CN116686969A - Preparation method of fish gravy - Google Patents
Preparation method of fish gravy Download PDFInfo
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- CN116686969A CN116686969A CN202310509326.6A CN202310509326A CN116686969A CN 116686969 A CN116686969 A CN 116686969A CN 202310509326 A CN202310509326 A CN 202310509326A CN 116686969 A CN116686969 A CN 116686969A
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- fermentation
- aspergillus oryzae
- fish
- lactobacillus plantarum
- preparing
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- 241000251468 Actinopterygii Species 0.000 title claims abstract description 80
- 235000013882 gravy Nutrition 0.000 title claims abstract description 47
- 238000002360 preparation method Methods 0.000 title claims abstract description 40
- 238000000855 fermentation Methods 0.000 claims abstract description 102
- 230000004151 fermentation Effects 0.000 claims abstract description 102
- 240000006439 Aspergillus oryzae Species 0.000 claims abstract description 75
- 235000002247 Aspergillus oryzae Nutrition 0.000 claims abstract description 75
- 240000006024 Lactobacillus plantarum Species 0.000 claims abstract description 47
- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims abstract description 47
- 229940072205 lactobacillus plantarum Drugs 0.000 claims abstract description 47
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims abstract description 31
- 239000000047 product Substances 0.000 claims abstract description 31
- 235000015067 sauces Nutrition 0.000 claims abstract description 31
- 108090000790 Enzymes Proteins 0.000 claims abstract description 17
- 102000004190 Enzymes Human genes 0.000 claims abstract description 17
- 108010007119 flavourzyme Proteins 0.000 claims abstract description 14
- 241000894006 Bacteria Species 0.000 claims abstract description 9
- 239000001963 growth medium Substances 0.000 claims description 42
- 239000002054 inoculum Substances 0.000 claims description 37
- 229920001817 Agar Polymers 0.000 claims description 35
- 239000008272 agar Substances 0.000 claims description 35
- 239000002994 raw material Substances 0.000 claims description 31
- 241000404975 Synchiropus splendidus Species 0.000 claims description 27
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 27
- 239000007858 starting material Substances 0.000 claims description 26
- 238000012258 culturing Methods 0.000 claims description 25
- 239000002504 physiological saline solution Substances 0.000 claims description 24
- 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 19
- 244000061456 Solanum tuberosum Species 0.000 claims description 19
- 235000002595 Solanum tuberosum Nutrition 0.000 claims description 19
- 239000007788 liquid Substances 0.000 claims description 19
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- 238000000034 method Methods 0.000 claims description 13
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- 241000235036 Debaryomyces hansenii Species 0.000 claims description 11
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- 239000012153 distilled water Substances 0.000 claims description 9
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- 150000003839 salts Chemical class 0.000 claims description 9
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 8
- 238000005520 cutting process Methods 0.000 claims description 8
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- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 4
- 235000015278 beef Nutrition 0.000 claims description 4
- 238000009835 boiling Methods 0.000 claims description 4
- 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 4
- 238000001816 cooling Methods 0.000 claims description 4
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 4
- 239000011521 glass Substances 0.000 claims description 4
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 4
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 4
- 229940099596 manganese sulfate Drugs 0.000 claims description 4
- 239000011702 manganese sulphate Substances 0.000 claims description 4
- 235000007079 manganese sulphate Nutrition 0.000 claims description 4
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 4
- 235000019319 peptone Nutrition 0.000 claims description 4
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 claims description 4
- 229920000053 polysorbate 80 Polymers 0.000 claims description 4
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- 235000015099 wheat brans Nutrition 0.000 claims description 4
- 239000000796 flavoring agent Substances 0.000 abstract description 15
- 150000001412 amines Chemical class 0.000 abstract description 13
- 235000019634 flavors Nutrition 0.000 abstract description 13
- 230000000035 biogenic effect Effects 0.000 abstract description 12
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- 239000004365 Protease Substances 0.000 abstract description 4
- 102000004169 proteins and genes Human genes 0.000 abstract description 4
- 108090000623 proteins and genes Proteins 0.000 abstract description 4
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 abstract description 3
- 150000001875 compounds Chemical class 0.000 abstract description 3
- 244000005700 microbiome Species 0.000 abstract description 3
- 108090000765 processed proteins & peptides Proteins 0.000 abstract 3
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 abstract 1
- 241000235035 Debaryomyces Species 0.000 abstract 1
- JVTAAEKCZFNVCJ-UHFFFAOYSA-M Lactate Chemical compound CC(O)C([O-])=O JVTAAEKCZFNVCJ-UHFFFAOYSA-M 0.000 abstract 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 abstract 1
- 102000035195 Peptidases Human genes 0.000 abstract 1
- 239000006227 byproduct Substances 0.000 abstract 1
- 239000008101 lactose Substances 0.000 abstract 1
- 150000003384 small molecules Chemical class 0.000 abstract 1
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 27
- OAKJQQAXSVQMHS-UHFFFAOYSA-N Hydrazine Chemical compound NN OAKJQQAXSVQMHS-UHFFFAOYSA-N 0.000 description 11
- NTYJJOPFIAHURM-UHFFFAOYSA-N Histamine Chemical compound NCCC1=CN=CN1 NTYJJOPFIAHURM-UHFFFAOYSA-N 0.000 description 8
- KIDHWZJUCRJVML-UHFFFAOYSA-N putrescine Chemical compound NCCCCN KIDHWZJUCRJVML-UHFFFAOYSA-N 0.000 description 8
- APJYDQYYACXCRM-UHFFFAOYSA-N tryptamine Chemical compound C1=CC=C2C(CCN)=CNC2=C1 APJYDQYYACXCRM-UHFFFAOYSA-N 0.000 description 8
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 6
- 150000001299 aldehydes Chemical class 0.000 description 5
- YGHRJJRRZDOVPD-UHFFFAOYSA-N 3-methylbutanal Chemical compound CC(C)CC=O YGHRJJRRZDOVPD-UHFFFAOYSA-N 0.000 description 4
- PAYRUJLWNCNPSJ-UHFFFAOYSA-N Aniline Chemical compound NC1=CC=CC=C1 PAYRUJLWNCNPSJ-UHFFFAOYSA-N 0.000 description 4
- 239000005700 Putrescine Substances 0.000 description 4
- 239000002253 acid Substances 0.000 description 4
- -1 aldehyde ketone compounds Chemical class 0.000 description 4
- HUMNYLRZRPPJDN-UHFFFAOYSA-N benzaldehyde Chemical compound O=CC1=CC=CC=C1 HUMNYLRZRPPJDN-UHFFFAOYSA-N 0.000 description 4
- VHRGRCVQAFMJIZ-UHFFFAOYSA-N cadaverine Chemical compound NCCCCCN VHRGRCVQAFMJIZ-UHFFFAOYSA-N 0.000 description 4
- 229960001340 histamine Drugs 0.000 description 4
- VSMOENVRRABVKN-UHFFFAOYSA-N oct-1-en-3-ol Chemical compound CCCCCC(O)C=C VSMOENVRRABVKN-UHFFFAOYSA-N 0.000 description 4
- 238000004321 preservation Methods 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 235000019640 taste Nutrition 0.000 description 4
- 241000228212 Aspergillus Species 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 150000001298 alcohols Chemical class 0.000 description 3
- 230000000052 comparative effect Effects 0.000 description 3
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- VSMOENVRRABVKN-MRVPVSSYSA-N 1-Octen-3-ol Natural products CCCCC[C@H](O)C=C VSMOENVRRABVKN-MRVPVSSYSA-N 0.000 description 2
- BWZVCCNYKMEVEX-UHFFFAOYSA-N 2,4,6-Trimethylpyridine Chemical compound CC1=CC(C)=NC(C)=C1 BWZVCCNYKMEVEX-UHFFFAOYSA-N 0.000 description 2
- IWTBVKIGCDZRPL-LURJTMIESA-N 3-Methylbutanol Natural products CC[C@H](C)CCO IWTBVKIGCDZRPL-LURJTMIESA-N 0.000 description 2
- YDXQPTHHAPCTPP-UHFFFAOYSA-N 3-Octen-1-ol Natural products CCCCC=CCCO YDXQPTHHAPCTPP-UHFFFAOYSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 235000013332 fish product Nutrition 0.000 description 2
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 description 2
- JARKCYVAAOWBJS-UHFFFAOYSA-N hexanal Chemical compound CCCCCC=O JARKCYVAAOWBJS-UHFFFAOYSA-N 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- PHTQWCKDNZKARW-UHFFFAOYSA-N isoamylol Chemical compound CC(C)CCO PHTQWCKDNZKARW-UHFFFAOYSA-N 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- FEMOMIGRRWSMCU-UHFFFAOYSA-N ninhydrin Chemical compound C1=CC=C2C(=O)C(O)(O)C(=O)C2=C1 FEMOMIGRRWSMCU-UHFFFAOYSA-N 0.000 description 2
- GYHFUZHODSMOHU-UHFFFAOYSA-N nonanal Chemical compound CCCCCCCCC=O GYHFUZHODSMOHU-UHFFFAOYSA-N 0.000 description 2
- NUJGJRNETVAIRJ-UHFFFAOYSA-N octanal Chemical compound CCCCCCCC=O NUJGJRNETVAIRJ-UHFFFAOYSA-N 0.000 description 2
- QNGNSVIICDLXHT-UHFFFAOYSA-N para-ethylbenzaldehyde Natural products CCC1=CC=C(C=O)C=C1 QNGNSVIICDLXHT-UHFFFAOYSA-N 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- 235000001674 Agaricus brunnescens Nutrition 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
- 208000035404 Autolysis Diseases 0.000 description 1
- 206010057248 Cell death Diseases 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
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 241000282414 Homo sapiens Species 0.000 description 1
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical class [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 230000009471 action Effects 0.000 description 1
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- 239000012620 biological material Substances 0.000 description 1
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- 238000002474 experimental method Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 235000019264 food flavour enhancer Nutrition 0.000 description 1
- 239000003205 fragrance Substances 0.000 description 1
- 239000013505 freshwater Substances 0.000 description 1
- 210000002816 gill Anatomy 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- PWSKHLMYTZNYKO-UHFFFAOYSA-N heptane-1,7-diamine Chemical compound NCCCCCCCN PWSKHLMYTZNYKO-UHFFFAOYSA-N 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 238000001819 mass spectrum Methods 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 235000010755 mineral Nutrition 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 230000000050 nutritive 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
- 239000008055 phosphate buffer solution Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000004007 reversed phase HPLC Methods 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 230000028043 self proteolysis Effects 0.000 description 1
- 238000002470 solid-phase micro-extraction Methods 0.000 description 1
- 238000009210 therapy by ultrasound Methods 0.000 description 1
- YNJBWRMUSHSURL-UHFFFAOYSA-N trichloroacetic acid Chemical compound OC(=O)C(Cl)(Cl)Cl YNJBWRMUSHSURL-UHFFFAOYSA-N 0.000 description 1
- 210000001835 viscera Anatomy 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 239000002699 waste material 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
- A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
- A23L27/50—Soya sauce
-
- 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
-
- 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
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/90—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in food processing or handling, e.g. food conservation
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Microbiology (AREA)
- Soy Sauces And Products Related Thereto (AREA)
Abstract
A preparation method of fish sauce comprises adding enzyme and bacteria for continuous fermentation to shorten fermentation period of fish sauce, fermenting to obtain flavored protease by Aspergillus oryzae, and removing bitter peptide chain of low hydrolysis degree product to specifically decompose macromolecular protein; the Aspergillus oryzae has multiple compound proteases, and can decompose the peptide chain which is not sheared by the flavourzyme into small molecule peptide or amino acid; the lactobacillus plantarum can inhibit microorganisms carried in byproducts, reduce biogenic amine in fish sauce and improve safety; debaryomyces has the ability to break down lactose, lactate and protein and metabolize the aroma, improving the flavor of fish gravy by adding yeast.
Description
Technical Field
The invention relates to the field of food processing, in particular to a method for preparing low-biogenic amine fish sauce by continuous fermentation of flavourzyme, aspergillus oryzae, lactobacillus plantarum and yeast.
Background
The fish sauce is produced with low-value leftover of aquatic product as main material and through natural fermentation, fermentation and decomposition of protein, fat and other components in fish material with the participation of various kinds of microbe. The food has delicious taste and unique flavor, and is often used as a flavor enhancer or a substitute salt in cooking foods. The preparation of fish gravy generally comprises a traditional fermentation process and a rapid fermentation process, wherein the traditional fermentation process is formed by fermenting raw materials by insolating the raw materials under the sun and utilizing microbial enzymes carried by the raw materials and other microorganisms in the air; the rapid fermentation process can be divided into heat preservation fermentation, enzyme fermentation, starter fermentation and mixed fermentation, wherein the heat preservation fermentation is carried out by utilizing the optimal temperature of autolysis of microorganisms in the raw materials; enzyme fermentation is to add proper protease into the raw materials for fermentation; the yeast adding fermentation is to add aspergillus species yeast for fermentation, and a large amount of protease is produced by aspergillus propagation for fermentation; the mixed fermentation is carried out by adding aspergillus on the basis of adding enzyme. The traditional fish sauce has longer fermentation period, lower production efficiency and high salt content, and is easy to produce harmful substances such as biogenic amine; along with the development of the fermentation process, the fermentation period is shortened by adding strain fermentation, and the product quality becomes a research hot spot.
The mandarin fish is a raw material for processing and producing the stink mandarin fish, the stink mandarin fish product is a produced by using a produced fermented mandarin fish in the Huizhou of a tunnel, the demand on the mandarin fish is large, the annual yield of the mandarin fish in the yellow mountain area of the 2022 year can reach 200 tons, the mandarin fish is mainly distributed in all main rivers and lakes in China, the mandarin fish belongs to the carnivorous freshwater fish, the action is quick, the movement capability is strong, the muscles are developed, the meat quality is fat, the taste is fresh and tender, and the mandarin fish has higher nutritive value and economic value. The production of stink mandarin fish requires a large amount of mandarin fish, so that a large amount of mandarin fish leftovers, such as viscera and gills of mandarin fish, are mostly thrown away directly in the process, which causes resource waste and environmental pollution. In addition, the mandarin fish leftovers contain amino acids, proteins, vitamins, minerals and other nutrients required by human beings, so that the mandarin fish leftovers have potential development value as fish sauce products.
With development of fish sauce products, consumers have higher requirements on the safety and flavor of fish sauce, and the differences of fermentation time, starter and fermentation conditions of the fish sauce can influence the flavor and safety of the fish sauce products, so that biogenic amine is a substance with larger influence on the safety of the fish sauce.
Disclosure of Invention
The invention aims to provide a preparation method of fish gravy.
To achieve the above and other related objects, the present invention provides the following technical solutions: a preparation method of fish sauce comprises the following steps:
step 1: preparing aspergillus oryzae starter propagation, lactobacillus plantarum inoculums and saccharomycetes inoculums;
step 2: pretreatment of raw materials: cleaning mandarin fish leftovers, and adding water to prepare minced meat;
step 3: adding flavourzyme into the meat emulsion for enzymolysis to obtain an enzymolysis product;
step 4: and (3) adding bacteria and fermenting: sequentially adding aspergillus oryzae starter propagation, lactobacillus plantarum inoculums and saccharomycetes inoculums into the enzymolysis products for fermentation to obtain fermentation products;
step 5: filtering and separating solid filter residues after fermentation is completed, and pasteurizing filtrate to obtain fish gravy.
The preferable technical scheme is as follows: the preparation of the aspergillus oryzae starter propagation comprises the following steps:
streaking the activated Aspergillus oryzae on a potato dextrose plate, and culturing for 65-80d at 28-30 ℃; preparing Aspergillus oryzae spore liquid with sterile physiological saline, transferring 1ml spore liquid into sterilized bran culture medium, culturing at 28-32deg.C for 36-48 hr, and collecting light yellow green Aspergillus oryzae AS a spare fermentation seed koji, wherein the Aspergillus oryzae is AS3.042 Aspergillus oryzae;
preparation of lactobacillus plantarum inoculum: inoculating lactobacillus plantarum on an inclined plane to an MRS agar culture medium for activation, culturing for 36-48h at 30-40 ℃, inoculating the obtained culture into an MRS liquid culture medium again, culturing for 24-28h at 30-40 ℃ to obtain an inoculum, centrifuging the liquid culture medium at a rotating speed of 3000-4000rpm/min, pouring out supernatant, washing the precipitate twice with sterile physiological saline, and preparing bacterial suspension with the sterile physiological saline;
preparation of yeast inoculum: streaking activated Debaryomyces hansenii, inoculating in PDA agar medium, culturing at 30-40deg.C for 45-50 hr, preparing into bacterial suspension with sterile physiological saline, and regulating bacterial concentration to about 8-9log CFU/m L.
The preferable technical scheme is as follows: the wheat bran culture medium comprises the following components: 45-55 parts of bran, 45-55 parts of water, 4-6 parts of crushed mandarin fish leftovers and sterilizing for 30min at 121 ℃;
the preferable technical scheme is as follows: the concentration of lactobacillus plantarum inoculum is 7-10log CFU/m L;
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 80, 15-20g of agar, pH of 6.5+/-0.2 and sterilizing at 121 ℃ for 20min; MRS liquid medium differs from MRS agar medium in that no agar is added; the PDA culture medium formula is as follows: 200g of potato, 20g of glucose, 15-20g of agar, 1000mL of distilled water and natural pH; the preparation method of the culture medium comprises the following steps: cleaning potato, peeling, weighing 200g potato, cutting into small pieces, boiling in water for 20-30 min, cutting with glass rod, filtering with 4 layers of gauze, adding glucose and agar into the filtrate, stirring, cooling, supplementing water to 1000mL, and sterilizing at 115deg.C for 20min.
The preferable technical scheme is as follows: the enzyme activity of the flavourzyme is more than or equal to 30000u/g, the effective temperature is 30-60 ℃, the optimal temperature is 50-55 ℃, and the optimal pH is 6-7; the enzyme is added according to 1-2% of the weight of the substrate, the enzymolysis time is 4-8h, and the enzymolysis temperature is 45-55 ℃.
The preferable technical scheme is as follows: before adding aspergillus oryzae starter propagation, adding 3-8% of salt by mass of the material into the enzymolysis product.
The preferable technical scheme is as follows: adding Aspergillus oryzae strain at 5-10% of the raw materials, mixing, fermenting for 3-5d at 28-32deg.C.
The preferable technical scheme is as follows: lactobacillus plantarum accounting for 1-2% of the mass of the raw materials is added for secondary fermentation, the fermentation time is 1.5-2.5d, and the fermentation temperature is 30-40 ℃.
The preferable technical scheme is as follows: adding yeast 1-2% of the material mass for the third fermentation, wherein the fermentation time is 2.5-3.5d, and the fermentation temperature is 30-40 ℃.
Due to the application of the technical scheme, compared with the prior art, the invention has the advantages that:
1. the content of amino nitrogen in the fish sauce added with the aspergillus oryzae starter for fermentation reaches more than the standard of the first-stage fish sauce, the content of amino nitrogen in each 100ml of fish sauce can reach 1.07g, and the amino nitrogen is improved by 44.59 percent compared with the fish sauce without aspergillus oryzae starter.
2. By adding aspergillus oryzae and lactobacillus plantarum for continuous fermentation, the biogenic amine content in fish sauce is obviously reduced, only three biogenic amines, namely tryptamine, putrescine and histamine, are detected, two biogenic amines are reduced compared with a control group, and the content of the three biogenic amines is reduced by 30.56%, 68.62% and 27.22% respectively.
3. The contents of amino acid and flavor substances in the fish sauce subjected to the composite fermentation of Aspergillus oryzae, lactobacillus plantarum and Debaryomyces hansenii are respectively increased by 5.91 percent and 12.87 percent.
Drawings
FIG. 1 is a flow chart of a fermentation process.
FIG. 2 shows fish gravy in different fermentation stages.
FIG. 3 is a graph comparing fish gravy after fermentation of three bacteria. (1) A fish sauce sample obtained after Aspergillus oryzae fermentation, filtration and sterilization; (2) fermenting Aspergillus oryzae and Lactobacillus plantarum, filtering, and sterilizing to obtain fish sauce sample; (3) aspergillus oryzae, lactobacillus plantarum and yeast, filtering, and sterilizing.
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. 1-3. 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 preserved in China Center for Type Culture Collection (CCTCC) for type number of times of preservation (CCTCC NO: M20221303) at month 8 of 2022.
The reagents and materials described in the examples below are commercially available unless otherwise indicated.
Example 1: preparation method of fish gravy
1. Preparation of aspergillus oryzae starter propagation: streaking the activated Aspergillus oryzae on a potato dextrose plate, and culturing for 3d at 28-30 ℃; preparation of Aspergillus oryzae spore liquid with sterile physiological saline, and removal of 1ml spore liquid (about 10 7 The sporophores) are cultured in a sterilized bran culture medium for 36 hours at the temperature of 30 ℃, and aspergillus oryzae growing light yellow green in the bran culture medium can be used as a spare fermentation starter.
2. Preparation of lactobacillus plantarum inoculum: inoculating Lactobacillus plantarum on an inclined plane, inoculating to MRS agar culture medium for activation, culturing at 35 ℃ for 36-48h, inoculating the obtained culture to MRS liquid culture medium again, culturing at 35 ℃ for 24-28h to obtain an inoculum, centrifuging the liquid culture medium at 3500rpm/min, pouring out the supernatant, washing the precipitate twice with sterile physiological saline, preparing a bacterial suspension with sterile physiological saline, and regulating the bacterial concentration to about 8-9log CFU/mL.
3. Preparation of debaryomyces hansenii inoculum: the activated Debaryomyces hansenii is streaked and inoculated in PDA agar medium, and cultured for 48 hours at 35 ℃, and then is prepared into bacterial suspension by sterile physiological saline, and the bacterial concentration is regulated to be about 8-9log CFU/mL.
4. Pretreatment of raw materials: cleaning mandarin fish leftovers, adding distilled water according to a ratio of 1:1, and homogenizing to obtain meat emulsion.
5. Adding 1.5% of flavourzyme into the meat emulsion sample, and carrying out enzymolysis for 6 hours at 50 ℃ and pH7 to obtain an enzymolysis product;
6. and (3) adding bacteria and fermenting: firstly, adding 6% of salt into an enzymolysis product, adding 6% of AS3.042 aspergillus oryzae (added by 6% of the mass of the raw materials), mixing, and then performing primary fermentation for 4 days at a fermentation temperature of 30 ℃; adding 1% lactobacillus plantarum (added in an amount of 1% of the mass of the raw materials) after the primary fermentation for secondary fermentation, wherein the fermentation time is 2d, and the fermentation temperature is 35 ℃; adding 2% of Debaryomyces hansenii (added in an amount of 2% of the mass of the raw material) to the mixture after the second fermentation for the third fermentation, wherein the fermentation time is 3d, and the fermentation temperature is 35 ℃.
7. After the three fermentations were completed, the solid filter residue was separated by filtration with 3 layers of 100 mesh gauze and sterilized to obtain fish gravy, which was designated as F3 group.
Comparative example 1: preparation method of fish gravy
1. Preparation of aspergillus oryzae starter propagation: streaking the activated Aspergillus oryzae on a potato dextrose plate, and culturing for 3d at 28-30 ℃; preparation of Aspergillus oryzae spore liquid with sterile physiological saline, and removal of 1ml spore liquid (about 10 7 The sporophores) are cultured in a sterilized bran culture medium for 36 hours at the temperature of 30 ℃, and aspergillus oryzae growing light yellow green in the bran culture medium can be used as a spare fermentation starter.
2. Preparation of lactobacillus plantarum inoculum: inoculating Lactobacillus plantarum on an inclined plane, inoculating to MRS agar culture medium for activation, culturing at 35 ℃ for 36-48h, inoculating the obtained culture to MRS liquid culture medium again, culturing at 35 ℃ for 24-28h to obtain an inoculum, centrifuging the liquid culture medium at 3500rpm/min, pouring out the supernatant, washing the precipitate twice with sterile physiological saline, preparing a bacterial suspension with sterile physiological saline, and regulating the bacterial concentration to about 8-9log CFU/mL.
3. Pretreatment of raw materials: cleaning mandarin fish leftovers, adding distilled water according to a ratio of 1:1, and homogenizing to obtain meat emulsion.
4. Adding 1.5% of flavourzyme into the meat emulsion sample, and carrying out enzymolysis for 6 hours at 50 ℃ and pH7 to obtain an enzymolysis product;
5. and (3) adding bacteria and fermenting: firstly, adding 6% of salt into an enzymolysis product, adding 6% of AS3.042 aspergillus oryzae (added by 6% of the mass of the raw materials), mixing, and then performing primary fermentation for 4 days at a fermentation temperature of 30 ℃; after the first fermentation, 1% lactobacillus plantarum (added in an amount of 1% of the mass of the raw materials) is added for the second fermentation, the fermentation time is 2d, and the fermentation temperature is 35 ℃.
6. After the secondary fermentation is completed, filtering and separating solid filter residues by using 3 layers of 100-mesh gauze, and sterilizing to obtain fish gravy, which is marked as F2 group.
Comparative example 2: preparation method of fish gravy
1. Preparation of aspergillus oryzae starter propagation: streaking the activated Aspergillus oryzae on a potato dextrose plate, and culturing for 3d at 28-30 ℃; preparation of Aspergillus oryzae spore liquid with sterile physiological saline, and removal of 1ml spore liquid (about 10 7 The sporophores) are cultured in a sterilized bran culture medium for 36 hours at the temperature of 30 ℃, and aspergillus oryzae growing light yellow green in the bran culture medium can be used as a spare fermentation starter.
2. Pretreatment of raw materials: cleaning mandarin fish leftovers, adding distilled water according to a ratio of 1:1, and homogenizing to obtain meat emulsion.
3. Adding 1.5% of flavourzyme into the meat emulsion sample, and carrying out enzymolysis for 6 hours at 50 ℃ and pH7 to obtain an enzymolysis product;
4. and (3) adding bacteria and fermenting: firstly adding 6% of salt into an enzymolysis product, adding 6% of AS3.042 Aspergillus oryzae (added by 6% of the mass of the raw materials), and fermenting for 4 days at a fermentation temperature of 30 ℃.
5. After fermentation, the solid filter residue is filtered and separated by 3 layers of 100-mesh gauze, and the fish gravy is obtained after sterilization and is marked as F1 group.
Comparative example 3: preparation method of fish gravy
1. Pretreatment of raw materials: cleaning mandarin fish leftovers, adding distilled water according to a ratio of 1:1, and homogenizing to obtain meat emulsion.
2. Adding 1.5% of flavourzyme into the meat emulsion sample, and carrying out enzymolysis for 6 hours at 50 ℃ and pH7 to obtain an enzymolysis product;
4. filtering the enzymolysis product with 3 layers of 100 mesh gauze to separate solid filter residues, sterilizing to obtain fish gravy, and marking as group C.
1. Determination of Ammonia-state nitrogen content: the amino nitrogen content of the fish gravy of the group C and the group F1 is measured, and the detection method adopts an ninhydrin colorimetric method, and the method is as follows:
taking 1mL of sample, diluting 500 times, dispersing uniformly, standing, taking 4mL of diluent, adding 1mL of ninhydrin solution and phosphate buffer solution, shaking uniformly, heating in water bath for 15min, standing for 15min, taking distilled water as blank control, measuring A570, and calculating by standard curve to obtain the amino nitrogen content of the sample.
The experimental results are as follows:
table 1 amino nitrogen content (unit: g/100 mL) of fish gravy of group C and group F1
| C | F1 | |
| Amino nitrogen content | 0.74±0.02 | 1.07±0.06 |
Note that: the C group is enzymolysis product, the F1 group is enzymolysis and Aspergillus oryzae fermentation group
The amino nitrogen content of fish gravy is an important index, and the amino nitrogen content is generally greater than or equal to 0.9g/100mL and is defined as first-class fish gravy; the amino nitrogen content in the fish sauce added with the aspergillus oryzae is increased by 0.33g/100mL compared with the control group, and the amino nitrogen content in the F1 group reaches the standard of the first-class fish sauce.
2. Determination of biogenic amine content: the biogenic amine content of the fish gravy of the F1 group and the F2 group is detected by adopting a High Performance Liquid Chromatography (HPLC), and the detection method is as follows:
sample treatment: taking 1mL of fish sauce, adding a proper amount of 0.6M HClO4 solution and 125 mu L of internal standard (1, 7-diaminoheptane), uniformly mixing, centrifuging, taking supernatant after centrifuging, repeating the steps, and finally merging the supernatant to a 10mL 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 2M NaOH, 60 mu L of saturated NaHCO3 and 400 mu L of 10mg/mL 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.
Table 2 biogenic amine content (unit: mg/L) of fermented fish gravy of groups F1 and F2
| F1 | F2 | |
| Tryptamine | 363.61±19.83 | 252.49±17.32 |
| Aniline | 12.62±1.38 | ND |
| Putrescine | 130.20±10.18 | 40.86±7.68 |
| Cadaverine | 12.58±0.71 | ND |
| Histamine | 153.22±5.10 | 111.51±5.62 |
Note that: the F1 group is zymolysis and Aspergillus oryzae fermentation, the F2 group is zymolysis and Aspergillus oryzae fermentation and Lactobacillus plantarum fermentation
The fish gravy fermented by lactobacillus plantarum can effectively reduce the content of biogenic amine, and the tryptamine, the aniline, the putrescine, the cadaverine and the histamine are detected in the F1 group (which is not connected with lactobacillus plantarum), and the tryptamine, the putrescine and the histamine are only detected in the F2 group (which is connected with lactobacillus plantarum), and the content of the biogenic amine is lower than that of the F1 group.
3. Determination of amino acid content: the amino acid content of the fish gravy of the F2 group and the F3 group is detected by adopting a full-automatic amino acid analyzer, and the detection method is as follows:
taking 5mL of a sample, adding 25mL of 5% trichloroacetic acid solution, carrying out ultrasonic treatment for 2 times each for 15min, standing for 1 hour, filtering by a needle filter with the thickness of 0.22 mu m, bottling, and carrying out on-machine analysis.
Table 3 amino acid content (unit: mg/kg) of fermented fish gravy of groups F2 and F3
Note that: the F2 group is zymolytic Aspergillus oryzae, lactobacillus plantarum and the F3 group is zymolytic Aspergillus oryzae, lactobacillus plantarum and Debaryomyces hansenii
The total content of free amino acids in the fish gravy added with yeast in the F3 group is increased by 5.9%, wherein the content of sweet amino acids is increased to the greatest extent, and is increased by 6.9% compared with that in the F2 group (without yeast), and the fresh taste and the bitter taste amino acids are respectively increased by 6.2% and 4.6%, which indicates that the amino acids beneficial to increasing the fish gravy after the yeast is added are improved, and the taste of the fish gravy is improved.
4. Determination of volatile flavour compounds: the volatile flavor compounds of the fish gravy of the F2 group and the F3 group are measured by adopting solid phase microextraction gas chromatography-mass spectrometry (HPMS-HS-GC-MS), and the measuring method is as follows:
each sample was taken in 5mL of a 20mL headspace bottle and 10. Mu.L of an internal standard (2, 4, 6-trimethylpyridine) was added at one ten-thousandth concentration. The extraction head was inserted into a headspace bottle at 70℃for 40min.
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.
The results were as follows:
table 4 volatile flavor compound content of fermented fish gravy of groups F2 and F3 (unit: μg/100 g)
Note that: the F2 group is zymolytic Aspergillus oryzae, lactobacillus plantarum and the F3 group is zymolytic Aspergillus oryzae, lactobacillus plantarum and Debaryomyces hansenii
The total flavor content of fish sauce is increased after yeast is added, wherein the contents of acids, esters, olefins and aromatic hydrocarbon compounds are increased and the contents of alcohols and aldehyde ketone compounds are reduced compared with the F2 group (not connected with yeast); the alcohol compounds are generally oxidized into acid, aldehyde and ketone compounds and esterified with acid into ester compounds after fermentation, and from the change, more acid and ester are generated by the F3 group added with yeast, the content of ester is increased more, and less aldehyde and ketone compounds with bad smell are generated by the F3 group (connected with yeast), so that the flavor of fish gravy is improved by adding yeast.
Table 5 alcohol content of fermented fish gravy of groups F2 and F3 (unit: μg/100 g)
Table 6 aldehyde content (unit: μg/100 g) of fermented fish gravy of groups F2 and F3
Note that: the F2 group is zymolytic Aspergillus oryzae, lactobacillus plantarum and the F3 group is zymolytic Aspergillus oryzae, lactobacillus plantarum and Debaryomyces hansenii
The alcohol compounds in the group of the grafted yeasts are reduced, but the content of 1-octen-3-ol is increased, the 1-octen-3-ol is a common flavor compound in fermented fish products, has mushroom fragrance, has a promoting effect on flavor, and 3-methyl-butanol has a pungent and unpleasant smell, and the content of 3-methyl-butanol in the group F3 (grafted yeasts) is reduced, so that the unpleasant smell of the flavor of the grafted yeasts is reduced. The straight-chain saturated aldehydes such as octanal, hexanal and nonanal have fishy smell, the content of the aldehydes in the yeast group F3 is reduced, the fishy smell in the fish sauce is reduced, the benzaldehyde and 3-methyl butyraldehyde have nut smell, the flavor of the fish sauce can be enhanced, and the content of the benzaldehyde and 3-methyl butyraldehyde in the F3 is increased compared with that in the yeast group F2 (not connected with yeast), so that the fish sauce flavor of the yeast fermentation group is better.
Example 2: preparation method of fish gravy
A preparation method of fish sauce comprises the following steps:
step 1: preparing aspergillus oryzae starter propagation, lactobacillus plantarum inoculums and saccharomycetes inoculums;
step 2: pretreatment of raw materials: cleaning mandarin fish leftovers, and adding water to prepare minced meat;
step 3: adding flavourzyme into the meat emulsion for enzymolysis to obtain an enzymolysis product;
step 4: and (3) adding bacteria and fermenting: sequentially adding aspergillus oryzae starter propagation, lactobacillus plantarum inoculums and saccharomycetes inoculums into the enzymolysis products for fermentation to obtain fermentation products;
step 5: filtering and separating solid filter residues after fermentation is completed, and pasteurizing filtrate to obtain fish gravy.
The preferable technical scheme is as follows: the preparation of the aspergillus oryzae starter propagation comprises the following steps:
streaking the activated aspergillus oryzae on a potato dextrose plate, and culturing for 65d at 28 ℃; preparing Aspergillus oryzae spore liquid with sterile physiological saline, transferring 1ml spore liquid into sterilized bran culture medium, culturing at 28deg.C for 36 hr, and taking Aspergillus oryzae AS a spare fermentation seed koji when light yellow green grows in the bran culture medium, wherein the Aspergillus oryzae is AS3.042 Aspergillus oryzae;
preparation of lactobacillus plantarum inoculum: inoculating lactobacillus plantarum on an inclined plane to an MRS agar culture medium for activation, culturing for 36 hours at 30 ℃, inoculating the obtained culture into an MRS liquid culture medium again by using the inoculating loop, culturing for 24 hours at 30 ℃ to obtain an inoculum, centrifuging the liquid culture medium at a rotating speed of 3000rpm/min, pouring out the supernatant, washing the precipitate twice by using sterile physiological saline, and preparing a bacterial suspension by using the sterile physiological saline;
preparation of yeast inoculum: the activated Debaryomyces hansenii is streaked and inoculated in PDA agar medium and cultured for 45 hours at 30 ℃, then the strain suspension is prepared by sterile physiological saline, and the strain concentration is regulated to about 8log CFU/m L.
The preferred embodiments are: the wheat bran culture medium comprises the following components: 45 parts of bran, 45 parts of water, 4 parts of crushed mandarin fish leftovers and sterilizing for 30min at 121 ℃;
the preferred embodiments are: the concentration of lactobacillus plantarum inoculum was 7log CFU/m L;
the preferred embodiments are: 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 80, 15-20g of agar, pH of 6.5+/-0.2 and sterilizing at 121 ℃ for 20min; MRS liquid medium differs from MRS agar medium in that no agar is added; the PDA culture medium formula is as follows: 200g of potato, 20g of glucose, 15g of agar and 1000mL of distilled water, and the natural pH; the preparation method of the culture medium comprises the following steps: cleaning potato, peeling, weighing 200g potato, cutting into small pieces, boiling in water for 20min, cutting with glass rod, filtering with 4 layers of gauze, adding glucose and agar into the filtrate, stirring, cooling, supplementing water to 1000mL, and sterilizing at 115deg.C for 20min.
The preferred embodiments are: the enzyme activity of the flavourzyme is more than or equal to 30000u/g, the effective temperature is 30 ℃, the optimal temperature is 50 ℃, and the optimal pH is 6; the enzyme is added according to 1% of the weight of the substrate, the enzyme hydrolysis time is 4 hours, and the enzyme hydrolysis temperature is 45 ℃.
The preferred embodiments are: before adding aspergillus oryzae starter propagation, adding 3% of salt by mass of the material into the enzymolysis product.
The preferred embodiments are: adding Aspergillus oryzae strain at 5% of the mass of the raw materials, mixing, fermenting for 3d at 28deg.C.
The preferred embodiments are: lactobacillus plantarum accounting for 1 percent of the mass of the raw materials is added for secondary fermentation, the fermentation time is 1.5d, and the fermentation temperature is 30 ℃.
The preferred embodiments are: adding saccharomycetes in an amount of 1% of the mass of the raw materials for third fermentation, wherein the fermentation time is 2.5d, and the fermentation temperature is 30 ℃.
Example 3: preparation method of fish gravy
A preparation method of fish sauce comprises the following steps:
step 1: preparing aspergillus oryzae starter propagation, lactobacillus plantarum inoculums and saccharomycetes inoculums;
step 2: pretreatment of raw materials: cleaning mandarin fish leftovers, and adding water to prepare minced meat;
step 3: adding flavourzyme into the meat emulsion for enzymolysis to obtain an enzymolysis product;
step 4: and (3) adding bacteria and fermenting: sequentially adding aspergillus oryzae starter propagation, lactobacillus plantarum inoculums and saccharomycetes inoculums into the enzymolysis products for fermentation to obtain fermentation products;
step 5: filtering and separating solid filter residues after fermentation is completed, and pasteurizing filtrate to obtain fish gravy.
The preferred embodiments are: the preparation of the aspergillus oryzae starter propagation comprises the following steps:
streaking the activated aspergillus oryzae on a potato dextrose plate, and culturing for 80d at 30 ℃; preparing Aspergillus oryzae spore liquid with sterile physiological saline, transferring 1ml spore liquid into sterilized bran culture medium, culturing at 32deg.C for 48 hr, and taking Aspergillus oryzae AS a spare fermentation seed koji when light yellow green grows in the bran culture medium, wherein the Aspergillus oryzae is AS3.042 Aspergillus oryzae;
preparation of lactobacillus plantarum inoculum: inoculating lactobacillus plantarum on an inclined plane to an MRS agar culture medium for activation, culturing for 48 hours at 40 ℃, inoculating the obtained culture into an MRS liquid culture medium again by using the inoculating loop, culturing for 28 hours at 40 ℃ to obtain an inoculum, centrifuging the liquid culture medium at the rotating speed of 4000rpm/min, pouring out the supernatant, washing the precipitate twice by using sterile physiological saline, and preparing bacterial suspension by using the sterile physiological saline;
preparation of yeast inoculum: the activated Debaryomyces hansenii is streaked and inoculated in PDA agar medium, and cultured for 50 hours at 40 ℃, and then the strain suspension is prepared by sterile physiological saline, and the strain concentration is regulated to be about 9log CFU/m L.
The preferred embodiments are: the wheat bran culture medium comprises the following components: 55 parts of bran, 55 parts of water, 6 parts of crushed mandarin fish leftovers and sterilizing for 30min at 121 ℃;
the preferred embodiments are: the concentration of lactobacillus plantarum inoculum was 10log CFU/m L;
the preferred embodiments are: 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 80, 15-20g of agar, pH of 6.5+/-0.2 and sterilizing at 121 ℃ for 20min; MRS liquid medium differs from MRS agar medium in that no agar is added; the PDA culture medium formula is as follows: 200g of potato, 20g of glucose, 20g of agar and 1000mL of distilled water, and the natural pH; the preparation method of the culture medium comprises the following steps: cleaning potato, peeling, weighing 200g potato, cutting into small pieces, boiling in water for 30min, cutting with glass rod, filtering with 4 layers of gauze, adding glucose and agar into the filtrate, stirring, cooling, supplementing water to 1000mL, and sterilizing at 115deg.C for 20min.
The preferred embodiments are: the enzyme activity of the flavourzyme is more than or equal to 30000u/g, the effective temperature is 60 ℃, the optimal temperature is 55 ℃, and the optimal pH is 7; the enzyme is added according to 2 percent of the weight of the substrate, the enzyme hydrolysis time is 8 hours, and the enzyme hydrolysis temperature is 55 ℃.
The preferred embodiments are: before aspergillus oryzae is added into the starter propagation, 8% of salt is added into the enzymolysis product according to the mass of the raw materials.
The preferred embodiments are: adding Aspergillus oryzae strain at a ratio of 10% of the mass of the raw materials, mixing, fermenting for 5 days at 32deg.C.
The preferred embodiments are: lactobacillus plantarum accounting for 2 percent of the mass of the raw materials is added for secondary fermentation, the fermentation time is 2.5d, and the fermentation temperature is 40 ℃.
The preferred embodiments are: adding yeast in an amount of 2% of the mass of the raw materials for the third fermentation, wherein the fermentation time is 3.5d, and the fermentation temperature is 40 ℃.
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 (10)
1. A preparation method of fish sauce is characterized by comprising the following steps: comprises the following steps:
step 1: preparing aspergillus oryzae starter propagation, lactobacillus plantarum inoculums and saccharomycetes inoculums;
step 2: pretreatment of raw materials: cleaning mandarin fish leftovers, and adding water to prepare minced meat;
step 3: adding flavourzyme into the meat emulsion for enzymolysis to obtain an enzymolysis product;
step 4: and (3) adding bacteria and fermenting: sequentially adding aspergillus oryzae starter propagation, lactobacillus plantarum inoculums and saccharomycetes inoculums into the enzymolysis products for fermentation to obtain fermentation products;
step 5: filtering and separating solid filter residues after fermentation is completed, and pasteurizing filtrate to obtain fish gravy.
2. The method for preparing fish gravy according to claim 1, wherein: the preparation of the aspergillus oryzae starter propagation comprises the following steps:
streaking the activated Aspergillus oryzae on a potato dextrose plate, and culturing for 65-80d at 28-30 ℃; preparing Aspergillus oryzae spore liquid with sterile physiological saline, transferring 1ml spore liquid into sterilized bran culture medium, culturing at 28-32deg.C for 36-48 hr, and collecting light yellow green Aspergillus oryzae AS a spare fermentation seed koji, wherein the Aspergillus oryzae is AS3.042 Aspergillus oryzae;
preparation of lactobacillus plantarum inoculum: inoculating lactobacillus plantarum on an inclined plane to an MRS agar culture medium for activation, culturing for 36-48h at 30-40 ℃, inoculating the obtained culture into an MRS liquid culture medium again, culturing for 24-28h at 30-40 ℃ to obtain an inoculum, centrifuging the liquid culture medium at a rotating speed of 3000-4000rpm/min, pouring out supernatant, washing the precipitate twice with sterile physiological saline, and preparing bacterial suspension with the sterile physiological saline;
preparation of yeast inoculum: streaking activated Debaryomyces hansenii, inoculating in PDA agar medium, culturing at 30-40deg.C for 45-50 hr, preparing into bacterial suspension with sterile physiological saline, and regulating bacterial concentration to about 8-9log CFU/m L.
3. The method for preparing fish gravy according to claim 2, wherein: the wheat bran culture medium comprises the following components: 45-55 parts of bran, 45-55 parts of water, 4-6 parts of crushed mandarin fish leftovers and sterilizing at 121 ℃ for 30min.
4. The method for preparing fish gravy according to claim 2, wherein: the concentration of Lactobacillus plantarum inoculum was 7-10log CFU/m L.
5. The method for preparing fish gravy according to claim 2, wherein: 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 80, 15-20g of agar, pH of 6.5+/-0.2 and sterilizing at 121 ℃ for 20min; MRS liquid medium differs from MRS agar medium in that no agar is added; the PDA culture medium formula is as follows: 200g of potato, 20g of glucose, 15-20g of agar, 1000mL of distilled water and natural pH; the preparation method of the culture medium comprises the following steps: cleaning potato, peeling, weighing 200g potato, cutting into small pieces, boiling in water for 20-30 min, cutting with glass rod, filtering with 4 layers of gauze, adding glucose and agar into the filtrate, stirring, cooling, supplementing water to 1000mL, and sterilizing at 115deg.C for 20min.
6. The method for preparing fish gravy according to claim 1, wherein: the enzyme activity of the flavourzyme is more than or equal to 30000u/g, the effective temperature is 30-60 ℃, the optimal temperature is 50-55 ℃, and the optimal pH is 6-7; the enzyme is added according to 1-2% of the weight of the substrate, the enzymolysis time is 4-8h, and the enzymolysis temperature is 45-55 ℃.
7. The method for preparing fish gravy according to claim 1, wherein: before adding aspergillus oryzae starter propagation, adding 3-8% of salt by mass of the material into the enzymolysis product.
8. The method for preparing fish gravy according to claim 1, wherein: adding Aspergillus oryzae strain at 5-10% of the raw materials, mixing, fermenting for 3-5d at 28-32deg.C.
9. The method for preparing fish gravy according to claim 1, wherein: lactobacillus plantarum accounting for 1-2% of the mass of the raw materials is added for secondary fermentation, the fermentation time is 1.5-2.5d, and the fermentation temperature is 30-40 ℃.
10. The method for preparing fish gravy according to claim 1, wherein: adding yeast 1-2% of the material mass for the third fermentation, wherein the fermentation time is 2.5-3.5d, and the fermentation temperature is 30-40 ℃.
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