CN116925953A - Fermentation method of stink mandarin fish and strain used by same - Google Patents
Fermentation method of stink mandarin fish and strain used by same Download PDFInfo
- Publication number
- CN116925953A CN116925953A CN202310217694.3A CN202310217694A CN116925953A CN 116925953 A CN116925953 A CN 116925953A CN 202310217694 A CN202310217694 A CN 202310217694A CN 116925953 A CN116925953 A CN 116925953A
- Authority
- CN
- China
- Prior art keywords
- mandarin fish
- fermentation
- inoculum
- staphylococcus
- agar
- 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
- 238000000855 fermentation Methods 0.000 title claims abstract description 92
- 230000004151 fermentation Effects 0.000 title claims abstract description 92
- 241000404975 Synchiropus splendidus Species 0.000 title claims abstract description 67
- 238000000034 method Methods 0.000 title claims abstract description 29
- 241000251468 Actinopterygii Species 0.000 claims abstract description 41
- 240000006024 Lactobacillus plantarum Species 0.000 claims abstract description 37
- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims abstract description 37
- 229940072205 lactobacillus plantarum Drugs 0.000 claims abstract description 37
- 238000011081 inoculation Methods 0.000 claims abstract description 30
- 235000013372 meat Nutrition 0.000 claims abstract description 30
- 241000191940 Staphylococcus Species 0.000 claims abstract description 25
- 239000002689 soil Substances 0.000 claims abstract description 23
- 241000191967 Staphylococcus aureus Species 0.000 claims abstract description 14
- 229920001817 Agar Polymers 0.000 claims description 31
- 239000008272 agar Substances 0.000 claims description 31
- 239000002054 inoculum Substances 0.000 claims description 31
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 22
- 239000001963 growth medium Substances 0.000 claims description 22
- 239000002609 medium Substances 0.000 claims description 19
- 238000012258 culturing Methods 0.000 claims description 18
- 239000011780 sodium chloride Substances 0.000 claims description 16
- 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
- 239000000843 powder Substances 0.000 claims description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 10
- 238000004321 preservation Methods 0.000 claims description 9
- 241001052560 Thallis Species 0.000 claims description 8
- 230000004913 activation Effects 0.000 claims description 7
- 239000007788 liquid Substances 0.000 claims description 7
- 235000015278 beef Nutrition 0.000 claims description 6
- 238000004140 cleaning Methods 0.000 claims description 6
- 210000001835 viscera Anatomy 0.000 claims description 6
- 239000001888 Peptone Substances 0.000 claims description 5
- 108010080698 Peptones Proteins 0.000 claims description 5
- 238000009630 liquid culture Methods 0.000 claims description 5
- 235000019319 peptone Nutrition 0.000 claims description 5
- 230000001954 sterilising effect Effects 0.000 claims description 5
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 4
- 239000012153 distilled water Substances 0.000 claims description 4
- 239000000284 extract Substances 0.000 claims description 4
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 3
- 229940099596 manganese sulfate Drugs 0.000 claims description 3
- 235000007079 manganese sulphate Nutrition 0.000 claims description 3
- 239000011702 manganese sulphate Substances 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
- 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 claims description 2
- 229930195725 Mannitol Natural products 0.000 claims description 2
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 2
- KLOIYEQEVSIOOO-UHFFFAOYSA-N carbocromen Chemical compound CC1=C(CCN(CC)CC)C(=O)OC2=CC(OCC(=O)OCC)=CC=C21 KLOIYEQEVSIOOO-UHFFFAOYSA-N 0.000 claims description 2
- 238000005119 centrifugation Methods 0.000 claims description 2
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 2
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 2
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 2
- 239000000594 mannitol Substances 0.000 claims description 2
- 235000010355 mannitol Nutrition 0.000 claims description 2
- 239000002504 physiological saline solution Substances 0.000 claims description 2
- 235000020245 plant milk Nutrition 0.000 claims description 2
- 229920000136 polysorbate Polymers 0.000 claims description 2
- 239000001632 sodium acetate Substances 0.000 claims description 2
- 235000017281 sodium acetate Nutrition 0.000 claims description 2
- 241000186660 Lactobacillus Species 0.000 claims 1
- 229940039696 lactobacillus Drugs 0.000 claims 1
- 150000001412 amines Chemical class 0.000 abstract description 30
- 230000000035 biogenic effect Effects 0.000 abstract description 26
- 239000000796 flavoring agent Substances 0.000 abstract description 11
- 235000019634 flavors Nutrition 0.000 abstract description 11
- 244000005700 microbiome Species 0.000 abstract description 9
- 238000000354 decomposition reaction Methods 0.000 abstract description 5
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 abstract description 4
- 150000001875 compounds Chemical class 0.000 abstract description 4
- 230000002829 reductive effect Effects 0.000 abstract description 4
- 239000011593 sulfur Substances 0.000 abstract description 4
- 229910052717 sulfur Inorganic materials 0.000 abstract description 4
- 102000004882 Lipase Human genes 0.000 abstract description 2
- 239000004367 Lipase Substances 0.000 abstract description 2
- 230000008859 change Effects 0.000 abstract description 2
- 235000019421 lipase Nutrition 0.000 abstract description 2
- 101710158368 Extracellular lipase Proteins 0.000 abstract 1
- 101710089384 Extracellular protease Proteins 0.000 abstract 1
- 108010028690 Fish Proteins Proteins 0.000 abstract 1
- 101710128940 Triacylglycerol lipase Proteins 0.000 abstract 1
- SIKJAQJRHWYJAI-UHFFFAOYSA-N Indole Chemical compound C1=CC=C2NC=CC2=C1 SIKJAQJRHWYJAI-UHFFFAOYSA-N 0.000 description 22
- 150000001413 amino acids Chemical class 0.000 description 18
- 238000002474 experimental method Methods 0.000 description 16
- 239000000047 product Substances 0.000 description 15
- 241000894006 Bacteria Species 0.000 description 14
- PZOUSPYUWWUPPK-UHFFFAOYSA-N indole Natural products CC1=CC=CC2=C1C=CN2 PZOUSPYUWWUPPK-UHFFFAOYSA-N 0.000 description 11
- RKJUIXBNRJVNHR-UHFFFAOYSA-N indolenine Natural products C1=CC=C2CC=NC2=C1 RKJUIXBNRJVNHR-UHFFFAOYSA-N 0.000 description 11
- 239000000523 sample Substances 0.000 description 11
- 206010018910 Haemolysis Diseases 0.000 description 10
- NTYJJOPFIAHURM-UHFFFAOYSA-N Histamine Chemical compound NCCC1=CN=CN1 NTYJJOPFIAHURM-UHFFFAOYSA-N 0.000 description 10
- 230000008588 hemolysis Effects 0.000 description 10
- 239000000126 substance Substances 0.000 description 10
- 241000768264 Staphylococcus edaphicus Species 0.000 description 9
- 238000004519 manufacturing process Methods 0.000 description 8
- KIDHWZJUCRJVML-UHFFFAOYSA-N putrescine Chemical compound NCCCCN KIDHWZJUCRJVML-UHFFFAOYSA-N 0.000 description 8
- 239000002994 raw material Substances 0.000 description 8
- GETQZCLCWQTVFV-UHFFFAOYSA-N trimethylamine Chemical compound CN(C)C GETQZCLCWQTVFV-UHFFFAOYSA-N 0.000 description 8
- 102000004169 proteins and genes Human genes 0.000 description 7
- 108090000623 proteins and genes Proteins 0.000 description 7
- 239000000243 solution Substances 0.000 description 7
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 6
- PAYRUJLWNCNPSJ-UHFFFAOYSA-N Aniline Chemical compound NC1=CC=CC=C1 PAYRUJLWNCNPSJ-UHFFFAOYSA-N 0.000 description 6
- 239000007858 starting material Substances 0.000 description 6
- 230000001580 bacterial effect Effects 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 229960001340 histamine Drugs 0.000 description 5
- 230000000813 microbial effect Effects 0.000 description 5
- 238000012216 screening Methods 0.000 description 5
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
- WSMYVTOQOOLQHP-UHFFFAOYSA-N Malondialdehyde Chemical compound O=CCC=O WSMYVTOQOOLQHP-UHFFFAOYSA-N 0.000 description 4
- 239000005700 Putrescine Substances 0.000 description 4
- 235000013305 food Nutrition 0.000 description 4
- 229940118019 malondialdehyde Drugs 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 108010006464 Hemolysin Proteins Proteins 0.000 description 3
- 238000009825 accumulation Methods 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 239000003925 fat Substances 0.000 description 3
- -1 fatty acid compounds Chemical class 0.000 description 3
- 239000007789 gas Substances 0.000 description 3
- 239000003228 hemolysin Substances 0.000 description 3
- 230000005764 inhibitory process Effects 0.000 description 3
- 239000007924 injection Substances 0.000 description 3
- 238000002347 injection Methods 0.000 description 3
- 230000003647 oxidation Effects 0.000 description 3
- 238000007254 oxidation reaction Methods 0.000 description 3
- 239000013641 positive control Substances 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- BWZVCCNYKMEVEX-UHFFFAOYSA-N 2,4,6-Trimethylpyridine Chemical compound CC1=CC(C)=NC(C)=C1 BWZVCCNYKMEVEX-UHFFFAOYSA-N 0.000 description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 150000001299 aldehydes Chemical class 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- VHRGRCVQAFMJIZ-UHFFFAOYSA-N cadaverine Chemical compound NCCCCCN VHRGRCVQAFMJIZ-UHFFFAOYSA-N 0.000 description 2
- 229910000019 calcium carbonate Inorganic materials 0.000 description 2
- 235000010216 calcium carbonate Nutrition 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 235000009508 confectionery Nutrition 0.000 description 2
- 238000005520 cutting process Methods 0.000 description 2
- 238000006114 decarboxylation reaction Methods 0.000 description 2
- 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 2
- 238000000605 extraction Methods 0.000 description 2
- 230000002349 favourable effect Effects 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 235000013332 fish product Nutrition 0.000 description 2
- 238000011010 flushing procedure Methods 0.000 description 2
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 230000015784 hyperosmotic salinity response Effects 0.000 description 2
- 230000000670 limiting effect Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 230000001766 physiological effect Effects 0.000 description 2
- NGVDGCNFYWLIFO-UHFFFAOYSA-N pyridoxal 5'-phosphate Chemical compound CC1=NC=C(COP(O)(O)=O)C(C=O)=C1O NGVDGCNFYWLIFO-UHFFFAOYSA-N 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- ATHGHQPFGPMSJY-UHFFFAOYSA-N spermidine Chemical compound NCCCCNCCCN ATHGHQPFGPMSJY-UHFFFAOYSA-N 0.000 description 2
- PFNFFQXMRSDOHW-UHFFFAOYSA-N spermine Chemical compound NCCCNCCCCNCCCN PFNFFQXMRSDOHW-UHFFFAOYSA-N 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 235000019640 taste Nutrition 0.000 description 2
- APJYDQYYACXCRM-UHFFFAOYSA-N tryptamine Chemical compound C1=CC=C2C(CCN)=CNC2=C1 APJYDQYYACXCRM-UHFFFAOYSA-N 0.000 description 2
- DZGWFCGJZKJUFP-UHFFFAOYSA-N tyramine Chemical compound NCCC1=CC=C(O)C=C1 DZGWFCGJZKJUFP-UHFFFAOYSA-N 0.000 description 2
- 235000019583 umami taste Nutrition 0.000 description 2
- FZRCKLPSHGTOAU-UHFFFAOYSA-N 6-amino-1,4-dimethylcyclohexa-2,4-diene-1-carbaldehyde Chemical compound CC1=CC(N)C(C)(C=O)C=C1 FZRCKLPSHGTOAU-UHFFFAOYSA-N 0.000 description 1
- 241000589291 Acinetobacter Species 0.000 description 1
- 206010067484 Adverse reaction Diseases 0.000 description 1
- USFZMSVCRYTOJT-UHFFFAOYSA-N Ammonium acetate Chemical compound N.CC(O)=O USFZMSVCRYTOJT-UHFFFAOYSA-N 0.000 description 1
- 239000005695 Ammonium acetate Substances 0.000 description 1
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 244000025254 Cannabis sativa Species 0.000 description 1
- 238000007400 DNA extraction Methods 0.000 description 1
- XPDXVDYUQZHFPV-UHFFFAOYSA-N Dansyl Chloride Chemical compound C1=CC=C2C(N(C)C)=CC=CC2=C1S(Cl)(=O)=O XPDXVDYUQZHFPV-UHFFFAOYSA-N 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 238000003794 Gram staining Methods 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 102000004157 Hydrolases Human genes 0.000 description 1
- 108090000604 Hydrolases Proteins 0.000 description 1
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 description 1
- HNDVDQJCIGZPNO-YFKPBYRVSA-N L-histidine Chemical compound OC(=O)[C@@H](N)CC1=CN=CN1 HNDVDQJCIGZPNO-YFKPBYRVSA-N 0.000 description 1
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 description 1
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 1
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 1
- 108090001060 Lipase Proteins 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- BHHGXPLMPWCGHP-UHFFFAOYSA-N Phenethylamine Chemical compound NCCC1=CC=CC=C1 BHHGXPLMPWCGHP-UHFFFAOYSA-N 0.000 description 1
- BELBBZDIHDAJOR-UHFFFAOYSA-N Phenolsulfonephthalein Chemical compound C1=CC(O)=CC=C1C1(C=2C=CC(O)=CC=2)C2=CC=CC=C2S(=O)(=O)O1 BELBBZDIHDAJOR-UHFFFAOYSA-N 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- 244000181025 Rosa gallica Species 0.000 description 1
- 235000000533 Rosa gallica Nutrition 0.000 description 1
- 241000264847 Siniperca chuatsi Species 0.000 description 1
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical class [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 1
- UCKMPCXJQFINFW-UHFFFAOYSA-N Sulphide Chemical compound [S-2] UCKMPCXJQFINFW-UHFFFAOYSA-N 0.000 description 1
- JZRWCGZRTZMZEH-UHFFFAOYSA-N Thiamine Natural products CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N JZRWCGZRTZMZEH-UHFFFAOYSA-N 0.000 description 1
- 241000607598 Vibrio Species 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 239000000853 adhesive Substances 0.000 description 1
- 230000001070 adhesive effect Effects 0.000 description 1
- 230000006838 adverse reaction Effects 0.000 description 1
- 235000013334 alcoholic beverage Nutrition 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 229940043376 ammonium acetate Drugs 0.000 description 1
- 235000019257 ammonium acetate Nutrition 0.000 description 1
- 235000011114 ammonium hydroxide Nutrition 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000005842 biochemical reaction Methods 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 239000012159 carrier gas Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 230000006835 compression Effects 0.000 description 1
- 238000007906 compression Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000000593 degrading effect Effects 0.000 description 1
- 238000001212 derivatisation Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 206010014665 endocarditis Diseases 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 210000003743 erythrocyte Anatomy 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 235000021107 fermented food Nutrition 0.000 description 1
- 235000003891 ferrous sulphate Nutrition 0.000 description 1
- 239000011790 ferrous sulphate Substances 0.000 description 1
- 235000021588 free fatty acids Nutrition 0.000 description 1
- 230000005802 health problem Effects 0.000 description 1
- 230000002949 hemolytic effect Effects 0.000 description 1
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 238000007912 intraperitoneal administration Methods 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 description 1
- 229910000359 iron(II) sulfate Inorganic materials 0.000 description 1
- 150000002576 ketones Chemical class 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 230000004130 lipolysis Effects 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 238000001819 mass spectrum Methods 0.000 description 1
- 230000018984 mastication Effects 0.000 description 1
- 238000010077 mastication Methods 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000013028 medium composition Substances 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 239000006872 mrs medium Substances 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 125000001477 organic nitrogen group Chemical group 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- VLTRZXGMWDSKGL-UHFFFAOYSA-N perchloric acid Chemical compound OCl(=O)(=O)=O VLTRZXGMWDSKGL-UHFFFAOYSA-N 0.000 description 1
- 239000012071 phase Substances 0.000 description 1
- 230000000704 physical effect Effects 0.000 description 1
- 238000005554 pickling Methods 0.000 description 1
- 231100000572 poisoning Toxicity 0.000 description 1
- 230000000607 poisoning effect Effects 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 230000017854 proteolysis Effects 0.000 description 1
- 235000007682 pyridoxal 5'-phosphate Nutrition 0.000 description 1
- 239000011589 pyridoxal 5'-phosphate Substances 0.000 description 1
- 229960001327 pyridoxal phosphate Drugs 0.000 description 1
- 238000003908 quality control method Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000004007 reversed phase HPLC Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 238000002470 solid-phase micro-extraction Methods 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 229940063673 spermidine Drugs 0.000 description 1
- 229940063675 spermine Drugs 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000009210 therapy by ultrasound Methods 0.000 description 1
- KYMBYSLLVAOCFI-UHFFFAOYSA-N thiamine Chemical compound CC1=C(CCO)SCN1CC1=CN=C(C)N=C1N KYMBYSLLVAOCFI-UHFFFAOYSA-N 0.000 description 1
- 229960003495 thiamine Drugs 0.000 description 1
- 235000019157 thiamine Nutrition 0.000 description 1
- 239000011721 thiamine Substances 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 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 description 1
- YNJBWRMUSHSURL-UHFFFAOYSA-N trichloroacetic acid Chemical compound OC(=O)C(Cl)(Cl)Cl YNJBWRMUSHSURL-UHFFFAOYSA-N 0.000 description 1
- 239000012137 tryptone Substances 0.000 description 1
- 229960003732 tyramine Drugs 0.000 description 1
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 1
- 235000019607 umami taste sensations Nutrition 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Classifications
-
- 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
- A23L17/00—Food-from-the-sea products; Fish products; Fish meal; Fish-egg substitutes; Preparation or treatment thereof
- A23L17/65—Addition of, or treatment with, microorganisms or enzymes
-
- 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
- A23L17/00—Food-from-the-sea products; Fish products; Fish meal; Fish-egg substitutes; Preparation or treatment thereof
- A23L17/10—Fish meal or powder; Granules, agglomerates or flakes
-
- 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
- A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
- A23L5/20—Removal of unwanted matter, e.g. deodorisation or detoxification
- A23L5/28—Removal of unwanted matter, e.g. deodorisation or detoxification using microorganisms
-
- 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
-
- 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
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
- C12R2001/25—Lactobacillus plantarum
-
- 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
- C12R2001/44—Staphylococcus
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Microbiology (AREA)
- Organic Chemistry (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Nutrition Science (AREA)
- Polymers & Plastics (AREA)
- Food Science & Technology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Medicinal Chemistry (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Marine Sciences & Fisheries (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention provides a fermentation method of mandarin fish and a strain used by the same, and provides a lactobacillus plantarum and a staphylococcus soil; the lactobacillus plantarum is added to change the community composition of microorganisms in the fermentation process of the mandarin fish, reduce the growth of other spoilage microorganisms and inhibit the spoilage of the mandarin fish; the staphylococcus aureus can produce extracellular protease and lipase to promote the decomposition of mandarin fish protein and fat; the two strains are mixed and inoculated to ferment the mandarin fish, so that not only can the growth of other spoilage microorganisms be inhibited, but also the decomposition of the fish meat can be promoted, biogenic amine of the mixed inoculation and fermentation fish meat can be inhibited, and the sulfur-containing compounds in the volatile flavor compounds of the mixed inoculation and fermentation fish meat can be reduced, thereby improving the flavor of the mandarin fish.
Description
Technical Field
The invention belongs to the technical field of microbial fermentation, and particularly relates to a fermentation method of stinky mandarin fish and a strain used by the fermentation method.
Background
The fermented mandarin fish is also called barren fresh fish, smelly mandarin fish, etc. and is one of the representative dishes of Huizhou dishes, and the traditional fermented mandarin fish is produced through pickling fresh or frozen mandarin fish, sealing and fermenting at room temperature for 7-15 days. The stink mandarin fish prepared by fermentation has the characteristics of smelling and smelling, and the stink-like and non-stink flavor is the important biochemical reaction in the fermentation maturation process because protein is decomposed under the action of endogenous enzyme and microbial enzyme, amino acid is released by proteolysis, free fatty acid is produced by lipolysis and the like. However, the complex microbial community in the fermentation system brings great challenges for identifying the fermentation process and realizing the quality control of the product.
In the traditional mandarin fish fermentation process, because of the existence of self-contained mandarin fish and complex microorganisms in the environment, uncontrollability and instability of products in the fermentation process can be caused, and the flavor and quality of the fermented products can be influenced. Research shows that the spoilage microorganisms such as psychrophilic bacillus, acinetobacter and vibrio in the traditional mandarin fish fermentation process can increase the decomposition of proteins and fats, so that the oxidation of proteins and lipids can be accelerated, the excessive oxidation of proteins can generate ammonia, amines and other alkaline nitrogen-containing substances, and the excessive decomposition and oxidation of fats can generate micromolecular aldehyde, ketone and fatty acid compounds, so that bad flavor is generated, commonly called rancidity.
Biogenic amine is one of main safety hazard factors of aquatic products, and is mainly divided into tryptamine, beta-phenethylamine, cadaverine, putrescine, tyramine, histamine, spermine, spermidine and the like, the biogenic amine in the aquatic products is mainly produced by degrading protein of the biogenic amine by microorganisms, the formation mechanism can be used as an important detection index for quality and safety of chilled fresh aquatic products such as minced fillet, fish block and the like, the biogenic amine content of salted or fermented fish products is determined by microbial flora, and spoilage bacteria are important causes for reducing the quality of the aquatic products. Some biogenic amines in trace amounts are normal active ingredients in the human body and have important physiological effects, but once the human body ingests excessively, serious harm and even poisoning are caused, biogenic Amines (BAs) are organic nitrogen-containing compounds, mainly formed by decarboxylation of corresponding amino acids. If the biogenic amine concentration reaches a threshold level, this can lead to mild to severe health problems in humans
Therefore, how to control spoilage microorganisms and reduce the biogenic amine content in fermented fish products is the key to fermenting stinky mandarin fish.
Disclosure of Invention
The invention aims to provide a saccule-covered yeast strain and application thereof in yellow wine brewing.
To achieve the above and other related objects, the present invention provides the following technical solutions: a lactobacillus plantarum strain is lactobacillus plantarum (Lactiplantibacillus plantarum) YR07 and is preserved in China Center for Type Culture Collection (CCTCC), the preservation date is 2022, 8 months and 18 days, and the strain preservation number is CCTCCNO: M20221303.
To achieve the above and other related objects, the present invention provides the following technical solutions: a staphylococcus aureus is staphylococcus aureus (Staphylococcus edaphicus) T-F, and is preserved in China Center for Type Culture Collection (CCTCC), the preservation date is 2022, 12 months and 19 days, and the strain preservation number is CCTCCNO: M20221987.
To achieve the above and other related objects, the present invention provides the following technical solutions: a fermentation method of stink mandarin fish comprises the following steps:
step 1: inoculating Lactobacillus plantarum (Lactiplantibacillus plantarum) YR07 of claim 1 to an MRS agar medium with an inoculating loop, activating, culturing at 30-37deg.C for 36-48 hr, inoculating the obtained culture with an inoculating loop, culturing at 30-37deg.C for 24-28 hr to obtain a first inoculum;
step 2: transferring the staphylococcus soil (Staphylococcus edaphicus) T-F of claim 1 from the inclined plane to the MSA agar culture medium by using an inoculating loop for activation, culturing at 30-37 ℃ for 36-48h, transferring the obtained culture to the MSA culture medium by using the inoculating loop, and culturing at 30-37 ℃ for 28-36h to obtain a second inoculum;
step 3: removing scales and viscera of fresh mandarin fish, cleaning, placing in a container, and adding sterile saline water;
step 4: adding the first inoculum and the second inoculum, capping, and fermenting at 8-12deg.C.
The preferable technical scheme is as follows: in the step 1, the obtained first inoculum was liquid, and the first inoculum was centrifuged at 3500rpm/min to obtain wet cells.
The preferable technical scheme is as follows: in step 2, the obtained second inoculum was washed with sterile physiological saline on an agar plate, followed by centrifugation at 3500rpm/min to obtain wet cells.
The preferable technical scheme is as follows: in the step 3, the mass fraction of the sterile saline is 5-10%, and the ratio of the sterile saline to the fish meat is as follows: 1L, 1.2-1.6kg.
The preferable technical scheme is as follows: in step 4, the inoculum size of the first inoculum and the second inoculum is 1.0-1.5g/kg according to the mass ratio of wet bacteria to fish meat. Namely: the inoculum size of the first inoculum is 1.0-1.5g/kg according to the ratio of wet thallus to fish meat mass, and the inoculum size of the second inoculum is the same as that of the first inoculum.
The preferable technical scheme is as follows: every liter of MRS agar medium contains: 10.0g of peptone, 8.0g of beef powder, 4.0g of yeast powder, 20.0g of glucose, 2.0g of dipotassium hydrogen phosphate, 2.0g of diammonium hydrogen citrate, 5.0g of sodium acetate, 0.2g of magnesium sulfate, 0.04g of manganese sulfate, 1.0g of tween and 15.0-20.0g of agar; sterilizing at 121deg.C for 20min at pH 6.5+ -0.2; MRS liquid medium differs from MRS agar medium in that no agar is added.
The preferable technical scheme is as follows: the formula of the MSA agar medium is as follows: 10g of peptone, 1g of beef extract powder, 10g of mannitol, 25g of sodium chloride, 15-20g of agar powder and 1L of distilled water; sterilizing at 121deg.C for 20min at pH 7.0+ -2.
Due to the application of the technical scheme, compared with the prior art, the invention has the advantages that:
1. the plant milk stalk can inhibit the increase of other putrefying bacteria, the staphylococcus aureus can generate protease and lipase, promote the decomposition of protein and fat to form good flavor, reduce the bad smell of sulfide, and the flavor of the final fermentation product is more easily accepted by consumers.
2. The strain does not produce amino acid decarboxylase in metabolism, controls the biogenic amine content of the product, and is beneficial to improving the safety of the product.
3. The mixed inoculation fermentation can improve the content of the delicious taste and sweet amino acid in the free amino acid of the mandarin fish flesh, reduce the content of bitter amino acid and improve the taste of the product.
Drawings
FIG. 1 morphology of Lactobacillus plantarum (left panel) and Staphylococcus aureus (right panel) under an optical microscope.
Fig. 2 glucose fermentation experiments, a staphylococcus soil group, a lactobacillus plantarum group and a control group are sequentially carried out from left to right.
The bacterial strain of FIG. 3 produces the biogenic amine experiment, which is a staphylococcus soil group, a lactobacillus plantarum group, a blank control group and a positive control group from left to right.
The hemolysin production experiment of the strain in figure 4 is carried out by lactobacillus plantarum and staphylococcus soil sequentially from left to right.
Fig. 5 shows indole production experiments of lactobacillus plantarum, staphylococcus soil, blank control and positive control groups from left to right.
FIG. 6 texture of fish after 7 days of fermentation between the inoculation group and the control group.
Detailed Description
Further advantages and effects of the present invention will be readily apparent to those skilled in the art from the following disclosure of the present invention by reference to the specific embodiments.
Please refer to fig. 1-6. It should be understood that the structures, proportions, sizes, etc. shown in the drawings are shown only in the drawings and should not be taken as limiting the invention to those having ordinary skill in the art, since modifications, changes in proportions, or adjustments of sizes, etc. could be made without departing from the spirit or essential characteristics of the invention. The following examples are provided for a better understanding of the present invention, but are not intended to limit the present invention. The experimental methods in the following examples are conventional methods unless otherwise specified. The experimental materials used in the examples described below were purchased from conventional biochemical reagent stores unless otherwise specified.
Preservation of biological material:
lactobacillus plantarum (Lactiplantibacillus plantarum) YR07 has been deposited with China Center for Type Culture Collection (CCTCC) at month 8 and 18 of 2022, with a deposit number of CCTCC NO: M20221303 and a strain of staphylococcus aureus (Staphylococcus edaphicus) T-F, and has been deposited with China Center for Type Culture Collection (CCTCC) at month 12 and 19 of 2022, with a deposit number of CCTCC NO: M20221987. Chinese typical culture collection address: no. 299 is eight paths in Wuchang district of Wuhan, hubei province.
The reagents and materials described in the examples below are commercially available unless otherwise indicated.
Example 1: fermentation method of stink mandarin fish and strain used by same
The screening procedure for Lactobacillus plantarum (Lactiplantibacillus plantarum) YR07 and Staphylococcus aureus (Staphylococcus edaphicus) T-F was as follows:
1. the method for separating and screening lactobacillus plantarum (Lactiplantibacillus plantarum) YR07 and staphylococcus soil (Staphylococcus edaphicus) T-F from natural fermentation stink mandarin fish comprises the following specific steps: 10g of fish meat is taken, homogenized for 10min in a sterile homogenizing bag filled with 90mL of sterile normal saline, 1mL of homogenized solution is taken for gradient dilution for 5 to 6 times, the final diluted solution is coated on an MRS agar medium plate containing light calcium carbonate and an MSA agar medium plate added with a phenol red indicator, and colonies with transparent rings in the MRS medium and yellowish colonies in the MSA medium are taken for further screening after culturing for 24 to 48h in an incubator. And purifying the bacterial colony obtained by primary screening for 3 times to obtain a pure bacterial colony, and then carrying out gram staining, microscopic examination, contact enzyme experiment and 6% salt tolerance experiment to carry out the next screening to obtain the bacterial strain with gram positive, contact enzyme negative and salt tolerance.
2. According to the microorganism identification technology, a physiological and biochemical experiment is carried out on the provided strain, and the physicochemical characteristics of the strain, such as a glucose fermentation experiment, a biogenic amine experiment, a hemolysis experiment and an indole experiment, which are favorable for the fermentation of the siniperca chuatsi are judged; the strain obtained by identification is subjected to DNA extraction and PCR amplification to obtain an amplified product, and then the amplified product is sent to a biological company to identify the sequence, and the sequence result is that: SEQ No.1 and SEQ No.2.
(1) Acid and gas production experiment of fermentation glucose
Inoculating Lactobacillus plantarum culture on MRS agar medium and Staphylococcus aureus culture on MSA agar medium to glucose fermentation tube, culturing a group of non-inoculated bacteria as control at 35deg.C for 24-48 hr, and observing the color of the culture medium and whether bubbles are generated in Du Shixiao tube; if the strain produces acid, the indicator in the culture medium changes from purple to yellow, if the strain ferments glucose to produce gas, bubbles are generated in the Du Shixiao tube, otherwise, the indicator does not exist. As a result, as shown in FIG. 2, lactobacillus plantarum and Staphylococcus aureus were found to ferment glucose but not produce gas.
Glucose fermentation medium composition: beef extract 5.0g, peptone 10.0g, naCl 3g and Na 2 HPO 4 ·12H 2 O2 g, 12mL of 0.2% bromocresol purple ethanol solution, 5g of glucose, 1L of distilled water to a constant volume, pH7.4; preparing hair: after the components are well distributed, the components are packaged in test tubes with inverted Du's small tubes, each tube is packaged with 10mL, and the tubes are sterilized for 15min at 121 ℃ for later use.
(2) Experiment of producing biogenic amine
Lactobacillus plantarum and Staphylococcus soil on agar medium are cultured in amine production liquid medium at 35deg.C for 24-48 hr, and the color change of indicator in the medium is observed, so that neither strain produces amine, as shown in figure 3.
Production of biogenic amine liquid medium: beef extract powder 5.0g, tryptone 5.0g, yeast extract powder 5.0g, glucose 0.5g, naCl2.5g, ammonium citrate 2.0g and K 2 HPO 4 2.0g, 0.1g CaCO3, 0.05g manganese sulfate, 0.04g ferrous sulfate, 0.01g thiamine, 0.05g pyridoxal phosphate, 1mL Tween-80, 0.06g bromocresol purple, amino acid (1 mL each of arginine, histidine, tyrosine and lysine), adding distilled water to 1L, adjusting pH to 5.3, and sterilizing at 121deg.C for 15min; the indicator in the culture medium is yellow in color at pH5.3, and if amine is produced after the growth of the inoculation, the pH of the culture medium can be increased, and the indicator in the culture medium can be changed from yellow to light purple.
(3) Hemolysis experiment
Some bacteria can generate hemolysin in the growth process, so that erythrocytes are broken and dissolved, transparent or semitransparent hemolytic rings are observed around colonies when the bacteria grow on a blood plate, alpha hemolysis (grass green), beta hemolysis (complete hemolysis) and gamma hemolysis (no hemolysis or hemolysis ring) can be formed on the plate due to different hemolysin generated by different bacteria, and many bacterial hemolysis can be pathogenic. Lactobacillus plantarum and Staphylococcus aureus on agar medium are streaked on blood plates, and cultured at 35deg.C for 24-48 hr, and both strains are observed to have no hemolysis, as shown in figure 4.
(4) Indole production experiment
Some bacteria have tryptophan hydrolase, and can decompose tryptophan in protein to produce indole, which may cause diseases such as intraperitoneal infection and endocarditis. Indole itself is not colored and cannot be seen directly, but by the addition of a para-dimethylaminobenzaldehyde reagent, it reacts with indole to give a red rose indole. The cultures on the culture medium were inoculated with a needle to obtain appropriate cultures in an indole kit, and the indole kit was cultured in an incubator at 35℃for 24-48 hours with non-inoculated bacteria as a negative control group and inoculated with E.coli as a positive control group, respectively, and it was found that Lactobacillus plantarum and Staphylococcus aureus did not produce indole, and the results are shown in FIG. 5.
Example 2: fermentation method of stink mandarin fish and strain used by same
The present example provides Lactobacillus plantarum (Lactiplantibacillus plantarum) YR07, which has been deposited at China Center for Type Culture Collection (CCTCC) at month 8 and 18 of 2022, with a deposit number of CCTCCNO: M20221303 and a strain of staphylococcus soil (Staphylococcus edaphicus) T-F, which has been deposited at China Center for Type Culture Collection (CCTCC) at month 12 and 19 of 2022, with a deposit number of CCTCC NO: M20221987.
The method is applied to the fermentation of the stinky mandarin fish, and comprises the following steps:
1. inoculating lactobacillus plantarum on an inclined plane, inoculating to an MRS agar culture medium for activation, culturing at 35 ℃ for 36-48h, inoculating the obtained culture to an MRS liquid culture medium again, culturing at 35 ℃ for 24-28h to obtain an inoculum, centrifuging the liquid culture medium at 3500rpm/min to obtain wet thalli, and inoculating as a starter.
2. Transferring staphylococcus soil from an inclined plane to an MSA agar culture medium for activation, culturing at 35 ℃ for 36-48h, transferring the obtained culture to the MSA culture medium again by using the inoculating loop, culturing at 35 ℃ for 28-36h to obtain an inoculum, flushing the obtained inoculum on an agar plate by using sterile normal saline, and centrifuging at 3500rpm/min to obtain wet thalli serving as a starter.
3. Preparing and treating mandarin fish raw materials: removing scales and viscera of fresh mandarin fish, cleaning, placing in a container, adding 6% sterile saline, and adding the sterile saline according to the water volume (L): the fish meat mass (kg) is 1:1.5.
4. Lactobacillus plantarum and staphylococcus are added into the prepared mandarin fish raw materials and marked as LS group, the inoculation amount of the fermentation strain is added according to the proportion of the mass of wet thalli and fish meat of 1.25g/kg, the addition amount of single strain is halved when the two strains are inoculated together, the container is covered after the fermentation agent is added, and the container is placed at 10+/-2 ℃ for fermentation for one week.
Example 3: fermentation method of stink mandarin fish and strain used by same
The present example provides Lactobacillus plantarum (Lactiplantibacillus plantarum) YR07, which has been deposited at China Center for Type Culture Collection (CCTCC) at month 8 and 18 of 2022, with a deposit number of CCTCCNO: M20221303 and a strain of staphylococcus soil (Staphylococcus edaphicus) T-F, which has been deposited at China Center for Type Culture Collection (CCTCC) at month 12 and 19 of 2022, with a deposit number of CCTCC NO: M20221987.
The method is applied to the fermentation of the stinky mandarin fish, and comprises the following steps:
1. inoculating lactobacillus plantarum on an inclined plane, inoculating to an MRS agar culture medium for activation, culturing at 35 ℃ for 36-48h, inoculating the obtained culture to an MRS liquid culture medium again, culturing at 35 ℃ for 24-28h to obtain an inoculum, centrifuging the liquid culture medium at 3500rpm/min to obtain wet thalli, and inoculating as a starter.
2. Preparing and treating mandarin fish raw materials: removing scales and viscera of fresh mandarin fish, cleaning, placing in a container, adding 6% sterile saline, and adding the sterile saline according to the water volume (L): the fish meat mass (kg) is 1:1.5.
3. Adding lactobacillus plantarum into the prepared mandarin fish raw material, marking the lactobacillus plantarum as L groups, adding the inoculum size of the fermentation strain according to the proportion of wet thalli and fish meat mass of 1.25g/kg, inoculating the two strains together, halving the addition size of a single strain, adding a starter, covering a container, and fermenting at 10+/-2 ℃ for one week.
Example 4: fermentation method of stink mandarin fish and strain used by same
The present example provides Lactobacillus plantarum (Lactiplantibacillus plantarum) YR07, which has been deposited at China Center for Type Culture Collection (CCTCC) at month 8 and 18 of 2022, with a deposit number of CCTCCNO: M20221303 and a strain of staphylococcus soil (Staphylococcus edaphicus) T-F, which has been deposited at China Center for Type Culture Collection (CCTCC) at month 12 and 19 of 2022, with a deposit number of CCTCC NO: M20221987.
The method is applied to the fermentation of the stinky mandarin fish, and comprises the following steps:
1. transferring staphylococcus soil from an inclined plane to an MSA agar culture medium for activation, culturing at 35 ℃ for 36-48h, transferring the obtained culture to the MSA culture medium again by using the inoculating loop, culturing at 35 ℃ for 28-36h to obtain an inoculum, flushing the obtained inoculum on an agar plate by using sterile normal saline, and centrifuging at 3500rpm/min to obtain wet thalli serving as a starter.
2. Preparing and treating mandarin fish raw materials: removing scales and viscera of fresh mandarin fish, cleaning, placing in a container, adding 6% sterile saline, and adding the sterile saline according to the water volume (L): the fish meat mass (kg) is 1:1.5.
3. Adding staphylococcus into the prepared mandarin fish raw material, marking as S group, adding the inoculation amount of the fermentation strain according to the proportion of wet thalli and fish meat mass of 1.25g/kg, inoculating two strains together, halving the addition amount of a single strain, adding a starter, covering a container, and fermenting at 10+/-2 ℃ for one week.
Example 5: fermentation method of stink mandarin fish and strain used by same
The present example provides Lactobacillus plantarum (Lactiplantibacillus plantarum) YR07, which has been deposited at China Center for Type Culture Collection (CCTCC) at month 8 and 18 of 2022, with a deposit number of CCTCCNO: M20221303 and a strain of staphylococcus soil (Staphylococcus edaphicus) T-F, which has been deposited at China Center for Type Culture Collection (CCTCC) at month 12 and 19 of 2022, with a deposit number of CCTCC NO: M20221987.
The method is applied to the fermentation of the stinky mandarin fish, and comprises the following steps:
1. preparing and treating mandarin fish raw materials: removing scales and viscera of fresh mandarin fish, cleaning, placing in a container, adding 6% sterile saline, and adding the sterile saline according to the water volume (L): the fish meat mass (kg) is 1:1.5.
2. Adding no fermentation strain into the prepared mandarin fish raw material, marking as group C, covering the container, and fermenting at 10+ -2deg.C for one week.
Example 6: fermentation method of stink mandarin fish and strain used by same
1. Determination of fish malondialdehyde content: the fish meat during fermentation was taken and the fish meat on day 1, day 3, day 5 and day 7 were taken as samples, and the content of malondialdehyde during fish fermentation was measured according to the second spectrophotometry in GB 5009.181-2016, "determination of malondialdehyde in food safety national Standard food", with the following results.
The malondialdehyde content (unit: mg/kg) of fish during fermentation of each group of stinky mandarin fish (note: group C is non-inoculating fermentation;
l group is single lactobacillus plantarum fermentation; s group is fermentation of staphylococcus soil singly; LS group is mixed inoculation fermentation
Group C | L group | S group | LS group | |
Day 1 of fermentation | 0.282±0.009 | 0.278±0.012 | 0.277±0.023 | 0.264±0.045 |
Day 3 of fermentation | 0.582±0.018 | 0.387±0.014 | 0.447±0.021 | 0.273±0.009 |
Day 5 of fermentation | 0.741±0.032 | 0.678±0.008 | 0.765±0.019 | 0.582±0.027 |
Day 7 of fermentation | 0.914±0.014 | 0.787±0.034 | 0.792±0.026 | 0.618±0.027 |
2. Determination of fish texture: taking fermented fish meat, peeling the back part near the head, and cutting into blocks with thickness of 10mm, length of 15mm and width of 15 mm. The samples were subjected to a texture multi-faceted analysis (TPA) mode test using a TA-XTPLUS physical property apparatus using a flat-bottomed cylindrical probe p36/R using a simulated human tooth to chew food, under conditions of 2mm/s pre-test rate, 1mm/s post-test rate, 50% compression, 5s dwell interval, 3 samples each, 6 times each, averaged, and texture results shown in FIG. 6.
Texture results: the hardness, the mastication degree and the elasticity of the fish meat of the three groups of bacteria groups are higher than those of the control group, the restoring force and the adhesive force of the fish meat of the single bacteria group (the L group and the S group) are lower than those of the mixed bacteria group, and the texture of the fish meat of the mixed bacteria group is best, which indicates that the mixed bacteria can promote the fermentation of mandarin fish and improve the texture of the fish meat.
3. Determination of fish biogenic amine: the method adopts High Performance Liquid Chromatography (HPLC) to measure the biogenic amine content in the fermentation process of the stinky mandarin fish, and comprises the following specific steps:
sample treatment: taking 2.5g of fish, cutting, adding 10mL of 0.6M HClO4 and 125 mu L of internal standard, homogenizing, centrifuging, taking supernatant after centrifuging, repeating the steps, and finally merging the supernatant to a 25mL volumetric flask to obtain a sample liquid;
sample and standard derivatization: adding 50 mu L of an internal standard into 0.2mL of a standard series, adding 40 mu L of 2MNaOH, 60 mu L of saturated NaHCO3 and 400 mu L of 10mg/mL of dansyl chloride, shaking uniformly, adding 20 mu L of NH4OH, carrying out water bath at 40 ℃ for 45min, reacting in the dark for 30min, adding acetonitrile to 1mL, filtering, and carrying out sample injection;
conditions for HPLC: reverse phase HPLC was used, ammonium acetate (0.1M; solvent A) and acetonitrile (solvent B) were used as mobile phases. Elution was performed using a gradient of 0min, 50% B;25 minutes, 90% B;35 minutes, 90% B;45,50% B. The flow rate was 0.8 ml/min and the temperature was 30 ℃. The sample was detected at 254nm and the sample volume was 10. Mu.L.
Only 3 biogenic amines, aniline, putrescine and histamine were detected during the fermentation of the stink mandarin fish, of which histamine is the most toxic biogenic amine common in aquatic products and its content is shown in Table 2.
TABLE 2 fish biogenic amine content (unit: mg/kg) during fermentation of various groups of stinky Mandarin fish
Note that: ND in the table indicates that no such material was detected.
Biogenic amine is a low molecular mass nitrogen-containing organic compound with biological activity, and biogenic amine in food is mainly formed by decarboxylation of amino acid by microbial amino acid decarboxylase. Production strains or environmental microorganisms in the production of fermented foods and fermented alcoholic beverages often cause the accumulation of biogenic amines in the products due to the activity of amino acid decarboxylase. Proper amounts of biogenic amine in the organism promote normal physiological activities, while excessive biogenic amine can cause adverse reactions in the organism. From the above table, it can be seen that aniline, putrescine and histamine are gradually accumulated as fermentation time is prolonged in the fermentation process of the mandarin fish samples, and aniline is detected on the 5 th day of group C and then on the 7 th day of group LS, in addition, the content of putrescine and histamine in the mixed inoculation LS group is obviously lower than that in the control group C and lower than that in the single inoculation L group and S group at the end of the 7 th day of fermentation, which indicates that the accumulation of biogenic amine is inhibited by the inoculated strain, and the mixed inoculation group is more favorable for inhibiting the accumulation of biogenic amine.
4. Determination of fish volatile compounds: the volatile flavor substances in the process of the fermentation of the stink mandarin fish are determined by adopting a solid phase microextraction gas chromatography-mass spectrometry (HPMS-HS-GC-MS), 5g of each sample is taken, the samples are crushed in a 20mL headspace bottle, and 10 mu L of an internal standard (2, 4, 6-trimethylpyridine) with the concentration of one ten thousandth is added. The extraction head was inserted into a headspace bottle at 70℃for 40min of extraction time, 3 replicates per sample.
The chromatographic column is DB-WAX capillary chromatographic column (30mx0.20 mm0.25 m), the temperature of the sample inlet is 250 ℃, the initial temperature of the programmed temperature is 30 DEG, and the temperature is kept for 1min. Raising the temperature to 92 ℃ at 4 ℃/min, and keeping the temperature for 2min, raising the temperature to 200 ℃ at 5 ℃/min, and raising the temperature to 240 ℃ at 6 ℃/min, and keeping the temperature for 6min. The carrier gas flow rate is 1mL/min, and the sample injection is not split. Mass spectrometry conditions ionization mode (EI); electron energy 70eV; the interface temperature is 250 ℃ and the ion source temperature is 250 ℃; the mass spectrum scanning range is 29-450m/z, and the acquisition mode is full scanning.
TABLE 3 content of various volatile flavor substances in fish meat after 7 days of fermentation of various groups of smelly Mandarin fish (unit: ng/g)
Note that: group C is non-inoculation fermentation; l group is single lactobacillus plantarum fermentation; s group is fermentation of staphylococcus soil singly; LS group is mixed inoculation fermentation.
TABLE 4 alcohol content in fish meat after 7 days of fermentation of the groups of stinky Mandarin fish (unit: ng/g)
Note that: ND in the table indicates that the substance was not detected; group C is non-inoculation fermentation; l group is single lactobacillus plantarum fermentation; s group is fermentation of staphylococcus soil singly; LS group is mixed inoculation fermentation.
TABLE 5 aldehyde and ketone Compound content in fish meat after 7 days of fermentation of the groups of stinky Mandarin fish (unit: ng/g)
Note that: ND in the table indicates that the substance was not detected; and (3) injection: group C is non-inoculation fermentation; l group is single lactobacillus plantarum fermentation; s group is fermentation of staphylococcus soil singly; LS group is mixed inoculation fermentation.
TABLE 6 content of Sulfur-containing and Nitrogen-containing Compounds in fish meat after 7 days of fermentation of the groups of stinky Mandarin fish (unit: ng/g)
Note that: ND in the table indicates that the substance was not detected; group C is non-inoculation fermentation; l group is single lactobacillus plantarum fermentation; s group is fermentation of staphylococcus soil singly; LS group is mixed inoculation fermentation.
The table shows that the total content of sulfur-containing and nitrogen-containing compounds after the fermentation of the inoculation group is obviously lower than that of the C group, the sulfur-containing and nitrogen-containing compounds are main substances which generate putrefactive flavor in the fermented mandarin fish, the substances of the mixed inoculation LS group have obvious inhibition effect, except indole, but the substances have smaller difference from the comparison group, wherein three groups of inoculation groups have inhibition effect on trimethylamine, the inhibition effect of the mixed inoculation group is more obvious, the trimethylamine content of the comparison C group is 415.61 +/-27.06 ng/g, the content of trimethylamine of the mixed inoculation group LS group is the lowest, and only 177.14 +/-35.56 ng/g, so that the content of trimethylamine is obviously reduced, and the content of the produced bad flavor substances is reduced.
5. Determination of free amino acids in fish: measuring free amino acid in fermentation process by using a full-automatic amino acid analyzer, taking 5.0g of fish meat of the control group and the inoculation group in 1, 3, 5 and 7 days of fermentation process, stirring, adding about 15mL of 5% trichloroacetic acid solution for homogenizing, fixing the volume to 25mL, carrying out ultrasonic treatment for 2 times for 15min each time, filtering by using a needle filter of 0.22 mu m, bottling, and carrying out on-machine analysis.
TABLE 7 content of Free Amino Acids (FAA) in fish meat after 7 days of fermentation of the groups of stinky Mandarin fish (unit: mg/100 g)
(note: group C is non-inoculating fermentation, group L is single inoculating Lactobacillus plantarum fermentation, group S is single inoculating Staphylococcus aureus fermentation;
LS group is mixed inoculation fermentation
/>
Note that: ND in the table indicates that no such material was detected.
From the above table, it can be seen that the umami amino acid and the sweet amino acid of the mixed inoculation group are obviously increased relative to the control group after fermentation is finished, and the bitter amino acid of the mixed inoculation group is reduced relative to the control group, which indicates that the inoculation of lactobacillus plantarum and staphylococcus soil can promote the formation of amino acid in the mandarin fish and improve the umami taste of the mandarin fish meat.
SEQ No.1 Lactobacillus plantarum molecular sequence:
CAGGGTTATGGCTGACAGCTCGTACATAACGGGTCGAACAGAACTCTGGTATTGATTGGTGCTTGCATCATGATTTACATTTGAGTGAGTGGCGAACTGGTGAGTAACACGTGGGAAACCTGCCCAGAAGCGGGGGATAACACCTGGAAACAGATGCTAATACCGCATAACAACTTGGACCGCATGGTCCGAGCTTGAAAGATGGCTTCGGCTATCACTTTTGGATGGTCCCGCGGCGTATTAGCTAGATGGTGGGGTAACGGCTCACCATGGCAATGATACGTAGCCGACCTGAGAGGGTAATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGACGAAAGTCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGGTTTCGGCTCGTAAAACTCTGTTGTTAAAGAAGAACATATCTGAGAGTAACTGTTCAGGTATTGACGGTATTTAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTTTTAAGTCTGATGTGAAAGCCTTCGGCTCAACCGAAGAAGTGCATCGGAAACTGGGAAACTTGAGTGCAGAAGAGGACAGTGGAACTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTGTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGTATGGGTAGCAAACAGGATTAGATACCCTGGTAGTCCATACCGTAAACGATGAATGCTAAGTGTTGGAGGGTTTCCGCCCTTCAGTGCTGCAGCTAACGCATTAAGCATTCCGCCTGGGGAGTACGGCCGCAAGGCTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCTACGCGAAGAACCTTACCAGGTCTTGACATACTATGCAAATCTAAGAGATTAGACGTTCCCTTCGGGGACATGGATACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATTATCAGTTGCCAGCATTAAGTTGGGCACTCTGGTGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGATGGTACAACGAGTTGCGAACTCGCGAGAGTAAGCTAATCTCTTAAAGCCATTCTCAGTTCGGATTGTAGGCTGCAACTCGCCTACATGAAGTCGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGAGAGTTTGTAACACCCAAAGTCGGTGGGGTAACCTTTGAGGAACCAGCCTACCTCATCGAGTTTCCCCCG。
SEQ No.2 Staphylococcus aureus molecular sequence:
ACCTTCGACGGCTAGCTCCATAAATGGTTACTCCACCGGCTTCGGGTGTTACAAACTCTCGTGGTGTGACGGGCGGTGTGTACAAGACCCGGGAACGTATTCACCGTAGCATGCTGATCTACGATTACTAGCGATTCCAGCTTCATGTAGTCGAGTTGCAGACTACAATCCGAACTGAGAACAACTTTATGGGATTTGCATGACCTCGCGGTTTAGCTGCCCTTTGTATTGTCCATTGTAGCACGTGTGTAGCCCAAATCATAAGGGGCATGATGATTTGACGTCATCCCCACCTTCCTCCGGTTTGTCACCGGCAGTCAACCTAGAGTGCCCAACTTAATGATGGCAACTAAGCTTAAGGGTTGCGCTCGTTGCGGGACTTAACCCAACATCTCACGACACGAGCTGACGACAACCATGCACCACCTGTCACTTTGTCCCCCGAAGGGGAAGGCTCTATCTCTAGAGTTTTCAAAGGATGTCAAGATTTGGTAAGGTTCTTCGCGTTGCTTCGAATTAAACCACATGCTCCACCGCTTGTGCGGGTCCCCGTCAATTCCTTTGAGTTTCAACCTTGCGGTCGTACTCCCCAGGCGGAGTGCTTAATGCGTTAGCTGCAGCACTAAGGGGCGGAAACCCCCTAACACTTAGCACTCATCGTTTACGGCGTGGACTACCAGGGTATCTAATCCTGTTTGATCCCCACGCTTTCGCACATCAGCGTCAGTTACAGACCAGAAAGTCGCCTTCGCCACTGGTGTTCCTCCATATCTCTGCGCATTTCACCGCTACACATGGAATTCCACTTTCCTCTTCTGCACTCAAGTTTCCCAGTTTCCAATGACCCTCCACGGTTGAGCCGTGGGCTTTCACATCAGACTTAAGAAACCGCCTACGCGCGCTTTACGCCCAATAATTCCGGATAACGCTTGCCACCTACGTATTACCGCGGCTGCTGGCACGTAGTTAGCCGTGGCTTTCTGATTAGGTACCGTCAAGACGTGCACAGTTACTTACACGTTTGTTCTTCCCTAATAACAGAGTTTTACGAGCCGAAACCCTTCATCACTCACGCGGCGTTGCTCCGTCAGGCTTTCGCCCATTGCGGAAGATTCCCTACTGCTGCCTCCCGTAGGAGTCTGGACCGTGTCTCAGTTCCAGTGTGGCCGATCACCCTCTCAGGTCGGCTACGTATCGTCGCCTTGGTAAGCCGTTACCTTACCAACTAGCTAATACGGCGCGGGTCCATCTATAAGTGATAGCAAAACCATCTTTCACTTTAGAACCATGCGGTTCCAAATGTTATCCGGTATTAGCTCCGGTTTCCCGAAGTTATCCCAGTCTTATAGGTAGGTTACCCACGTGTTACTCACCCGTCCGCCGCTAACGTCAAAGGAGCAAGCTCCTTATCTGTTCGCTCGACTtGCAtGtAtTAG。
the foregoing description of the preferred embodiment of the invention is not intended to be limiting in any way, but rather, it is intended to cover all modifications or variations of the invention which fall within the spirit and scope of the invention.
Claims (9)
1. A lactobacillus plantarum, characterized in that: the lactobacillus plantarum is lactobacillus plantarumLactiplantibacillus plantarum) YR07 is preserved in China Center for Type Culture Collection (CCTCC), the preservation date is 2022, 8 months and 18 days, and the strain preservation number is CCTCCNO: M20221303.
2. A staphylococcus soil strain, characterized in that: the staphylococcus soil is staphylococcus soilStaphylococcus edaphicus) T-F is preserved in China Center for Type Culture Collection (CCTCC), the preservation date is 2022, 12 months and 19 days, and the strain preservation number is CCTCC NO: M20221987.
3. A fermentation method of smelly mandarin fish is characterized in that: comprises the following steps:
step 1: plant milk as claimed in claim 1Bacillus [(s) ]Lactiplantibacillus plantarum) Transferring one loop of YR07 from the inclined plane to an MRS agar culture medium for activation by using an inoculating loop, culturing for 36-48h at 30-37 ℃, transferring the obtained culture to an MRS liquid culture medium by using the inoculating loop again, and culturing for 24-28h at 30-37 ℃ to obtain a first inoculum;
step 2: the staphylococcus aureus of claim 1Staphylococcus edaphicus) Transferring the T-F into an MSA agar culture medium from an inclined plane by using an inoculating loop for activation, culturing at 30-37 ℃ for 36-48h, transferring the obtained culture into the MSA agar culture medium by using the inoculating loop, and culturing at 30-37 ℃ for 28-36h to obtain a second inoculum;
step 3: removing scales, cheeks and viscera of fresh mandarin fish, cleaning, placing in a container, and adding sterile saline;
step 4: adding the first inoculum and the second inoculum, capping, and fermenting at 8-12deg.C.
4. A method of fermenting a stinky mandarin fish according to claim 3, characterized in that: in the step 1, the obtained first inoculum was liquid, and the first inoculum was centrifuged at 3500rpm/min to obtain wet cells.
5. A method of fermenting a stinky mandarin fish according to claim 3, characterized in that: in step 2, the obtained second inoculum was washed with sterile physiological saline on an agar plate, followed by centrifugation at 3500rpm/min to obtain wet cells.
6. A method of fermenting a stinky mandarin fish according to claim 3, characterized in that: in the step 3, the mass fraction of the sterile saline is 5-10%, and the ratio of the sterile saline to the fish meat is as follows: 1 L:1.2-1.6. 1.6kg.
7. A method of fermenting a stinky mandarin fish according to claim 3, characterized in that: in the step 4, the inoculation amount of the first inoculum and the second inoculum is 1.0-1.5g/kg according to the mass ratio of the wet thalli to the fish meat.
8. A method of fermenting a stinky mandarin fish according to claim 3, characterized in that: every liter of MRS agar medium contains: 10.0g of peptone, 8.0g of beef powder, 4.0g of yeast powder, 20.0g of glucose, 2.0g of dipotassium hydrogen phosphate, 2.0g of diammonium hydrogen citrate, 5.0g of sodium acetate, 0.2g of magnesium sulfate, 0.04g of manganese sulfate, 1.0g of tween and 15.0-20.0g of agar; sterilizing at 121deg.C for 20min at pH 6.5+ -0.2; MRS liquid medium differs from MRS agar medium in that no agar is added.
9. A method of fermenting a stinky mandarin fish according to claim 3, characterized in that: the formula of the MSA agar medium is as follows: 10g of peptone, 1g of beef extract powder, 10g of mannitol, 25g of sodium chloride, 15-20g of agar powder and 1L of distilled water; sterilizing at 121deg.C for 20min at pH 7.0+ -2.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202310217694.3A CN116925953A (en) | 2023-03-08 | 2023-03-08 | Fermentation method of stink mandarin fish and strain used by same |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202310217694.3A CN116925953A (en) | 2023-03-08 | 2023-03-08 | Fermentation method of stink mandarin fish and strain used by same |
Publications (1)
Publication Number | Publication Date |
---|---|
CN116925953A true CN116925953A (en) | 2023-10-24 |
Family
ID=88386803
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202310217694.3A Pending CN116925953A (en) | 2023-03-08 | 2023-03-08 | Fermentation method of stink mandarin fish and strain used by same |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN116925953A (en) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN116622567A (en) * | 2023-05-19 | 2023-08-22 | 合肥工业大学 | Probiotics and application thereof in preparation of heme peptide iron |
-
2023
- 2023-03-08 CN CN202310217694.3A patent/CN116925953A/en active Pending
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN116622567A (en) * | 2023-05-19 | 2023-08-22 | 合肥工业大学 | Probiotics and application thereof in preparation of heme peptide iron |
CN116622567B (en) * | 2023-05-19 | 2024-01-23 | 合肥工业大学 | Probiotics and application thereof in preparation of heme peptide iron |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN108587983B (en) | Lactobacillus plantarum and application thereof in preparation of Sichuan sausage through fermentation | |
CN109182188B (en) | Lactobacillus plantarum incapable of producing tyramine or histamine and application thereof | |
Choudhury et al. | Formation of histamine and tyramine by lactic acid bacteria in decarboxylase assay medium | |
CN109136129B (en) | Lactobacillus acidophilus NCU426 | |
JP2023515791A (en) | Saccharopolyspora and use thereof in reducing biogenic amines | |
CN111979148B (en) | Saccharopolyspora composition and application thereof in food | |
CN116925953A (en) | Fermentation method of stink mandarin fish and strain used by same | |
CN113999793A (en) | Lactobacillus plantarum with good fermentation characteristics and fragrance production function and screening method thereof | |
CN114107099A (en) | Lactobacillus plantarum capable of improving flavor quality of fermented sausage and application thereof | |
Kim et al. | Characterization of the production of biogenic amines and gamma-aminobutyric acid in the soybean pastes fermented by Aspergillus oryzae and Lactobacillus brevis | |
CN108977391B (en) | Lactic acid bacterial strain with color development and corrosion prevention functions on meat products | |
CN110760456A (en) | Lactobacillus plantarum La1 for degrading cholesterol and application thereof | |
CN113736694B (en) | Lactococcus lactis capable of degrading butter and application thereof | |
CN113604385B (en) | Lactobacillus delbrueckii with butter degrading capability and application thereof | |
CN112458003B (en) | Diacetyl-producing lactobacillus plantarum and application thereof in pickled vegetables | |
CN114591860A (en) | Direct vat set starter for high-lactic acid bacteria-carrying meat and preparation method and application thereof | |
JP4434927B2 (en) | Method for producing γ-aminobutyric acid-containing food, and yeast having high γ-aminobutyric acid production ability | |
CN114107085A (en) | Lactobacillus plantarum ING8 and application thereof | |
JP3957132B2 (en) | Separation medium for low turbidity soy sauce lactic acid bacteria, separation method for low turbidity soy sauce lactic acid bacteria using the same medium, and method for producing highly clear soy sauce using the same lactic acid bacteria | |
CN115305217B (en) | Lactobacillus plantarum for degrading nitrite, resisting oxidization and producing fragrance and application thereof | |
CN112094763A (en) | Saccharomyces cerevisiae and application thereof in fattening pig feed | |
CN111454858A (en) | Method for inhibiting biogenic amine accumulation of amine-producing bacteria by using hurdle technology | |
CN116064277B (en) | Lactobacillus plantarum at low temperature and application thereof in production of dry cooked meat products | |
CN117223808B (en) | Bigeminal live bacteria fermented beverage for high yield of gamma-aminobutyric acid | |
WO2022033010A1 (en) | Saccharopolyspora composition and application thereof in food |
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 |