CN114591860A - 一种高载乳酸菌肉用直投式发酵剂及其制备方法和应用 - Google Patents
一种高载乳酸菌肉用直投式发酵剂及其制备方法和应用 Download PDFInfo
- Publication number
- CN114591860A CN114591860A CN202210278576.9A CN202210278576A CN114591860A CN 114591860 A CN114591860 A CN 114591860A CN 202210278576 A CN202210278576 A CN 202210278576A CN 114591860 A CN114591860 A CN 114591860A
- Authority
- CN
- China
- Prior art keywords
- meat
- direct vat
- vat set
- lactobacillus plantarum
- starter
- 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.)
- Pending
Links
- 235000013372 meat Nutrition 0.000 title claims abstract description 37
- 239000007858 starting material Substances 0.000 title claims abstract description 36
- 238000002360 preparation method Methods 0.000 title claims abstract description 19
- 239000004310 lactic acid Substances 0.000 title claims abstract description 16
- 240000006024 Lactobacillus plantarum Species 0.000 claims abstract description 28
- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims abstract description 28
- 229940072205 lactobacillus plantarum Drugs 0.000 claims abstract description 28
- 238000004321 preservation Methods 0.000 claims abstract description 13
- 235000013622 meat product Nutrition 0.000 claims abstract description 12
- 244000005700 microbiome Species 0.000 claims abstract description 6
- 230000001580 bacterial effect Effects 0.000 claims description 30
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 20
- 239000000843 powder Substances 0.000 claims description 19
- 239000007788 liquid Substances 0.000 claims description 18
- 241000894006 Bacteria Species 0.000 claims description 17
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims description 16
- 239000001963 growth medium Substances 0.000 claims description 15
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 11
- 239000008103 glucose Substances 0.000 claims description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 11
- 239000012153 distilled water Substances 0.000 claims description 9
- 235000014655 lactic acid Nutrition 0.000 claims description 8
- 239000003223 protective agent Substances 0.000 claims description 8
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 7
- 239000002131 composite material Substances 0.000 claims description 7
- 235000020183 skimmed milk Nutrition 0.000 claims description 7
- 239000004475 Arginine Substances 0.000 claims description 6
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 6
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 claims description 6
- 238000000855 fermentation Methods 0.000 claims description 6
- 230000004151 fermentation Effects 0.000 claims description 6
- 238000000034 method Methods 0.000 claims description 6
- 239000010802 sludge Substances 0.000 claims description 6
- LPXPTNMVRIOKMN-UHFFFAOYSA-M sodium nitrite Chemical compound [Na+].[O-]N=O LPXPTNMVRIOKMN-UHFFFAOYSA-M 0.000 claims description 6
- 239000012137 tryptone Substances 0.000 claims description 6
- 239000011734 sodium Substances 0.000 claims description 5
- 239000007836 KH2PO4 Substances 0.000 claims description 4
- 238000004108 freeze drying Methods 0.000 claims description 4
- 239000002054 inoculum Substances 0.000 claims description 4
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 4
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 claims description 4
- 229920000053 polysorbate 80 Polymers 0.000 claims description 4
- 150000003839 salts Chemical class 0.000 claims description 4
- 240000004160 Capsicum annuum Species 0.000 claims description 3
- 235000008534 Capsicum annuum var annuum Nutrition 0.000 claims description 3
- 244000203593 Piper nigrum Species 0.000 claims description 3
- 235000008184 Piper nigrum Nutrition 0.000 claims description 3
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 3
- 229930006000 Sucrose Natural products 0.000 claims description 3
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 3
- 235000013614 black pepper Nutrition 0.000 claims description 3
- 239000001511 capsicum annuum Substances 0.000 claims description 3
- 238000005119 centrifugation Methods 0.000 claims description 3
- 235000020379 cucumber juice Nutrition 0.000 claims description 3
- 238000004898 kneading Methods 0.000 claims description 3
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 3
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 3
- 229940099596 manganese sulfate Drugs 0.000 claims description 3
- 239000011702 manganese sulphate Substances 0.000 claims description 3
- 235000007079 manganese sulphate Nutrition 0.000 claims description 3
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 3
- 239000001931 piper nigrum l. white Substances 0.000 claims description 3
- 238000005096 rolling process Methods 0.000 claims description 3
- 239000001632 sodium acetate Substances 0.000 claims description 3
- 235000017281 sodium acetate Nutrition 0.000 claims description 3
- 235000010352 sodium erythorbate Nutrition 0.000 claims description 3
- 239000004320 sodium erythorbate Substances 0.000 claims description 3
- 235000010288 sodium nitrite Nutrition 0.000 claims description 3
- 235000019832 sodium triphosphate Nutrition 0.000 claims description 3
- RBWSWDPRDBEWCR-RKJRWTFHSA-N sodium;(2r)-2-[(2r)-3,4-dihydroxy-5-oxo-2h-furan-2-yl]-2-hydroxyethanolate Chemical compound [Na+].[O-]C[C@@H](O)[C@H]1OC(=O)C(O)=C1O RBWSWDPRDBEWCR-RKJRWTFHSA-N 0.000 claims description 3
- YWYZEGXAUVWDED-UHFFFAOYSA-N triammonium citrate Chemical compound [NH4+].[NH4+].[NH4+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O YWYZEGXAUVWDED-UHFFFAOYSA-N 0.000 claims description 3
- 239000001393 triammonium citrate Substances 0.000 claims description 3
- 235000011046 triammonium citrate Nutrition 0.000 claims description 3
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 abstract description 18
- 239000002253 acid Substances 0.000 abstract description 7
- 238000002474 experimental method Methods 0.000 description 13
- 238000012258 culturing Methods 0.000 description 11
- 238000012360 testing method Methods 0.000 description 11
- 108090000623 proteins and genes Proteins 0.000 description 9
- 235000018102 proteins Nutrition 0.000 description 8
- 102000004169 proteins and genes Human genes 0.000 description 8
- 239000000243 solution Substances 0.000 description 8
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 7
- 230000000694 effects Effects 0.000 description 7
- 235000010855 food raising agent Nutrition 0.000 description 7
- 238000005259 measurement Methods 0.000 description 7
- 230000000052 comparative effect Effects 0.000 description 6
- 239000000047 product Substances 0.000 description 6
- 239000003153 chemical reaction reagent Substances 0.000 description 5
- 239000000796 flavoring agent Substances 0.000 description 5
- 235000019634 flavors Nutrition 0.000 description 5
- 235000011187 glycerol Nutrition 0.000 description 5
- 238000011081 inoculation Methods 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 4
- 229920002472 Starch Polymers 0.000 description 4
- 235000009697 arginine Nutrition 0.000 description 4
- 238000009630 liquid culture Methods 0.000 description 4
- 239000000523 sample Substances 0.000 description 4
- 238000012216 screening Methods 0.000 description 4
- 235000019698 starch Nutrition 0.000 description 4
- 239000008107 starch Substances 0.000 description 4
- 230000001954 sterilising effect Effects 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 229920001817 Agar Polymers 0.000 description 3
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 239000008272 agar Substances 0.000 description 3
- 238000003556 assay Methods 0.000 description 3
- 230000015556 catabolic process Effects 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 238000005520 cutting process Methods 0.000 description 3
- 238000006731 degradation reaction Methods 0.000 description 3
- 238000010790 dilution Methods 0.000 description 3
- 239000012895 dilution Substances 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 238000012163 sequencing technique Methods 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 108020004465 16S ribosomal RNA Proteins 0.000 description 2
- HVBSAKJJOYLTQU-UHFFFAOYSA-N 4-aminobenzenesulfonic acid Chemical compound NC1=CC=C(S(O)(=O)=O)C=C1 HVBSAKJJOYLTQU-UHFFFAOYSA-N 0.000 description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 108090000489 Carboxy-Lyases Proteins 0.000 description 2
- RWSOTUBLDIXVET-UHFFFAOYSA-N Dihydrogen sulfide Chemical compound S RWSOTUBLDIXVET-UHFFFAOYSA-N 0.000 description 2
- 229910019142 PO4 Inorganic materials 0.000 description 2
- UYXTWWCETRIEDR-UHFFFAOYSA-N Tributyrin Chemical compound CCCC(=O)OCC(OC(=O)CCC)COC(=O)CCC UYXTWWCETRIEDR-UHFFFAOYSA-N 0.000 description 2
- 235000001014 amino acid Nutrition 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 235000015278 beef Nutrition 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 239000007789 gas Substances 0.000 description 2
- 239000003102 growth factor Substances 0.000 description 2
- 229910000037 hydrogen sulfide Inorganic materials 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 230000000670 limiting effect Effects 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- CEQFOVLGLXCDCX-WUKNDPDISA-N methyl red Chemical compound C1=CC(N(C)C)=CC=C1\N=N\C1=CC=CC=C1C(O)=O CEQFOVLGLXCDCX-WUKNDPDISA-N 0.000 description 2
- 235000013336 milk Nutrition 0.000 description 2
- 239000008267 milk Substances 0.000 description 2
- 210000004080 milk Anatomy 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 239000006872 mrs medium Substances 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 238000005457 optimization Methods 0.000 description 2
- 150000007524 organic acids Chemical class 0.000 description 2
- 238000005554 pickling Methods 0.000 description 2
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Chemical compound [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 230000001953 sensory effect Effects 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 238000009777 vacuum freeze-drying Methods 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- NWCHELUCVWSRRS-SECBINFHSA-N (2r)-2-hydroxy-2-phenylpropanoic acid Chemical compound OC(=O)[C@@](O)(C)C1=CC=CC=C1 NWCHELUCVWSRRS-SECBINFHSA-N 0.000 description 1
- ZCUQOPGIJRGJDA-UHFFFAOYSA-N 1-naphthalen-1-ylethane-1,2-diamine Chemical compound C1=CC=C2C(C(N)CN)=CC=CC2=C1 ZCUQOPGIJRGJDA-UHFFFAOYSA-N 0.000 description 1
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 1
- LVSQXDHWDCMMRJ-UHFFFAOYSA-N 4-hydroxybutan-2-one Chemical compound CC(=O)CCO LVSQXDHWDCMMRJ-UHFFFAOYSA-N 0.000 description 1
- 108010062877 Bacteriocins Proteins 0.000 description 1
- 102000016938 Catalase Human genes 0.000 description 1
- 108010053835 Catalase Proteins 0.000 description 1
- 240000008067 Cucumis sativus Species 0.000 description 1
- 235000010799 Cucumis sativus var sativus Nutrition 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 241000192125 Firmicutes Species 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 238000003794 Gram staining Methods 0.000 description 1
- 101000666730 Homo sapiens T-complex protein 1 subunit alpha Proteins 0.000 description 1
- ODKSFYDXXFIFQN-BYPYZUCNSA-N L-arginine Chemical compound OC(=O)[C@@H](N)CCCN=C(N)N ODKSFYDXXFIFQN-BYPYZUCNSA-N 0.000 description 1
- 229930064664 L-arginine Natural products 0.000 description 1
- 235000014852 L-arginine Nutrition 0.000 description 1
- 241000186660 Lactobacillus Species 0.000 description 1
- 239000004367 Lipase Substances 0.000 description 1
- 102000004882 Lipase Human genes 0.000 description 1
- 108090001060 Lipase Proteins 0.000 description 1
- 235000007688 Lycopersicon esculentum Nutrition 0.000 description 1
- 108010048581 Lysine decarboxylase Proteins 0.000 description 1
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 1
- 108090000913 Nitrate Reductases Proteins 0.000 description 1
- IOVCWXUNBOPUCH-UHFFFAOYSA-M Nitrite anion Chemical compound [O-]N=O IOVCWXUNBOPUCH-UHFFFAOYSA-M 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 108010080032 Pediocins Proteins 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 240000001462 Pleurotus ostreatus Species 0.000 description 1
- 235000001603 Pleurotus ostreatus Nutrition 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- 235000016998 Ribes burejense Nutrition 0.000 description 1
- 241000735566 Ribes burejense Species 0.000 description 1
- 240000003768 Solanum lycopersicum Species 0.000 description 1
- 244000046109 Sorghum vulgare var. nervosum Species 0.000 description 1
- 102100038410 T-complex protein 1 subunit alpha 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
- 230000009471 action Effects 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 239000011543 agarose gel Substances 0.000 description 1
- 238000007605 air drying Methods 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 235000011130 ammonium sulphate Nutrition 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 230000002368 bacteriocinic effect Effects 0.000 description 1
- 230000003385 bacteriostatic effect Effects 0.000 description 1
- 238000010009 beating Methods 0.000 description 1
- 238000010256 biochemical assay Methods 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 235000013339 cereals Nutrition 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 230000001112 coagulating effect Effects 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 230000006835 compression Effects 0.000 description 1
- 238000007906 compression Methods 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 239000002577 cryoprotective agent Substances 0.000 description 1
- 235000013365 dairy product Nutrition 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 229910000397 disodium phosphate Inorganic materials 0.000 description 1
- RSEOPLGIFDMYKN-UHFFFAOYSA-N ethanol;naphthalen-1-ol Chemical compound CCO.C1=CC=C2C(O)=CC=CC2=C1 RSEOPLGIFDMYKN-UHFFFAOYSA-N 0.000 description 1
- XJRPTMORGOIMMI-UHFFFAOYSA-N ethyl 2-amino-4-(trifluoromethyl)-1,3-thiazole-5-carboxylate Chemical compound CCOC(=O)C=1SC(N)=NC=1C(F)(F)F XJRPTMORGOIMMI-UHFFFAOYSA-N 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 1
- 230000015784 hyperosmotic salinity response Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 229910017053 inorganic salt Inorganic materials 0.000 description 1
- 229940039696 lactobacillus Drugs 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 235000019421 lipase Nutrition 0.000 description 1
- 229940057995 liquid paraffin Drugs 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 244000000010 microbial pathogen Species 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 239000008055 phosphate buffer solution Substances 0.000 description 1
- 235000021110 pickles Nutrition 0.000 description 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 1
- 239000004323 potassium nitrate Substances 0.000 description 1
- 235000010333 potassium nitrate Nutrition 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 230000017854 proteolysis Effects 0.000 description 1
- 239000007671 pyg medium Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 238000009938 salting Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 230000001932 seasonal effect Effects 0.000 description 1
- 239000013049 sediment Substances 0.000 description 1
- 238000013207 serial dilution Methods 0.000 description 1
- 238000004904 shortening Methods 0.000 description 1
- 235000013599 spices Nutrition 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 229950000244 sulfanilic acid Drugs 0.000 description 1
- 230000019086 sulfide ion homeostasis Effects 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 238000009461 vacuum packaging Methods 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 238000004383 yellowing Methods 0.000 description 1
Images
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
- C12N1/20—Bacteria; Culture media therefor
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
- A23B4/00—General methods for preserving meat, sausages, fish or fish products
- A23B4/14—Preserving with chemicals not covered by groups A23B4/02 or A23B4/12
- A23B4/18—Preserving with chemicals not covered by groups A23B4/02 or A23B4/12 in the form of liquids or solids
- A23B4/20—Organic compounds; Microorganisms; Enzymes
- A23B4/22—Microorganisms; Enzymes; Antibiotics
-
- 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
- A23L13/00—Meat products; Meat meal; Preparation or treatment thereof
- A23L13/70—Tenderised or flavoured meat pieces; Macerating or marinating solutions specially adapted therefor
- A23L13/72—Tenderised or flavoured meat pieces; Macerating or marinating solutions specially adapted therefor using additives, e.g. by injection of solutions
- A23L13/74—Tenderised or flavoured meat pieces; Macerating or marinating solutions specially adapted therefor using additives, e.g. by injection of solutions using microorganisms or enzymes
-
- 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
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/169—Plantarum
-
- 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
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/90—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in food processing or handling, e.g. food conservation
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Microbiology (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Polymers & Plastics (AREA)
- Food Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Virology (AREA)
- Medicinal Chemistry (AREA)
- Nutrition Science (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
本发明公开了一种高载乳酸菌肉用直投式发酵剂及其制备方法和应用,属于发酵剂技术领域。该高载乳酸菌肉用直投式发酵剂中包含植物乳杆菌SJ‑4,所述植物乳杆菌SJ‑4已在中国工业微生物菌种保藏中心保藏,保藏编号为CICC NO.11119s,保藏地址为北京市朝阳区酒仙桥中路24号院6号楼,保藏日期为2020年12月22日。在本发明的直投式发酵剂中,植物乳杆菌SJ‑4具有较好的产酸能力和生长速度,能满足作为肉用发酵剂的安全性、生产适应性等条件,可提高肉制品品质,缩短生产周期,易于推广应用。
Description
技术领域
本发明涉及发酵剂技术领域,特别是涉及一种高载乳酸菌肉用直投式发酵剂及其制备方法和应用。
背景技术
乳酸菌是一类革兰氏阳性菌,需氧或兼性厌氧,可以通过多种途径抑制某些腐败菌的生长,如产生苯乳酸、乙酸或乳酸等有机酸而导致环境pH值降低,从而抑制不耐酸细菌的生长;其次乳酸菌能产生抑菌活性的代谢产物,即细菌素(尼森、罗氏菌素、片球菌素、链球菌素等)以及类细菌素,同时其产生的有机酸既可以延长产品货架期还可以增加肉制品的风味,广泛应用于发酵工业。
发酵肉制品是指在微生物或酶的作用下,自然或人为控制条件形成的一类具有特殊风味、质地和色泽,且能保存较长时间的高档肉制品。我国传统发酵肉制品制作周期长,生产季节性、区域性较强,标准化生产不足,因此,开发直投式发酵剂不仅有助于缩短生产周期,实现规模化生产,而且可以提升产品的安全性,抑制腐败微生物和病原微生物生长,提高产品的稳定性、感官质量和保存性。
国外对发酵剂的研究从上世纪80年代开始,我国起步较晚,且市售发酵剂多用于发酵泡菜、乳制品,肉用发酵剂较为罕见。
发明内容
本发明的目的是提供一种高载乳酸菌肉用直投式发酵剂及其制备方法和应用,以解决上述现有技术存在的问题,该发酵剂中植物乳杆菌SJ-4具有较高的密度和活性,可用于制备获得质地和色泽较好且风味独特的发酵肉制品。
为实现上述目的,本发明提供了如下方案:
本发明提供一种高载乳酸菌肉用直投式发酵剂,包含植物乳杆菌SJ-4,所述植物乳杆菌SJ-4已在中国工业微生物菌种保藏中心保藏,保藏编号为CICC NO.11119s,保藏地址为北京市朝阳区酒仙桥中路24号院6号楼,保藏日期为2020年12月22日。
进一步地,所述高载乳酸菌肉用直投式发酵剂中植物乳杆菌SJ-4活菌数≥1011CFU/g。
本发明还提供一种上述的高载乳酸菌肉用直投式发酵剂的制备方法,包括如下步骤:
植物乳杆菌SJ-4经高密度培养后离心收集菌泥,并向其中加入由脱脂乳、精氨酸和甘油组成的复合保护剂,之后进行冷冻干燥,即得所述高载乳酸菌肉用直投式发酵剂。
进一步地,所述高密度培养具体为:将植物乳杆菌SJ-4按体积百分比5%接种量接种于优化培养基,37℃100r/min摇床培养16h。
进一步地,所述优化培养基包括2%w/v葡萄糖、1.5%w/v酵母粉、1.5%w/v胰蛋白胨、4%w/v黄瓜汁、0.2%w/v 0.2mol/L Na2HPO4-0.2mol/LKH2PO4、0.5%w/v乙酸钠、0.2%w/v柠檬酸三胺、0.1%w/v吐温80、0.02%w/v硫酸镁和0.005%w/v硫酸锰,余量为水。
进一步地,所述复合保护剂通过以下步骤获得:先将11g脱脂乳和6g精氨酸溶于100mL蒸馏水中115℃灭菌15min,再加入6g甘油121℃灭菌20min。
进一步地,所述复合保护剂加入量与所述植物乳杆菌SJ-4经高密度培养后得到的菌液的体积比是1:10。
本发明还提供一种上述的高载乳酸菌肉用直投式发酵剂在制备发酵肉制品中的应用。
进一步地,所述发酵肉制品的制备方法为:
向处理后的肉样中加入2.5wt%食盐、0.01wt%亚硝酸钠、0.15wt%异抗坏血酸钠、0.8wt%花椒粉、0.5wt%三聚磷酸钠、0.8wt%白糖、0.5wt%白胡椒和0.5wt%五香粉,0-4℃发酵24h后加入所述高载乳酸菌肉用直投式发酵剂,真空滚揉30min后于30℃发酵48h,结束发酵后50℃烘烤3h。
进一步地,所述高载乳酸菌肉用直投式发酵剂的加入量为0.001-0.002%w/v。
本发明公开了以下技术效果:
1.在本发明的直投式发酵剂中,植物乳杆菌SJ-4具有较好的产酸能力和生长速度,能满足作为肉用发酵剂的安全性、生产适应性等条件,可提高肉制品品质,缩短生产周期。
2.在本发明的直投式发酵剂中,植物乳杆菌SJ-4经高密度培养后制成冻干粉,其中冷冻保护剂可以保护该菌株在低温条件下避免损伤,使菌种具有较高的密度和活性。
3.采用本发明制得的直投式发酵剂用于发酵里脊火腿,可得到质地和色泽较好,且风味独特的发酵肉制品。
附图说明
为了更清楚地说明本发明实施例或现有技术中的技术方案,下面将对实施例中所需要使用的附图作简单地介绍,显而易见地,下面描述中的附图仅仅是本发明的一些实施例,对于本领域普通技术人员来讲,在不付出创造性劳动的前提下,还可以根据这些附图获得其他的附图。
图1为本发明基于16S rDNA基因序列建立的植物乳杆菌SJ-4系统发育树;
图2为本发明植物乳杆菌SJ-4生长曲线及产酸曲线;
图3为本发明直投式发酵剂冻干粉图片;
图4为实施例4、对比例1和对比例2的发酵剂对发酵里脊火腿电子舌味觉响应雷达图。
具体实施方式
现详细说明本发明的多种示例性实施方式,该详细说明不应认为是对本发明的限制,而应理解为是对本发明的某些方面、特性和实施方案的更详细的描述。
应理解本发明中所述的术语仅仅是为描述特别的实施方式,并非用于限制本发明。另外,对于本发明中的数值范围,应理解为还具体公开了该范围的上限和下限之间的每个中间值。在任何陈述值或陈述范围内的中间值以及任何其他陈述值或在所述范围内的中间值之间的每个较小的范围也包括在本发明内。这些较小范围的上限和下限可独立地包括或排除在范围内。
除非另有说明,否则本文使用的所有技术和科学术语具有本发明所述领域的常规技术人员通常理解的相同含义。虽然本发明仅描述了优选的方法和材料,但是在本发明的实施或测试中也可以使用与本文所述相似或等同的任何方法和材料。本说明书中提到的所有文献通过引用并入,用以公开和描述与所述文献相关的方法和/或材料。在与任何并入的文献冲突时,以本说明书的内容为准。
在不背离本发明的范围或精神的情况下,可对本发明说明书的具体实施方式做多种改进和变化,这对本领域技术人员而言是显而易见的。由本发明的说明书得到的其他实施方式对技术人员而言是显而易见得的。本发明说明书和实施例仅是示例性的。
关于本文中所使用的“包含”、“包括”、“具有”、“含有”等等,均为开放性的用语,即意指包含但不限于。
实施例1菌株的分离纯化、鉴定与保藏
1.菌株分离
以贵州农家自制酸肉作为分离源,将肉样剪碎成米粒大小或黄豆大小的肉沫,称取10g肉沫,向其中加入90mL无菌生理盐水,经无菌均质机拍打后作10倍系列稀释,选取3个适当稀释浓度分别吸取200μL涂布于MSR琼脂培养基,37℃恒温培养48h,选取白色,圆形,表面湿润不透明,边缘整齐的单个菌落反复划线直至革兰氏染色镜检结果显示没有杂菌,停止分离纯化,将纯化的革兰氏阳性菌株4℃斜面保存,同时保存于-80℃的40%甘油管中。
2.菌株初筛
2.1以提取的肌原纤维蛋白和肌浆蛋白作为菌株初筛的原料,其制备方法分述如下:
肌原纤维蛋白的制备:称取10g肉样于离心管中,加入200mL 50mmol/L pH 7.5预冷的磷酸盐缓冲液,12000r/min匀浆3次后于4℃8000r/min离心15min,重复3次收集沉淀并冷冻干燥成粉末。
肌浆蛋白的制备:将新鲜碎肉分散在十倍体积的磷酸盐缓冲溶液(40mmol/LNaH2PO4/Na2HPO4,50mmol/L NaCl pH 6.0)中,12000rpm均质30s后12000r/min 4℃离心15min,用纱布快速过滤收集上清液,加入硫酸铵至饱和,连续搅拌析出肌浆蛋白,10000r/min 4℃离心15min收集沉淀的蛋白质并冷冻干燥成粉末。
2.2蛋白质降解实验:在MRS培养基中加入1wt%上述预先提取的肌原纤维蛋白和肌浆蛋白,115℃灭菌后使用7mm打孔器打孔,加入100μL菌液,37℃培养24h选出出现降解圈的菌株。
2.3产酸实验:吸取上述实验筛选的400μL菌液至20mL MRS液体培养基,37℃恒温培养24h,选出pH 6.0以下菌株。
2.4发酵葡萄糖产气实验:在1L PY培养基中加入30g葡萄糖、0.5mL吐温80、1.4mL1.6g/100mL溴甲酚紫,分装10mL试管后放入倒置的杜氏小管,灭菌后分别接入产酸实验选出的200μL菌液,使用无菌石蜡液封,37℃恒温培养24h,选出杜氏小管中无气泡的阴性菌株。
3.菌株复筛
3.1菌株生产适应性
3.1.1耐盐性实验:将2.4筛选出的活菌数为107-108CFU/mL的菌株按2%接种量活化3代后接入含有6%NaCl的MRS肉汤培养基中37℃培养24h测定OD600,选择OD600值偏高的菌株。
3.1.2耐亚硝酸盐实验:将3.1.1筛选出的活菌数为107-108CFU/mL的菌株按2%接种量活化3代后接入含150mg/kg NaNO2的MRS肉汤培养基中,37℃培养24小时测定OD600,选择OD600值偏高的菌株。
3.2耐酸实验:将3.1.2筛选的活菌数为107-108CFU/mL的菌株按2%接种量活化3代后接入pH 5.0的MRS肉汤培养基,37培培养24h测定其OD600,选择OD600值偏高的菌株。
3.3菌株安全性
3.3.1氨基酸脱羧酶活性实验:分别吸取3.2筛选的300μL菌液于细菌微量生化鉴定管(赖氨酸脱羧酶肉汤、L-精氨酸双水解酶分别为实验组、氨基酸脱羧酶为对照组)中,接种后覆盖300μL无菌液体石蜡,37℃培养24h,选出试验管和对照管均为黄色的阴性菌株。
3.4菌株作为肉用发酵剂的评定
3.4.1硝酸盐还原酶实验:在MRS培养基中加入0.2%硝酸钾,将3.3.1筛选的菌株的纯菌落划线后37℃培养48h,滴入Griess试剂A液(150mL20%盐酸中加入0.5g对氨基苯磺酸)、B液(0.2g盐酸萘乙二胺溶于100mL蒸馏水),菌落周围未出现红圈。
3.4.2产氨实验:将0.5g NaCl、0.5g精氨酸溶于100mL脱脂牛乳中,接入0.2%的3.4.1选出的菌株菌液,37℃培养24h后滴入1滴纳赛尔试剂,选出无沉淀的阴性菌株。
3.4.3产粘性实验:将3.4.2选出的菌株的菌落接种于MRS培养基,37℃培养24h,挑取单个菌落进行观察,选出不产粘性的菌株。
3.4.4产硫化氢实验:将3.4.3选出的菌落穿刺于硫化氢细菌微量生化鉴定管,37℃培养24h,选出穿刺线清晰无变化的阴性菌株。
3.4.5蛋白酶降解活性实验:见2.菌株初筛
3.4.6脂肪酶分解活性实验:MRS琼脂培养基121℃灭菌20min后冷却至80℃,加入1%灭菌的三丁酸甘油油脂,挑取3.4.5选出的单个菌落接种于培养基中,37℃培养48h,菌落周围未出现降解圈。
3.5生理生化鉴定
3.5.1发酵葡萄糖产气实验:见2.菌株初筛
3.5.2石蕊牛奶实验:在100mL脱脂牛乳中加入4mL 25g/mL石蕊试剂,分装10mL试管灭菌后接入0.2%3.5.1选出的菌液37℃培养2d选出使石蕊牛奶凝固的菌株。
3.5.3接触酶实验(过氧化氢实验):挑取3.5.2筛选的单个菌落至PYG培养基中,滴加3%过氧化氢,选出无气泡的阴性菌株。
3.5.4甲基红实验:接种2%3.5.3筛选的菌液至PYG培养基培养24h后滴入甲基红指示剂,选出变黄的阴性菌株。
3.5.5乙酰甲基甲醇实验(V-P实验):挑取3.5.4选出的单个菌落穿刺V-P半固体琼脂细菌微量生化鉴定管,37℃培养24h,滴入试剂A(α-萘酚乙醇溶液)6滴,试剂B(40%氢氧化钠水溶液)2滴,静置30min后选出穿刺线无明显变化的阴性菌株。
4.菌株SJ-4的鉴定
菌株SJ-4 16S rDNA分子生物学鉴定:将3.5.5筛选出来的菌液使用细菌通用引物27F(5′-AGAGTTTGATCMTGGCTCAG-3′)、1492R(5′-TACGGYTACCTTGTTACGACTT-3′)扩增细菌16S rDNA,使用1%浓度琼脂糖凝胶,电泳20min,送至武汉天一辉远生物科技有限公司进行测序,测序结果如下(SEQ ID NO:1):
GAGTGAGTGGCGAACTGGTGAGTAACACGTGGGAAACCTGCCCAGAAGCGGGGGATAACACCTGGAAACAGATGCTAATACCGCATAACAACTTGGACCGCATGGTCCGAGTTTGAAAGATGGCTTCGGCTATCACTTTTGGATGGTCCCGCGGCGTATTAGCTAGATGGTGGGGTAACGGCTCACCATGGCAATGATACGTAGCCGACCTGAGAGGGTAATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGACGAAAGTCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGGTTTCGGCTCGTAAAACTCTGTTGTTAAAGAAGAACATATCTGAGAGTAACTGTTCAGGTATTGACGGTATTTAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTTTTAAGTCTGATGTGAAAGCCTTCGGCTCAACCGAAGAAGTGCATCGGAAACTGGGAAACTTGAGTGCAGAAGAGGACAGTGGAACTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTGTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGTATGGGTAGCAAACAGGATTAGATACCCTGGTAGTCCATACCGTAAACGATGAATGCTAAGTGTTGGAGGGTTTCCGCCCTTCAGTGCTGCAGCTAACGCATTAAGCATTCCGCCTGGGGAGTACGGCCGCAAGGCTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCTACGCGAAGAACCTTACCAGGTCTTGACATACTATGCAAATCTAAGAGATTAGACGTTCCCTTCGGGGACATGGATACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATTATCAGTTGCCAGCATTAAGTTGGGCACTCTGGTGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGATGGTACAACGAGTTGCGAACTCGCGAGAGTAAGCTAATCTCTTAAAGCCATTCTCAGTTCGGATTGTAGGCTGCAACTCGCCTACATGAAGTCGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGAGAGTTTGTAACACCCAAAGTCGGTGGGGTAACCTTTTAGGAACCAGCCGCCTA
将测序结果进入NCBI网站中BLAST程序进行同源性比对,使用MEGA-X软件构建系统发育树,如图1,明确菌株SJ-4为植物乳杆菌(Lactobacillus plantarum)。
5.菌株SJ-4的保藏
菌株SJ-4的分类命名为植物乳杆菌(Lactobacillus plantarum),已于2020年12月22日保藏于中国工业微生物菌种保藏中心(简称CICC,地址为:北京市朝阳区酒仙桥中路24号院6号楼),保藏编号:CICC NO.11119s。
6.菌株SJ-4生长条件的优化
6.1菌株SJ-4最适生长温度的确定:将菌液以4%接种量接种于pH=6.0的MRS液体培养基中,并分别在17、22、27、32、37、42、47℃下恒温培养24h后测定菌液OD600,结果表明,菌株SJ-4在37℃条件下有最高OD600为1.584,即该菌株最适生长温度为37℃。
6.2菌株SJ-4最适生长pH的测定:将菌液以4%接种量分别接种于pH 1.0、2.0、3.0、4.0、5.0、6.0、7.0、8.0、9.0的MRS液体培养基中,37℃培养48h,每12h测定一次菌液OD600,结果表明,菌株SJ-4在pH6.0条件下有最高OD600为1.611,即该菌株最适生长pH为6.0。
6.3菌株SJ-4生长曲线和产酸曲线的测定:将4%菌液接种于MRS液体培养基中37℃培养24h,每隔2h测定一次OD600和pH值,如图2。
实施例2菌株SJ-4高密度培养条件的优化
以MRS液体培养基为参照,选择不同的碳源(1.00%淀粉、1.00%麦芽糖、1.00%葡萄糖、0.33%淀粉+0.67%葡萄糖、0.33%葡萄糖+0.67%淀粉、0.50%淀粉+0.50%葡萄糖)、氮源(1.00%酵母粉、1.00%牛肉膏、1.00%胰蛋白胨、0.50%酵母粉+0.50%牛肉膏、0.50%酵母粉+0.50%胰蛋白胨)、无机盐(0.2mol/L Na2HPO4-0.1mol/L柠檬酸、0.2mol/LNaAc-0.2mol/L HAc、0.2mol/L Na2HPO4-0.2mol/L NaH2PO4、0.2mol/L Na2HPO4-0.2mol/LKH2PO4)、生长因子(平菇、玉米、番茄、黄瓜榨汁后过滤),培养条件(摇床转速80r/min,90r/min,100r/min,110r/min,120r/min)进行优化。以上%代表质量体积百分浓度。
经实验结果显示优选以下成分:葡萄糖、酵母粉+胰蛋白胨、0.2mol/LNa2HPO4-0.2mol/L KH2PO4、黄瓜汁;再经浓度优化,最终确定最佳碳源为2%w/v葡萄糖,最佳氮源为1.5%w/v酵母粉+1.5%w/v胰蛋白胨,最佳生长因子为4%w/v黄瓜汁,最佳无机盐为0.2%w/v 0.2mol/LNa2HPO4-0.2mol/L KH2PO4稀释4倍,其余含量为:0.5%w/v乙酸钠、0.2%w/v柠檬酸三胺、0.1%w/v吐温80、0.02%w/v硫酸镁、0.005%w/v硫酸锰,再向培养基中添加蒸馏水至1000mL。将甘油管中保存的植物乳杆菌按5%接种量接种于该优化培养基,37℃100r/min摇床培养16h,活化3代后活菌数达109CFU/mL。
实施例3直投式发酵剂的制备
该直投式发酵剂中冷冻保护剂的原料组成包括11g/100mL脱脂乳、6g/100mL精氨酸溶于蒸馏水中115℃灭菌15min,6g/100mL甘油溶于蒸馏水中121℃灭菌20min,植物乳杆菌SJ-4经高密度培养后8000r/min 4℃离心10min收集菌泥,向菌泥中加入离心前菌液1/10体积的复合保护剂,-80℃预冷冻6h,放入真空冷冻干燥箱48h后,得到植物乳杆菌SJ-4冻干粉如图3,菌株存活率为70.72%,活菌数达1011CFU/g。
实施例4直投式发酵剂在发酵里脊火腿中的应用
1.发酵里脊火腿的制备
1.1将市售里脊使用2wt%高粱酒去腥,风干后加入2.5wt%食盐、0.01wt%亚硝酸钠、0.15wt%异抗坏血酸钠、0.8wt%花椒粉、0.5wt%三聚磷酸钠、0.8wt%白糖、0.5wt%白胡椒、0.5wt%五香粉,0-4℃发酵24h。
1.2加入0.001-0.002%w/v直投式发酵剂,加入少量蒸馏水稀释后注射腌制,真空滚揉30min。
1.3 30℃发酵48h,结束后50℃烘烤3h,冷却至室温后真空包装即得发酵里脊火腿产品。
2.发酵里脊火腿质构的测定
将火腿表面切除后,切成2×2×2cm的立方体,使用TA/36探头,测前速率1mm/s,测后速率1mm/s,压缩率50%,触发力5g,间隔时间2s,测定硬度、弹性、粘聚性、咀嚼性。
3.发酵里脊火腿色泽的测定
4.发酵里脊火腿电子舌五味的测定
将火腿表面切除后,取10g加入100mL蒸馏水使用高压均质机搅碎,30℃浸提3h后于冷冻离心机4℃6000r/min离心15min后取上清液20mL,加入60mL蒸馏水稀释,倒入电子舌样品杯中稀释,取中间测定数据进行分析。
对比例1
与实施例4的区别在于,选择0.05%w/v,107CFU/mL的植物乳杆菌SJ-4菌液注射腌制。
对比例2
与实施例4的区别在于,选择由植物乳杆菌SJ-4菌液不添加任何保护剂直接在-80℃预冷冻6h,放入真空冷冻干燥箱48h后得到的冻干粉腌制,加入0.001-0.002%w/v该冻干粉。
效果验证
统计实施例4、对比例1、对比例2制得的里脊火腿的质构、色泽、电子舌五味值测定结果,如表1、图4所示,结果表明直投式发酵剂制得的里脊火腿促进了色泽的形成和稳定,接种后有利于火腿的切片,且添加发酵剂后火腿滋味更丰富,较好地改善了产品的品质。
表1各组发酵产品的品质测定结果
以上所述的实施例仅是对本发明的优选方式进行描述,并非对本发明的范围进行限定,在不脱离本发明设计精神的前提下,本领域普通技术人员对本发明的技术方案做出的各种变形和改进,均应落入本发明权利要求书确定的保护范围内。
序列表
<110> 贵州大学
<120> 一种高载乳酸菌肉用直投式发酵剂及其制备方法和应用
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1384
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 1
gagtgagtgg cgaactggtg agtaacacgt gggaaacctg cccagaagcg ggggataaca 60
cctggaaaca gatgctaata ccgcataaca acttggaccg catggtccga gtttgaaaga 120
tggcttcggc tatcactttt ggatggtccc gcggcgtatt agctagatgg tggggtaacg 180
gctcaccatg gcaatgatac gtagccgacc tgagagggta atcggccaca ttgggactga 240
gacacggccc aaactcctac gggaggcagc agtagggaat cttccacaat ggacgaaagt 300
ctgatggagc aacgccgcgt gagtgaagaa gggtttcggc tcgtaaaact ctgttgttaa 360
agaagaacat atctgagagt aactgttcag gtattgacgg tatttaacca gaaagccacg 420
gctaactacg tgccagcagc cgcggtaata cgtaggtggc aagcgttgtc cggatttatt 480
gggcgtaaag cgagcgcagg cggtttttta agtctgatgt gaaagccttc ggctcaaccg 540
aagaagtgca tcggaaactg ggaaacttga gtgcagaaga ggacagtgga actccatgtg 600
tagcggtgaa atgcgtagat atatggaaga acaccagtgg cgaaggcggc tgtctggtct 660
gtaactgacg ctgaggctcg aaagtatggg tagcaaacag gattagatac cctggtagtc 720
cataccgtaa acgatgaatg ctaagtgttg gagggtttcc gcccttcagt gctgcagcta 780
acgcattaag cattccgcct ggggagtacg gccgcaaggc tgaaactcaa aggaattgac 840
gggggcccgc acaagcggtg gagcatgtgg tttaattcga agctacgcga agaaccttac 900
caggtcttga catactatgc aaatctaaga gattagacgt tcccttcggg gacatggata 960
caggtggtgc atggttgtcg tcagctcgtg tcgtgagatg ttgggttaag tcccgcaacg 1020
agcgcaaccc ttattatcag ttgccagcat taagttgggc actctggtga gactgccggt 1080
gacaaaccgg aggaaggtgg ggatgacgtc aaatcatcat gccccttatg acctgggcta 1140
cacacgtgct acaatggatg gtacaacgag ttgcgaactc gcgagagtaa gctaatctct 1200
taaagccatt ctcagttcgg attgtaggct gcaactcgcc tacatgaagt cggaatcgct 1260
agtaatcgcg gatcagcatg ccgcggtgaa tacgttcccg ggccttgtac acaccgcccg 1320
tcacaccatg agagtttgta acacccaaag tcggtggggt aaccttttag gaaccagccg 1380
ccta 1384
Claims (10)
1.一种高载乳酸菌肉用直投式发酵剂,其特征在于,包含植物乳杆菌SJ-4,所述植物乳杆菌SJ-4已在中国工业微生物菌种保藏中心保藏,保藏编号为CICC NO.11119s,保藏地址为北京市朝阳区酒仙桥中路24号院6号楼,保藏日期为2020年12月22日。
2.根据权利要求1所述的一种高载乳酸菌肉用直投式发酵剂,其特征在于,所述高载乳酸菌肉用直投式发酵剂中植物乳杆菌SJ-4活菌数≥1011CFU/g。
3.一种根据权利要求1或权利要求2所述的一种高载乳酸菌肉用直投式发酵剂的制备方法,其特征在于,包括如下步骤:
植物乳杆菌SJ-4经高密度培养后离心收集菌泥,并向其中加入由脱脂乳、精氨酸和甘油组成的复合保护剂,之后进行冷冻干燥,即得所述高载乳酸菌肉用直投式发酵剂。
4.根据权利要求3所述的制备方法,其特征在于,所述高密度培养具体为:将植物乳杆菌SJ-4按体积百分比5%接种量接种于优化培养基,37℃100r/min摇床培养16h。
5.根据权利要求4所述的制备方法,其特征在于,所述优化培养基包括2%w/v葡萄糖、1.5%w/v酵母粉、1.5%w/v胰蛋白胨、4%w/v黄瓜汁、0.2%w/v 0.2mol/L Na2HPO4-0.2mol/L KH2PO4、0.5%w/v乙酸钠、0.2%w/v柠檬酸三胺、0.1%w/v吐温80、0.02%w/v硫酸镁和0.005%w/v硫酸锰,余量为水。
6.根据权利要求3所述的制备方法,其特征在于,所述复合保护剂通过以下步骤获得:先将11g脱脂乳和6g精氨酸溶于100mL蒸馏水中115℃灭菌15min,再加入6g甘油121℃灭菌20min。
7.根据权利要求3所述的制备方法,其特征在于,所述复合保护剂加入量与所述植物乳杆菌SJ-4经高密度培养后得到的菌液的体积比是1:10。
8.一种根据权利要求1或权利要求2所述的一种高载乳酸菌肉用直投式发酵剂在制备发酵肉制品中的应用。
9.根据权利要求8所述的应用,其特征在于,所述发酵肉制品的制备方法为:
向处理后的肉样中加入2.5wt%食盐、0.01wt%亚硝酸钠、0.15wt%异抗坏血酸钠、0.8wt%花椒粉、0.5wt%三聚磷酸钠、0.8wt%白糖、0.5wt%白胡椒和0.5wt%五香粉,0-4℃发酵24h后加入所述高载乳酸菌肉用直投式发酵剂,真空滚揉30min后于30℃发酵48h,结束发酵后50℃烘烤3h。
10.根据权利要求9所述的应用,其特征在于,所述高载乳酸菌肉用直投式发酵剂的加入量为0.001-0.002%w/v。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210278576.9A CN114591860A (zh) | 2022-03-17 | 2022-03-17 | 一种高载乳酸菌肉用直投式发酵剂及其制备方法和应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210278576.9A CN114591860A (zh) | 2022-03-17 | 2022-03-17 | 一种高载乳酸菌肉用直投式发酵剂及其制备方法和应用 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN114591860A true CN114591860A (zh) | 2022-06-07 |
Family
ID=81820248
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202210278576.9A Pending CN114591860A (zh) | 2022-03-17 | 2022-03-17 | 一种高载乳酸菌肉用直投式发酵剂及其制备方法和应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN114591860A (zh) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN115720993A (zh) * | 2022-11-25 | 2023-03-03 | 贵州大学 | 一种山梨糖醇介导腌制里脊火腿品质改善的方法 |
Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2009093900A1 (en) * | 2008-01-25 | 2009-07-30 | Stichting Top Institute Food And Nutrition | Mid-log phase lactic acid bacteria for inducing immune tolerance in a subject |
CN101993845A (zh) * | 2010-09-25 | 2011-03-30 | 浙江工业大学 | 植物乳杆菌zy-40及其在发酵制备鱼肉香肠中的应用 |
CN102994420A (zh) * | 2012-11-09 | 2013-03-27 | 黑龙江八一农垦大学 | 一株具有ace抑制活性的耐低温乳酸菌 |
CN104560802A (zh) * | 2014-12-24 | 2015-04-29 | 中国肉类食品综合研究中心 | 一种肉灌制品发酵菌株及发酵剂 |
CN104928215A (zh) * | 2015-06-23 | 2015-09-23 | 贵州大学 | 降胆固醇、耐氧动物双歧杆菌乳亚种bz11的发酵培养基 |
AU2016314014A1 (en) * | 2015-08-31 | 2018-03-22 | Biobalance Co., Ltd. | Method for producing meat capable of reducing saturated fatty acid intake |
-
2022
- 2022-03-17 CN CN202210278576.9A patent/CN114591860A/zh active Pending
Patent Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2009093900A1 (en) * | 2008-01-25 | 2009-07-30 | Stichting Top Institute Food And Nutrition | Mid-log phase lactic acid bacteria for inducing immune tolerance in a subject |
CN101993845A (zh) * | 2010-09-25 | 2011-03-30 | 浙江工业大学 | 植物乳杆菌zy-40及其在发酵制备鱼肉香肠中的应用 |
CN102994420A (zh) * | 2012-11-09 | 2013-03-27 | 黑龙江八一农垦大学 | 一株具有ace抑制活性的耐低温乳酸菌 |
CN104560802A (zh) * | 2014-12-24 | 2015-04-29 | 中国肉类食品综合研究中心 | 一种肉灌制品发酵菌株及发酵剂 |
CN104928215A (zh) * | 2015-06-23 | 2015-09-23 | 贵州大学 | 降胆固醇、耐氧动物双歧杆菌乳亚种bz11的发酵培养基 |
AU2016314014A1 (en) * | 2015-08-31 | 2018-03-22 | Biobalance Co., Ltd. | Method for producing meat capable of reducing saturated fatty acid intake |
Non-Patent Citations (2)
Title |
---|
JIE ZHOU等: "Exploration on method of lyophilized Lactobacillus plantarum SJ-4 and its population succession pattern in fermented tenderloin ham", FOOD SCIENCE OF ANIMAL PRODUCTS * |
周洁: "发酵里脊火腿的乳酸菌筛选和直投式发酵剂的研发", 中国优秀硕士学位论文全文数据库 工程科技I辑, no. 2 * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN115720993A (zh) * | 2022-11-25 | 2023-03-03 | 贵州大学 | 一种山梨糖醇介导腌制里脊火腿品质改善的方法 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN108949645B (zh) | 植物乳杆菌cq02-108及其在发酵香肠制备中的应用 | |
CN108587983B (zh) | 一种植物乳杆菌及其在发酵制备四川香肠中的应用 | |
CN110724651B (zh) | 凝结芽孢杆菌l-h7及其应用 | |
CN106754472B (zh) | 一株果蔬发酵用植物乳杆菌grx15及其应用 | |
CN109306332B (zh) | 发酵乳杆菌cd110及其在发酵香肠制备中的应用 | |
CN104745501B (zh) | 一种表皮葡萄球菌及其应用 | |
CN114231473B (zh) | 一株益生植物乳杆菌及其在低盐发酵肉食品制备中的应用 | |
CN109735461B (zh) | 一株植物乳杆菌及其在降低鱼茶生物胺含量中的应用 | |
CN103421704A (zh) | 一株淡水鱼发酵制品用的植物乳杆菌及应用 | |
CN113068782B (zh) | 一种褐色活性乳酸菌饮品及其制备方法 | |
CN111944731A (zh) | 一种基于多菌共酵技术改善发酵果蔬风味的方法 | |
CN112655863A (zh) | 一种唾液乳杆菌发酵液在作为饲料防霉剂中的应用 | |
CN113969250A (zh) | 干酪乳杆菌dy13及其产品和应用 | |
CN113999793A (zh) | 一株发酵特性良好且具有产香功能的植物乳杆菌及其筛选方法 | |
CN109504636B (zh) | 一种植物乳杆菌p12及其用途 | |
CN114591860A (zh) | 一种高载乳酸菌肉用直投式发酵剂及其制备方法和应用 | |
CN106119166B (zh) | 一株瑞士乳酸菌及其应用 | |
CN113980853A (zh) | 一株高产乳酸的格氏乳球菌wbt0008及其应用 | |
CN109097312B (zh) | 戊糖乳杆菌hn127及其在发酵香肠制备中的应用 | |
CN114350557B (zh) | 一株发酵乳杆菌及其应用 | |
CN112195119B (zh) | 一种能降解生物胺且耐盐的植物乳杆菌及其应用 | |
CN112715758B (zh) | 一株布氏乳杆菌bl4及其制备青贮饲料的方法 | |
CN109456922B (zh) | 一株提高鱼酱发酵品质的中度嗜盐菌菌株花津滩芽孢杆菌 | |
CN112458003A (zh) | 一株产双乙酰植物乳杆菌及其在泡菜中的应用 | |
CN105483039A (zh) | 一株植物乳杆菌及其在生产发酵鹅肉中的应用 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
RJ01 | Rejection of invention patent application after publication |
Application publication date: 20220607 |