.0X 109cfu/ml
The preparation method of the selenium-rich fermented soybean meal comprises the following steps:
(1) selenium-enriched strain breeding
The detailed description is the same as example 1.
(2) Preparation of seed culture solution
Candida utilis C.utilis cas6 is inoculated in a slant culture medium and cultured for 32h at the temperature of 28 ℃ to obtain a slant strain, and the slant culture medium comprises the following components: 20g/L of glucose, 20g/L of peptone, 10g/L of yeast extract, 30mg/L of sodium selenite and 18g/L of agar.
Inoculating the slant strain into 80mL seed culture medium, and performing shake culture at 28 deg.C and 200r/min for 36h to obtain seed culture solution. The seed culture medium comprises the following components: 20g/L of malt extract, 15g/L of peptone, 10g/L of yeast extract and 60mg/L of sodium selenite.
(3) Fermentation culture
Inoculating the seed culture solution into a 50L fermentation tank containing a fermentation culture medium according to the inoculation amount of 12% (volume percentage content) (the liquid loading amount is 50%), controlling dissolved oxygen at 28 ℃, maintaining pH at 6.0, fermenting for 14h to logarithmic phase, supplementing sodium selenite with the final concentration of 200mg/L into the fermentation product, and continuing to ferment and culture for 24h to obtain a zymophyte solution.
The fermentation medium comprises the following components: 40g/L of fermentable reducing sugar, 0.05g/L of copper sulfate, 0.04g/L of manganese sulfate, 0.45g/L of ferrous sulfate, 7.0g/L of magnesium sulfate, 3.5g/L of zinc sulfate, 5.5g/L of calcium chloride, 7g/L of ammonium sulfate, 11g/L of sodium chloride, 11g/L of glutamic acid and 13g/L of L-cysteine, and the balance of the components is made up to 1L by using distilled water.
(4) Fermenting with single strain
Bacillus licheniformis B.Linchenoforms cas58 was cultured in LB medium (tryptone 10g, yeast extract 5g, sodium chloride 10g, purified water 1000ml) for 36h with an inoculum size of 3% and a shaker speed of 200 rpm.
Pediococcus pentosaceus P.penttosaceus cas138 was cultured in MRS medium (peptone 10.0g, beef powder 5.0g, yeast powder 4.0g, glucose 20.0g, Tween 801.0 mL, dipotassium hydrogen phosphate 2.0g, sodium acetate 5.0g, triammonium citrate 2.0g, magnesium sulfate 0.2g, manganese sulfate 0.05g, pure water 1000mL) for 36h with an inoculum size of 3% and a shaker rotation speed of 200 rpm.
Lactic streptococci S.lactis cas168 was cultured in a medium (10 g sucrose, 20g peptone, 10g yeast extract, 30g potassium dihydrogen phosphate, 2g sodium chloride, 0.2g magnesium sulfate, 1000ml pure water) for 36h with an inoculum size of 3% and a shaker rotation speed of 200 rpm.
(5) Preparation of probiotic composite bacterial liquid
After the fermentation of the single strains is finished, four strains of fresh fermentation liquor of yeast C.utilis cas6, Bacillus licheniformis B.Lincheniforms cas58, Pediococcus pentosaceus P.pentosaceus cas138 and Streptococcus lactis S.lactis 168 are mixed according to the volume ratio of 5:3:3:2 to prepare the probiotic compound bacterial liquid, wherein the effective viable count is 1.3 multiplied by 1010cfu/ml。
(6) Fresh media addition
Adding fresh culture medium-cane molasses (containing more than 50% of sugar and more than 85% of brix) into the composite bacterial liquid according to the mass concentration of 3%.
(7) Preparation of high-activity selenium-rich bean pulp
100 kg (dry weight) of soybean meal is weighed, 30 kg of feather meal and 20 kg of bran are added, 50 kg of tap water is placed overnight, and materials are mixed. And (3) after uniformly stirring, adding 0.6 kg of the probiotic compound bacterial liquid prepared in the step (4) according to the proportion of 3 per mill, uniformly mixing and stirring, fermenting for 48 hours at the constant temperature of 42 ℃, and drying by adopting a roller type process (the airflow temperature does not exceed 80 ℃) to prepare the selenium-enriched bean pulp rich in high activity.
The content detection of the high-activity selenium-enriched bean pulp prepared by the embodiment: according to GB/T6432-1994, detecting the content of crude protein in the prepared soybean meal by adopting a Kjeldahl apparatus; and (4) determining the selenium content in the prepared bean pulp by adopting a conventional detection method. The detection result shows that the selenium-enriched soybean meal contains 46.5% of crude protein, 6.8% of small peptide, 7.3% of water and 8.3mg/kg of organic selenium.
The high-activity selenium-rich soybean meal prepared in the embodiment 1-3 is applied to a certain pig factory, a group of control groups and a group of test groups, wherein each group of piglets comprises 50 piglets, and test results show that the high-activity selenium-rich soybean meal can improve the growth performance and the nutrient digestibility of weaned piglets, reduce intestinal injury and relieve diarrhea.
The incidence rate of the yellow and white Li of the piglets eating the high-activity selenium-rich bean pulp is reduced by 50 percent, and no antibiotic is used in the breeding process, thereby greatly reducing the use cost of the antibiotic and reducing the environmental pollution caused by the abuse of the antibiotic. Meanwhile, the growth speed of the pigs fed with the high-activity selenium-rich soybean meal prepared by the embodiment is obviously higher than that of a control group, the pigs can be slaughtered 5-10 days in advance, and the feed cost of breeders is greatly reduced.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention are intended to be included within the scope of the present invention.
Claims (10)
1. The preparation method of the selenium-rich fermented soybean meal is characterized by comprising the following steps of:
(1) selenium-enriched strain breeding
Performing nitrogen ion implantation mutagenesis on Candida utilis, and performing multiple mutagenesis screening to obtain a yeast mutant strain tolerant to high-concentration sodium selenite, namely C. utilis cas 6;
(2) preparation of seed culture solution
Inoculating a candida utilis mutant strain C, utilis cas6 to a slant culture medium to obtain a slant strain, and then inoculating the slant strain to a seed culture medium to obtain a seed culture solution;
(3) fermentation culture
Inoculating the seed culture solution into a fermentation culture medium according to the inoculation amount of 8-12% of the volume percentage, supplementing sodium selenite with the final concentration of 150-200 mg/L into a fermentation product, and continuing fermentation culture for 20-24 hours to obtain a zymophyte solution;
(4) probiotic single strain fermentation
Separately fermenting three strains of Bacillus licheniformis Lincheniforms cas58, pediococcus pentosaceus cas138 and Streptococcus lactis cas168 to obtain respective probiotic zymocyte liquid;
(5) preparation of probiotic composite bacterial liquid
Fermenting bacterial liquids of four strains of selenium-enriched yeast C, utilis cas6, Bacillus licheniformis B, Lincheniforms cas58, Pediococcus pentosaceus P, pentusaceus cas138 and Streptococcus lactis cas168 are mixed according to the ratio of 3-5: 2-3: 2-3: 1-2, and obtaining the probiotic composite bacterial liquid, wherein the effective viable count is 8.0 multiplied by 109～1.3×1010cfu/ml；
(6) Fresh media addition
Adding a fresh culture medium-cane molasses into the composite bacterial liquid according to the mass concentration of 2-3%;
(7) preparation of high-activity selenium-rich bean pulp
Mixing bean pulp, feather meal, bran and water according to a weight ratio of 10: 2-3: 1.5-2: and 4-5, adding the probiotic compound bacterial liquid prepared in the step (6) according to the weight ratio of 2-3 per mill after mixing, uniformly mixing and stirring, fermenting for 36-48 hours at the constant temperature of 36-42 ℃, and drying to obtain the selenium-enriched soybean meal.
2. The preparation method of the selenium-enriched fermented soybean meal as claimed in claim 1, wherein the step (1) comprises the following steps:
(11) culturing Candida utilis on a YEPD culture plate for 48-72h, picking a single colony to a culture bottle containing a YEPD liquid culture medium, culturing at 180-200 rpm for 48-72h by shaking, centrifuging and washing a bacterial liquid to prepare a single-cell suspension, and ensuring the cell concentration to be 108～109Per mL;
(12) uniformly coating the bacterial suspension in a sterile culture dish, and air-drying to prepare a bacterial membrane;
(13) in N+Performing ion implantation in an implanter with energy of 14-18 KeV;
(14) 6 gradient doses per strain, two plates per dose, N+The implantation doses were 4.0X 10 respectively14、6.0×1014、8.0×1014、1.0×1015、1.2×1015、1.4×1015ion/cm2And making a vacuum control flat plate;
(15) after mutagenesis is finished, adding sterile water to wash the thalli to prepare bacterial suspension;
(16) diluting the bacterial suspension 104～108The yeast mutant strain which is tolerant to the high-concentration sodium selenite is obtained by multiple mutagenesis and screening after being coated on a screening culture medium containing the sodium selenite in times, and the fermentation biomass of the yeast mutant strain is improved by more than 2 times compared with the original strain under the same condition, so that the yeast mutant strain is named as C. utilis cas 6.
3. The method for preparing the selenium-enriched fermented soybean meal as claimed in claim 1, wherein in the step (2), the candida utilis mutant strain c. utilis cas6 is inoculated in a slant culture medium at 25-30 ℃ for 24-32 h to obtain a slant strain, and then the slant strain is inoculated in a seed culture medium at 25-30 ℃ and 180-200 r/min for shaking culture for 24-36 h to obtain a seed culture solution.
4. The method for preparing the selenium-enriched fermented soybean meal according to claim 3, wherein the slant culture medium comprises the following components: 15-20 g/L of glucose, 15-20 g/L of peptone, 5-10 g/L of yeast extract, 20-30 mg/L of sodium selenite and 12-18 g/L of agar.
5. The method for preparing the selenium-enriched fermented soybean meal according to claim 3, wherein the seed culture medium comprises the following components: 10-20 g/L of malt extract, 10-15 g/L of peptone, 5-10 g/L of yeast extract and 30-60 mg/L of sodium selenite.
6. The method for preparing selenium-enriched fermented soybean meal according to claim 1, wherein in the step (3), the fermentation medium comprises the following components: 30-40 g/L of fermentable reducing sugar, 0.035-0.05 g/L of copper sulfate, 0.03-0.04 g/L of manganese sulfate, 0.35-0.45 g/L of ferrous sulfate, 5.5-7.0 g/L of magnesium sulfate, 2.5-3.5 g/L of zinc sulfate, 3.5-5.5 g/L of calcium chloride, 5-7 g/L of ammonium sulfate, 8.5-11 g/L of sodium chloride, 7-11 g/L of glutamic acid, 11-13 g/L of L-cysteine, and the balance of the total amount of the components is 1L of distilled water.
7. The method for preparing selenium-enriched fermented soybean meal according to claim 1, wherein in the step (3), the fermentation process is as follows: and (3) fermenting for 12-14 h to a logarithmic phase at 25-30 ℃ with dissolved oxygen of 50-70% and pH of 5.5-6.0, supplementing sodium selenite with final concentration of 150-200 mg/L into the fermentation product, and continuing fermentation culture for 20-24 h to obtain zymogen liquid.
8. The preparation method of the selenium-rich fermented soybean meal as claimed in claim 1, wherein in the step (4), the Bacillus licheniformis Lincheniforms cas58 is cultured for 24-36 h by adopting an LB culture medium, the inoculation amount is 2-3%, and the rotation speed of a shaking table is 180-200 rpm; the Pediococcus pentosaceus pentacoccus pentosaceus cas138 is cultured for 24-36 hours by adopting an MRS culture medium, the inoculation amount is 2-3%, and the rotating speed of a shaking table is 180-200 rpm; the Streptococcus lactis cas168 is cultured for 24-36 hours by adopting a culture medium, the inoculation amount is 2-3%, and the rotating speed of a shaking table is 180-200 rpm.
9. The selenium-enriched fermented soybean meal prepared by the preparation method of the selenium-enriched fermented soybean meal according to any one of claims 1 to 8, wherein the selenium-enriched fermented soybean meal contains 45-48% of crude protein, 6-8% of small peptides, 7-8% of water and 8-10 mg/kg of organic selenium.
10. The use of the selenium enriched fermented soybean meal of claim 9 in the preparation of animal feed.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101792720A (en) * | 2009-05-12 | 2010-08-04 | 广州市博善生物饲料有限公司 | Production method of selenium-enriched yeast culture |
| CN103027192A (en) * | 2012-12-17 | 2013-04-10 | 宁波天邦股份有限公司 | Method for preparing selenium-enriched fermented soybean meal |
| CN105441363A (en) * | 2015-12-30 | 2016-03-30 | 北京资源亚太饲料科技有限公司 | Strain, method for preparing fermented soybean meal from strain and prepared fermented soybean meal |
-
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101792720A (en) * | 2009-05-12 | 2010-08-04 | 广州市博善生物饲料有限公司 | Production method of selenium-enriched yeast culture |
| CN103027192A (en) * | 2012-12-17 | 2013-04-10 | 宁波天邦股份有限公司 | Method for preparing selenium-enriched fermented soybean meal |
| CN105441363A (en) * | 2015-12-30 | 2016-03-30 | 北京资源亚太饲料科技有限公司 | Strain, method for preparing fermented soybean meal from strain and prepared fermented soybean meal |
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