CN113999780B - Selenium-enriched yeast powder and preparation method and application thereof - Google Patents
Selenium-enriched yeast powder and preparation method and application thereof Download PDFInfo
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- BUGBHKTXTAQXES-UHFFFAOYSA-N Selenium Chemical compound [Se] BUGBHKTXTAQXES-UHFFFAOYSA-N 0.000 title claims abstract description 77
- 239000011669 selenium Substances 0.000 title claims abstract description 77
- 229910052711 selenium Inorganic materials 0.000 title claims abstract description 77
- 240000004808 Saccharomyces cerevisiae Species 0.000 title claims abstract description 62
- 239000000843 powder Substances 0.000 title claims abstract description 36
- 238000002360 preparation method Methods 0.000 title claims abstract description 14
- 241000512259 Ascophyllum nodosum Species 0.000 claims abstract description 35
- 239000001963 growth medium Substances 0.000 claims abstract description 27
- 239000000413 hydrolysate Substances 0.000 claims abstract description 19
- 238000000034 method Methods 0.000 claims abstract description 19
- 238000009630 liquid culture Methods 0.000 claims abstract description 13
- 238000012258 culturing Methods 0.000 claims abstract description 12
- 238000001694 spray drying Methods 0.000 claims abstract description 10
- 238000000855 fermentation Methods 0.000 claims abstract description 9
- 230000004151 fermentation Effects 0.000 claims abstract description 9
- 238000004519 manufacturing process Methods 0.000 claims abstract 2
- 229940091258 selenium supplement Drugs 0.000 claims description 69
- BVTBRVFYZUCAKH-UHFFFAOYSA-L disodium selenite Chemical compound [Na+].[Na+].[O-][Se]([O-])=O BVTBRVFYZUCAKH-UHFFFAOYSA-L 0.000 claims description 16
- 239000007788 liquid Substances 0.000 claims description 16
- 229960001471 sodium selenite Drugs 0.000 claims description 16
- 239000011781 sodium selenite Substances 0.000 claims description 16
- 235000015921 sodium selenite Nutrition 0.000 claims description 16
- 102000004190 Enzymes Human genes 0.000 claims description 10
- 108090000790 Enzymes Proteins 0.000 claims description 10
- 229940088598 enzyme Drugs 0.000 claims description 10
- 108091005658 Basic proteases Proteins 0.000 claims description 6
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 6
- 102000057297 Pepsin A Human genes 0.000 claims description 6
- 108090000284 Pepsin A Proteins 0.000 claims description 6
- 102000004142 Trypsin Human genes 0.000 claims description 6
- 108090000631 Trypsin Proteins 0.000 claims description 6
- 229940111202 pepsin Drugs 0.000 claims description 6
- 239000012588 trypsin Substances 0.000 claims description 6
- 239000003674 animal food additive Substances 0.000 claims description 4
- 244000005700 microbiome Species 0.000 claims description 4
- 239000006228 supernatant Substances 0.000 claims description 4
- 230000003301 hydrolyzing effect Effects 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- 241001465754 Metazoa Species 0.000 abstract description 6
- 230000036039 immunity Effects 0.000 abstract description 4
- 150000004676 glycans Chemical class 0.000 abstract description 2
- 229920001282 polysaccharide Polymers 0.000 abstract description 2
- 239000005017 polysaccharide Substances 0.000 abstract description 2
- 239000000126 substance Substances 0.000 abstract 1
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 46
- 238000012360 testing method Methods 0.000 description 12
- 231100000350 mutagenesis Toxicity 0.000 description 10
- 238000002703 mutagenesis Methods 0.000 description 9
- 230000001580 bacterial effect Effects 0.000 description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 8
- 241000282887 Suidae Species 0.000 description 7
- 230000008569 process Effects 0.000 description 7
- GGLZPLKKBSSKCX-YFKPBYRVSA-N L-ethionine Chemical compound CCSCC[C@H](N)C(O)=O GGLZPLKKBSSKCX-YFKPBYRVSA-N 0.000 description 5
- 239000002609 medium Substances 0.000 description 5
- 238000012216 screening Methods 0.000 description 5
- 239000001888 Peptone Substances 0.000 description 4
- 108010080698 Peptones Proteins 0.000 description 4
- 229940041514 candida albicans extract Drugs 0.000 description 4
- 235000019319 peptone Nutrition 0.000 description 4
- 238000004321 preservation Methods 0.000 description 4
- 239000008223 sterile water Substances 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- 238000005406 washing Methods 0.000 description 4
- 239000012138 yeast extract Substances 0.000 description 4
- 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 description 3
- 241000235342 Saccharomycetes Species 0.000 description 3
- 230000009471 action Effects 0.000 description 3
- 238000000576 coating method Methods 0.000 description 3
- 230000000052 comparative effect Effects 0.000 description 3
- 238000007865 diluting Methods 0.000 description 3
- 239000012153 distilled water Substances 0.000 description 3
- 239000008103 glucose Substances 0.000 description 3
- 235000016709 nutrition Nutrition 0.000 description 3
- 238000012408 PCR amplification Methods 0.000 description 2
- 238000004820 blood count Methods 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 239000011248 coating agent Substances 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000002708 enhancing effect Effects 0.000 description 2
- 230000035764 nutrition Effects 0.000 description 2
- 229920001542 oligosaccharide Polymers 0.000 description 2
- 150000002482 oligosaccharides Chemical class 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 230000001737 promoting effect Effects 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 239000011573 trace mineral Substances 0.000 description 2
- 235000013619 trace mineral Nutrition 0.000 description 2
- DBTMGCOVALSLOR-DEVYUCJPSA-N (2s,3r,4s,5r,6r)-4-[(2s,3r,4s,5r,6r)-3,5-dihydroxy-6-(hydroxymethyl)-4-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyoxan-2-yl]oxy-6-(hydroxymethyl)oxane-2,3,5-triol Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](CO)O[C@H](O)[C@@H]2O)O)O[C@H](CO)[C@H]1O DBTMGCOVALSLOR-DEVYUCJPSA-N 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 102000006587 Glutathione peroxidase Human genes 0.000 description 1
- 108700016172 Glutathione peroxidases Proteins 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 208000012659 Joint disease Diseases 0.000 description 1
- 208000019926 Keshan disease Diseases 0.000 description 1
- 229920001543 Laminarin Polymers 0.000 description 1
- 239000005717 Laminarin Substances 0.000 description 1
- 206010039921 Selenium deficiency Diseases 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 238000010241 blood sampling Methods 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000003651 drinking water Substances 0.000 description 1
- 235000020188 drinking water Nutrition 0.000 description 1
- 230000002526 effect on cardiovascular system Effects 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 210000003608 fece Anatomy 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 235000012041 food component Nutrition 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 239000006481 glucose medium Substances 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 239000010871 livestock manure Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000033116 oxidation-reduction process Effects 0.000 description 1
- 238000005375 photometry Methods 0.000 description 1
- 244000144977 poultry Species 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 125000003748 selenium group Chemical group *[Se]* 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- AKHNMLFCWUSKQB-UHFFFAOYSA-L sodium thiosulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=S AKHNMLFCWUSKQB-UHFFFAOYSA-L 0.000 description 1
- 235000019345 sodium thiosulphate Nutrition 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 238000011282 treatment Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
-
- 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
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
-
- 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
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/30—Feeding-stuffs specially adapted for particular animals for swines
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/70—Feeding-stuffs specially adapted for particular animals for birds
- A23K50/75—Feeding-stuffs specially adapted for particular animals for birds for poultry
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/80—Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
Abstract
The invention relates to the technical field of selenium-enriched yeast, in particular to selenium-enriched yeast powder and a preparation method and application thereof, wherein the preparation method of the selenium-enriched yeast powder comprises the following steps: (1) Adding kelp hydrolysate into the liquid culture medium to prepare fermentation culture solution; (2) Inoculating the selenium-enriched yeast strain into the fermentation culture solution according to 1% -10%, and culturing for 12-36h to obtain selenium-enriched yeast solution; and (3) spray drying to obtain the selenium-enriched yeast powder. According to the method, the kelp hydrolysate is added into the liquid culture medium, and substances such as polysaccharide in the kelp hydrolysate can cooperate with yeast selenium to improve the immunity of animal organisms and improve the production performance.
Description
Technical Field
The invention relates to the technical field of selenium-enriched yeast, in particular to selenium-enriched yeast powder and a preparation method and application thereof.
Background
Selenium is an essential trace element for human and animals, is an important component of various enzymes, is an important component of glutathione peroxidase, and protects a biological membrane from damage through oxidation reduction, and maintains normal functions of cells. Selenium also has immunity enhancing, cardiovascular protecting, growth promoting, antitumor, toxic materials removing, and metabolism regulating effects. The local diseases caused by selenium deficiency in China include keshan disease and large bone joint disease.
The yeast has high selenium-rich capability and can convert inorganic selenium into organic selenium, so that the selenium-rich yeast is widely used as a source for supplementing trace element selenium in the industries of foods, health-care products and feeds. The selenium-enriched yeast is a microbial fermentation product obtained by adding inorganic selenium in the form of sodium selenite and the like in the process of culturing the yeast, converting the inorganic selenium into an organic selenium form and enriching the organic selenium in the selenium-enriched yeast.
The existing selenium-enriched yeast powder is obtained by expanding culture, centrifugation, washing and freeze-drying of selenium-enriched yeast strains, mainly comprises saccharomycetes and has limited nutritional ingredients.
Disclosure of Invention
Therefore, the selenium-enriched yeast powder and the preparation method and the application thereof are needed to be provided, and the problem of single nutrition component of the existing selenium-enriched yeast powder is solved.
In order to achieve the above purpose, the invention provides a preparation method of selenium-enriched yeast powder, which comprises the following steps:
(1) Adding kelp hydrolysate into the liquid culture medium to prepare fermentation culture solution;
(2) Inoculating the selenium-enriched yeast strain into the fermentation culture solution according to 1% -10%, and culturing for 12-36h to obtain selenium-enriched yeast solution;
(3) Spray drying to obtain selenium-enriched yeast powder;
the selenium-enriched yeast strain is preserved in the microorganism strain collection of Guangdong province, the preservation date is 2021, 9 and 9 days, and the preservation number is GDMCC No.61733.
Further, the kelp hydrolysate is prepared by the following method:
(1) Mixing the kelp powder and hydrochloric acid with the concentration of 0.1-0.5mol/L at the weight ratio of 1:10-1:20, fully dissolving, hydrolyzing at 45-60 ℃ for 2-6h, centrifuging at 6000-8000r/min for 5-10min, and obtaining supernatant as kelp pretreatment liquid;
(2) Adjusting pH of the kelp pretreatment liquid to 6.0-7.0, adding mixed enzyme with the addition amount of 0.2-3% of the weight of the kelp pretreatment liquid, and performing enzymolysis at 35-60deg.C for 2-8 hr;
(3) And after the enzymolysis is finished, obtaining kelp hydrolysate.
Further, the mixed enzyme is alkaline protease, pepsin and trypsin, and the ratio of the alkaline protease to the pepsin to the trypsin is 1:1:1.
Further, the kelp hydrolysate accounts for 5% -15% of the total weight of the liquid culture medium.
Further, the liquid culture medium comprises a YEPD culture medium and a sodium selenite solution with the volume of 0.9% of that of the YEPD culture medium, and the concentration of the sodium selenite solution is 100mg/ml.
Further, the air inlet temperature of spray drying is 130-150 ℃ and the air outlet temperature is 80-85 ℃.
A selenium-rich yeast powder prepared by the above preparation method.
A feed additive comprises the selenium-rich yeast powder.
The technical scheme has the following beneficial effects:
the yeast powder is obtained by jointly carrying out spray drying on yeast liquid and culture liquid for culture, the yeast powder contains a large amount of nutritional components, the nutrition is sufficient, meanwhile, kelp hydrolysate contains a large amount of laminarin and oligosaccharide, after spray drying, the components are mixed to form the yeast powder, the used yeast strain is a selenium-enriched yeast strain obtained by self-induced mutation screening, and the polysaccharide and oligosaccharide hydrolyzed by the kelp can play a role of synergistically enhancing immunity with the yeast selenium, so that the immunity of animal organisms can be effectively improved, the survival rate can be improved, and the animal growth can be promoted when the kelp is mixed in livestock and poultry or aquatic feeds.
Drawings
FIG. 1 is a graph showing the growth of yeasts described in example 1.
Detailed Description
In order to describe the technical content, constructional features, achieved objects and effects of the technical solution in detail, the following description is made in connection with the specific embodiments in conjunction with the accompanying drawings.
Example 1
The mutagenesis screening method of the selenium-enriched yeast strain comprises the following steps:
(1) Commercial Strain activation
Inoculating strain Saccharomyces cerevisiae (accession number: CICC 33033) into 100ml YEPD at 28deg.C and 200rpm for about 24-48 hr, culturing to logarithmic phase,
yeast extract peptone glucose medium (YEPD medium) component content: 20g of glucose, 20g of peptone and 10g of yeast extract powder, distilled water is added to 1L, and the mixture is autoclaved at the natural pH and 105 ℃ for 30min.
(2) Isolation and identification of strains
Diluting the activated bacterial liquid into 10 -1 、10 -2 、10 -3 、10 -4 And (3) respectively absorbing 200 mu L of concentration gradient, coating the concentration gradient on a selenium-enriched culture medium, performing 3 parallel coatings on each concentration, culturing the culture medium in a constant temperature incubator at 28 ℃ for 48 hours, streaking and purifying to obtain single colonies, performing PCR amplification identification on the single colonies, and accurately and then placing the single colonies in a refrigerator at-20 ℃ for later use.
The selenium-enriched culture medium comprises the following components: 20g of glucose, 20g of peptone, 10g of yeast extract powder, xmg of sodium selenite, distilled water to 1L, natural pH and autoclaving at 121 ℃ for 20min.
Xmg sodium selenite preparation gradient concentration selenium-rich culture medium, in this embodiment, the selenium content of the selenium-rich culture medium is 2×10 in sequence -4 molSe 4+ /L、4×10 -4 molSe 4+ /L、8×10 -4 molSe 4+ /L、1×10 -3 molSe 4+ /L、2×10 - 3 molSe 4+ /L。
And (3) streaking and purifying the single colony on the selenium-enriched culture medium with gradient concentration, selecting a strain capable of growing in the selenium-enriched culture medium with higher concentration, and carrying out PCR amplification identification.
(3) Determination of Strain growth Curve
Inoculating the activated bacteria liquid into 100mL YEPD culture medium according to 4%, drawing a growth curve of saccharomycetes by adopting a light absorption photometry, shake culturing at 28 ℃ at 180r/min, taking the curved line as a starting point, measuring the number of saccharomycetes by using a blood cell counting plate, sampling every 2h (taking 2mL by using a pipette) in the culturing process, measuring and recording the absorbance at an OD600nm, measuring for 48h in total, and drawing a strain growth curve, wherein the growth curve is shown in figure 1.
(4) Preparation of mutant Strain suspension
Culturing activated strain to logarithmic phase, collecting 5ml of bacterial liquid 4800r/min, centrifuging for 5min, washing with sterile water for 2 times, discarding supernatant, adding sterile water again, shaking on vortex mixer to obtain bacterial suspension, measuring the number of bacterial strain in the bacterial suspension with blood cell counting plate, gradient diluting with sterile water, and keeping the number of bacterial strain at 10 6 About, the prepared bacterial suspension is used for mutagenesis.
(5) DES mutagenesis
Adding 0.8% DES, shaking at 25deg.C and 180r/min for 80min, adding 6% sodium thiosulfate solution, stopping reaction, centrifuging, washing with sterile water for 2 times, adding 5mLYEPD culture medium, and culturing overnight.
(6) Screening of sodium selenite resistant plates
Centrifugally collecting thallus, washing 2 times, diluting and coating 2X 10 -3 molSe 4+ L sodium selenite resistant plates.
(7) Screening of ethionine resistant plates
Colonies growing on sodium selenite resistant plates were subjected to liquid culture, DES mutagenesis was again performed as described above in step (5), cultured overnight, and after centrifugation to collect the cells, which were washed 2 times with water and plated on 3.0g/L ethionine resistant plates.
Ethionine resistant plates: the ethionine solution filter sterilized with the filter was added to the sterilized YNB medium. The selenium-enriched amount of the yeast is better than that of the yeast.
(8) UV mutagenesis and screening of sodium selenite resistant plates
And (3) streaking a colony plate growing on the ethionine plate on a sodium selenite plate, placing the plate on an 8w UV light source for mutagenesis after bacterial colony is grown, wherein the mutagenesis distance is 20cm, and the mutagenesis time is 90s. And (3) continuously culturing the plate after mutagenesis, picking out a monoclonal with large growth in light orange yellow, and inoculating the monoclonal into a YEPD culture medium for expansion culture.
The screened selenium-enriched yeast is preserved in the microorganism strain collection of Guangdong province at 2021, 9 and 9 days, address: building 59 of Guangzhou Mr. first, china, no. 100 university, with deposit number GDMCC No.61733.
Example 2
The embodiment provides a preparation method of selenium-enriched yeast powder, which comprises the following steps:
(1) 1L of kelp hydrolysate is added into 10L of liquid culture medium to prepare fermentation culture solution, wherein the liquid culture medium comprises YEPD culture medium and sodium selenite solution with the volume of 0.9% of that of the YEPD culture medium, the concentration of the sodium selenite solution is 100mg/ml, and the YEPD culture medium is as follows: 20g of glucose, 20g of peptone, 10g of yeast extract powder, 1L of distilled water, natural pH and high-pressure sterilization at 105 ℃ for 30min;
wherein the preparation steps of the kelp hydrolysate are as follows;
(1.1) 40-60 meshes of kelp powder, mixing hydrochloric acid with the concentration of 0.2mol/L according to the weight ratio of 1:10, fully dissolving, hydrolyzing at 60 ℃ for 2-6h, centrifuging at 8000r/min for 10min, and obtaining supernatant as kelp pretreatment liquid;
(1.2) regulating the pH of the kelp pretreatment liquid to be 6.0-7.0, adding mixed enzyme, wherein the addition amount of the mixed enzyme is 1% of the weight of the kelp pretreatment liquid, the enzymolysis temperature is 50 ℃, the enzymolysis time is 6 hours, the mixed enzyme is alkaline protease, pepsin and trypsin, and the weight ratio of the alkaline protease to the pepsin to the trypsin is 1:1:1;
(1.3) filtering residues by a screen after the enzymolysis is finished, and obtaining kelp hydrolysate.
(2) Inoculating a selenium-enriched yeast strain into a fermentation culture solution according to 4%, and culturing for 24 hours to obtain a selenium-enriched yeast solution, wherein the selenium-enriched yeast strain is preserved in the Guangdong province microorganism strain preservation center, the preservation date is 2021, 9 and 9, and the preservation number is GDMCC No.61733;
(3) Spray drying to obtain selenium-enriched yeast powder with total selenium content of 1000mg/kg, wherein air inlet temperature of spray drying is 130-150deg.C, and air outlet temperature is 80-85deg.C.
Comparative example 1
Unlike example 1, the preparation of kelp hydrolysate was not added to the liquid medium, which was a YEPD medium and a 0.9% sodium selenite solution in volume of the YEPD medium, followed by inoculation of the selenium-enriched yeast strain for cultivation, and after completion of cultivation, spray-drying to obtain selenium-enriched yeast powder having a total selenium content of 1000 mg/kg.
Animal experiment test was performed on the samples of example 2 and comparative example 1
1. Experimental Effect of sample applied to pig
1.1 test animals
80 growing-finishing pigs, with a weight of 24+/-1.8 kg, were randomly divided into 4 groups, and the initial weight of each group of pigs was recorded.
1.2 test treatments
Control group: the total selenium content of the basic feed is 0.07mg/kg;
test group 1: the selenium-enriched yeast powder of example 2 was mixed with a basal feed with a total selenium content of 0.4mg/kg.
Test group 2: the selenium-enriched yeast powder of comparative example 1 was mixed with a basal feed with a total selenium content of 0.4mg/kg.
Test group 3, the feed additive sodium selenite is mixed with basic feed, and the total selenium content is 0.4mg/kg.
In the experimental process, feeding is carried out for 2 times a day, automatic drinking water and manual manure cleaning are carried out, the experimental time is 50d, and the feeding amount and the residual amount of each group of pigs are accurately recorded every day.
1.3 detection
On the day of the end of the test, blood sampling was performed and the end of the test for each group of pigs was accurately recorded.
1.4 detection results
The test results are shown in Table 1 below
TABLE 1 influence of different selenium-enriched feeds on growth of growing-finishing pigs and blood selenium concentration
As can be seen from the above table 1, the selenium-enriched feed can reduce the feed-meat ratio of growing-finishing pigs, and the growth promoting effect of the selenium-enriched yeast on the growing-finishing pigs is remarkably higher than that of the sodium Yu Yaxi acid feed additive compared with the comparison of the test group 1, the test group 2 and the test group 3.
Example 3
A method for preparing selenium-rich yeast powder. The difference from example 2 is that the kelp hydrolysate is 5% by weight of the liquid culture substrate.
Example 4
A method for preparing selenium-rich yeast powder. The difference from example 2 is that the kelp hydrolysate is 15% by weight of the liquid culture substrate.
Example 5
A method for preparing selenium-rich yeast powder. The difference from example 2 is that the addition amount of the mixed enzyme is 0.2% of the weight of the kelp pretreatment solution when the kelp hydrolysate is prepared, the enzymolysis temperature is 60 ℃, and the enzymolysis time is 8 hours.
Example 6
A method for preparing selenium-rich yeast powder. The difference from example 2 is that the addition amount of the mixed enzyme is 3% of the weight of the kelp pretreatment liquid, the enzymolysis temperature is 35 ℃ and the enzymolysis time is 2 hours.
It is noted that relational terms such as first and second, and the like are used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Moreover, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or terminal that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or terminal. Without further limitation, an element defined by the statement "comprising … …" or "comprising … …" does not exclude the presence of additional elements in a process, method, article or terminal device comprising the element. Further, herein, "greater than," "less than," "exceeding," and the like are understood to not include the present number; "above", "below", "within" and the like are understood to include this number.
While the embodiments have been described above, other variations and modifications will occur to those skilled in the art once the basic inventive concepts are known, and it is therefore intended that the foregoing description and drawings illustrate only embodiments of the invention and not limit the scope of the invention, and it is therefore intended that the invention not be limited to the specific embodiments described, but that the invention may be practiced with their equivalent structures or with their equivalent processes or with their use directly or indirectly in other related fields.
Claims (4)
1. The preparation method of the selenium-enriched yeast powder is characterized by comprising the following steps:
(1) Adding kelp hydrolysate into the liquid culture medium to prepare fermentation culture solution; the kelp hydrolysate accounts for 5% -15% of the total weight of the liquid culture medium; the liquid culture medium comprises a YEPD culture medium and a sodium selenite solution with the volume of 0.9% of that of the YEPD culture medium, and the concentration of the sodium selenite solution is 100 mg/ml;
(2) Inoculating the selenium-enriched yeast strain into the fermentation culture solution according to 1% -10%, and culturing for 12-36h to obtain selenium-enriched yeast solution;
(3) Spray drying to obtain selenium-enriched yeast powder;
the selenium-enriched yeast strainSaccharomyces cerevisiaeHJC 01001 is deposited in the microorganism strain collection center of Guangdong province, the date of deposit is 2021, 9 and 9, and the deposit number is GDMCC No.61733;
wherein the kelp hydrolysate is prepared by the following method:
(1) Mixing the kelp powder and hydrochloric acid with the concentration of 0.1-0.5mol/L at the weight ratio of 1:10-1:20, fully dissolving, hydrolyzing at 45-60 ℃ for 2-6h, centrifuging at 6000-8000r/min for 5-10min, and obtaining supernatant as kelp pretreatment liquid;
(2) Adjusting pH of the kelp pretreatment liquid to 6.0-7.0, adding mixed enzyme with the addition amount of 0.2-3% of the weight of the kelp pretreatment liquid, and performing enzymolysis at 35-60deg.C for 2-8 hr; the mixed enzyme is alkaline protease, pepsin and trypsin, and the ratio of the alkaline protease to the pepsin to the trypsin is 1:1:1;
(3) And after the enzymolysis is finished, obtaining kelp hydrolysate.
2. The method for preparing selenium-enriched yeast powder of claim 1, wherein the spray-drying inlet air temperature is 130-150 ℃ and the outlet air temperature is 80-85 ℃.
3. A selenium-enriched yeast powder produced by the production process of any one of claims 1-2.
4. A feed additive comprising the selenium-enriched yeast powder of claim 3.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20180015437A (en) * | 2016-08-03 | 2018-02-13 | 네이처퓨어코리아 주식회사 | Composition for Tableting Comprising Natural Excipient |
| CN110801012A (en) * | 2019-10-25 | 2020-02-18 | 运鸿集团股份有限公司 | Active polysaccharide compound nutrient for enhancing immunity and resisting fatigue and preparation method thereof |
| CN112931877A (en) * | 2019-11-26 | 2021-06-11 | 威海紫光科技园有限公司 | Laminarin preparation with antitumor and immunity enhancing effects, and its preparation method |
| KR20210109301A (en) * | 2020-02-27 | 2021-09-06 | 서울대학교산학협력단 | Animal feed additives for substituting antibiotics and preparation method thereof |
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| KR20180015437A (en) * | 2016-08-03 | 2018-02-13 | 네이처퓨어코리아 주식회사 | Composition for Tableting Comprising Natural Excipient |
| CN110801012A (en) * | 2019-10-25 | 2020-02-18 | 运鸿集团股份有限公司 | Active polysaccharide compound nutrient for enhancing immunity and resisting fatigue and preparation method thereof |
| CN112931877A (en) * | 2019-11-26 | 2021-06-11 | 威海紫光科技园有限公司 | Laminarin preparation with antitumor and immunity enhancing effects, and its preparation method |
| KR20210109301A (en) * | 2020-02-27 | 2021-09-06 | 서울대학교산학협력단 | Animal feed additives for substituting antibiotics and preparation method thereof |
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