CN111685223A - Feed containing yarrowia lipolytica, preparation method and application thereof - Google Patents
Feed containing yarrowia lipolytica, preparation method and application thereof Download PDFInfo
- Publication number
- CN111685223A CN111685223A CN201910188061.8A CN201910188061A CN111685223A CN 111685223 A CN111685223 A CN 111685223A CN 201910188061 A CN201910188061 A CN 201910188061A CN 111685223 A CN111685223 A CN 111685223A
- Authority
- CN
- China
- Prior art keywords
- yarrowia lipolytica
- fermentation
- product
- feed
- yeast
- 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
- 241000235015 Yarrowia lipolytica Species 0.000 title claims abstract description 65
- 238000002360 preparation method Methods 0.000 title claims abstract description 11
- 238000000855 fermentation Methods 0.000 claims abstract description 90
- 230000004151 fermentation Effects 0.000 claims abstract description 90
- 241001465754 Metazoa Species 0.000 claims abstract description 31
- 239000000203 mixture Substances 0.000 claims abstract description 20
- 239000000047 product Substances 0.000 claims description 54
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 47
- 239000000843 powder Substances 0.000 claims description 33
- 239000000758 substrate Substances 0.000 claims description 30
- 238000000034 method Methods 0.000 claims description 22
- 240000008042 Zea mays Species 0.000 claims description 19
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 19
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 19
- 235000005822 corn Nutrition 0.000 claims description 19
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 claims description 18
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 18
- 235000014113 dietary fatty acids Nutrition 0.000 claims description 18
- 229930195729 fatty acid Natural products 0.000 claims description 17
- 239000000194 fatty acid Substances 0.000 claims description 17
- 150000004665 fatty acids Chemical class 0.000 claims description 15
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 14
- 229910052799 carbon Inorganic materials 0.000 claims description 14
- 238000002156 mixing Methods 0.000 claims description 14
- 229940041514 candida albicans extract Drugs 0.000 claims description 13
- 239000012138 yeast extract Substances 0.000 claims description 13
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical group [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 12
- 235000019764 Soybean Meal Nutrition 0.000 claims description 12
- 239000004455 soybean meal Substances 0.000 claims description 12
- 239000011573 trace mineral Substances 0.000 claims description 12
- 235000013619 trace mineral Nutrition 0.000 claims description 12
- 239000001632 sodium acetate Substances 0.000 claims description 10
- 235000017281 sodium acetate Nutrition 0.000 claims description 10
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 9
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 9
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 9
- 238000001035 drying Methods 0.000 claims description 9
- 235000013312 flour Nutrition 0.000 claims description 9
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 9
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 9
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 9
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 9
- 238000004519 manufacturing process Methods 0.000 claims description 8
- 239000003674 animal food additive Substances 0.000 claims description 7
- 235000013325 dietary fiber Nutrition 0.000 claims description 7
- 235000012054 meals Nutrition 0.000 claims description 7
- 239000002244 precipitate Substances 0.000 claims description 7
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 6
- 238000012258 culturing Methods 0.000 claims description 6
- 235000005911 diet Nutrition 0.000 claims description 6
- 230000000378 dietary effect Effects 0.000 claims description 6
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 5
- 239000004519 grease Substances 0.000 claims description 5
- 239000003337 fertilizer Substances 0.000 claims description 4
- 239000000463 material Substances 0.000 claims description 3
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims description 3
- USFZMSVCRYTOJT-UHFFFAOYSA-N Ammonium acetate Chemical compound N.CC(O)=O USFZMSVCRYTOJT-UHFFFAOYSA-N 0.000 claims description 2
- 239000005695 Ammonium acetate Substances 0.000 claims description 2
- 235000011054 acetic acid Nutrition 0.000 claims description 2
- 235000019257 ammonium acetate Nutrition 0.000 claims description 2
- 229940043376 ammonium acetate Drugs 0.000 claims description 2
- 235000016709 nutrition Nutrition 0.000 abstract description 6
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 40
- 240000000073 Achillea millefolium Species 0.000 description 11
- 235000007754 Achillea millefolium Nutrition 0.000 description 11
- 239000002609 medium Substances 0.000 description 11
- 235000018102 proteins Nutrition 0.000 description 10
- 108090000623 proteins and genes Proteins 0.000 description 10
- 102000004169 proteins and genes Human genes 0.000 description 10
- 241000282887 Suidae Species 0.000 description 8
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 8
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 6
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 6
- 229910021578 Iron(III) chloride Inorganic materials 0.000 description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- 229910000365 copper sulfate Inorganic materials 0.000 description 6
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 description 6
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 description 6
- 230000004130 lipolysis Effects 0.000 description 6
- 229940099596 manganese sulfate Drugs 0.000 description 6
- 239000011702 manganese sulphate Substances 0.000 description 6
- 235000007079 manganese sulphate Nutrition 0.000 description 6
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 6
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 6
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 description 6
- 229910000368 zinc sulfate Inorganic materials 0.000 description 6
- 229960001763 zinc sulfate Drugs 0.000 description 6
- 235000019733 Fish meal Nutrition 0.000 description 5
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 5
- 230000029087 digestion Effects 0.000 description 5
- 235000020776 essential amino acid Nutrition 0.000 description 5
- 239000003797 essential amino acid Substances 0.000 description 5
- 239000003925 fat Substances 0.000 description 5
- 239000004467 fishmeal Substances 0.000 description 5
- 239000004615 ingredient Substances 0.000 description 5
- -1 organic acid salt Chemical class 0.000 description 5
- 150000001413 amino acids Chemical class 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 4
- 235000019197 fats Nutrition 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 238000011081 inoculation Methods 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 235000015097 nutrients Nutrition 0.000 description 4
- 239000003921 oil Substances 0.000 description 4
- 235000019198 oils Nutrition 0.000 description 4
- 230000001954 sterilising effect Effects 0.000 description 4
- 238000003756 stirring Methods 0.000 description 4
- 239000013589 supplement Substances 0.000 description 4
- 235000021122 unsaturated fatty acids Nutrition 0.000 description 4
- 150000004670 unsaturated fatty acids Chemical class 0.000 description 4
- 238000005406 washing Methods 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- WRIDQFICGBMAFQ-UHFFFAOYSA-N (E)-8-Octadecenoic acid Natural products CCCCCCCCCC=CCCCCCCC(O)=O WRIDQFICGBMAFQ-UHFFFAOYSA-N 0.000 description 3
- LQJBNNIYVWPHFW-UHFFFAOYSA-N 20:1omega9c fatty acid Natural products CCCCCCCCCCC=CCCCCCCCC(O)=O LQJBNNIYVWPHFW-UHFFFAOYSA-N 0.000 description 3
- QSBYPNXLFMSGKH-UHFFFAOYSA-N 9-Heptadecensaeure Natural products CCCCCCCC=CCCCCCCCC(O)=O QSBYPNXLFMSGKH-UHFFFAOYSA-N 0.000 description 3
- 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
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 description 3
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 3
- AYFVYJQAPQTCCC-GBXIJSLDSA-N L-threonine Chemical compound C[C@@H](O)[C@H](N)C(O)=O AYFVYJQAPQTCCC-GBXIJSLDSA-N 0.000 description 3
- OYHQOLUKZRVURQ-HZJYTTRNSA-N Linoleic acid Chemical compound CCCCC\C=C/C\C=C/CCCCCCCC(O)=O OYHQOLUKZRVURQ-HZJYTTRNSA-N 0.000 description 3
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 3
- 239000004472 Lysine Substances 0.000 description 3
- 239000005642 Oleic acid Substances 0.000 description 3
- ZQPPMHVWECSIRJ-UHFFFAOYSA-N Oleic acid Natural products CCCCCCCCC=CCCCCCCCC(O)=O ZQPPMHVWECSIRJ-UHFFFAOYSA-N 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 description 3
- 239000004473 Threonine Substances 0.000 description 3
- 239000000654 additive Substances 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 235000004626 essential fatty acids Nutrition 0.000 description 3
- 239000008103 glucose Substances 0.000 description 3
- QXJSBBXBKPUZAA-UHFFFAOYSA-N isooleic acid Natural products CCCCCCCC=CCCCCCCCCC(O)=O QXJSBBXBKPUZAA-UHFFFAOYSA-N 0.000 description 3
- 235000020778 linoleic acid Nutrition 0.000 description 3
- OYHQOLUKZRVURQ-IXWMQOLASA-N linoleic acid Natural products CCCCC\C=C/C\C=C\CCCCCCCC(O)=O OYHQOLUKZRVURQ-IXWMQOLASA-N 0.000 description 3
- 229930182817 methionine Natural products 0.000 description 3
- ICYJJTNLBFMCOZ-UHFFFAOYSA-J molybdenum(4+);disulfate Chemical compound [Mo+4].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O ICYJJTNLBFMCOZ-UHFFFAOYSA-J 0.000 description 3
- ZQPPMHVWECSIRJ-KTKRTIGZSA-N oleic acid Chemical compound CCCCCCCC\C=C/CCCCCCCC(O)=O ZQPPMHVWECSIRJ-KTKRTIGZSA-N 0.000 description 3
- 239000011684 sodium molybdate Substances 0.000 description 3
- 235000015393 sodium molybdate Nutrition 0.000 description 3
- TVXXNOYZHKPKGW-UHFFFAOYSA-N sodium molybdate (anhydrous) Chemical compound [Na+].[Na+].[O-][Mo]([O-])(=O)=O TVXXNOYZHKPKGW-UHFFFAOYSA-N 0.000 description 3
- 235000012424 soybean oil Nutrition 0.000 description 3
- 239000003549 soybean oil Substances 0.000 description 3
- 239000002699 waste material Substances 0.000 description 3
- 238000005303 weighing Methods 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 235000019750 Crude protein Nutrition 0.000 description 2
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 239000001888 Peptone Substances 0.000 description 2
- 108010080698 Peptones Proteins 0.000 description 2
- 229920000392 Zymosan Polymers 0.000 description 2
- IKHGUXGNUITLKF-XPULMUKRSA-N acetaldehyde Chemical compound [14CH]([14CH3])=O IKHGUXGNUITLKF-XPULMUKRSA-N 0.000 description 2
- 239000004480 active ingredient Substances 0.000 description 2
- 230000000996 additive effect Effects 0.000 description 2
- 238000005273 aeration Methods 0.000 description 2
- 150000001298 alcohols Chemical class 0.000 description 2
- 150000001299 aldehydes Chemical class 0.000 description 2
- 150000001336 alkenes Chemical class 0.000 description 2
- 239000010775 animal oil Substances 0.000 description 2
- 239000012141 concentrate Substances 0.000 description 2
- 238000009833 condensation Methods 0.000 description 2
- 230000005494 condensation Effects 0.000 description 2
- 229960000355 copper sulfate Drugs 0.000 description 2
- 235000019797 dipotassium phosphate Nutrition 0.000 description 2
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 2
- 238000004821 distillation Methods 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 229940032296 ferric chloride Drugs 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- 235000011187 glycerol Nutrition 0.000 description 2
- 239000010806 kitchen waste Substances 0.000 description 2
- 230000002366 lipolytic effect Effects 0.000 description 2
- 235000013372 meat Nutrition 0.000 description 2
- 239000002480 mineral oil Substances 0.000 description 2
- 235000010446 mineral oil Nutrition 0.000 description 2
- JRZJOMJEPLMPRA-UHFFFAOYSA-N olefin Natural products CCCCCCCC=C JRZJOMJEPLMPRA-UHFFFAOYSA-N 0.000 description 2
- 150000007524 organic acids Chemical class 0.000 description 2
- 235000019319 peptone Nutrition 0.000 description 2
- 239000003208 petroleum Substances 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 235000015112 vegetable and seed oil Nutrition 0.000 description 2
- 239000008158 vegetable oil Substances 0.000 description 2
- 238000009423 ventilation Methods 0.000 description 2
- MKNQNPYGAQGARI-UHFFFAOYSA-N 4-(bromomethyl)phenol Chemical compound OC1=CC=C(CBr)C=C1 MKNQNPYGAQGARI-UHFFFAOYSA-N 0.000 description 1
- 241000251468 Actinopterygii Species 0.000 description 1
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 241000235646 Cyberlindnera jadinii Species 0.000 description 1
- 241000287828 Gallus gallus Species 0.000 description 1
- 108010068370 Glutens Proteins 0.000 description 1
- 241000235058 Komagataella pastoris Species 0.000 description 1
- 241000081271 Phaffia rhodozyma Species 0.000 description 1
- 241000235070 Saccharomyces Species 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 241001052560 Thallis Species 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 208000026935 allergic disease Diseases 0.000 description 1
- 235000011114 ammonium hydroxide Nutrition 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 description 1
- 239000004327 boric acid Substances 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 235000013330 chicken meat Nutrition 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 229910000366 copper(II) sulfate Inorganic materials 0.000 description 1
- JZCCFEFSEZPSOG-UHFFFAOYSA-L copper(II) sulfate pentahydrate Chemical compound O.O.O.O.O.[Cu+2].[O-]S([O-])(=O)=O JZCCFEFSEZPSOG-UHFFFAOYSA-L 0.000 description 1
- 230000018044 dehydration Effects 0.000 description 1
- 238000006297 dehydration reaction Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 239000002270 dispersing agent Substances 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 235000021312 gluten Nutrition 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 210000000936 intestine Anatomy 0.000 description 1
- 230000007794 irritation Effects 0.000 description 1
- 235000020997 lean meat Nutrition 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 235000010755 mineral Nutrition 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 235000021049 nutrient content Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- OTYBMLCTZGSZBG-UHFFFAOYSA-L potassium sulfate Chemical compound [K+].[K+].[O-]S([O-])(=O)=O OTYBMLCTZGSZBG-UHFFFAOYSA-L 0.000 description 1
- 229910052939 potassium sulfate Inorganic materials 0.000 description 1
- 235000011151 potassium sulphates Nutrition 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 239000004460 silage Substances 0.000 description 1
- 108010027322 single cell proteins Proteins 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-N sulfuric acid Substances OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 1
- 238000004448 titration Methods 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
Classifications
-
- 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
- A23K10/30—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
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
- A23K10/37—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms from waste material
-
- 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
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
Abstract
The invention relates to a feed containing yarrowia lipolytica, a preparation method and application thereof. The present invention discloses for the first time a novel fermentation product applicable to animal feed, which is obtained mainly by fermentation of yarrowia lipolytica under appropriate conditions, which has a good nutritional composition. The invention also discloses a preparation method and application of the fermentation product.
Description
Technical Field
The invention belongs to the field of animal feed, and particularly relates to feed containing yarrowia lipolytica, a preparation method and application thereof.
Background
Animal feed is indispensable in animal husbandry production, and animal feed with good safety is one of the factors for promoting the benign development of animal husbandry production. Animal feed typically includes protein feed, energy feed, roughage, greenfeed, silage, mineral feed, and feed additives.
Animal feeds can be classified into: complete feed, concentrated feed and premix. The complete feed is a mixed batch consisting of four parts, namely protein feed, energy feed, coarse feed and an additive, is a granular feed which is mainly granulated by machine processing and sold on the market, and is an expanded granular feed, and can be directly used for feeding objects. The concentrated feed is prepared by premixing a protein raw material and an additive, and energy needs to be supplemented when feeding, so that the concentrated feed has the advantage of convenient use, is suitable for large-scale farmers, particularly farmers who own energy feeds such as corn and the like, and has at least 50 concentrated feeds in the market at present. The premix is an important component of a complete formula feed, and is an intermediate type formula feed product prepared by uniformly mixing one or more trace components (including various trace mineral elements, various vitamins, synthetic amino acids, certain medicaments and other additives) with a diluent or a mixture according to a required proportion.
With the improvement of living standard of people, the meat quality of some farm animals (such as pigs and the like) is required to be higher and higher, and the demand in the aspect can be solved by improving the formula of the feed, so that the unbalanced and reasonable nutrition supply is improved for the animals.
There are currently some animal feeds in the art that are formulated with yeast. However, yeasts commonly used in feeds in the market at present are mainly saccharomyces cerevisiae, pichia pastoris, candida utilis, phaffia rhodozyma and the like, and thalli of the yeasts can contain abundant proteins and essential amino acids, but the contents of fat and essential fatty acid are obviously insufficient, so that fish meal, grease and the like are further added when the yeasts are used as the feeds to ensure the nutritional requirements of feeding animals, the addition increases the feeding cost and limits the large-scale application of the yeasts.
There is therefore a need in the art to develop improved feeds, to provide more nutritionally complete feeds,
disclosure of Invention
The invention aims to provide a feed containing yarrowia lipolytica, a preparation method and application thereof.
In a first aspect of the invention, there is provided a method of making a yarrowia lipolytica fermentation product, the method comprising:
(1) fermenting and culturing yarrowia lipolytica, and adding a carbon source substrate into a fermentation system in the fermentation process; wherein, the carbon source substrate in the step (a) is a grease substrate, and the total amount of fatty acid is controlled to be 0.05-2% (w/V); or, (b) the carbon source substrate is a substrate for generating acetate, and the concentration of acetate ions is controlled to be 0.4-2 g/L;
(2) after fermentation, the fermentation precipitate is collected and dried to obtain the yarrowia lipolytica fermentation product.
In a preferred embodiment, in (a), the total amount of fatty acid is controlled to be 0.08-1%; preferably 0.1 to 0.8 percent.
In another preferred embodiment, (b) the acetate concentration is controlled to be 1 +/-0.5 g/L; preferably 1 + -0.3 g/L.
In another preferred embodiment, when yarrowia lipolytica is fermentatively cultured, the initial medium comprises: yeast extract (or yeast extract), magnesium sulfate, ammonium sulfate, potassium dihydrogen phosphate and/or dipotassium hydrogen phosphate; preferably, the fertilizer also comprises trace elements; and/or said acetate producing substrate is sodium acetate, ammonium acetate, acetic acid.
In another preferred embodiment, the compound comprises: 0.5-5 g/L of yeast extract (or yeast extract), 0.5-5 g/L of magnesium sulfate, 1-30 g/L of ammonium sulfate, 1-5 g/L of monopotassium phosphate and/or 1-5 g/L of dipotassium phosphate.
In another preferred embodiment, the trace elements include: ferric chloride, manganese sulfate, zinc sulfate, copper sulfate and molybdenum sulfate.
In another preferred embodiment, the yarrowia lipolytica is yarrowia lipolytica CICC 32291.
In another preferred embodiment, the fermentation product of yarrowia lipolytica comprises a product selected from the group consisting of: wet yeast obtained by partially dehydrating yarrowia lipolytica fermentation liquor, yeast powder obtained by drying and dehydrating, or yeast meal obtained by extracting oil.
In another preferred embodiment, (a) the pH is maintained at 7.5 ± 0.3; preferably, 7.5 + -0.2 or 7.5 + -0.1.
In another preferred embodiment, (b) the pH is maintained at 6.5 ± 0.3; preferably 6.5 + -0.2 or 6.5 + -0.1.
In another aspect of the invention, there is provided a yarrowia lipolytica fermentation product produced by the method described above; preferably, the yarrowia lipolytica fermentation product comprises a product selected from the group consisting of: wet yeast obtained by partially dehydrating yarrowia lipolytica fermentation liquor, yeast powder obtained by drying and dehydrating, or yeast meal obtained by extracting oil.
In a preferred embodiment, the yarrowia lipolytica fermentation product is rich in protein, fat, yeast polysaccharide and amino acids, wherein lysine, methionine and threonine are particularly abundant.
In another preferred embodiment, the yarrowia lipolytica fermentation product is rich in fatty acids, wherein the content of unsaturated fatty acids such as oleic acid and linoleic acid is particularly rich.
In another preferred embodiment, the fermentation duration is 24-120 hours, such as 36, 48, 60, 72, 84, 96, 108 hours, etc.
In another aspect of the invention, there is provided the use of said yarrowia lipolytica fermentation product for the preparation of an animal feed or feed additive.
In another aspect of the invention, there is provided an animal feed or feed additive comprising said yarrowia lipolytica fermentation product, and a dietetically or feedetically acceptable mix; preferably, the dietetic or feedlot acceptable mix includes (but is not limited to) materials selected from the group consisting of: corn flour, pig feed premix, corn, bran and soybean meal.
In another aspect of the present invention, there is provided a method of preparing an animal feed, the method comprising: (i) mixing corn flour and feed premix; (ii) mixing the yarrowia lipolytica fermentation product obtained by said method with the product mixture of (i); (iii) (iii) mixing the mixed product of (ii) with a roughage.
In a preferred embodiment, (i) when the corn flour and the feed premix are mixed, the corn flour and the feed premix are mixed according to the weight ratio of (1-10): 1, preferably according to (2-8): 1, and more preferably according to (3-7): 1 or (4-6): 1.
In another preferred embodiment, (ii) the yarrowia lipolytica fermentation product is mixed with the mixed product of (i) at a weight ratio of (1-5) to (1-5), preferably at a weight ratio of (1-3) to (1-3), more preferably at a weight ratio of (1-3) to (1-3) or at a weight ratio of (1-2) to (1-2).
In another preferred embodiment, (iii), the mixed product of (ii) is mixed with the roughage according to the weight ratio of (2-15): 1, preferably (4-10): 1, more preferably (5-8): 1 or (6-7): 1.
In another preferred embodiment, the roughage includes, but is not limited to: corn and bran.
In another preferred embodiment, the feed premix includes, but is not limited to: commercially available premixes for pigs, cattle, chickens, fish, etc.
Other aspects of the invention will be apparent to those skilled in the art in view of the disclosure herein.
Detailed Description
The present inventors have made intensive studies to disclose for the first time a novel fermentation product applicable to animal feeds, which is obtained mainly by fermenting Yarrowia Lipolytica (Yarrowia Lipolytica or saccharomyces Lipolytica) under appropriate conditions, and which has a good nutritional composition.
Based on the novel findings of the present invention, there is provided a method for producing a fermentation product of yarrowia lipolytica, comprising: (1) fermenting and culturing yarrowia lipolytica, and adding a carbon source substrate into a fermentation system in the fermentation process; wherein the carbon source substrate is a grease substrate, and the total amount of fatty acid is controlled to be 0.05-2%; or the carbon source substrate is a substrate for generating acetate, and the concentration of the acetate is controlled to be 0.4-2 g/L; (2) after fermentation, the fermentation precipitate is collected and dried to obtain the yarrowia lipolytica fermentation product.
The invention determines an ideal method for fermenting by using yarrowia lipolytica, and in a preferred embodiment of the invention, the yarrowia lipolytica is a strain with the preservation number CICC32291 in China center for Industrial culture Collection of microorganisms, and a fermentation product of the yarrowia lipolytica has good nutritional composition and is suitable for preparing feeds.
Feeding a carbon source substrate after the fermentation is started, wherein the carbon source substrate can be: organic acid or organic acid salt, volatile fatty acid or fatty acid salt, alcohols such as methanol and ethanol, aldehydes such as formaldehyde and acetaldehyde, glycerin, vegetable oil, animal oil, waste kitchen waste, petroleum, mineral oil, olefin and the like. Some substrates that are discarded or difficult to utilize by conventional bacteria may also be used.
In a preferred embodiment of the present invention, the carbon source substrate is a lipid substrate or an acetate-producing substrate. When the oil substrate is used for fermentation, the total amount of fatty acid is preferably controlled to be 0.05-2%, more preferably 0.08-1%, and further preferably 0.1-0.6%. When the acetate substrate is used for fermentation, the concentration of acetate is preferably controlled to be 1-2 g/L, and more preferably, the concentration of acetate is controlled to be 1 +/-0.5 g/L; further, it may be 1. + -. 0.3g/L or 1. + -. 0.2 g/L.
As a preferred mode of the invention, the yarrowia lipolytica fermentation product comprises but is not limited to: wet yeast obtained by partially dehydrating yarrowia lipolytica fermentation liquor, yeast powder obtained by drying and dehydrating, or yeast meal obtained by extracting oil. The yeast powder is preferably obtained after dehydration by drying from the viewpoint of easy storage or processing.
At the beginning of the fermentation, the fermenter is filled with the initial fermentation medium. The initial fermentation medium may be a conventional yeast medium suitable for yeast growth, capable of providing the necessary components for growth of the strain. In a preferred embodiment of the present invention, when yarrowia lipolytica is fermentatively cultured, the initial medium comprises: yeast extract (or yeast extract), magnesium sulfate, ammonium sulfate, potassium dihydrogen phosphate and/or dipotassium hydrogen phosphate; preferably, the fertilizer also comprises trace elements. For example, the medium may be a formulation as shown in Table 1, but is not limited thereto.
TABLE 1
In a preferred embodiment of the present invention, trace elements are further added to the initial fermentation medium, preferably the trace elements include: ferric chloride, manganese sulfate, zinc sulfate, copper sulfate and molybdenum sulfate. For example, the trace elements may be the formulations shown in table 2, but are not limited thereto.
TABLE 2
| General scope | Preferred range | Preferred range | |
| Ferric chloride | 0.002～0.05g/L | 0.005～0.04g/L | 0.008～0.02g/L |
| Manganese sulfate | 0.005～0.1g/L | 0.008～0.08g/L | 0.01～0.04g/L |
| Zinc sulfate | 0.005～0.1g/L | 0.008～0.08g/L | 0.01～0.04g/L |
| Copper sulfate | 0.5～10mg/L | 0.8～8mg/L | 1～4mg/L |
| Molybdenum sulfate | 0.5～10mg/L | 0.8～8mg/L | 1～4mg/L |
In the method, the fermentation duration is 24-120 hours, such as 36, 48, 60, 72, 84, 96, 108 hours and the like.
In the present invention, the pH varies depending on the substrate, and the pH is adjusted depending on the use of the substrate on the basis of the optimum of the strain itself. When the substrate is grease, the pH is properly increased for better mutual solubility under weak alkalinity.
The invention also includes the fermentation product obtained by fermentation using the above-described method. The fermentation product has rich protein, fat, zymosan, amino acids and essential amino acids, and especially rich lysine, methionine and threonine. Meanwhile, the fermentation product is rich in fatty acid, wherein the unsaturated fatty acid such as oleic acid and linoleic acid is particularly rich.
In the invention, the enrichment is obvious and has the characteristics of statistical significance, and generally means that the content of corresponding nutrient components is more than 5 percent of that of similar products sold in the market; more preferably more than 10%, such as more than 15%, 20%, 30%, 50%, 70%, 90% or more.
The concentrated fermentation product has a high content of active ingredients, such as 30-100 wt%, while the diluted fermentation product and the practically used fermentation product have a low content of active ingredients, typically 0.00005-30 wt%. In addition, other suitable trace elements, stabilizers, binders, dispersants, emulsifiers, penetrants, solvents, fillers, and other common ingredients may be included.
The fermentation product can be used as a feed additive for feeding animals or as concentrated feed after being processed properly; alternatively, it can be further mixed with a mixture or excipient acceptable in dietetics or feedology to prepare an animal feed. The animal feed is rich in nutritional ingredients. Such as concentration, drying, etc.
Accordingly, the present invention also provides an animal feed or feed additive comprising an effective amount of the fermentation product as hereinbefore described, together with a sitologically or feedstuffs (including fertilizer or agro-pharmaceutical) acceptable mix.
In the present invention, the term "comprising" means that various ingredients can be applied together in the mixture or fermentation product of the present invention. Thus, the terms "consisting essentially of and" consisting of are encompassed by the term "comprising.
In the context of the present invention, a "dietetic or feedstuffs-acceptable" ingredient is a substance which is suitable for dietetic or feedstuffs use without undue adverse side effects (such as toxicity, irritation and allergic response) to the animal, i.e. with a reasonable benefit/risk ratio.
In the present invention, some "dietetic or feedlot acceptable mixes" are exemplified by, but not limited to: corn flour, pig feed premix, corn, bran, soybean meal and the like.
The present invention also provides a method of preparing an animal feed, the method comprising: (i) mixing corn flour and feed premix; (ii) mixing the yarrowia lipolytica fermentation product obtained by the preparation method with the mixed product of (i); and (iii) mixing the mixed product of (ii) with roughage.
The fermentation product of the invention, animal feed containing the fermentation product and a dietetically or dietetically acceptable mix, can also be contained in a package (e.g., a box or a bag) for use in production or feed supply.
The invention has the beneficial effects that:
the explanation of the present invention reveals that Yarrowia lipolytica (Yarrowia lipolytica) is a yeast rich in proteins and essential amino acids, as well as fats and essential fatty acids, and the content of proteins and essential amino acids in the fermentation product is comparable to that of the yeast species commonly used in feeds on the market at present, but the content of fats and essential fatty acids, especially unsaturated fatty acids, is far more than that.
The method of the invention can utilize production raw materials with wide sources and low price, and can use waste or difficult-to-utilize organic acid or organic acid salt, volatile fatty acid or fatty acid salt, alcohols such as methanol and ethanol, aldehydes such as formaldehyde and acetaldehyde, glycerol, vegetable oil, animal oil, waste kitchen waste, petroleum, mineral oil, olefin and the like besides using conventional fermentation substrates such as glucose, xylose, cellulose and the like as substrates for fermentation. Has great potential for feed application.
Meanwhile, the feed provided by the invention can provide nutrient substances required by feeding animals, can regulate intestines and stomach of the animals and enhance immunity of the animals, is a natural component, cannot cause tolerance to the animals, and has the advantage that other feed components are incomparable.
The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. The experimental procedures, for which specific conditions are not noted in the following examples, are generally performed according to conventional conditions such as those described in J. SammBruk et al, molecular cloning protocols, third edition, scientific Press, 2002, or according to the manufacturer's recommendations.
EXAMPLE 1 Single cell protein feed Yeast powder production by fermentation of Soybean oil
Yarrowia lipolytica CICC32291 (Yarrowia lipolytica) was inoculated into 100mL of seed medium for fermentation to prepare a seed solution. The seed medium is typically a glucose yeast peptone medium, typically consisting of: 20% of glucose, 10% of yeast extract and 20% of peptone. The fermentation is carried out in a shake flask, the rotation speed of a shaking table is usually 100-200 rpm, and the temperature is 28 ℃.
After the seed solution is obtained, inoculating the seed solution into a 5L fermentation tank according to the inoculation concentration of 5% (v/v) for culturing, wherein the temperature is 28 ℃, the initial stirring rotation speed is 200rpm, the initial ventilation volume is 1vvm, and the stirring rotation speed and the ventilation volume are set in the whole fermentation process to ensure that the dissolved oxygen is between 20 and 60 percent. The incubation period was 72 hours. Wherein the culture medium of 5L fermenter is 2L fermentation broth sterilized at 121 deg.C for 30 min, and has pH of 7.5. The fermentation liquor comprises the following components: 1g/L of yeast extract, 1g/L of magnesium sulfate, 20g/L of ammonium sulfate, 3g/L of monopotassium phosphate and 3g/L of dipotassium phosphate, and trace elements: the content of ferric chloride is 0.01g/L, the content of manganese sulfate is 0.02g/L, the content of zinc sulfate is 0.02g/L, the content of copper sulfate is 2mg/L, and the content of sodium molybdate is 2 mg/L.
And (3) adding sterile soybean oil into the fermentation tank according to the proportion of 10g/L at the beginning of fermentation, feeding the soybean oil when the total fatty acid content is lower than 0.1% by sampling and detecting, controlling the total fatty acid content to be between 0.1% and 0.3%, and automatically feeding and regulating by using 25% concentrated ammonia water after the fermentation is started to keep the pH value to be 7.5.
Determination of the dry cell weight: weighing a fermentation liquid sample, centrifuging at the rotation speed of 4000rpm for 5 minutes, washing the obtained precipitate with water once, centrifuging at the rotation speed of 4000rpm for 5 minutes, drying the obtained precipitate at 80 ℃ overnight, and weighing to obtain yarrowia lipolytica yeast powder, wherein the dry weight of cells in the fermentation liquid is measured to be 68 g/kg.
Determination of crude protein:
1. digestion of the sample: weighing 0.5-1 g of sample (the nitrogen content is 5-80mg), placing the sample into a digestion tube, adding 0.4g of mixed catalyst of blue vitriol and 6g of potassium sulfate and 20ml of concentrated sulfuric acid, digesting the sample on a digestion furnace for about 1h at 420 ℃ until the solution is clear, taking out the solution, cooling the solution, transferring the solution onto a digestion tube tray of a Kjeldahl azotometer, inserting a steam tube into the digestion tube, abutting a tube opening against a rubber head, setting the acid adding amount (boric acid) to be 25ml, the alkali adding amount (sodium hydroxide) to be 50ml, the distilled water amount to be 20ml and the distillation time to be 400s (the absorption volume to be 100 ml), after the distillation is finished, washing the tail end of the condensation tube by using distilled water, enabling the washing liquid to flow into a conical flask, taking down the conical flask at the tail end of the condensation tube, and adding about 5 drops of bromocresol green-methyl.
2. Titration: the solution was titrated with 0.1mol/L of a standard hydrochloric acid solution, and the end point was the change from blue-green to gray-red.
A blank was also prepared using 0.5g of sucrose as described above. According to calculation, 71.2% of crude protein can be obtained in 1g of dry cell weight.
Example 2 preparation of yarrowia lipolytica powder 2
Preparing lipolysis yarrow yeast powder according to the following steps:
1) inoculating Yarrowia lipolytica CICC32291 bacterial liquid (Yarrowia lipolytica CICC 32291) into a 5L seed shake flask according to the inoculation amount of 10% for culturing at the temperature of 30 ℃ and the rotation speed of a shaking table of 200rpm for 16 hours, wherein the composition of 800g of culture medium of the 5L seed shake flask is as follows: 20g/L of sodium acetate, 2g/L of ammonium sulfate, 2g/L of monopotassium phosphate, 1g/L of yeast extract, 2g/L of magnesium sulfate, 6.5 of pH value and high-temperature steam sterilization.
2) Then transferring the mixture into a 10L fermentation tank for culture according to the inoculation amount of 10%, wherein the temperature is 30 ℃, the stirring speed is 200rpm, the aeration is 1vvm, and the culture is carried out for 24 hours, wherein 7kg of culture medium of the 10L fermentation tank comprises the following components: 20g/L of sodium acetate, 2g/L of ammonium sulfate, 2g/L of monopotassium phosphate, 1g/L of yeast extract, 2g/L of magnesium sulfate, 6.5 of pH value and high-temperature steam sterilization.
3) Then transferring the mixture into a 100L fermentation tank according to the inoculation amount of 10 percent for culturing at the temperature of 30 ℃, the stirring speed of 300rpm and the aeration speed of 1.5vvm for 48 hours, wherein 50kg of culture medium of the 100L fermentation tank comprises the following components: 20g/L of sodium acetate, 2g/L of ammonium sulfate, 2g/L of monopotassium phosphate, 1g/L of yeast extract and 2g/L of magnesium sulfate; and trace elements: the content of ferric chloride is 0.01g/L, the content of manganese sulfate is 0.02g/L, the content of zinc sulfate is 0.02g/L, the content of copper sulfate is 2mg/L, and the content of sodium molybdate is 2 mg/L. pH 6.5, and high-temperature steam sterilizing. In the fermentation process, sodium acetate (CH) is fed3COONa) to supplement a carbon source necessary for yeast growth, measuring the concentration of acetate to determine the supplement amount, and feeding a sodium acetate solution when the concentration of acetate ions is less than 1 g/L.
4) And after the fermentation is finished, centrifuging the fermentation liquor at the rotating speed of 4000rpm for 2min, washing the obtained precipitate with water once, centrifuging at the rotating speed of 4000rpm for 2min, directly drying the obtained precipitate overnight at 80 ℃, and then crushing to obtain the yarrowia lipolytica yeast powder.
Example 3 preparation of yarrowia lipolytica powder 3
The main steps are according to the method described in example 3, varying only in that:
50kg of medium in a 100L fermenter consisted of: 15g/L of sodium acetate, 3g/L of ammonium sulfate, 3g/L of monopotassium phosphate, 0.6g/L of yeast extract and 1g/L of magnesium sulfate; and trace elements: the content of ferric chloride is 0.02g/L, the content of manganese sulfate is 0.015g/L, the content of zinc sulfate is 0.015g/L, the content of copper sulfate is 1.8mg/L, and the content of sodium molybdate is 2.5 mg/L. pH 6.5, and high-temperature steam sterilizing.
In the fermentation process, sodium acetate (CH) is fed3COONa) to supplement a carbon source necessary for yeast growth, measuring the concentration of acetate to determine the supplement amount, and feeding a sodium acetate solution when the concentration of acetate is less than 0.9 g/L.
Analysis of nutrient composition of yarrow lipolytica powder in example 4, examples 1, 2 and 3
The analysis of the nutrient contents of the purchased fish meal (domestic fish meal, Shandong shun-Yuan feed), soybean meal, corn gluten meal and beer yeast powder, together with the self-made yarrow yeast powder, is shown in Table 3.
TABLE 3 analysis of nutrients in homemade lipolysis yarrow yeast powder and several commercial feed ingredients
As can be seen from Table 3, the self-made lipolytic yarrow yeast powder has rich protein content, which is superior to commercially available beer yeast powder and soybean meal and is comparable to commercially available fish meal and corn protein powder.
As can be seen from Table 3, the self-made lipolytic yarrow yeast powder has rich fat content, which is superior to commercially available soybean meal, corn protein powder and beer yeast powder, and is comparable to commercially available fish meal.
As can be seen from Table 3, the self-made yarrowia lipolytica yeast powder has rich content of zymosan; the amino acid and essential amino acid are rich, wherein the content of lysine, methionine and threonine is particularly rich.
Further, the fatty acid content of the self-made yarrowia lipolytica powder was analyzed, and the results are shown in table 4.
TABLE 4 analysis of fatty acid content of self-made yarrow lipolytica powder
According to Table 4, the self-made yarrowia lipolytica powder is rich in fatty acids, wherein the content of unsaturated fatty acids such as oleic acid and linoleic acid is particularly rich.
Example 5 animal feeding study
The method comprises the following steps of feeding 30-60 kg of Chinese lean-type growing-finishing pigs by using self-made lipolysis yarrow yeast powder and commercially available soybean meal (Shandong Shunyuan feed) in a contrast manner:
1) the experimental groups for feeding the prepared lipolysis yarrow yeast powder are as follows:
mixing commercially available corn flour and 30-60 kg of Chinese lean type commercial premix (Shandong Shuyuan feed) of growing-finishing pigs according to a weight ratio of 5: 1 to prepare dry powder after uniformly mixing; mixing the prepared yarrowia lipolytica yeast powder and the dry powder according to the weight ratio of 1: 1 to prepare the yarrowia lipolytica concentrate; the yarrowia lipolytica concentrate is mixed with commercial coarse feed (prepared by uniformly mixing commercial corn and commercial bran according to the weight ratio of 45: 7) according to the weight ratio of 48: 52 to prepare the final daily ration of the experimental group.
2) Control group fed with commercial soybean meal: according to the formula of the daily ration of the experimental group, all the self-made lipolysis yarrow yeast powder is replaced by the soybean meal sold in the market to prepare the final daily ration of the control group.
3) About 20 growing-finishing pigs of 35kg are selected as experimental animals, are randomly divided into an experimental group and a control group, 10 pigs in each group are respectively fed with the daily ration of the experimental group and the daily ration of the control group, the pre-feeding period is 5 days, the experimental period is 35 days, and the total period is 40 days, and the experimental results are shown in table 3.
The experimental results of the comparison of the self-made lipolysis yarrow yeast powder (experimental group) and the commercial soybean meal (control group) on the feeding of the growing-finishing pigs with the lean meat type of 30-60 kg in China are shown in the table 5.
TABLE 5
| Average daily gain (g/head/day) | Meat ratio of materials | |
| Experimental group | 708.6±20.8 | 2.11：1 |
| Control group | 653.3±17.5 | 2.29：1 |
According to the table 5, the comparative feeding experiment shows that, compared with soybean meal, the yarrowia lipolytica can obviously improve the daily gain of growing-finishing pigs, reduce the feed conversion ratio and can partially or completely replace the soybean meal.
All documents referred to herein are incorporated by reference into this application as if each were individually incorporated by reference. Furthermore, it should be understood that various changes and modifications of the present invention can be made by those skilled in the art after reading the above teachings of the present invention, and these equivalents also fall within the scope of the present invention as defined by the appended claims.
Claims (10)
1. A method of making a yarrowia lipolytica fermentation product, comprising:
(1) fermenting and culturing yarrowia lipolytica, and adding a carbon source substrate into a fermentation system in the fermentation process; wherein, the carbon source substrate in the step (a) is a grease substrate, and the total amount of fatty acid is controlled to be 0.05-2% (w/V); or, (b) the carbon source substrate is a substrate for generating acetate, and the concentration of acetate ions is controlled to be 0.4-2 g/L;
(2) after fermentation, the fermentation precipitate is collected and dried to obtain the yarrowia lipolytica fermentation product.
2. The method of claim 1, wherein in (a), the total amount of fatty acids is controlled to be 0.08% to 1%; preferably 0.1% -0.8%; or (b), controlling the concentration of acetate at 1 +/-0.5 g/L; preferably 1 + -0.3 g/L.
3. The method of claim 1, wherein the initial medium comprises, in the fermentative culture of yarrowia lipolytica: yeast extract, magnesium sulfate, ammonium sulfate, potassium dihydrogen phosphate and/or dipotassium hydrogen phosphate; preferably, the fertilizer also comprises trace elements; and/or
The substrate for generating acetate is sodium acetate, ammonium acetate and acetic acid.
4. The method of claim 1, wherein the yarrowia lipolytica fermentation product comprises a product selected from the group consisting of: wet yeast obtained by partially dehydrating yarrowia lipolytica fermentation liquor, yeast powder obtained by drying and dehydrating, or yeast meal obtained by extracting oil.
5. The method of claim 1, wherein in (a), the pH is maintained at 7.5 ± 0.3; preferably, 7.5 ± 0.2 or 7.5 ± 0.1; or in (b), the pH is maintained at 6.5. + -. 0.3; preferably 6.5 + -0.2 or 6.5 + -0.1.
6. A yarrowia lipolytica fermentation product produced by the method of any one of claims 1-5; preferably, the yarrowia lipolytica fermentation product comprises a product selected from the group consisting of: wet yeast obtained by partially dehydrating yarrowia lipolytica fermentation liquor, yeast powder obtained by drying and dehydrating, or yeast meal obtained by extracting oil.
7. Use of the yarrowia lipolytica fermentation product of claim 6 for the preparation of an animal feed or feed additive.
8. An animal feed or feed additive comprising the yarrowia lipolytica fermentation product of claim 6, and a dietetically or feedetically acceptable mix; preferably, the dietetic or feedlot acceptable mix comprises a material selected from the group consisting of: corn flour, pig feed premix, corn, bran and soybean meal.
9. A method of preparing an animal feed, the method comprising:
(i) mixing corn flour and feed premix;
(ii) mixing a yarrowia lipolytica fermentation product produced by the method of any one of claims 1-5 with the product mixture of (i);
(iii) (iii) mixing the mixed product of (ii) with a roughage.
10. The method according to claim 9, wherein in (i), the corn meal and the feed premix are mixed according to the weight ratio of (1-10): 1, preferably according to (2-8): 1, more preferably according to (3-7): 1 or (4-6): 1; or
(ii) In (i), the yarrowia lipolytica fermentation product and the mixed product of (i) are mixed in a weight ratio of (1-5) to (1-5), preferably (1-3) to (1-3), more preferably (1-3) to (1-3) or (1-2) to (1-2); or
(iii) In (iii), the mixed product of (ii) is mixed with the roughage in a weight ratio of (2-15): 1, preferably (4-10): 1, more preferably (5-8): 1 or (6-7): 1.
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| CN201910188061.8A Active CN111685223B (en) | 2019-03-13 | 2019-03-13 | Feed containing yarrowia lipolytica, preparation method and application thereof |
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