CN116925938A - Yeast protein powder and preparation method thereof - Google Patents
Yeast protein powder and preparation method thereof Download PDFInfo
- Publication number
- CN116925938A CN116925938A CN202310959726.7A CN202310959726A CN116925938A CN 116925938 A CN116925938 A CN 116925938A CN 202310959726 A CN202310959726 A CN 202310959726A CN 116925938 A CN116925938 A CN 116925938A
- Authority
- CN
- China
- Prior art keywords
- yeast
- protein
- washing
- saccharomyces cerevisiae
- yeast protein
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 108010058643 Fungal Proteins Proteins 0.000 title claims abstract description 85
- 239000000843 powder Substances 0.000 title claims abstract description 46
- 238000002360 preparation method Methods 0.000 title claims abstract description 14
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims abstract description 112
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 claims abstract description 112
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 54
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 43
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 43
- 238000000855 fermentation Methods 0.000 claims abstract description 41
- 230000004151 fermentation Effects 0.000 claims abstract description 41
- 238000000034 method Methods 0.000 claims abstract description 41
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 29
- 150000003839 salts Chemical class 0.000 claims abstract description 14
- 238000001914 filtration Methods 0.000 claims abstract description 13
- 238000004062 sedimentation Methods 0.000 claims abstract description 11
- 102000039446 nucleic acids Human genes 0.000 claims abstract description 10
- 108020004707 nucleic acids Proteins 0.000 claims abstract description 10
- 150000007523 nucleic acids Chemical class 0.000 claims abstract description 10
- 238000004519 manufacturing process Methods 0.000 claims abstract description 8
- 238000001179 sorption measurement Methods 0.000 claims abstract description 6
- 238000005406 washing Methods 0.000 claims description 48
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 38
- 235000013336 milk Nutrition 0.000 claims description 33
- 239000008267 milk Substances 0.000 claims description 33
- 210000004080 milk Anatomy 0.000 claims description 33
- 239000007787 solid Substances 0.000 claims description 30
- 239000007788 liquid Substances 0.000 claims description 27
- 239000001963 growth medium Substances 0.000 claims description 21
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical group [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 18
- 230000001580 bacterial effect Effects 0.000 claims description 17
- 239000000287 crude extract Substances 0.000 claims description 15
- 230000008569 process Effects 0.000 claims description 14
- 239000002244 precipitate Substances 0.000 claims description 13
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 12
- 230000001105 regulatory effect Effects 0.000 claims description 12
- 239000002609 medium Substances 0.000 claims description 11
- 238000011218 seed culture Methods 0.000 claims description 11
- 210000002421 cell wall Anatomy 0.000 claims description 9
- 239000000284 extract Substances 0.000 claims description 9
- 239000003513 alkali Substances 0.000 claims description 8
- 238000012258 culturing Methods 0.000 claims description 8
- 238000001035 drying Methods 0.000 claims description 8
- 239000000706 filtrate Substances 0.000 claims description 8
- 238000010438 heat treatment Methods 0.000 claims description 8
- 238000005119 centrifugation Methods 0.000 claims description 7
- 239000003153 chemical reaction reagent Substances 0.000 claims description 7
- 238000000605 extraction Methods 0.000 claims description 7
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 6
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 6
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical group [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 claims description 6
- LFVGISIMTYGQHF-UHFFFAOYSA-N ammonium dihydrogen phosphate Chemical compound [NH4+].OP(O)([O-])=O LFVGISIMTYGQHF-UHFFFAOYSA-N 0.000 claims description 6
- 229910000387 ammonium dihydrogen phosphate Inorganic materials 0.000 claims description 6
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 6
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 6
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 6
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 6
- 230000000694 effects Effects 0.000 claims description 6
- 238000011068 loading method Methods 0.000 claims description 6
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 6
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 6
- 235000013379 molasses Nutrition 0.000 claims description 6
- 235000019837 monoammonium phosphate Nutrition 0.000 claims description 6
- 239000006012 monoammonium phosphate Substances 0.000 claims description 6
- 238000001694 spray drying Methods 0.000 claims description 6
- 238000009423 ventilation Methods 0.000 claims description 6
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 claims description 6
- 229910000368 zinc sulfate Inorganic materials 0.000 claims description 6
- 229960001763 zinc sulfate Drugs 0.000 claims description 6
- 108090000790 Enzymes Proteins 0.000 claims description 5
- 102000004190 Enzymes Human genes 0.000 claims description 5
- 238000004321 preservation Methods 0.000 claims description 5
- 238000003756 stirring Methods 0.000 claims description 5
- 229940041514 candida albicans extract Drugs 0.000 claims description 4
- 239000012141 concentrate Substances 0.000 claims description 4
- 239000013049 sediment Substances 0.000 claims description 4
- 239000012138 yeast extract Substances 0.000 claims description 4
- RBCOYOYDYNXAFA-UHFFFAOYSA-L (5-hydroxy-4,6-dimethylpyridin-3-yl)methyl phosphate Chemical compound CC1=NC=C(COP([O-])([O-])=O)C(C)=C1O RBCOYOYDYNXAFA-UHFFFAOYSA-L 0.000 claims description 3
- 229930003270 Vitamin B Natural products 0.000 claims description 3
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 3
- 229910000365 copper sulfate Inorganic materials 0.000 claims description 3
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 claims description 3
- 238000009629 microbiological culture Methods 0.000 claims description 3
- JZRWCGZRTZMZEH-UHFFFAOYSA-N thiamine Chemical compound CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N JZRWCGZRTZMZEH-UHFFFAOYSA-N 0.000 claims description 3
- 235000019156 vitamin B Nutrition 0.000 claims description 3
- 239000011720 vitamin B Substances 0.000 claims description 3
- GNFTZDOKVXKIBK-UHFFFAOYSA-N 3-(2-methoxyethoxy)benzohydrazide Chemical compound COCCOC1=CC=CC(C(=O)NN)=C1 GNFTZDOKVXKIBK-UHFFFAOYSA-N 0.000 claims description 2
- FGUUSXIOTUKUDN-IBGZPJMESA-N C1(=CC=CC=C1)N1C2=C(NC([C@H](C1)NC=1OC(=NN=1)C1=CC=CC=C1)=O)C=CC=C2 Chemical compound C1(=CC=CC=C1)N1C2=C(NC([C@H](C1)NC=1OC(=NN=1)C1=CC=CC=C1)=O)C=CC=C2 FGUUSXIOTUKUDN-IBGZPJMESA-N 0.000 claims description 2
- YTAHJIFKAKIKAV-XNMGPUDCSA-N [(1R)-3-morpholin-4-yl-1-phenylpropyl] N-[(3S)-2-oxo-5-phenyl-1,3-dihydro-1,4-benzodiazepin-3-yl]carbamate Chemical compound O=C1[C@H](N=C(C2=C(N1)C=CC=C2)C1=CC=CC=C1)NC(O[C@H](CCN1CCOCC1)C1=CC=CC=C1)=O YTAHJIFKAKIKAV-XNMGPUDCSA-N 0.000 claims description 2
- 239000002585 base Substances 0.000 claims description 2
- 238000002156 mixing Methods 0.000 claims description 2
- 239000005909 Kieselgur Substances 0.000 claims 1
- 241000894006 Bacteria Species 0.000 abstract description 5
- 235000013305 food Nutrition 0.000 abstract description 4
- 238000002703 mutagenesis Methods 0.000 abstract description 4
- 231100000350 mutagenesis Toxicity 0.000 abstract description 4
- 239000002994 raw material Substances 0.000 abstract description 4
- 238000012216 screening Methods 0.000 abstract description 4
- 235000019643 salty taste Nutrition 0.000 abstract description 3
- 230000001953 sensory effect Effects 0.000 abstract description 3
- 239000000049 pigment Substances 0.000 abstract description 2
- 238000005352 clarification Methods 0.000 abstract 1
- 235000018102 proteins Nutrition 0.000 description 37
- 239000000047 product Substances 0.000 description 13
- 239000000243 solution Substances 0.000 description 13
- 235000019640 taste Nutrition 0.000 description 9
- 235000001014 amino acid Nutrition 0.000 description 7
- 150000001413 amino acids Chemical class 0.000 description 7
- 230000001276 controlling effect Effects 0.000 description 7
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 6
- 239000006228 supernatant Substances 0.000 description 5
- 238000004806 packaging method and process Methods 0.000 description 4
- 238000007670 refining Methods 0.000 description 4
- 239000008223 sterile water Substances 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 241001465754 Metazoa Species 0.000 description 3
- 239000007864 aqueous solution Substances 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 3
- 235000013339 cereals Nutrition 0.000 description 3
- 239000011248 coating agent Substances 0.000 description 3
- 238000000576 coating method Methods 0.000 description 3
- 150000004676 glycans Chemical class 0.000 description 3
- 230000007935 neutral effect Effects 0.000 description 3
- 229910052757 nitrogen Inorganic materials 0.000 description 3
- 229920001282 polysaccharide Polymers 0.000 description 3
- 239000005017 polysaccharide Substances 0.000 description 3
- 230000001954 sterilising effect Effects 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- 239000012224 working solution Substances 0.000 description 3
- WTLKTXIHIHFSGU-UHFFFAOYSA-N 2-nitrosoguanidine Chemical compound NC(N)=NN=O WTLKTXIHIHFSGU-UHFFFAOYSA-N 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 229920000392 Zymosan Polymers 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- 235000021120 animal protein Nutrition 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 238000007865 diluting Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000001704 evaporation Methods 0.000 description 2
- 230000008020 evaporation Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000011049 filling Methods 0.000 description 2
- 239000000796 flavoring agent Substances 0.000 description 2
- 235000019634 flavors Nutrition 0.000 description 2
- 239000005556 hormone Substances 0.000 description 2
- 229940088597 hormone Drugs 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 239000000447 pesticide residue Substances 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- FYGDTMLNYKFZSV-URKRLVJHSA-N (2s,3r,4s,5s,6r)-2-[(2r,4r,5r,6s)-4,5-dihydroxy-2-(hydroxymethyl)-6-[(2r,4r,5r,6s)-4,5,6-trihydroxy-2-(hydroxymethyl)oxan-3-yl]oxyoxan-3-yl]oxy-6-(hydroxymethyl)oxane-3,4,5-triol Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1[C@@H](CO)O[C@@H](OC2[C@H](O[C@H](O)[C@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O FYGDTMLNYKFZSV-URKRLVJHSA-N 0.000 description 1
- 241000251468 Actinopterygii Species 0.000 description 1
- 229920002498 Beta-glucan Polymers 0.000 description 1
- 239000002028 Biomass Substances 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 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 1
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 201000005569 Gout Diseases 0.000 description 1
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 1
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 1
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-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
- 108010084695 Pea Proteins Proteins 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 1
- 108010073771 Soybean Proteins Proteins 0.000 description 1
- 241000209140 Triticum Species 0.000 description 1
- 235000021307 Triticum Nutrition 0.000 description 1
- LEHOTFFKMJEONL-UHFFFAOYSA-N Uric Acid Chemical compound N1C(=O)NC(=O)C2=C1NC(=O)N2 LEHOTFFKMJEONL-UHFFFAOYSA-N 0.000 description 1
- TVWHNULVHGKJHS-UHFFFAOYSA-N Uric acid Natural products N1C(=O)NC(=O)C2NC(=O)NC21 TVWHNULVHGKJHS-UHFFFAOYSA-N 0.000 description 1
- 108010046377 Whey Proteins Proteins 0.000 description 1
- 102000007544 Whey Proteins Human genes 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- 235000004279 alanine Nutrition 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 235000003704 aspartic acid Nutrition 0.000 description 1
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 229940098773 bovine serum albumin Drugs 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 239000010949 copper Substances 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 230000002354 daily effect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 235000019621 digestibility Nutrition 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 235000013601 eggs Nutrition 0.000 description 1
- 239000003797 essential amino acid Substances 0.000 description 1
- 235000020776 essential amino acid Nutrition 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
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- 239000012467 final product Substances 0.000 description 1
- 235000019688 fish Nutrition 0.000 description 1
- 235000011194 food seasoning agent Nutrition 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 235000013922 glutamic acid Nutrition 0.000 description 1
- 239000004220 glutamic acid Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
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- 230000031700 light absorption Effects 0.000 description 1
- 239000012263 liquid product Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 208000030159 metabolic disease Diseases 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
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- 230000000050 nutritive effect Effects 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 235000019702 pea protein Nutrition 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 230000002980 postoperative effect Effects 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 238000000751 protein extraction Methods 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 239000010802 sludge Substances 0.000 description 1
- 235000019710 soybean protein Nutrition 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
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- 229940116269 uric acid Drugs 0.000 description 1
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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
- C12N1/18—Baker's yeast; Brewer's yeast
- C12N1/185—Saccharomyces isolates
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
- A23J1/18—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from yeasts
-
- 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
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/195—Proteins from microorganisms
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/145—Extraction; Separation; Purification by extraction or solubilisation
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/34—Extraction; Separation; Purification by filtration, ultrafiltration or reverse osmosis
-
- 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
- C12N1/18—Baker's yeast; Brewer's yeast
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
-
- 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
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
- C12R2001/85—Saccharomyces
- C12R2001/865—Saccharomyces cerevisiae
-
- 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
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/10—Biofuels, e.g. bio-diesel
Abstract
The invention discloses yeast protein powder and a preparation method thereof, and belongs to the technical field of protein preparation. The invention obtains a strain of Saccharomyces cerevisiae with high protein yield through mutagenesis screening, the concentration of dry bacteria after fermentation culture is more than 50g/L, the protein content is more than 60%, and a very good strain is provided for the preparation of the yeast protein powder. Then removing yeast nucleic acid by a concentrated salt method, improving the content of yeast protein by an enzymolysis-thermokalite extraction-isoelectric point sedimentation combined method, improving the yield, reducing the cost, and removing peculiar smell and pigment by diatomite filtration-activated carbon adsorption clarification solution; the fresh salty taste is removed by the high-speed disc separator, and the yeast protein powder product with high protein content, good sensory state and low production cost is finally obtained, thereby providing a new protein raw material for the fields of food and health-care food.
Description
Technical Field
The invention belongs to the technical field of protein preparation, and particularly relates to yeast protein powder and a preparation method thereof.
Background
Proteins are important components of all cells and tissues of a human body, and play a role in the whole life process. The types of proteins are very large, and the proteins can be divided into complete proteins and incomplete proteins by whether the proteins contain essential amino acids, wherein the complete proteins comprise milk, eggs, fish, meat, yeast, soybean and the like, and the proteins are eaten by a plurality of people, so that the proteins are beneficial to the growth and development of teenagers, the prenatal and postnatal care of pregnant women and the health and longevity of the old. The incomplete protein includes cereal, wheat, corn, gelatin in animal skin bone, etc., and plays an indispensable role in maintaining human life.
People can ensure the normal and healthy operation of the body only by taking a certain amount of protein every day in daily diet. With the gradual improvement of living standard, people have higher and higher demands on protein types and quality, especially for some fitness and weight-losing people and some postoperative patients. At present, most of protein powder in the market mainly comprises animal raw materials or plant raw materials, and microbial protein products are fresh. The digestibility and the nutritive value of plant source proteins such as soybean protein and pea protein are not as good as those of animal proteins such as whey protein, and the safety risks such as pesticide residues and the like exist at the same time, while the animal proteins can not meet the requirements of vegetarian people, and the residual risks such as antibiotics and hormone also exist at the same time. So many scientific teams now focus on yeast proteins.
Yeast contains a complete amino acid group, including 8 amino acids essential to the human body, especially lysine, which is less in cereal proteins, and higher in yeasts. In addition, the proportion of amino acid in yeast is close to the ideal amino acid composition value recommended by the national grain and agricultural organization (FAO), and the digestion utilization rate can reach more than 90%, so that the nutritional value is higher. Meanwhile, the high-density liquid fermentation technology used by the yeast protein is adopted, the production process is carried out in a modern factory, the occupied cultivated land is small, the emission is small, the production efficiency is high, and the novel protein is environment-friendly and sustainable. In addition, yeast protein is a safe protein product which is not of animal origin and has no risk of pesticide residue and hormone residue. Although some yeast protein powder has been disclosed in the prior art, the existing yeast protein powder has the problems of low purity of yeast protein, heavy taste, bad smell, poor solubility, uneven quality and the like, and research and development of high-quality yeast protein products become an important subject to be researched at present.
Disclosure of Invention
In view of the above, the invention aims to provide a yeast protein powder and a preparation method thereof, which solve the problems of low purity, heavy taste, bad smell, bad solubility and the like of the existing yeast protein powder, obtain the yeast protein powder with high protein content and good sensory state, and provide a new protein raw material for the fields of food and health-care food.
The invention aims at realizing the following steps:
the invention provides a saccharomyces cerevisiae (Saccharomyces cerevisiae) ZAB 02 which is preserved in China general microbiological culture Collection center (CGMCC No. 27288) in the 05 th month 08 of 2023, wherein the preservation number is the third national institute of advanced technology, north Chen, west Lu, beijing, and the preservation unit address is the North Chan of the Korean region.
The invention also provides a preparation method of the yeast protein powder, which mainly comprises the following steps:
(1) The saccharomyces cerevisiae ZAB 02 is subjected to expansion culture in a seed culture medium, inoculated into a fermentation culture medium and fermented to obtain the saccharomyces cerevisiae with high protein yield;
(2) Removing yeast nucleic acid from the high-protein saccharomyces cerevisiae obtained in the step (1) by a concentrated salt method, performing enzymolysis on cell walls, extracting proteins, performing isoelectric point sedimentation, filtering by diatomite, adsorbing by activated carbon, and drying to obtain yeast protein powder.
Based on the above technical solution, further, step (1) includes the following steps:
1) Inoculating Saccharomyces cerevisiae ZAB 02 into a liquid seed culture medium, and culturing at 25-35 ℃ for 10-30 h under ventilation and stirring to obtain seed bacterial liquid;
2) Inoculating the seed bacterial liquid obtained in the step 1) into a fermentation culture medium, fermenting at 25-35 ℃, and carrying out ventilation stirring fermentation to obtain the saccharomyces cerevisiae with high protein yield.
Based on the technical scheme, the pH value in the step 1) and the step 2) is further controlled to be 4.5-5.5.
Based on the technical scheme, the liquid seed culture medium further comprises the following components: 70-90g/L of molasses, 2-3g/L of ammonium sulfate, 0.1-0.5g/L of monoammonium phosphate, 0.05-0.3g/L of magnesium sulfate, 0.1-0.5g/L of copper sulfate, 0.01-0.1g/L of zinc sulfate and 0.5-5g/L of yeast extract; the fermentation medium comprises the following components: 150-300g/L molasses, 15-30g/L ammonium sulfate, 2-8g/L monoammonium phosphate, 0.5-3g/L magnesium sulfate, 0.05-0.3g/L zinc sulfate, vitamin B 1 0.01-0.1g/L vitamin B 6 0.01-0.1g/L vitamin B 7 0.001-0.01g/L。
Based on the technical scheme, further, the fermentation mode in the step 2) is any one of batch fermentation, semi-continuous fermentation and continuous fermentation.
Based on the above technical solution, further, step (2) includes the following steps:
a. preparing high-protein yeast milk: preparing Saccharomyces cerevisiae ZAB 02 obtained in the step (1) into yeast milk with a solid content of 12-18% by using process water;
b. and (3) removing RNA by a concentrated salt method: adding edible salt accounting for 8-14% of the weight of the yeast milk, heating to 80-95 ℃ and preserving heat for 2-6h;
c. separating and washing: centrifuging to separate yeast milk, washing the obtained precipitate with water, centrifuging, and collecting heavy phase;
d. cell wall enzymolysis: preparing yeast milk with 6-10% of solid content by using process water, adjusting pH to 4.0-6.0, adding yeast glucanase with 0.1-2.0% of yeast dry weight, and performing enzymolysis for 4-24h at 30-65deg.C;
e. and (3) hot alkali extraction: adding strong alkali accounting for 0.2% -1.0% of the weight of the yeast milk, heating to 85-95 ℃ and preserving heat for 1-5h;
f. separating and washing: centrifuging to separate yeast milk, collecting light phase, washing heavy phase with water, centrifuging, collecting light phase, washing heavy phase for 1-5 times, and mixing all light phases;
g. isoelectric point sedimentation: regulating the pH of the combined light phases to 4.0-5.0, and standing for 1-5h;
h. separating and washing: centrifuging to separate the precipitate, washing the precipitate with water, centrifuging to obtain heavy phase, washing the heavy phase for 1-5 times, and collecting the washed heavy phase to obtain yeast protein crude extract;
i. and (3) diatomite filtration: regulating the solid content of the yeast protein crude extract to 6% -10%, regulating the pH value to 7.5-9.0, precoating diatomite on a diatomite filter, and filtering the yeast protein crude extract;
j. activated carbon column adsorption: loading the diatomite filtered filtrate to an activated carbon column to obtain a yeast protein refined extract;
k. concentrating: concentrating the yeast protein refined extract to a solid content of 20-40% to obtain a yeast protein concentrated solution;
and I, drying: drying the yeast protein concentrate to obtain yeast protein powder.
Based on the technical scheme, further, the rotating speed of centrifugation in the step c is 5000-8000 rpm, and the mass ratio of the yeast milk to the washing water is 1:1.5-1:2.
Based on the technical scheme, the enzymolysis condition in the step d is that the enzymolysis is carried out for 6-12 hours at 40-60 ℃, and the enzyme activity of the yeast glucanase is 10000-120000U/g.
Based on the above technical scheme, further, the strong base in step e is sodium hydroxide or potassium hydroxide.
Based on the technical scheme, further, the rotating speed of the centrifugation in the step f is 5000-8000 rpm, and the mass ratio of the heavy phase to the washing water is 1:0.5-1:1.
Based on the technical scheme, the reagent used for adjusting the pH in the step g is any one of hydrochloric acid, phosphoric acid and citric acid.
Based on the technical scheme, further, the rotational speed of centrifugation in the step h is 5000-8000 rpm, and the mass ratio of the precipitate to the washing water is 1:0.2-1:1.5.
Based on the technical scheme, the diatomite dosage in the step i is 0.1-0.5 times of the solid weight of the yeast protein crude extract.
Based on the technical proposal, the dosage of the activated carbon in the step j is 0.5 to 2 times of the weight of the solid matters in the filtrate filtered by the diatomite; the diameter-to-height ratio of the activated carbon column is 1:1-1:5, and the flow rate of the activated carbon column per hour is 20-60% of the effective activated carbon volume.
Based on the technical scheme, further, the concentration in the step k is performed by using a triple effect evaporator.
Based on the above technical solution, further, the drying in step l is performed by using pressure spray drying.
The invention also provides the yeast protein powder prepared by the preparation method.
Compared with the prior art, the invention has the following beneficial effects:
1. the invention obtains a strain of Saccharomyces cerevisiae with high protein yield through mutagenesis screening, the concentration of dry bacteria after fermentation culture is more than 50g/L, the protein content is more than 60%, and a very good strain is provided for the preparation of the yeast protein powder.
2. The method adopts a concentrated salt method to remove the nucleic acid in the yeast, the nucleic acid removal rate can reach more than 90 percent, and the nucleic acid content in the yeast protein powder is less than or equal to 0.5 percent after further refining, so that the method is suitable for ventilation people to eat.
3. The invention adopts the methods of enzymolysis, thermokalite extraction, isoelectric point sedimentation and the like to ensure that the protein extraction rate reaches more than 85 percent, the protein content reaches more than 85 percent, and the invention has larger promotion than the common 70 percent yeast protein powder on the market.
4. According to the invention, diatomite is adopted for filtration and large-particle activated carbon adsorption, so that the yeast smell of the product is greatly weakened, and meanwhile, insoluble impurities are removed, so that the product is completely soluble, and a 5% solution state is clear and transparent, and the problems of turbidity and a large amount of precipitation after dissolution are solved.
5. The invention adopts the high-speed disc separator to wash for multiple times, so that the amino acid nitrogen of the high-protein yeast is less than or equal to 0.8 percent, the ash content is less than or equal to 5 percent, the product takes macromolecular protein as the main component, and the whole product has neutral taste, thereby solving the problem of heavy fresh salty taste of the yeast protein powder product.
Drawings
In order to more clearly illustrate the embodiments of the present invention, the drawings to which the embodiments relate will be briefly described.
FIG. 1 is an evaluation of stability of 6 strains of Saccharomyces cerevisiae having a high protein content obtained in example 1.
FIG. 2 is a photograph of Saccharomyces cerevisiae ZAB 02 after slant culture in YPD solid medium.
FIG. 3 is a photograph of the yeast morphology of Saccharomyces cerevisiae ZAB 02 after culture in liquid medium.
FIG. 4 is a graph showing the concentration of cells and the protein content during fermentation in a 500L fermenter.
FIG. 5 is a photograph of 5% aqueous solution of yeast protein powder in example 3.
Detailed Description
The following detailed description of the invention is provided in connection with examples, but the implementation of the invention is not limited thereto, and it is obvious that the examples described below are only some examples of the invention, and that it is within the scope of protection of the invention to those skilled in the art to obtain other similar examples without inventive faculty.
Unless otherwise indicated, the instruments, reagents, materials, etc. according to the present embodiment are conventional instruments, reagents, materials, etc. existing in the prior art, and are commercially available.
The formulation of the medium used in the examples is as follows:
(1) YPD solid medium: 20g/L peptone, 20g/L glucose, 10g/L yeast extract and 20g/L agar powder, sterilizing at 115deg.C for 15 min under high pressure, cooling, and pouring into a plate under aseptic condition.
(2) Liquid seed medium: molasses 78.7g/L, ammonium sulfate 2.6g/L, monoammonium phosphate 0.3g/L, magnesium sulfate 0.15g/L, copper sulfate 0.31g/L, zinc sulfate 0.03g/L, yeast extract 1g/L; sterilizing at 115 deg.c for 15 min.
(3) Liquid fermentation medium: 215g/L molasses, 20g/L ammonium sulfate, 4g/L monoammonium phosphate, 1g/L magnesium sulfate, 0.1g/L zinc sulfate and vitamin B 1 0.06g/L vitamin B 6 0.03g/L, vitamin B 7 0.003g/L; sterilizing at 115 deg.c for 15 min.
The analytical method for protein content in Saccharomyces cerevisiae in the examples is as follows:
preparing a standard curve: protein content determination is carried out by adopting a BCA protein concentration determination kit, BCA working solution is prepared according to the quantity of standard substances and samples according to the ratio of BCA reagent A to BCA reagent B (Cu reagent) of 50:1 by volume, and is fully and uniformly mixed, 20 mu L of 5mg/mL BSA standard substance is diluted to 100 mu L by PBS buffer solution, so that the final concentration is 1mg/mL; adding standard substances into centrifuge tubes according to 0, 20, 40, 60, 80 and 100 mu L, adding PBS to make up to 100 mu L, taking 30 mu L to sample holes of 96-well plates, respectively adding 200 mu L BCA working solution into each hole, standing at 37deg.C for 30min, and measuring OD with enzyme-labeled instrument 562 Linear fitting is carried out by taking the protein concentration (mg/mL) as an abscissa (x) and the light absorption value as an ordinate (y) to obtain the proteinStandard curve y=0.2517x+0.1058 (based on bovine serum albumin BSA), linear correlation coefficient R 2 ＝0.9996。
Protein content determination in Saccharomyces cerevisiae: taking 5mL of fermentation bacteria liquid, centrifuging for 10min at 5000r/min, discarding supernatant, weighing wet weight of bacteria, suspending bacteria with 5mL of physiological saline, performing ultrasonic crushing, centrifuging for 10min at 5000r/min, taking supernatant, diluting 100 times, adding 20 mu L of supernatant into a sample hole of a 96-well plate, adding 200 mu LBCA working solution, standing for 30min at 37 ℃, and measuring OD by an enzyme-labeling instrument 562 And calculating the protein concentration according to a standard curve, and further calculating the protein content in the saccharomyces cerevisiae. The protein content in Saccharomyces cerevisiae is calculated as mass fraction omega in mg/g, and repeated six times according to the following formula.
ω＝m*v*100/(M*R)
m-calculating the protein concentration in the sample measurement solution from a standard curve, wherein the unit is mg/mL;
v-total volume of supernatant in mL;
100-dilution times;
m is the wet weight of the thallus, and the unit is g;
r-dry-wet weight ratio of yeasts.
Example 1
The original strain in this embodiment is Saccharomyces cerevisiae used in a production workshop, and Saccharomyces cerevisiae capable of producing protein with high yield is obtained by mutagenesis and screening, and the main process is as follows:
(1) Inoculating the original strain to liquid seed culture medium, shake culturing at 28-32deg.C and 150-180 rpm for 18 hr, centrifuging to obtain bacterial precipitate, washing with PBS, re-suspending with sterile water, and diluting the bacterial solution to 1×10 7 Adding 200 mu L of CFU/mL into a sterile plate, uniformly coating, carrying out ultraviolet irradiation mutagenesis treatment on the bacterial suspension under a 30W ultraviolet lamp (the distance is 30 cm), controlling the death rate to be about 90%, then coating the bacterial suspension on a solid culture medium under a sterile environment, placing the solid culture medium in a dark environment, culturing the solid culture medium in an incubator at 28-32 ℃ for 48 hours in an inversion way, and selecting a bacterial strain with good growth;
(2) Inoculating the strains selected in step (1) with good growthInoculating to liquid seed culture medium, waiting for bacterial liquid OD 600 After reaching about 2, the bacterial precipitate is obtained by centrifugation, PBS is used for washing, sterile water is used for resuspension, and the bacterial suspension is diluted to 1 multiplied by 10 10 Placing CFU/mL into a centrifuge tube, adding nitrosoguanidine to make the final concentration of nitrosoguanidine be 0.6mg/mL, then oscillating for 25min, 35min and 45min at 28-32 ℃ and 150-180 rpm, washing with PBS after sampling at each time point, resuspending with an equal volume of sterile water, taking 200 mu L of the solution under a sterile environment, adding the solution into a YPD solid culture medium plate, uniformly coating, inversely culturing for 48h in an incubator at 28-32 ℃, and selecting a strain with good growth;
(3) Inoculating the strains selected in the step (2) with good growth into 10mL of liquid seed culture medium, culturing for 24 hours at the temperature of 28-32 ℃ and at the speed of 150-180 rpm, transferring the strains into 50mL of liquid fermentation culture medium with the inoculum size of 10%, culturing for 24 hours at the temperature of 28-32 ℃ and at the speed of 150-180 rpm, detecting the protein content in the fermented saccharomyces cerevisiae, and reserving 6 strains with higher protein content, wherein the strains are respectively numbered as S1, S2, S3, S4, S5 and S6;
(4) The strains with higher protein content obtained by screening in the step (3) are respectively transferred into a liquid fermentation culture medium according to the inoculation amount of 10 percent, the culture is carried out for 24 hours under the conditions of 28-32 ℃ and 150-180 rpm, the obtained bacterial liquid is transferred into a new liquid fermentation culture medium, the operation is repeated for 10 times, the protein content in the saccharomyces cerevisiae obtained by each fermentation is detected, the result is shown in figure 1 (three groups of parallel experiments are carried out), and the highest protein content and stable content of the strain S5 can be seen from figure 1.
The strain D is named ZAB 02 and is preserved in China general microbiological culture Collection center (CGMCC) at the date 08 of 2023, 05 month, and the preservation number is CGMCC No.27288 and the preservation unit address is Beichen Xiyun one No. three in the Chaiyang area of Beijing city.
Saccharomyces cerevisiae ZAB 02 has the following characteristics:
on the wort agar slope, the colonies were milky, round, with clean edges, flat and glossy, as shown in FIG. 2.
After 24 hours of culture in liquid medium (30 ℃), the cells were oval or spherical in shape and had a size of 2 to 6. Mu.m.times.5 to 15. Mu.m, as shown in FIG. 3.
Example 2
The Saccharomyces cerevisiae ZAB 02 obtained in example 1 was used for fermentation culture, and the specific procedure was as follows:
(1) Inoculating the Saccharomyces cerevisiae ZAB 02 screened in the embodiment 1 into 1000mL of liquid seed culture medium after enlarging culture, carrying out shaking culture for 24 hours at 28-32 ℃ under the condition of 150-180 rpm, transferring into a 30L fermentation tank filled with the liquid seed culture medium, controlling the fermentation temperature to be 28-32 ℃, controlling the tank pressure to be 0.5-1kPa, controlling the pH value to be 4.6-5.0, and carrying out ventilation stirring culture for 24 hours to obtain seed bacterial liquid;
(2) Inoculating the seed bacterial liquid obtained in the step (1) to a 500L fermentation tank filled with 300L liquid fermentation medium, controlling the fermentation temperature to be 28-32 ℃, controlling the pressure of the fermentation tank to be 5-10kPa, controlling the pH value to be 4.6-5.0, and stopping fermentation after culturing for 20 hours.
In the fermentation process in a 500L fermentation tank, 100mL is sampled every 2h, 5000r/min is centrifuged for 10min, supernatant is discarded, wet bacterial sludge is resuspended by 100mL sterile water, 5000r/min is centrifuged for 10min, the obtained precipitate is dried in a far infrared oven at 120 ℃ for 2h, biomass is measured, the detection results of the thallus concentration of saccharomyces cerevisiae ZAB 02 and the protein content in saccharomyces cerevisiae ZAB 02 in the fermentation process are shown in figure 4 (three groups of parallel experiments), according to the growth curve of saccharomyces cerevisiae shown in figure 4, the propagation speed of saccharomyces cerevisiae ZAB 02 is obviously accelerated in the incubation period at 0-4h at the beginning of fermentation, the fermentation is in the logarithmic growth period at 8-16h, the growth speed of saccharomyces cerevisiae ZAB 02 is obviously accelerated in the slow period at 16-20h, the dry bacterial concentration is up to 52.9g/L, and the protein content of saccharomyces cerevisiae ZAB 02 is up to 60.7%.
Example 3
The embodiment provides a preparation method of yeast protein powder, which adopts Saccharomyces cerevisiae ZAB 02 prepared by fermentation in the embodiment 2, and the specific process is as follows:
a. preparing high-protein yeast milk: the Saccharomyces cerevisiae ZAB 02 collected in example 2 was prepared into a yeast milk with a solids content of 12% using process water;
b. and (3) removing RNA by a concentrated salt method: adding edible salt accounting for 8% of the weight of the yeast milk, heating to 95 ℃ and preserving heat for 4 hours;
c. separating and washing: separating yeast milk by using a high-speed disc centrifuge (6000 rpm), adding water for washing, wherein the mass ratio of the yeast milk to the washing water is 1:1.5, centrifuging, taking a separated heavy phase, and refining yeast RNA by a light phase;
d. cell wall enzymolysis: preparing yeast milk with 6% solid content by using process water, adjusting pH to 4.0, adding yeast glucanase with 1.0% dry weight of yeast (enzyme activity is 100000U/g, and performing enzymolysis at 40deg.C for 10 hr;
e. and (3) hot alkali extraction: adding sodium hydroxide accounting for 0.2 percent of the weight of the yeast milk, heating to 95 ℃ and preserving heat for 3 hours;
f. separating and washing: separating yeast milk by using a high-speed disc centrifuge (6000 rpm), taking a light phase, washing a heavy phase by adding water, wherein the mass ratio of the heavy phase to the washing water is 1:0.5, centrifuging, taking the light phase, combining all the light phases, and producing zymosan by the heavy phase;
g. isoelectric point sedimentation: adjusting the pH of the combined light phases to 4.0 by hydrochloric acid, and standing for 2h;
h. separating and washing: separating the settled solution by using a high-speed disc centrifuge (6000 rpm), adding water for washing, wherein the mass ratio of the precipitate to the washing water is 1:0.5, centrifuging, taking a heavy phase, washing the obtained heavy phase for 1 time, the mass ratio of the heavy phase to the washing water is 1:1.5, centrifuging, collecting the washed heavy phase to obtain a yeast protein crude extract, and carrying out the next procedure;
i. and (3) diatomite filtration: regulating the solid content of the yeast protein crude extract to 6%, regulating the pH to 7.5 by using sodium hydroxide, precoating diatomite on a diatomite filter, filtering the yeast protein crude extract, wherein the diatomite dosage is 0.15 times of the solid weight of the yeast protein crude extract;
j. activated carbon column adsorption: loading the diatomite filtered filtrate into an activated carbon column (2-4 columns with the same specification are connected in series), wherein the column loading flow rate per hour is 20% of the volume of the effective activated carbon; the dosage of the activated carbon is 0.5 time of the weight of the solid matters in the diatomite filtrate; the diameter-to-height ratio of the activated carbon after column filling is 1:2, and the yeast protein refined extract is obtained;
k. three-effect evaporation: concentrating the yeast protein refined extract by using a triple-effect evaporator until the solid content reaches 25%;
and l, spray drying: processing the yeast protein concentrate into yeast protein powder with a pressure spray drying tower, wherein the moisture is less than or equal to 8%.
And m, packaging: and packaging the yeast protein powder according to the required specification in a clean area.
According to the consumption of the saccharomyces cerevisiae ZAB 02 and the yield of the obtained yeast protein powder, the protein yield of the method for preparing the yeast protein powder of the embodiment is calculated to be as high as 85.7%, the protein content in the yeast protein powder is as high as 87.8%, the content of RNA is only 0.5%, the aqueous solution containing the 5% yeast protein powder prepared by the method is clear and transparent, no precipitation occurs (figure 5), the amino acid nitrogen in the yeast protein powder is less than or equal to 0.8%, the ash content is less than or equal to 5%, the product takes macromolecular protein as the main component, and the whole product has neutral taste.
Example 4
The embodiment provides a preparation method of yeast protein powder, which adopts Saccharomyces cerevisiae ZAB 02 prepared by fermentation in the embodiment 2, and the specific process is as follows:
a. preparing high-protein yeast milk: the Saccharomyces cerevisiae ZAB 02 collected in example 2 was prepared into a yeast milk with a solids content of 18% using process water;
b. and (3) removing RNA by a concentrated salt method: adding edible salt accounting for 14% of the weight of the yeast milk, heating to 90 ℃, and preserving heat for 3 hours;
c. separating and washing: separating yeast milk by using a high-speed disc centrifuge (6000 rpm), adding water for washing, wherein the mass ratio of the yeast milk to the washing water is 1:2, centrifuging, taking a separated heavy phase, and refining yeast RNA by using a light phase;
d. cell wall enzymolysis: preparing yeast milk with solid content of 10% by weight with process water, adjusting pH to 4.5, adding yeast glucanase with dry weight of 0.5% (enzyme activity of 100000U/g), and performing enzymolysis at 60deg.C for 8 hr;
e. and (3) hot alkali extraction: adding sodium hydroxide accounting for 0.4 percent of the weight of the yeast milk, heating to 85 ℃ and preserving heat for 2 hours;
f. separating and washing: separating yeast milk by using a high-speed disc centrifuge (6000 rpm), taking a light phase, washing a heavy phase by adding water, wherein the mass ratio of the heavy phase to the washing water is 1:1, centrifuging, taking the light phase, washing the heavy phase for 3 times, combining all the light phases, and producing zymosan by the heavy phase;
g. isoelectric point sedimentation: regulating the pH of the combined light phases to 5.0 by phosphoric acid, and standing for 3h;
h. separating and washing: separating the settled solution by using a high-speed disc centrifuge (6000 rpm), adding water for washing, wherein the mass ratio of the precipitate to the washing water is 1:0.2, centrifuging, taking a heavy phase, washing the obtained heavy phase for 3 times, the mass ratio of the heavy phase to the washing water is 1:1.0, centrifuging, collecting the washed heavy phase to obtain a yeast protein crude extract, and carrying out the next procedure;
i. and (3) diatomite filtration: regulating the solid content of the yeast protein crude extract to 10%, regulating the pH to 9.0 by using sodium hydroxide, precoating diatomite on a diatomite filter, filtering the yeast protein crude extract, wherein the diatomite dosage is 0.15 times of the solid weight of the yeast protein crude extract;
j. activated carbon column adsorption: loading the diatomite filtered filtrate into an activated carbon column (2-4 columns with the same specification are connected in series), wherein the column loading flow rate per hour is 60% of the volume of the effective activated carbon; the dosage of the activated carbon is 2 times of the weight of the solid matters in the diatomite filtrate; the diameter-to-height ratio of the activated carbon after column filling is 1:5, and the yeast protein refined extract is obtained;
k. three-effect evaporation: concentrating the yeast protein refined extract by using a triple-effect evaporator until the solid content reaches 35%;
and l, spray drying: processing the yeast protein concentrate into yeast protein powder with a pressure spray drying tower, wherein the moisture is less than or equal to 5%.
And m, packaging: and packaging the yeast protein powder according to the required specification in a clean area.
According to the consumption of the saccharomyces cerevisiae ZAB 02 and the yield of the obtained yeast protein powder, the protein yield of the method for preparing the yeast protein powder is calculated to be 86.3 percent, the protein content in the yeast protein powder is up to 87.1 percent, the RNA content is only 0.4 percent, the aqueous solution dissolved with 5 percent of the yeast protein powder is clear and transparent, no sediment exists, the amino acid nitrogen in the yeast protein powder is less than or equal to 0.8 percent, the ash content is less than or equal to 5 percent, the product takes macromolecular protein as the main component, and the whole product has neutral taste.
In conclusion, the yeast nucleic acid is removed by using a concentrated salt method, the nucleic acid removal rate can reach more than 90%, and the final product has the RNA content of less than 0.5% after further refining, so that metabolic disorder caused by the increase of uric acid level after the nucleic acid substances are decomposed in vivo after eating is avoided, and the yeast protein powder disclosed by the invention is suitable for gout people.
The existing yeast protein powder has low protein purity, mainly contains more yeast polysaccharide components, and the yeast polysaccharide is mainly yeast beta-glucan and yeast mannooligosaccharide, which are high molecular polysaccharide, and have poor solubility, so that the yeast protein is in a turbid state after being dissolved, and a large amount of precipitate exists at the same time, so that the yeast protein powder is limited to be used in liquid products such as drinks. The invention adopts the combined treatment method of enzymolysis cell wall, hot alkali extraction and isoelectric point sedimentation, firstly, the cell wall structure of the yeast is destroyed by enzymolysis, then the cell wall components are removed, and then the hot alkali extraction and isoelectric point sedimentation method are carried out, so that the protein in the yeast is directly extracted, the yield and the purity of the protein are greatly improved, the protein content can reach more than 85 percent, and the method has no sediment and is completely soluble.
The yeast has strong characteristic smell and darker color, and the smell of the yeast is not completely removed after the yeast protein is extracted because the yeast generally has obvious characteristic flavor, so that the flavor of the yeast protein is poor, the subsequent seasoning trouble of the end product is caused, and the use of the yeast protein is influenced.
The yeast contains rich glutamic acid, aspartic acid, alanine and other taste amino acids and taste peptides, so that the yeast has heavy taste and strong fresh, salty, astringent and other tastes.
The invention removes yeast nucleic acid by a concentrated salt method, improves the content of yeast protein by an enzymolysis-thermokalite extraction-isoelectric point sedimentation combined method, improves the yield, reduces the cost and removes the sediment; filtering with diatomite, adsorbing with activated carbon to clarify the solution, and removing odor and pigment; the fresh salty taste is removed by a high-speed disc separator, and the yeast protein powder product with high protein content, good sensory state and low production cost is finally obtained.
Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present invention, and not for limiting the same; although the invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some or all of the technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit of the invention.
Claims (10)
1. A strain of Saccharomyces cerevisiae (Saccharomyces cerevisiae) ZAB 02 is characterized in that the strain is preserved in China general microbiological culture Collection center (CGMCC No. 27288) in the 05 th month 08 of 2023, and the preservation unit address is the first national institute of advanced North Chen, west Lu, beijing city.
2. The preparation method of the yeast protein powder is characterized by mainly comprising the following steps:
(1) Inoculating the saccharomyces cerevisiae ZAB 02 of claim 1 into a fermentation medium after being subjected to expansion culture in a seed culture medium, and fermenting to obtain the saccharomyces cerevisiae with high protein yield;
(2) Removing yeast nucleic acid from the high-protein saccharomyces cerevisiae obtained in the step (1) by a concentrated salt method, performing enzymolysis on cell walls, extracting proteins, performing isoelectric point sedimentation, filtering by diatomite, adsorbing by activated carbon, and drying to obtain yeast protein powder.
3. The method of manufacturing according to claim 2, wherein step (1) comprises the steps of:
1) Inoculating Saccharomyces cerevisiae ZAB 02 into a liquid seed culture medium, and culturing at 25-35 ℃ for 10-30 h under ventilation and stirring to obtain seed bacterial liquid;
2) Inoculating the seed bacterial liquid obtained in the step 1) into a fermentation culture medium, fermenting at 25-35 ℃, and carrying out ventilation stirring fermentation to obtain the saccharomyces cerevisiae with high protein yield.
4. The method according to claim 3, wherein the pH value in step 1) and step 2) is controlled to be 4.5 to 5.5; the liquid seed culture medium comprises the following components: 70-90g/L of molasses, 2-3g/L of ammonium sulfate, 0.1-0.5g/L of monoammonium phosphate, 0.05-0.3g/L of magnesium sulfate, 0.1-0.5g/L of copper sulfate, 0.01-0.1g/L of zinc sulfate and 0.5-5g/L of yeast extract; the fermentation medium comprises the following components: 150-300g/L molasses, 15-30g/L ammonium sulfate, 2-8g/L monoammonium phosphate, 0.5-3g/L magnesium sulfate, 0.05-0.3g/L zinc sulfate, vitamin B 1 0.01-0.1g/L vitamin B 6 0.01-0.1g/L vitamin B 7 0.001-0.01g/L。
5. The process according to claim 3, wherein the fermentation in step 2) is performed by any one of batch fermentation, semi-continuous fermentation and continuous fermentation.
6. The method of manufacturing according to claim 2, wherein step (2) comprises the steps of:
a. preparing high-protein yeast milk: preparing Saccharomyces cerevisiae ZAB 02 obtained in the step (1) into yeast milk with a solid content of 12-18% by using process water;
b. and (3) removing RNA by a concentrated salt method: adding edible salt accounting for 8-14% of the weight of the yeast milk, heating to 80-95 ℃ and preserving heat for 2-6h;
c. separating and washing: centrifuging to separate yeast milk, washing the obtained precipitate with water, centrifuging, and collecting heavy phase;
d. cell wall enzymolysis: preparing yeast milk with 6-10% of solid content by using process water, adjusting pH to 4.0-6.0, adding yeast glucanase with 0.1-2.0% of yeast dry weight, and performing enzymolysis for 4-24h at 30-65deg.C;
e. and (3) hot alkali extraction: adding strong alkali accounting for 0.2% -1.0% of the weight of the yeast milk, heating to 85-95 ℃ and preserving heat for 1-5h;
f. separating and washing: centrifuging to separate yeast milk, collecting light phase, washing heavy phase with water, centrifuging, collecting light phase, washing heavy phase for 1-5 times, and mixing all light phases;
g. isoelectric point sedimentation: regulating the pH of the combined light phases to 4.0-5.0, and standing for 1-5h;
h. separating and washing: centrifuging to separate the precipitate, washing the precipitate with water, centrifuging to obtain heavy phase, washing the heavy phase for 1-5 times, and collecting the washed heavy phase to obtain yeast protein crude extract;
i. and (3) diatomite filtration: regulating the solid content of the yeast protein crude extract to 6% -10%, regulating the pH value to 7.5-9.0, precoating diatomite on a diatomite filter, and filtering the yeast protein crude extract;
j. activated carbon column adsorption: loading the diatomite filtered filtrate to an activated carbon column to obtain a yeast protein refined extract;
k. concentrating: concentrating the yeast protein refined extract to a solid content of 20-40% to obtain a yeast protein concentrated solution;
and I, drying: drying the yeast protein concentrate to obtain yeast protein powder.
7. The method according to claim 6, wherein the rotational speed of centrifugation in step c is 5000-8000 rpm, and the mass ratio of the yeast milk to the amount of washing water is 1:1.5-1:2; the enzymolysis condition in the step d is that the enzymolysis is carried out for 6 to 12 hours at the temperature of 40 to 60 ℃, and the enzyme activity of the yeast glucanase is 10000 to 120000U/g; the strong base in the step e is sodium hydroxide or potassium hydroxide.
8. The method according to claim 6, wherein the rotational speed of centrifugation in step f is 5000-8000 rpm, and the mass ratio of heavy phase to washing water amount is 1:0.5-1:1; the reagent used for regulating the pH in the step g is any one of hydrochloric acid, phosphoric acid and citric acid; the rotational speed of centrifugation in the step h is 5000-8000 rpm, and the mass ratio of the sediment to the washing water is 1:0.2-1:1.5.
9. The process according to claim 6, wherein the amount of diatomaceous earth used in step i is 0.1 to 0.5 times the weight of solids of the crude yeast protein extract; the dosage of the activated carbon in the step j is 0.5-2 times of the weight of the solid matters in the filtrate filtered by the diatomite; the diameter-to-height ratio of the activated carbon column is 1:1-1:5, and the flow rate of the activated carbon column per hour is 20-60% of the volume of the effective activated carbon; concentrating in the step k by using a triple effect evaporator; the drying in step l is carried out using pressure spray drying.
10. The yeast protein powder produced by the production method according to any one of claims 2 to 9.
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