CN106434441A - Lactococcus lactis subsp.cremoris and application thereof in cheese production - Google Patents
Lactococcus lactis subsp.cremoris and application thereof in cheese production Download PDFInfo
- Publication number
- CN106434441A CN106434441A CN201610840578.7A CN201610840578A CN106434441A CN 106434441 A CN106434441 A CN 106434441A CN 201610840578 A CN201610840578 A CN 201610840578A CN 106434441 A CN106434441 A CN 106434441A
- Authority
- CN
- China
- Prior art keywords
- lactococcus lactis
- cremoris
- lactis subsp
- culture
- medium
- 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
- 241000194034 Lactococcus lactis subsp. cremoris Species 0.000 title claims abstract description 108
- 235000014962 Streptococcus cremoris Nutrition 0.000 title claims abstract description 108
- 235000013351 cheese Nutrition 0.000 title claims abstract description 56
- 238000004519 manufacturing process Methods 0.000 title abstract description 11
- 235000013336 milk Nutrition 0.000 claims abstract description 67
- 239000008267 milk Substances 0.000 claims abstract description 67
- 210000004080 milk Anatomy 0.000 claims abstract description 67
- 239000002609 medium Substances 0.000 claims description 51
- 239000001963 growth medium Substances 0.000 claims description 49
- 238000005238 degreasing Methods 0.000 claims description 41
- 239000002054 inoculum Substances 0.000 claims description 28
- 230000004913 activation Effects 0.000 claims description 24
- 238000011218 seed culture Methods 0.000 claims description 22
- 235000020183 skimmed milk Nutrition 0.000 claims description 18
- 239000000843 powder Substances 0.000 claims description 15
- 238000000034 method Methods 0.000 claims description 14
- 239000000203 mixture Substances 0.000 claims description 11
- 230000001954 sterilising effect Effects 0.000 claims description 11
- 239000006071 cream Substances 0.000 claims description 10
- 244000005700 microbiome Species 0.000 claims description 10
- 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 9
- 239000008103 glucose Substances 0.000 claims description 9
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 claims description 8
- 229910052564 epsomite Inorganic materials 0.000 claims description 7
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 6
- 239000001888 Peptone Substances 0.000 claims description 6
- 108010080698 Peptones Proteins 0.000 claims description 6
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Natural products OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 6
- -1 citric acid hydrogen diamine Chemical class 0.000 claims description 6
- 235000019319 peptone Nutrition 0.000 claims description 6
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 5
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 5
- 239000001632 sodium acetate Substances 0.000 claims description 5
- 235000017281 sodium acetate Nutrition 0.000 claims description 5
- CATWCRUATKKAAQ-UHFFFAOYSA-N 2,3-dihydroxypropyl dihydrogen phosphate;sodium Chemical compound [Na].[Na].OCC(O)COP(O)(O)=O CATWCRUATKKAAQ-UHFFFAOYSA-N 0.000 claims description 4
- 108010076039 Polyproteins Proteins 0.000 claims description 4
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 4
- VLSOAXRVHARBEQ-UHFFFAOYSA-N [4-fluoro-2-(hydroxymethyl)phenyl]methanol Chemical compound OCC1=CC=C(F)C=C1CO VLSOAXRVHARBEQ-UHFFFAOYSA-N 0.000 claims description 4
- AIUDWMLXCFRVDR-UHFFFAOYSA-N dimethyl 2-(3-ethyl-3-methylpentyl)propanedioate Chemical compound CCC(C)(CC)CCC(C(=O)OC)C(=O)OC AIUDWMLXCFRVDR-UHFFFAOYSA-N 0.000 claims description 4
- 229910000396 dipotassium phosphate Inorganic materials 0.000 claims description 4
- WHWDWIHXSPCOKZ-UHFFFAOYSA-N hexahydrofarnesyl acetone Natural products CC(C)CCCC(C)CCCC(C)CCCC(C)=O WHWDWIHXSPCOKZ-UHFFFAOYSA-N 0.000 claims description 4
- 239000001257 hydrogen Substances 0.000 claims description 4
- 229910052739 hydrogen 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
- 235000010323 ascorbic acid Nutrition 0.000 claims description 3
- 239000011668 ascorbic acid Substances 0.000 claims description 3
- 229960005070 ascorbic acid Drugs 0.000 claims description 3
- 229940041514 candida albicans extract Drugs 0.000 claims description 3
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 3
- 229910000357 manganese(II) sulfate Inorganic materials 0.000 claims description 3
- 239000001509 sodium citrate Substances 0.000 claims description 3
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 claims description 3
- 239000012137 tryptone Substances 0.000 claims description 3
- 239000012138 yeast extract Substances 0.000 claims description 3
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 2
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 2
- 229940099596 manganese sulfate Drugs 0.000 claims description 2
- 239000011702 manganese sulphate Substances 0.000 claims description 2
- 235000007079 manganese sulphate Nutrition 0.000 claims description 2
- 229920000136 polysorbate Polymers 0.000 claims description 2
- 238000002360 preparation method Methods 0.000 abstract description 25
- 238000000855 fermentation Methods 0.000 abstract description 22
- 230000004151 fermentation Effects 0.000 abstract description 22
- 239000000796 flavoring agent Substances 0.000 abstract description 17
- 235000019634 flavors Nutrition 0.000 abstract description 17
- 239000002253 acid Substances 0.000 abstract description 15
- 238000004321 preservation Methods 0.000 abstract description 10
- 239000003795 chemical substances by application Substances 0.000 abstract description 8
- 235000021107 fermented food Nutrition 0.000 abstract description 2
- 238000009629 microbiological culture Methods 0.000 abstract 1
- 230000001580 bacterial effect Effects 0.000 description 34
- 241000194041 Lactococcus lactis subsp. lactis Species 0.000 description 33
- 235000014969 Streptococcus diacetilactis Nutrition 0.000 description 33
- 230000000694 effects Effects 0.000 description 22
- 241000196324 Embryophyta Species 0.000 description 16
- 240000006024 Lactobacillus plantarum Species 0.000 description 16
- 235000013965 Lactobacillus plantarum Nutrition 0.000 description 16
- 241000194035 Lactococcus lactis Species 0.000 description 16
- 235000014897 Streptococcus lactis Nutrition 0.000 description 16
- 229940072205 lactobacillus plantarum Drugs 0.000 description 16
- 235000013365 dairy product Nutrition 0.000 description 15
- 239000012467 final product Substances 0.000 description 12
- 238000002156 mixing Methods 0.000 description 12
- 235000020185 raw untreated milk Nutrition 0.000 description 12
- 230000000052 comparative effect Effects 0.000 description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 11
- 239000000126 substance Substances 0.000 description 9
- 210000000481 breast Anatomy 0.000 description 8
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 8
- 239000004310 lactic acid Substances 0.000 description 7
- 241000894006 Bacteria Species 0.000 description 6
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 6
- 108090000746 Chymosin Proteins 0.000 description 6
- 108091005804 Peptidases Proteins 0.000 description 6
- 102000035195 Peptidases Human genes 0.000 description 6
- 229940080701 chymosin Drugs 0.000 description 6
- GNOLWGAJQVLBSM-UHFFFAOYSA-N n,n,5,7-tetramethyl-1,2,3,4-tetrahydronaphthalen-1-amine Chemical compound C1=C(C)C=C2C(N(C)C)CCCC2=C1C GNOLWGAJQVLBSM-UHFFFAOYSA-N 0.000 description 6
- 235000019833 protease Nutrition 0.000 description 6
- 238000011160 research Methods 0.000 description 6
- 238000003756 stirring Methods 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- 238000005520 cutting process Methods 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 108090000623 proteins and genes Proteins 0.000 description 5
- 238000012549 training Methods 0.000 description 5
- 108020004465 16S ribosomal RNA Proteins 0.000 description 4
- 229920001817 Agar Polymers 0.000 description 4
- 239000005862 Whey Substances 0.000 description 4
- 102000007544 Whey Proteins Human genes 0.000 description 4
- 108010046377 Whey Proteins Proteins 0.000 description 4
- 239000008272 agar Substances 0.000 description 4
- 229940024606 amino acid Drugs 0.000 description 4
- 235000001014 amino acid Nutrition 0.000 description 4
- 150000001413 amino acids Chemical class 0.000 description 4
- 125000003118 aryl group Chemical group 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 4
- 238000001914 filtration Methods 0.000 description 4
- 239000003205 fragrance Substances 0.000 description 4
- 235000014655 lactic acid Nutrition 0.000 description 4
- 239000012528 membrane Substances 0.000 description 4
- 230000002906 microbiologic effect Effects 0.000 description 4
- 235000018102 proteins Nutrition 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 4
- 238000009938 salting Methods 0.000 description 4
- 239000000523 sample Substances 0.000 description 4
- 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 3
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 3
- 229920001202 Inulin Polymers 0.000 description 3
- 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 3
- 229930195725 Mannitol Natural products 0.000 description 3
- WQZGKKKJIJFFOK-PHYPRBDBSA-N alpha-D-galactose Chemical compound OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-PHYPRBDBSA-N 0.000 description 3
- 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 3
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 description 3
- 229910000019 calcium carbonate Inorganic materials 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 3
- 239000012530 fluid Substances 0.000 description 3
- 229930182830 galactose Natural products 0.000 description 3
- 150000004676 glycans Chemical class 0.000 description 3
- 238000011081 inoculation Methods 0.000 description 3
- JYJIGFIDKWBXDU-MNNPPOADSA-N inulin Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)OC[C@]1(OC[C@]2(OC[C@]3(OC[C@]4(OC[C@]5(OC[C@]6(OC[C@]7(OC[C@]8(OC[C@]9(OC[C@]%10(OC[C@]%11(OC[C@]%12(OC[C@]%13(OC[C@]%14(OC[C@]%15(OC[C@]%16(OC[C@]%17(OC[C@]%18(OC[C@]%19(OC[C@]%20(OC[C@]%21(OC[C@]%22(OC[C@]%23(OC[C@]%24(OC[C@]%25(OC[C@]%26(OC[C@]%27(OC[C@]%28(OC[C@]%29(OC[C@]%30(OC[C@]%31(OC[C@]%32(OC[C@]%33(OC[C@]%34(OC[C@]%35(OC[C@]%36(O[C@@H]%37[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O%37)O)[C@H]([C@H](O)[C@@H](CO)O%36)O)[C@H]([C@H](O)[C@@H](CO)O%35)O)[C@H]([C@H](O)[C@@H](CO)O%34)O)[C@H]([C@H](O)[C@@H](CO)O%33)O)[C@H]([C@H](O)[C@@H](CO)O%32)O)[C@H]([C@H](O)[C@@H](CO)O%31)O)[C@H]([C@H](O)[C@@H](CO)O%30)O)[C@H]([C@H](O)[C@@H](CO)O%29)O)[C@H]([C@H](O)[C@@H](CO)O%28)O)[C@H]([C@H](O)[C@@H](CO)O%27)O)[C@H]([C@H](O)[C@@H](CO)O%26)O)[C@H]([C@H](O)[C@@H](CO)O%25)O)[C@H]([C@H](O)[C@@H](CO)O%24)O)[C@H]([C@H](O)[C@@H](CO)O%23)O)[C@H]([C@H](O)[C@@H](CO)O%22)O)[C@H]([C@H](O)[C@@H](CO)O%21)O)[C@H]([C@H](O)[C@@H](CO)O%20)O)[C@H]([C@H](O)[C@@H](CO)O%19)O)[C@H]([C@H](O)[C@@H](CO)O%18)O)[C@H]([C@H](O)[C@@H](CO)O%17)O)[C@H]([C@H](O)[C@@H](CO)O%16)O)[C@H]([C@H](O)[C@@H](CO)O%15)O)[C@H]([C@H](O)[C@@H](CO)O%14)O)[C@H]([C@H](O)[C@@H](CO)O%13)O)[C@H]([C@H](O)[C@@H](CO)O%12)O)[C@H]([C@H](O)[C@@H](CO)O%11)O)[C@H]([C@H](O)[C@@H](CO)O%10)O)[C@H]([C@H](O)[C@@H](CO)O9)O)[C@H]([C@H](O)[C@@H](CO)O8)O)[C@H]([C@H](O)[C@@H](CO)O7)O)[C@H]([C@H](O)[C@@H](CO)O6)O)[C@H]([C@H](O)[C@@H](CO)O5)O)[C@H]([C@H](O)[C@@H](CO)O4)O)[C@H]([C@H](O)[C@@H](CO)O3)O)[C@H]([C@H](O)[C@@H](CO)O2)O)[C@@H](O)[C@H](O)[C@@H](CO)O1 JYJIGFIDKWBXDU-MNNPPOADSA-N 0.000 description 3
- 229940029339 inulin Drugs 0.000 description 3
- 239000000594 mannitol Substances 0.000 description 3
- 235000010355 mannitol Nutrition 0.000 description 3
- 230000004060 metabolic process Effects 0.000 description 3
- 230000000877 morphologic effect Effects 0.000 description 3
- 230000001766 physiological effect Effects 0.000 description 3
- 229920001282 polysaccharide Polymers 0.000 description 3
- 239000005017 polysaccharide Substances 0.000 description 3
- 238000001556 precipitation Methods 0.000 description 3
- 230000002797 proteolythic effect Effects 0.000 description 3
- 239000002994 raw material Substances 0.000 description 3
- 238000012216 screening Methods 0.000 description 3
- GWHJZXXIDMPWGX-UHFFFAOYSA-N 1,2,4-trimethylbenzene Chemical compound CC1=CC=C(C)C(C)=C1 GWHJZXXIDMPWGX-UHFFFAOYSA-N 0.000 description 2
- FERIUCNNQQJTOY-UHFFFAOYSA-N Butyric acid Chemical compound CCCC(O)=O FERIUCNNQQJTOY-UHFFFAOYSA-N 0.000 description 2
- QSJXEFYPDANLFS-UHFFFAOYSA-N Diacetyl Chemical group CC(=O)C(C)=O QSJXEFYPDANLFS-UHFFFAOYSA-N 0.000 description 2
- 241000192125 Firmicutes Species 0.000 description 2
- 241000186660 Lactobacillus Species 0.000 description 2
- 241000194036 Lactococcus Species 0.000 description 2
- 241000192130 Leuconostoc mesenteroides Species 0.000 description 2
- AMQJEAYHLZJPGS-UHFFFAOYSA-N N-Pentanol Chemical compound CCCCCO AMQJEAYHLZJPGS-UHFFFAOYSA-N 0.000 description 2
- CTQNGGLPUBDAKN-UHFFFAOYSA-N O-Xylene Chemical compound CC1=CC=CC=C1C CTQNGGLPUBDAKN-UHFFFAOYSA-N 0.000 description 2
- 241000233855 Orchidaceae Species 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 238000012300 Sequence Analysis Methods 0.000 description 2
- 239000011324 bead Substances 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- YFHFHLSMISYUAQ-UHFFFAOYSA-N farnesane Chemical compound CCC(C)CCCC(C)CCCC(C)C YFHFHLSMISYUAQ-UHFFFAOYSA-N 0.000 description 2
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 description 2
- PHTQWCKDNZKARW-UHFFFAOYSA-N isoamylol Chemical compound CC(C)CCO PHTQWCKDNZKARW-UHFFFAOYSA-N 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 229920001184 polypeptide Polymers 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 239000007858 starting material Substances 0.000 description 2
- POAXRNROGRXJKW-UHFFFAOYSA-N C(CCC)OC(=O)CCCC.C1=CC=CC=C1 Chemical group C(CCC)OC(=O)CCCC.C1=CC=CC=C1 POAXRNROGRXJKW-UHFFFAOYSA-N 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 description 1
- 238000001712 DNA sequencing Methods 0.000 description 1
- 241000287828 Gallus gallus Species 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- PDSNLYSELAIEBU-UHFFFAOYSA-N Longifolene Chemical compound C1CCC(C)(C)C2C3CCC2C1(C)C3=C PDSNLYSELAIEBU-UHFFFAOYSA-N 0.000 description 1
- ZPUKHRHPJKNORC-UHFFFAOYSA-N Longifolene Natural products CC1(C)CCCC2(C)C3CCC1(C3)C2=C ZPUKHRHPJKNORC-UHFFFAOYSA-N 0.000 description 1
- 244000131316 Panax pseudoginseng Species 0.000 description 1
- 235000005035 Panax pseudoginseng ssp. pseudoginseng Nutrition 0.000 description 1
- 235000003140 Panax quinquefolius Nutrition 0.000 description 1
- 241000194020 Streptococcus thermophilus Species 0.000 description 1
- 229930003268 Vitamin C Natural products 0.000 description 1
- NIXOWILDQLNWCW-UHFFFAOYSA-N acrylic acid group Chemical group C(C=C)(=O)O NIXOWILDQLNWCW-UHFFFAOYSA-N 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 235000004251 balanced diet Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 238000005138 cryopreservation Methods 0.000 description 1
- 239000012531 culture fluid Substances 0.000 description 1
- JHIVVAPYMSGYDF-UHFFFAOYSA-N cyclohexanone Chemical group O=C1CCCCC1 JHIVVAPYMSGYDF-UHFFFAOYSA-N 0.000 description 1
- KCIDZIIHRGYJAE-YGFYJFDDSA-L dipotassium;[(2r,3r,4s,5r,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl] phosphate Chemical compound [K+].[K+].OC[C@H]1O[C@H](OP([O-])([O-])=O)[C@H](O)[C@@H](O)[C@H]1O KCIDZIIHRGYJAE-YGFYJFDDSA-L 0.000 description 1
- 235000021113 dry cheese Nutrition 0.000 description 1
- 239000003797 essential amino acid Substances 0.000 description 1
- 235000020776 essential amino acid Nutrition 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 235000019197 fats Nutrition 0.000 description 1
- 235000021105 fermented cheese Nutrition 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 235000008434 ginseng Nutrition 0.000 description 1
- 125000004435 hydrogen atom Chemical class [H]* 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 239000010985 leather Substances 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000002932 luster Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 235000010755 mineral Nutrition 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 230000020477 pH reduction Effects 0.000 description 1
- 239000000575 pesticide Substances 0.000 description 1
- 230000008092 positive effect Effects 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000005180 public health Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 239000013049 sediment Substances 0.000 description 1
- 230000001953 sensory effect Effects 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 238000012807 shake-flask culturing Methods 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 210000001082 somatic cell Anatomy 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000005728 strengthening Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 230000009897 systematic effect Effects 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 235000019154 vitamin C Nutrition 0.000 description 1
- 239000011718 vitamin C Substances 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 239000002023 wood Substances 0.000 description 1
- 235000013618 yogurt Nutrition 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C19/00—Cheese; Cheese preparations; Making thereof
- A23C19/02—Making cheese curd
- A23C19/032—Making cheese curd characterised by the use of specific microorganisms, or enzymes of microbial origin
- A23C19/0323—Making cheese curd characterised by the use of specific microorganisms, or enzymes of microbial origin using only lactic acid bacteria, e.g. Pediococcus and Leuconostoc species; Bifidobacteria; Microbial starters in general
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/21—Streptococcus, lactococcus
- A23V2400/231—Lactis
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/46—Streptococcus ; Enterococcus; Lactococcus
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Microbiology (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Polymers & Plastics (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Food Science & Technology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Dairy Products (AREA)
Abstract
The invention discloses lactococcus lactis subsp.cremoris CGMC No.10746 and application thereof in cheese production. The lactococcus lactis subsp.cremoris is preserved in the China General Microbiological Culture Collection Center, and the preservation number is CGMCC No.10746. The lactococcus lactis subsp.cremoris is good in growth performance in milk, good in phage resistance and moderate in acid production speed, and is used for producing cheese, the flavor is good, and the yield is high. Therefore, the lactococcus lactis subsp.cremoris can be used for preparing a fermentation agent and is applied to production of the cheese and other fermented food. The cheese made through lactococcus lactis subsp.cremoris strains is fragrant, good in flavor and reasonable in texture, and it is clear that the strains have wide application prospects in industrial cheese preparation.
Description
Technical field
The present invention relates to dairy product processing field is and in particular to a kind of lactococcus lactis subsp.cremoris and its in cheesemaking
In application.
Background technology
The invention of cheese makes milk be able to long-term preservation, and different cheese has diversified local flavor, quality structure
And character, be one of advantage source of protein, fat and mineral, wherein also contain essential amino acids and vitamin such as VA,
VB2, VB6 and VB12.Aged cheese be a kind of hard, semi-rigid by Chymosin make protein cross formed milk acidification dense
Contracting product.Cheese is one of optimum selection of balanced diet.
Cheesy flavor is that impact consumer spending selects one of key factor with acceptance, and a lot of western countries receive greatly vigorously
The cheese varieties met but rarely have in China makes inquiries.Cheesy flavor depends on the complicated chemical composition of cheese itself and by wherein micro-
Biological impact is very big.The formation of cheesy flavor is through series of complex microorganism, biochemistry and chemical action process.Though
So cheese microorganism composition major part is derived from the environment of local milk supply and locality, but enterprise produces cheese and remains a need for adding newly
Strain adjusting the local flavor of the cheese needs to adapt to consuming public, therefore excellent to cheesy flavor contribution bacterial strain screening
Imperative.
At present China is more rare to the research of novel species fermented cheese, research and develop suitable Chinese taste, there is China spy
The Flavor Cheese of color is the historic responsibility of the Chinese government, research worker and Dairy Enterprise.
Content of the invention
The technical problem to be solved is to fill up prior art blank, provides a kind of lactococcus lactis butterfat sub-
Plant (Lactococcus lactis subsp.cremoris) and its application in cheesemaking.Described lactococcus lactis
Subsp. cremoris can prepare leaven, thus being applied to cheesemaking.
One of technical scheme is：A kind of lactococcus lactis subsp.cremoris (Lactococcus lactis
Subsp.cremoris), it is deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and preserving number is：
CGMCC No.10746.
In the present invention, lactococcus lactis subsp.cremoris CGMCC No.10746 is isolatable from Inner Mongolia Autonomous Region Lun Beier city
The self-control milk product of Chenbarhu Banner east crow bead Lignum Sappan, its well-grown in breast, acid production speed is moderate, for producing cheese
Excellent flavor, yield is high.This bacterial strain is identified, result is lactococcus lactis subsp.cremoris (Lactococcus lactis
Subsp.cremoris), it is named as E11.This bacterial strain was preserved in Chinese microorganism strain preservation management on April 27th, 2015
Committee's common micro-organisms center, and receive collection and register on the books numbering CGMCC No.10746.
Lactococcus lactis subsp.cremoris CGMCC No.10746 has following Microbiological Characteristics：
1st, morphologic characteristic
Well-grown in M17 culture medium, it is gram positive bacteria, and cell grows in pairs in oval, milky, no
Sprout spore, do not move, no pod membrane.
2nd, the characteristic that culture is learned
In MRS fluid medium, culture forms precipitation；Transparent ring can be formed on the agar culture medium containing Calcium Carbonate；
Amphimicrobian；Chemoheterotrophy, homofermentation metabolism, produce acid with L (+) based on-lactic acid, not aerogenesis, being capable of extracellular polysaccharide.?<
Well-grown under the conditions of 45 DEG C, 30 DEG C of optimum growth temperature.
3rd, physiological property
Acid producing ability is strong, and acid production speed is very fast；Curdled milk speed, curdled milk viscosity is high；Fermenting characteristic aspect, can separate out breast
Clearly, curdled milk structural state is good, give off a strong fragrance；Proteolytic activity is stronger, breaks down proteins can be polypeptide, aminoacid or its
Its inorganic, small molecule organic compound.Glucose, galactose, maltose and Mannitol can be utilized it is impossible to utilize inulin.
The two of technical scheme are：A kind of prepare described lactococcus lactis subsp.cremoris CGMCC No.10746
Method, it comprises the following steps, cultivates described lactococcus lactis subsp.cremoris CGMCC No.10746 in the medium.
Wherein, described culture medium is the conventional culture medium in this area, can grow described lactococcus lactis butterfat sub-
Plant CGMCC No.10746, preferably MRS culture medium, M17 culture medium, degreasing milk medium or culture medium A；More preferably
For culture medium A, described culture medium A by glucose 23g/L, soy peptone 11g/L, Carnis Bovis seu Bubali cream 11g/L, tryptone 5g/L,
Sodium acetate 1.8g/L, K2HPO41.2g/L, sodium citrate 1.2g/L, MgSO4·7H2O 0.4g/L、MnSO4·5H2O 54mg/L、
L-cysteine hydrochloride 0.5g/L and Tween 80 1g/L composition, pH 7.
In the present invention, described MRS culture medium is the conventional MRS culture medium in this area, it is preferred that its by peptone 10g/L,
Carnis Bovis seu Bubali cream 10g/L, yeast extract 5g/L, citric acid hydrogen diamine 2g/L, glucose 20g/L, tween 1ml/L, sodium acetate 5g/L, phosphoric acid
Hydrogen dipotassium 2g/L, magnesium sulfate 0.58g/L and manganese sulfate 0.25g/L composition, pH6.6.
In the present invention, described M17 culture medium is the conventional M17 culture medium in this area, it is preferred that it is by phytone
5g/L, yeast powder 5g/L, polyprotein peptone 5g/L, ascorbic acid 0.5g/L, Carnis Bovis seu Bubali cream 2.5g/L, MgSO4·7H2O 0.01g/L and
Phosphoglycerol disodium 19g/L forms, pH7.
In the present invention, described degreasing milk medium is the conventional degreasing milk medium in this area, preferably 10～14%
Degreasing milk medium.The skimmed milk powder mixing that is dissolved in the water is obtained final product described degreasing milk medium, described percentage ratio is described de-
Fat milk powder accounts for described skimmed milk powder and the mass percent of described water gross mass.
Wherein, the temperature of described culture is the conventional temperature in this area, can grow described lactococcus lactis butterfat sub-
Kind CGMCC No.10746, preferably 26～34 DEG C, more preferably for 30 DEG C.The time of described culture is that this area is conventional
Time, preferably 12～48 hours, more preferably for 48 hours.The pH of described culture is the conventional pH, Neng Gousheng in this area
Long described lactococcus lactis subsp.cremoris CGMCC No.10746, preferably 7.
It is preferred that also including the step carrying out seed culture using seed culture medium before described culture.Described seed
Cultivate as the conventional seed culture in this area.Described seed culture medium is the conventional seed culture medium in this area, preferably
Degreasing milk medium, more preferably for 12% degreasing milk medium, described percentage ratio is mass percent.Described skimmed milk training
Foster base is through 115～120 DEG C, 15～20min sterilizing.The time of described seed culture is this area conventional time, preferably
For 12～48 hours.The temperature of described seed culture is the conventional temperature in this area, preferably 26～34 DEG C.Described seed training
Foster inoculum concentration is the conventional inoculum concentration in this area, preferably 1～3%, described percentage ratio is percent by volume.Described seed
The algebraically of the activation of culture is the conventional algebraically in this area, preferably 2～3 generations.
Wherein, the method for described culture is the method for the conventional culture in this area, preferably shake-flask culture or fermentation tank
Culture.
The technical scheme that the present invention provides three be：Described lactococcus lactis subsp.cremoris CGMCC No.10746 is dry
Application in cheese production.
Described cheese is the conventional cheese in this area, preferably Ida nurse cheese, Gouda cheese, Cheddar and handkerchief
The gloomy cheese of horse.It is more preferably Ida nurse cheese.
Closely, elasticity is moderate for obtained cheese quality, and color and luster is micro- yellow and uniform, has aged cheese characteristic chicken flavor and taste
Taste, the cheese yield with the preparation of this strain is high, can deposit 6～24 months no whey and separate out or become sour under non-refrigerated conditions.
On the basis of meeting common sense in the field, above-mentioned each optimum condition, can combination in any, obtain final product each preferable reality of the present invention
Example.
Agents useful for same of the present invention and raw material are all commercially available.
The positive effect of the present invention is：The invention provides a kind of lactococcus lactis subsp.cremoris CGMCC
No.10746 and its application in cheesemaking.This bacterial strain growth performance in breast is good, and phage resistance is good, acid production speed
Moderate, produced cheesy flavor is good, and yield is high, therefore can be used for preparing leaven, thus being applied to cheese or others
In the production of fermented food.The cheese fragrance being obtained using this lactococcus lactis subsp.cremoris bacterial strain is pleasant, excellent flavor, quality structure
Rationally.It can be seen that this bacterial strain has broad application prospects in preparation of industrialization cheese.
Biomaterial preservation information
The lactococcus lactis subsp.cremoris E11 of the present invention, was deposited in Chinese microorganism strain and protects on April 27th, 2015
Hide administration committee's common micro-organisms center (CGMCC), preservation address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, postcode：
100101, deposit number is：CGMCC No.10746, culture title is lactococcus lactis subsp.cremoris, and Classification And Nomenclature is
Lactococcus lactis subsp.cremoris.
The Lactococcus lactis subsp.lactis C24 of the present invention, was deposited in Chinese microorganism strain and protects on April 27th, 2015
Hide administration committee's common micro-organisms center (CGMCC), preservation address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, postcode：
100101, deposit number is：CGMCC No.10744, culture title is Lactococcus lactis subsp.lactis, and Classification And Nomenclature is
Lactococcus lactis subsp.lactis.
The Lactococcus lactis subsp.lactis C45 of the present invention, was deposited in Chinese microorganism strain and protects on April 27th, 2015
Hide administration committee's common micro-organisms center (CGMCC), preservation address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, postcode：
100101, deposit number is：CGMCC No.10745, culture title is Lactococcus lactis subsp.lactis, and Classification And Nomenclature is
Lactococcus lactis subsp.lactis.
The Lactobacillus plantarum G42 of the present invention, was deposited in Chinese microorganism strain preservation management committee on April 27th, 2015
Member's meeting common micro-organisms center (CGMCC), preservation address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, postcode：100101,
Deposit number is：CGMCC No.10747, culture title is Lactobacillus plantarum, and Classification And Nomenclature is Lactobacillus
plantarum.
Brief description
Fig. 1 is the colonial morphology of lactococcus lactis subsp.cremoris CGMCC No.10746.
Fig. 2 is the growth curve of lactococcus lactis subsp.cremoris CGMCC No.10746.
Fig. 3 is the radar obtaining cheese fermentation later stage main flavor of effect example 2 and comparative example 2 preparation
Figure.
Specific embodiment
Further illustrate the present invention below by the mode of embodiment, but therefore do not limit the present invention to described reality
Apply among a scope.The experimental technique of unreceipted actual conditions in the following example, conventionally and condition, or according to business
Product description selects.
The acquisition of embodiment 1 lactococcus lactis subsp.cremoris CGMCC No.10746
(1), by local self-control milk product, (collection is from Inner Mongolia Autonomous Region Lun Beier city Chenbarhu Banner east crow bead Soviet Union
Wood) take out from cryopreservation tube, carry out gradient dilution using sterilized water, 10 times of gradient dilution liquid are spread evenly across and have passed through 121
DEG C, 15 minutes sterilizing M17 broth bouillon in (M17 broth bouillon is the M17Agar culture medium not containing agar, is purchased from
OXOID company of Britain) 30 DEG C of cultures 48 hours, isolated and purified, carry out microbiologic propertiess and physiological property screening, choose full
Bacterium colony enough to lower condition：Somatic cells are creamy white, oval grows in pairs, spore of not sprouting, and do not move, no pod membrane, thus
According to above-mentioned condition, isolate suitable bacterial strain；These suitable bacterial strains are cultivated under the conditions of degreasing milk medium, selects 48
After hour, pH drops to 3.87, and viscosity is 0.215, and total peptidase activity is calculated as 1.23 bacterial strain (specific operating condition ginseng with Leu
See the embodiment of the present invention 3～4), as bacterial strain E11, inoculating loop picking bacterial strain E11, continuous liquid cultivated for 3 generations, obtained activation
Strain.
(2), the strain of the activation of step (1) gained is inoculated in 12% skimmed milk of pH7 with the inoculum concentration of 2% (v/v)
(skimmed milk powder is purchased from New Zealand Westland cooperation dairy industry company limited to culture medium, and the skimmed milk powder mixing that is dissolved in the water is
Obtain degreasing milk medium, described percentage ratio is that described skimmed milk powder accounts for described skimmed milk powder and the percent mass of described water gross mass
Than) in, 30 DEG C of cultures obtain culture fluid in 16 hours.
Lactococcus lactis subsp.cremoris E11 is deposited in Chinese microorganism strain preservation management committee on April 27th, 2015
Member understands common micro-organisms center (CGMCC), and deposit number is：CGMCC No.10746, culture title is lactococcus lactis breast
Fat subspecies, Classification And Nomenclature is Lactococcus lactis subsp.cremoris.
The morphological characteristic of embodiment 2 lactococcus lactis subsp.cremoris CGMCC No.10746
After 30 DEG C of culture lactococcus lactis subsp.cremoris CGMCC No.10746 of M17 culture medium 48 hours, seen
Examine, colonial morphology is as shown in Figure 1.
Wherein, described M17 culture medium is by phytone 5g/L, yeast powder 5g/L, polyprotein peptone 5g/L, ascorbic acid
0.5g/L, Carnis Bovis seu Bubali cream 2.5g/L, MgSO4·7H2O 0.01g/L and phosphoglycerol disodium 19g/L composition, pH7.
It was found that lactococcus lactis subsp.cremoris CGMCC No.10746 well-grown in M17 culture medium, it is leather
Lan Shi positive bacteria, cell grows in pairs in oval, milky, spore of not sprouting, and does not move, no pod membrane.
The culture characteristic of embodiment 3 lactococcus lactis subsp.cremoris CGMCC No.10746
Lactococcus lactis subsp.cremoris CGMCC No.10746 are cultivated 48 hours for 30 DEG C using M17 culture medium.
Now the growth curve of lactococcus lactis subsp.cremoris CGMCC No.10746 is as shown in Fig. 2 wherein vertical coordinate is
OD600nm, abscissa be culture hourage.Fig. 2 illustrates, prolongation over time, and this bacterial strain constantly rises in value, little to culture 22
Constantly enter stable phase.
Research finds, it is heavy that lactococcus lactis subsp.cremoris CGMCC No.10746 cultivates formation in MRS fluid medium
Form sediment；Transparent ring is all formed on the agar culture medium containing Calcium Carbonate；Its amphimicrobian；Chemoheterotrophy, homofermentation metabolism, produce
Acid with L (+) based on-lactic acid, not aerogenesis, being capable of extracellular polysaccharide.?<Well-grown under the conditions of 45 DEG C, optimum growth temperature 30
℃.
The characteristic of embodiment 4 lactococcus lactis subsp.cremoris CGMCC No.10746
A), product acid activity
Lactococcus lactis subsp.cremoris CGMCC No.10746 is inoculated in equipped with sterilizing by the inoculum concentration of 1% (v/v)
In the triangular flask of degreasing milk medium, it is placed in 30 DEG C of constant incubator cultures, takes out a triangular flask, not inoculate within every 6 hours
The degreasing milk medium of lactococcus lactis subsp.cremoris CGMCC No.10746, as comparison, surveys pH value, and result is as shown in table 1.
The rate of producing acid of table 1 lactococcus lactis subsp.cremoris CGMCC No.10746
| 0 hour | 6 hours | 12 hours | 18 hours | 24 hours | 30 hours | 36 hours | 48 hours | |
| pH | 7 | 5.5 | 5.0 | 4.88 | 4.48 | 4.12 | 3.90 | 3.87 |
As shown in Table 1, lactococcus lactis subsp.cremoris CGMCC No.10746 acid production speed is very fast, and after 48 hours, pH is only
For 3.87, it is stronger to produce sour power, can be used for the pre-acidified speed of raw material milk, and the formation of acid has the function beneficial to strengthening Chymosin
In effect and breast, the dissolving of calcium, is easy to the formation of ziega.
B), the curdled milk time in skimmed milk and curdled milk viscosity measurement
Lactococcus lactis subsp.cremoris CGMCC No.10746 is inoculated in equipped with sterilizing by the inoculum concentration of 1% (v/v)
In the triangular flask of degreasing milk medium, it is placed in 30 DEG C of constant incubator cultures, until its curdled milk, keeps 48 hours under 4 DEG C of environment.
Measure the viscosity (m Pa/s) of each fermentation yogurt respectively with rotational viscometer (purchased from German proRheo company) after curdled milk.Measure
No. 2 rotors of Shi Xuanyong, speed is 64r/s.During mensure, once, minute is 3 minutes to every 15 seconds values, and result is as shown in table 2.
As shown in Table 2, lactococcus lactis subsp.cremoris CGMCC No.10746 curdled milk speed is fast, and curdled milk viscosity is higher.
The curdled milk time of table 2 lactococcus lactis subsp.cremoris CGMCC No.10746 and curdled milk viscosity
| Bacterial strain | The curdled milk time/h | Viscosity m Pa/s |
| Lactococcus lactis subsp.cremoris CGMCC No.10746 | 24 | 215 |
C), fermentation character test
Lactococcus lactis subsp.cremoris CGMCC No.10746 is inoculated in equipped with sterilizing by the inoculum concentration of 1% (v/v)
In the triangular flask of degreasing milk medium, it is placed in 30 DEG C of constant incubator cultures, until its curdled milk, investigates its curd taste, milk surum
Precipitation situation and the quality of grumeleuse, result is as shown in table 3.
The fermentation character of table 3 lactococcus lactis subsp.cremoris CGMCC No.10746
D), total peptidase activity test of thalline cell-free extract
Lactococcus lactis subsp.cremoris CGMCC No.10746 cell-free extract inoculation by 100 μ g protein content volumes
In 100 μ L substrates, cultivate 23 hours at 13 DEG C, measure the concentration of total free amino acid in solution, weigh each bacterial strain according to this
The height of total peptidase activity, (concrete operation step is referring to Zhao wherein to adopt cadmium-ninhydrin method to measure total free amino acid content
Build, etc. the screening of peptidase superior strain and its biological characteristic research [J]. dairy industry science and technology, 2007,123 (2):69-
72.).
Total peptidase activity (being counted with Leu) that result records lactococcus lactis subsp.cremoris CGMCC No.10746 is 1.23.
This explanation lactococcus lactis subsp.cremoris CGMCC No.10746 has certain proteolytic activity, and total peptidase activity value is got over
Greatly, its proteoclastic ability is stronger.
E), other physiological features test
According to《Primary Jie Shi systematic bacteriology handbook》The record of (the 8th edition, 1984, Science Press), to lactococcus lactis
Subsp. cremoris CGMCC No.10746 carries out physiological feature test, it was found that it can utilize glucose, galactose, maltose
With Mannitol it is impossible to utilize inulin.
F), 16S rDNA sequence analysis
16S rDNA sequence analysis：16S using primer amplification lactococcus lactis subsp.cremoris CGMCC No.10746
RDNA fragment, purification, reclaims, (the upper Shanghai's style gloomy promise biotechnology is limited then to carry out DNA sequencing using sequenator ABI3730-XL
Company).It was found that lactococcus lactis subsp.cremoris CGMCC No.10746 is carried out alignment in ncbi database,
It is 99% with lactococcus lactis subsp.cremoris sequence homology.Due to when 16S rDNA sequence homology be higher than 97% when, can
To be considered of the same race in belonging to, therefore lactococcus lactis subsp.cremoris CGMCC No.10746 belongs to lactococcus lactis subsp.cremoris
(Lactococcus lactis subsp.cremoris).
Wherein, the result such as SEQ ID of the 16S rRNA gene sequencing of lactococcus lactis subsp.cremoris CGMCC No.10746
Shown in NO.1.
To sum up, embodiment 2～4 data explanation, described lactococcus lactis subsp.cremoris CGMCC No.10746 have with
Under Microbiological Characteristics：
1st, morphologic characteristic
Well-grown in M17 culture medium, it is gram positive bacteria, and cell grows in pairs in oval, milky, no
Sprout spore, do not move, no pod membrane.
2nd, the characteristic that culture is learned
In MRS fluid medium, culture forms precipitation；Transparent ring can be formed on the agar culture medium containing Calcium Carbonate；
Amphimicrobian；Chemoheterotrophy, homofermentation metabolism, produce acid with L (+) based on-lactic acid, not aerogenesis, being capable of extracellular polysaccharide.?<
Well-grown under the conditions of 45 DEG C, 30 DEG C of optimum growth temperature.
3rd, physiological property
Acid producing ability is strong, and acid production speed is very fast；Curdled milk speed, curdled milk viscosity is high；Fermenting characteristic aspect, can separate out breast
Clearly, curdled milk structural state is good, give off a strong fragrance；Proteolytic activity is stronger, breaks down proteins can be polypeptide, aminoacid or its
Its inorganic, small molecule organic compound.Glucose, galactose, maltose and Mannitol can be utilized it is impossible to utilize inulin.
The fermentation of embodiment 5 lactococcus lactis subsp.cremoris CGMCC No.10746
Seed culture medium：(skimmed milk powder is purchased from the limited public affairs of New Zealand Westland cooperation dairy industry to 12% degreasing milk medium
Department, the skimmed milk powder mixing that is dissolved in the water is obtained final product degreasing milk medium, and described percentage ratio is that described skimmed milk powder accounts for described taking off
Fat milk powder and the mass percent of described water gross mass), described percentage ratio is mass percent.Culture bacterium passes through this skimmed milk
115 DEG C, sterilizing in 15 minutes.
28 DEG C of the cultivation temperature of seed culture medium in seed bottle, cultivation cycle 32 hours, inoculum concentration is 1%, described hundred
Divide than the percent by volume for accounting for fermentation medium.Activated for 2 generations in seed culture medium.
Fermentation medium：By glucose 23g/L, soy peptone 11g/L, Carnis Bovis seu Bubali cream 11g/L, tryptone 5g/L, second
Sour sodium 1.8g/L, K2HPO41.2g/L, sodium citrate 1.2g/L, MgSO4·7H2O0.4g/L、MnSO4·5H2O 54mg/L、L-
Cysteine hydrochloride 0.5g/L and Tween 80 1g/L composition, pH 7.
Cultivation temperature is 30 DEG C, cultivation cycle 48 hours, and culture pH is 7.
The fermentation of embodiment 6 lactococcus lactis subsp.cremoris CGMCC No.10746
Seed culture medium：12% degreasing milk medium, described percentage ratio is mass percent.This skimmed milk cultivates bacterium warp
Cross 120 DEG C, sterilizing in 20 minutes.
26 DEG C of the cultivation temperature of seed culture medium in seed bottle, cultivation cycle 48 hours, inoculum concentration is 3%, described hundred
Divide than the percent by volume for accounting for fermentation medium.Activated for 3 generations in seed culture medium.
Fermentation medium (M17 culture medium)：By phytone 5g/L, yeast powder 5g/L, polyprotein peptone 5g/L, Vitamin C
Sour 0.5g/L, Carnis Bovis seu Bubali cream 2.5g/L, MgSO4·7H2O 0.01g/L and phosphoglycerol disodium 19g/L composition, pH7.
Cultivation temperature is 26 DEG C, cultivation cycle 48 hours, and culture pH is 7.
The fermentation of embodiment 7 lactococcus lactis subsp.cremoris CGMCC No.10746
Seed culture medium：10% degreasing milk medium, described percentage ratio is mass percent.This skimmed milk cultivates bacterium warp
Cross 115 DEG C, sterilizing in 15 minutes.
34 DEG C of the cultivation temperature of seed culture medium in seed bottle, cultivation cycle 12 hours, inoculum concentration is 2%, described hundred
Divide than the percent by volume for accounting for fermentation medium.Activated for 2 generations in seed culture medium.
Fermentation medium (MRS culture medium)：By 20g/L glucose, 10g/L soy peptone, 10g/L Carnis Bovis seu Bubali cream, 5g/L
Yeast extract, 5g/L sodium acetate, 2g/L K2HPO4, 2g/L citric acid hydrogen diamine, 0.58g/L MgSO4·7H2O、0.25g/
LMnSO4·5H2O, 0.5g/L L-cysteine hydrochloride and 1ml/L Tween 80 composition, pH6.6.
Cultivation temperature is 34 DEG C, cultivation cycle 12 hours, and culture pH is 6.6.
The preparation of the leaven containing lactococcus lactis subsp.cremoris CGMCC No.10746 for the embodiment 8
Lactococcus lactis subsp.cremoris E11 is inoculated in, by the inoculum concentration of 2% (v/v), the skimmed milk that 12% (w/v) sterilizes
In culture medium (purchased from New Zealand Westland cooperation dairy industry company limited), it is placed in 30 DEG C of constant incubators and cultivates 24 hours, live
Changed for 2 generations, obtain the bacterial strain of activation.The bacterial strain of activation is inoculated in, by the inoculum concentration of 2% (v/v), the skimmed milk that 12% (w/v) sterilizes
In culture medium, it is placed in 30 DEG C of constant incubator cultures and is enlarged within 24 hours cultivating, be repeated 3 times, obtain final product the leaven containing E11
1.
The preparation of the leaven containing lactococcus lactis subsp.cremoris CGMCC No.10746 for the embodiment 9
(1), lactococcus lactis subsp.cremoris E11, Lactococcus lactis subsp.lactis C24 are connect by the inoculum concentration of 2% (v/v)
Plant in the degreasing milk medium (purchased from New Zealand Westland cooperation dairy industry company limited) that 12% (w/v) sterilizes, be placed in 30
DEG C constant incubator is cultivated 24 hours, activated for 2 generations, obtains the bacterial strain of activation.The bacterial strain of activation is connect by the inoculum concentration of 2% (v/v)
Plant in the degreasing milk medium that 10% (w/v) sterilizes, be placed in 30 DEG C of constant incubators cultures and be enlarged within 24 hours cultivating, weight
Multiple 3 times.
(2), will be sub- to the lactococcus lactis subsp.cremoris E11 of the culture acquisition of step (1) gained, Lactococcus lactis
Plant C24, the viable count according to described lactococcus lactis subsp.cremoris E11, Lactococcus lactis subsp.lactis C24 is 1:1 ratio
Mixing, obtains final product the leaven 2 containing E11 and C24.
The preparation of the leaven containing lactococcus lactis subsp.cremoris CGMCC No.10746 for the embodiment 10
(1), lactococcus lactis subsp.cremoris E11, Lactococcus lactis subsp.lactis C45 are connect by the inoculum concentration of 2% (v/v)
Plant in the degreasing milk medium (purchased from New Zealand Westland cooperation dairy industry company limited) that 12% (w/v) sterilizes, be placed in 30
DEG C constant incubator is cultivated 24 hours, activated for 2 generations, obtains the bacterial strain of activation.The bacterial strain of activation is connect by the inoculum concentration of 2% (v/v)
Plant in the degreasing milk medium that 10% (w/v) sterilizes, be placed in 30 DEG C of constant incubators cultures and be enlarged within 24 hours cultivating, weight
Multiple 3 times.
(2), will be sub- to the lactococcus lactis subsp.cremoris E11 of the culture acquisition of step (1) gained, Lactococcus lactis
Plant C45, the viable count according to described lactococcus lactis subsp.cremoris E11, Lactococcus lactis subsp.lactis C45 is 1:1 ratio
Mixing, obtains final product the leaven 3 containing E11 and C45.
The preparation of the leaven containing lactococcus lactis subsp.cremoris CGMCC No.10746 for the embodiment 11
(1), lactococcus lactis subsp.cremoris E11, Lactobacillus plantarum G42 are inoculated in 12% by the inoculum concentration of 2% (v/v)
(w/v), in the degreasing milk medium (purchased from New Zealand Westland cooperation dairy industry company limited) sterilizing, it is placed in 30 DEG C of constant temperature trainings
Foster case is cultivated 24 hours, activated for 2 generations, obtains the bacterial strain of activation.The bacterial strain of activation is inoculated in 10% by the inoculum concentration of 2% (v/v)
(w/v), in the degreasing milk medium sterilizing, it is placed in 30 DEG C of constant incubator cultures and is enlarged within 24 hours cultivating, be repeated 3 times.
(2), lactococcus lactis subsp.cremoris E11, the Lactobacillus plantarum G42 obtaining the culture of step (1) gained, presses
Viable count according to described lactococcus lactis subsp.cremoris E11, Lactobacillus plantarum G42 is 1:1 ratio mixing, obtains final product containing E11
Leaven 4 with G42.
The preparation of the leaven containing lactococcus lactis subsp.cremoris CGMCC No.10746 for the embodiment 12
(1), by lactococcus lactis subsp.cremoris E11, Lactococcus lactis subsp.lactis C24 and Lactococcus lactis subsp.lactis
C45 is inoculated in the degreasing milk medium that 12% (w/v) sterilize (purchased from New Zealand Westland cooperation by the inoculum concentration of 2% (v/v)
Dairy industry company limited) in, it is placed in 30 DEG C of constant incubators and cultivates 24 hours, activated for 2 generations, obtain the bacterial strain of activation.Bacterium by activation
Strain is inoculated in the degreasing milk medium that 10% (w/v) sterilizes by the inoculum concentration of 2% (v/v), is placed in 30 DEG C of constant incubator trainings
Support and be enlarged within 24 hours cultivating, be repeated 3 times.
(2), will be sub- to the lactococcus lactis subsp.cremoris E11 of the culture acquisition of step (1) gained, Lactococcus lactis
Plant C24 and Lactococcus lactis subsp.lactis C45, according to described lactococcus lactis subsp.cremoris E11, Lactococcus lactis subsp.lactis
The viable count of C24 and Lactococcus lactis subsp.lactis C45 is 6:2:4 ratio mixing, obtains final product sending out containing E11, C24 and C45
Ferment agent 5.
The preparation of the leaven containing lactococcus lactis subsp.cremoris CGMCC No.10746 for the embodiment 13
(1), lactococcus lactis subsp.cremoris E11, Lactococcus lactis subsp.lactis C24 and Lactobacillus plantarum G42 are pressed 2%
(v/v) it is (limited purchased from New Zealand Westland cooperation dairy industry that inoculum concentration is inoculated in the degreasing milk medium that 12% (w/v) sterilize
Company) in, it is placed in 30 DEG C of constant incubators and cultivates 24 hours, activated for 2 generations, obtain the bacterial strain of activation.The bacterial strain of activation is pressed 2%
(v/v) inoculum concentration is inoculated in the degreasing milk medium that 10% (w/v) sterilizes, and is placed in 30 DEG C of constant incubators and cultivates 24 hours
It is enlarged cultivating, be repeated 3 times.
(2), will be sub- to the lactococcus lactis subsp.cremoris E11 of the culture acquisition of step (1) gained, Lactococcus lactis
Plant C24 and Lactobacillus plantarum G42, according to described lactococcus lactis subsp.cremoris E11, Lactococcus lactis subsp.lactis C24 and plant
The viable count of thing lactobacilluss G42 is 2:1:1 ratio mixing, obtains final product the leaven 6 containing E11, C24 and G42.
The preparation of the leaven containing lactococcus lactis subsp.cremoris CGMCC No.10746 for the embodiment 14
(1), lactococcus lactis subsp.cremoris E11, Lactococcus lactis subsp.lactis C45 and Lactobacillus plantarum G42 are pressed 2%
(v/v) it is (limited purchased from New Zealand Westland cooperation dairy industry that inoculum concentration is inoculated in the degreasing milk medium that 12% (w/v) sterilize
Company) in, it is placed in 30 DEG C of constant incubators and cultivates 24 hours, activated for 2 generations, obtain the bacterial strain of activation.The bacterial strain of activation is pressed 2%
(v/v) inoculum concentration is inoculated in the degreasing milk medium that 10% (w/v) sterilizes, and is placed in 30 DEG C of constant incubators and cultivates 24 hours
It is enlarged cultivating, be repeated 3 times.
(2), will be sub- to the lactococcus lactis subsp.cremoris E11 of the culture acquisition of step (1) gained, Lactococcus lactis
Plant C45 and Lactobacillus plantarum G42, according to described lactococcus lactis subsp.cremoris E11, Lactococcus lactis subsp.lactis C45 and plant
The viable count of thing lactobacilluss G42 is 3:1:1 ratio mixing, obtains final product the leaven 7 containing E11, C45 and G42.
The preparation of the leaven containing lactococcus lactis subsp.cremoris CGMCC No.10746 for the embodiment 15
(1), by lactococcus lactis subsp.cremoris E11, Lactococcus lactis subsp.lactis C24, Lactococcus lactis subsp.lactis
C45 and Lactobacillus plantarum G42 is inoculated in the degreasing milk medium that 12% (w/v) sterilize (purchased from new by the inoculum concentration of 2% (v/v)
West orchid Westland cooperation dairy industry company limited) in, it is placed in 30 DEG C of constant incubators and cultivates 24 hours, activated for 2 generations, obtain activation
Bacterial strain.The bacterial strain of activation is inoculated in the degreasing milk medium that 10% (w/v) sterilizes by the inoculum concentration of 2% (v/v), is placed in 30
The culture of DEG C constant incubator is enlarged cultivating for 24 hours, is repeated 3 times.
(2), will be sub- to the lactococcus lactis subsp.cremoris E11 of the culture acquisition of step (1) gained, Lactococcus lactis
Plant C24, Lactococcus lactis subsp.lactis C45 and Lactobacillus plantarum G42, according to described lactococcus lactis subsp.cremoris E11, lactic acid
The viable count of Lactococcus lactic acid subspecies C24, Lactococcus lactis subsp.lactis C45 and Lactobacillus plantarum G42 is 20:1:2:2 ratio
Example mixing, obtains final product the leaven 8 containing E11, C24, C45 and G42.
The preparation of the leaven containing lactococcus lactis subsp.cremoris CGMCC No.10746 for the embodiment 16
(1), by lactococcus lactis subsp.cremoris E11, Lactococcus lactis subsp.lactis C24, Lactococcus lactis subsp.lactis
C45 and Lactobacillus plantarum G42 is inoculated in the degreasing milk medium that 12% (w/v) sterilize (purchased from new by the inoculum concentration of 2% (v/v)
West orchid Westland cooperation dairy industry company limited) in, it is placed in 30 DEG C of constant incubators and cultivates 24 hours, activated for 2 generations, obtain activation
Bacterial strain.The bacterial strain of activation is inoculated in the degreasing milk medium that 10% (w/v) sterilizes by the inoculum concentration of 2% (v/v), is placed in 30
The culture of DEG C constant incubator is enlarged cultivating for 24 hours, is repeated 3 times.
(2), will be sub- to the lactococcus lactis subsp.cremoris E11 of the culture acquisition of step (1) gained, Lactococcus lactis
Plant C24, Lactococcus lactis subsp.lactis C45 and Lactobacillus plantarum G42, according to described lactococcus lactis subsp.cremoris E11, lactic acid
The viable count of Lactococcus lactic acid subspecies C24, Lactococcus lactis subsp.lactis C45 and Lactobacillus plantarum G42 is 5:2:1:2 ratio
Mixing, obtains final product the leaven 9 containing E11, C24, C45 and G42.
Effect example 1
(1) by 50L raw milk (purchased from Kingsoft pasture) through filtering, stir, at 72 DEG C, cool down after 2min pasteurize
To 31 DEG C.The leaven 1 that embodiment 8 is obtained is poured in the raw milk after pasteurize, makes the dense of leaven 1 in raw milk
Spend for 2%, described percentage ratio is percent by volume.0.5g/50L Chymosin (Fromase is added while adding leaven
750XLG, purchased from Chr. Hansen A/S), cultivate under 31 DEG C of constant temperature to pH 6.5.
2) by step 1) to become volume be 1cm for the curd cutting that is obtained3Grumeleuse, discharge all breasts after being slowly stirred 10min
Clearly；The Sal salting of addition 2.5% 20 hours, described percentage ratio is mass percent.Enter square dies afterwards, mould is put
One keg water (about 1.0 kilograms) simultaneously periodically overturns, and the time continues 12h.10 DEG C afterwards, ripe 4 months, obtain Cheddar.
Effect example 2
(1) by 50L raw milk through filtering, stir, at 75 DEG C, after 2min pasteurize, be cooled to 28 DEG C.To implement
The leaven 5 that example 12 is obtained is poured in the raw milk after pasteurize, and the concentration making the leaven 5 in raw milk is 1%, described
Percentage ratio is percent by volume.(Fromase 750XLG, is purchased to add 0.8g/50L Chymosin while adding leaven
Chr. Hansen A/S), cultivate under 28 DEG C of constant temperature to pH 6.5.
2) by step 1) to become volume be 1.2cm for the curd cutting that is obtained3Grumeleuse, be slowly stirred 20min.Then exist
In 45min, temperature is increased to 40 DEG C from 28 DEG C.It is then discharged out all whey；The Sal salting of addition 2% 20 hours, described
Percentage ratio is mass percent.Enter square dies afterwards, a keg water (about 1.0 kilograms) is put on mould and periodically overturns, when
Between continue 15 hours.Then 15 DEG C, ripe 3 months, obtain Ida nurse cheese.
Effect example 3
(1) by 50L raw milk through filtering, stir, at 75 DEG C, after 3min pasteurize, be cooled to 32 DEG C.To implement
The leaven 8 that example 15 is obtained is poured in the raw milk after pasteurize, and the concentration making the leaven 8 in raw milk is 1%, described
Percentage ratio is percent by volume.(Fromase 750XLG, is purchased to add 0.3g/50L Chymosin while adding leaven
Chr. Hansen A/S), cultivate under 32 DEG C of constant temperature to pH 6.5.
2) by step 1) to become volume be 1.4cm for the curd cutting that is obtained3Grumeleuse, be slowly stirred 20min.Use at 35 DEG C
Water washing grumeleuse, and it is kept stirring for 25min.It is then discharged out all whey.Add square dies, squeeze 75min, be acidified 1 hour.
Add 2% Sal salting 4 days, described percentage ratio is mass percent.Then 12 DEG C, ripe 6 months, obtain cutting reach dry
Cheese.
Effect example 4
(1) by 50L raw milk through filtering, stir, at 73 DEG C, after 3min pasteurize, be cooled to 30 DEG C.To implement
The leaven 9 that example 16 is obtained is poured in the raw milk after pasteurize, and the concentration making the leaven 9 in raw milk is 1%, described
Percentage ratio is percent by volume.(Fromase 750XLG, is purchased to add 0.7g/50L Chymosin while adding leaven
Chr. Hansen A/S), cultivate under 30 DEG C of constant temperature to pH 6.5.
2) by step 1) to become volume be 1.0cm for the curd cutting that is obtained3Grumeleuse.Then in 30min, by temperature from
30 DEG C are increased to 39 DEG C, and are kept stirring for 1 hour.It is then discharged out all whey.Add square dies, dry salt salting 2 days.Then
12 DEG C, ripe 5 months, obtain Gouda cheese.
Comparative example 1
Cheddar is made using commercial fermentation agent CHOOZITTM RM 32 (DANISCO company).Contain in this leaven
Lactococcus lactis subsp.cremoris, lactococcus lactis subsp.cremoris, lactococcus lactis biacetyl subspecies and streptococcus thermophiluss.
Remaining preparation method is identical with the preparation method of effect example 1.
Comparative example 2
Ida nurse cheese is made using commercial fermentation agent FD-DVS R-704 (Chr. Hansen A/S).In this leaven
Containing lactococcus lactis subsp.cremoris and lactococcus lactis subsp.cremoris.
Remaining preparation method is identical with the preparation method of effect example 2.
Comparative example 3
Commercial fermentation agent FD-DVS CHN-22 (Chr. Hansen A/S) makes Cheddar.Contain in this leaven
Lactococcus lactis subsp.cremoris, lactococcus lactis subsp.cremoris, lactococcus lactis biacetyl subspecies and leukonid.
Remaining preparation method is identical with the preparation method of effect example 3.
Comparative example 4
(1) by Lactococcus lactis subsp.lactis BD164, (deposit number is：CGMCC No.10751), lactococcus lactis breast
(deposit number is for sour subspecies BD2263：CGMCC No.10749), (deposit number is Lactococcus lactis subsp.lactis BD401：
CGMCC No.10752) and Leuconostoc mesenteroides LM79 (deposit number is：CGMCC No.10750) by 2% (v/v) inoculation
Amount is inoculated in the degreasing milk medium (purchased from New Zealand Westland cooperation dairy industry company limited) that 10% (w/v) sterilizes, and puts
Cultivate 24 hours in 30 DEG C of constant incubators, activated for 2 generations, obtain the bacterial strain of activation.The bacterial strain of activation is pressed the inoculation of 2% (v/v)
Amount is inoculated in the degreasing milk medium that 10% (w/v) sterilizes, and is placed in 30 DEG C of constant incubator cultures and is enlarged within 24 hours training
Support, be repeated 3 times, obtain culture.
(2) culture of step (1) gained is obtained Lactococcus lactis subsp.lactis BD164, Lactococcus lactis Asia
Plant BD2263, Lactococcus lactis subsp.lactis BD401 and Leuconostoc mesenteroides LM79, then according to described Lactococcus lactis
Subspecies BD401, described Lactococcus lactis subsp.lactis BD2263, described Lactococcus lactis subsp.lactis BD164 and described goldbeater's skin
The viable count of leukonid LM79 is 1:1:1:1 ratio mixing, obtains cheese starter A.
(3) cheese starter A is adopted to make Cheddar.Remaining preparation method is complete with the preparation method of effect example 1
Exactly the same.
The mensure of flavor substance in Application Example 1 cheese
Using low-polarity components (GC-MS) (referring to Wu Gu equality, gas chromatograph-mass spectrometer method measure 7 in vegetable in amino first
Ester pesticide, Chinese public health, 1999,15 (6):534), measure effect example 1,4 and comparative example 1,4 is made
The flavor substance of standby cheese.Result is as shown in table 4.
Table 4 cheese fermentation initial stage main flavor and its content
Wherein, the implication of "-" represents under content as little as minimum detection value, can estimate and not contain this material.Can by table 4
To find out, compare with the Ida cheese (i.e. the cheese of comparative example 1 preparation) of commercial fermentation agent preparation and comparative example 4,
It is different that the aged cheese that the leaven of the special bacterial strain gained that the present invention selects makes has special aromatic substance at the fermentation initial stage
Amylalcohol, isoamyl alcohol mainly has calvados fragrance and acid.And other key aroma compositions and commercial fermentation agent preparation
Ida cheese is compared content and is had significant difference.
Application Example 2
Using low-polarity components (GC-MS), measure the cheese prepared by effect example 2,3 and comparative example 2,4
Aromatic substance.Result is as shown in table 5.Fig. 3 is the radar map of these aromatic substance contents.
The mensure of aromatic substance content in table 5 cheese
Wherein, the implication of "-" represents under content as little as minimum detection value, can estimate and not contain this material.By table 5 He
Fig. 3 can be seen that in cheese prepared by effect example and also detects butanoic acid, 1,2,4- trimethylbenzene, amylalcohol, Ketohexamethylene, 1- second
Base -2,3 dimethyl benzene, 3- methyltridec, 2,6,10- trimethyldodecane, acrylic acid-2-ethyl caproite, longifolene, adjacent benzene
Dibutyl carboxylic acid, and do not detect above-mentioned substance in the cheese of comparative example's preparation.
Application Example 3
Using Texture instrument (purchased from Stable Micro Systems company of Britain), will be real to effect example 1,2 and contrast
Apply the mensure that the cheese prepared by example 3 grade carries out texture characteristic.Wherein, metering system is to push, the test speed 5.00mm/ second；
Testing time is 5 seconds；Probe type is P/5 probe, every group of sample parallel assay 5 times.Result is as shown in table 6.
The mensure of table 6 cheese texture characteristic
| Embodiment | Hardness/g | Viscosity | Elasticity |
| Effect example 1 | 1301.66±38.32 | -40.00±7.64 | 0.25±0.04 |
| Effect example 2 | 1206.85±55.36 | -39.74±5.46 | 0.32±0.08 |
| Comparative example 3 | 1026.34±215.84 | -40.50±6.76 | 0.21±0.06 |
Application Example 4
Subjective appreciation is tested
This subjective appreciation personnel include 12 personnel being engaged in food research, are all familiar with subjective appreciation points for attention and comment
Minute mark is accurate.Total score is 50 points, and fraction is evaluated in accordance with table 7.Each assessment officer is independently evaluated, and changes every time during sample using clear water
Gargle, result is as shown in table 8.
Table 7 cheese subjective appreciation method
Table 8 cheese results of sensory evaluation
The result explanation of table 8, the cheese that the present invention is obtained is substantially better than contrast at aspects such as characteristic odor, plasticity to be implemented
Cheese obtained by example, overall scoring is also of a relatively high.
It should be understood that after the above having read the present invention, those skilled in the art can make various to the present invention
Change or change, these equivalent form of values equally fall within the application appended claims limited range.
Claims (10)
1. a kind of lactococcus lactis subsp.cremoris (Lactococcus lactis subsp.cremoris) it is characterised in that its
It is deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preserving number is：CGMCC No.10746.
2. a kind of method preparing lactococcus lactis subsp.cremoris CGMCC No.10746 as claimed in claim 1, its feature
It is, it comprises the following steps, cultivate described lactococcus lactis subsp.cremoris CGMCC No.10746 in the medium.
3. method as claimed in claim 2 is it is characterised in that described culture medium is MRS culture medium, M17 culture medium, skimmed milk
Culture medium or culture medium A, described culture medium A is by glucose 23g/L, soy peptone 11g/L, Carnis Bovis seu Bubali cream 11g/L, tryptone
5g/L, sodium acetate 1.8g/L, K2HPO41.2g/L, sodium citrate 1.2g/L, MgSO4·7H2O 0.4g/L、MnSO4·5H2O
54mg/L, L-cysteine hydrochloride 0.5g/L and Tween 80 1g/L composition；It is preferred that described MRS culture medium is by peptone
10g/L, Carnis Bovis seu Bubali cream 10g/L, yeast extract 5g/L, citric acid hydrogen diamine 2g/L, glucose 20g/L, tween 1ml/L, sodium acetate 5g/
L, dipotassium hydrogen phosphate 2g/L, magnesium sulfate 0.58g/L and manganese sulfate 0.25g/L composition；Described M17 culture medium is by phytone
5g/L, yeast powder 5g/L, polyprotein peptone 5g/L, ascorbic acid 0.5g/L, Carnis Bovis seu Bubali cream 2.5g/L, MgSO4·7H2O 0.01g/L and
Phosphoglycerol disodium 19g/L forms；Described degreasing milk medium is 10～14% degreasing milk mediums, and described percentage ratio is quality
Percentage ratio；And/or, it is preferred that described culture medium is described culture medium A.
4. method as claimed in claim 2 it is characterised in that described culture temperature be 26～34 DEG C, preferably 30 DEG C；
And/or, the time of described culture is 12～48 hours.
5. method as claimed in claim 2 is it is characterised in that also include being planted using seed culture medium before described culture
The step of son culture；Described seed culture is degreasing milk medium.
6. method as claimed in claim 5 it is characterised in that described degreasing milk medium be 12% degreasing milk medium, institute
The percentage ratio stated is mass percent；And/or, described degreasing milk medium is through 115～120 DEG C, 15～20min sterilizing.
7. method as claimed in claim 5 is it is characterised in that the temperature of described seed culture is 26～34 DEG C；And/or, institute
The time stating seed culture is 12～48 hours.
8. method as claimed in claim 5 it is characterised in that described seed culture inoculum concentration be 1～3%, described percentage
Than for percent by volume；And/or, the algebraically of the activation of described seed culture was 2～3 generations.
9. a kind of lactococcus lactis subsp.cremoris CGMCC No.10746 as claimed in claim 1 answering in cheesemaking
With.
10. application as claimed in claim 9 is it is characterised in that described cheese is Ida nurse cheese, Gouda cheese and Qie Da
Cheese.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610840578.7A CN106434441B (en) | 2016-09-21 | 2016-09-21 | A kind of lactococcus lactis subsp and its application in cheesemaking |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610840578.7A CN106434441B (en) | 2016-09-21 | 2016-09-21 | A kind of lactococcus lactis subsp and its application in cheesemaking |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106434441A true CN106434441A (en) | 2017-02-22 |
| CN106434441B CN106434441B (en) | 2019-07-26 |
Family
ID=58166185
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610840578.7A Active CN106434441B (en) | 2016-09-21 | 2016-09-21 | A kind of lactococcus lactis subsp and its application in cheesemaking |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106434441B (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109105491A (en) * | 2018-09-12 | 2019-01-01 | 内蒙古农业大学 | One plant produces the glutinous Lactococcus lactis IMAU11823 for producing junket fragrance and its application |
| CN112574928A (en) * | 2021-01-05 | 2021-03-30 | 东北林业大学 | Cold region straw decomposition microbial inoculum and preparation method and application thereof |
| CN114806949A (en) * | 2022-05-06 | 2022-07-29 | 河北一然生物科技股份有限公司 | Lactococcus lactis S133 and application thereof in improving intestinal health and enhancing immunity |
| CN117062905A (en) * | 2021-03-02 | 2023-11-14 | 富吉高株式会社 | Novel lactic acid bacteria strain and composition comprising the same |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101961054A (en) * | 2009-07-22 | 2011-02-02 | 内蒙古伊利实业集团股份有限公司 | Cheese and preparation method thereof |
| CN102952758A (en) * | 2011-08-17 | 2013-03-06 | 北京德青源农业科技股份有限公司 | Lactoccous lactis subsp. cremoris strain and application thereof |
| CN104946566A (en) * | 2015-07-01 | 2015-09-30 | 光明乳业股份有限公司 | Lactococcus lactis subsp.lactis strain and application thereof in cheese production |
-
2016
- 2016-09-21 CN CN201610840578.7A patent/CN106434441B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101961054A (en) * | 2009-07-22 | 2011-02-02 | 内蒙古伊利实业集团股份有限公司 | Cheese and preparation method thereof |
| CN102952758A (en) * | 2011-08-17 | 2013-03-06 | 北京德青源农业科技股份有限公司 | Lactoccous lactis subsp. cremoris strain and application thereof |
| CN104946566A (en) * | 2015-07-01 | 2015-09-30 | 光明乳业股份有限公司 | Lactococcus lactis subsp.lactis strain and application thereof in cheese production |
Non-Patent Citations (2)
| Title |
|---|
| *** 等: "乳酸乳球菌对切达干酪成熟过程中质构和风味的影响", 《食品科学》 * |
| 陈雪 等: "切达干酪发酵菌种的特性测定及发酵干酪的评价", 《食品工业科技》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109105491A (en) * | 2018-09-12 | 2019-01-01 | 内蒙古农业大学 | One plant produces the glutinous Lactococcus lactis IMAU11823 for producing junket fragrance and its application |
| CN109105491B (en) * | 2018-09-12 | 2021-07-23 | 内蒙古农业大学 | Lactococcus lactis IMAU11823 capable of producing sticky cheese fragrance and application thereof |
| CN112574928A (en) * | 2021-01-05 | 2021-03-30 | 东北林业大学 | Cold region straw decomposition microbial inoculum and preparation method and application thereof |
| CN117062905A (en) * | 2021-03-02 | 2023-11-14 | 富吉高株式会社 | Novel lactic acid bacteria strain and composition comprising the same |
| CN114806949A (en) * | 2022-05-06 | 2022-07-29 | 河北一然生物科技股份有限公司 | Lactococcus lactis S133 and application thereof in improving intestinal health and enhancing immunity |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106434441B (en) | 2019-07-26 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106148253B (en) | A kind of lactobacillus plantarum and its application in cheesemaking | |
| Heenan et al. | Survival and sensory acceptability of probiotic microorganisms in a nonfermented frozen vegetarian dessert | |
| CN103421704B (en) | Lactobacillus plantarum for freshwater fish fermentation product and application thereof | |
| CN104911134B (en) | A kind of Leuconostoc mesenteroides and its application in cheesemaking | |
| CN104585333B (en) | A kind of monascus cheese and a kind of purplish red bent and its cultural method | |
| CN104962483B (en) | A kind of monascus purpureus bacterial strain and application thereof | |
| CN101485473A (en) | Fish meat emulsion fermented with mixed bacteria based on lactobacillus and preparation method thereof | |
| CN106434441A (en) | Lactococcus lactis subsp.cremoris and application thereof in cheese production | |
| CN106190931B (en) | A kind of Lactococcus lactis subsp. lactis and its application in cheesemaking | |
| CN106148252B (en) | A kind of Lactococcus lactis subsp. lactis and its application in cheesemaking | |
| CN101691551A (en) | Lactobacillus plantarum for food fermentation and applications thereof | |
| CN103589658B (en) | One strain bacteriocinogeny bifidus bacillus and the application in suppression post-acidification of yoghurt thereof | |
| MALAKA et al. | Quality and storage time of traditional dangke cheese inoculated with indigenous lactic acid bacteria isolated from Enrekang District, South Sulawesi, Indonesia | |
| CN104894037B (en) | A kind of Lactococcus lactis subsp. lactis and its purposes in cheesemaking | |
| CN104894038B (en) | A kind of Lactococcus lactis subsp. lactis and its application in cheesemaking | |
| CN104946566B (en) | A kind of Lactococcus lactis subsp. lactis and its application in cheesemaking | |
| CN102753026A (en) | Improved soy milk fermentation | |
| CN110331104A (en) | A kind of lactobacillus plantarum CV10D1 and its application | |
| CN105505789B (en) | One plant of monascus purpureus bacterial strain and its preparing the application in food | |
| CN105505788B (en) | One plant of monascus purpureus bacterial strain and its preparing the application in food | |
| CN101333507B (en) | Enterococcus faecium Grx28 with flame resistance and its use | |
| CN106399173B (en) | A kind of cheese leavening and preparation method thereof | |
| CN104877952B (en) | A kind of Lactococcus lactis subsp. lactis and its application in preparation of cheese | |
| CN110055189A (en) | A kind of lactobacillus plantarum YL15 and its application in red wine flavor yoghourt | |
| CN104099256A (en) | Strain of Monaacus sp. and use thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |