CN111534451B - Planococcus fermentation agent for improving fermentation quality of low-salt fish gravy - Google Patents

Planococcus fermentation agent for improving fermentation quality of low-salt fish gravy Download PDF

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CN111534451B
CN111534451B CN202010025134.4A CN202010025134A CN111534451B CN 111534451 B CN111534451 B CN 111534451B CN 202010025134 A CN202010025134 A CN 202010025134A CN 111534451 B CN111534451 B CN 111534451B
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planococcus
fish sauce
fish
cgmcc
sauce
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CN111534451A (en
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高瑞昌
周晶
袁丽
周越
王柱梅
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Jiangsu University
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Jiangsu University
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Priority to CN202110920482.2A priority patent/CN113528396B/en
Priority to CN202110919121.6A priority patent/CN113684145B/en
Priority to US17/256,008 priority patent/US11337447B2/en
Priority to PCT/CN2020/092490 priority patent/WO2021143006A1/en
Priority to JP2021513414A priority patent/JP7060902B2/en
Priority to EP20841648.7A priority patent/EP3878945A4/en
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    • 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
    • A23L17/70—Comminuted, e.g. emulsified, fish products; Processed products therefrom such as pastes, reformed or compressed products
    • 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
    • A23L27/20—Synthetic spices, flavouring agents or condiments
    • A23L27/24—Synthetic spices, flavouring agents or condiments prepared by fermentation
    • 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
    • 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
    • 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
    • Y02A40/963—Off-grid food refrigeration
    • Y02A40/966—Powered by renewable energy sources

Abstract

The invention discloses a planococcus fermenting agent for improving the fermentation quality of low-salt fish gravy, and relates to the field of application of food microbial technology. The 4 strains of the Planococcus is preserved in the China general microbiological culture Collection center (CGMCC) with the serial numbers of CGMCC NO.17057, CGMCC NO.17058, CGMCC NO.17059 and CGMCC NO.17060 respectively, and the suggested classification names are as follows: planococcus maritimus, Planococcus platortidis, Planococcus decalingnsis and Planococcus rieitoensis. The production period of the fish gravy fermented by utilizing the planococcus is greatly shortened, and the fermentation temperature is reduced. The fish sauce obtained by the method is transparent brownish red, has the special fragrance of aquatic products, and has no suspended or flocculent impurities and the like.

Description

Planococcus fermentation agent for improving fermentation quality of low-salt fish gravy
Technical Field
The invention relates to the field of application of food microbial technology, in particular to a planococcus fermenting agent for improving the fermentation quality of low-salt fish gravy.
Background
China is a big fishery country, and freshwater aquaculture has become an important industry pillar of fishery in China. The total output of freshwater aquaculture in 2017 is about 2905 ten thousand tons, but the processing utilization rate is less than 15%, so that the problems of resource waste, environmental pollution and the like are caused. Therefore, how to solve the problem of comprehensive utilization of freshwater fish is a great challenge in the aquatic product processing industry in China. Fish sauce (fish sauce) is a traditional aquatic product fermentation seasoning, also called fish sauce, is one of the most popular seasonings in southeast Asia, is clear and brownish red in color, has unique flavor and aroma, and becomes part of daily diet of people. The traditional fish sauce is produced by taking low-value fish and shrimp or aquatic product processing wastes (fish heads, internal organs and the like) as raw materials and utilizing self enzymes or microorganisms for natural fermentation. The fish sauce is rich in nutrition, contains various essential amino acids, and mineral elements such as calcium, magnesium, zinc, iron, etc. beneficial to human body, and is a low-fat high-protein fermented seasoning. Meanwhile, the fish sauce is also a product for high-value utilization of low-value fish, and has important practical significance for promoting the healthy development of freshwater fish industry. Protease is mainly used for catalyzing protein hydrolysis in the fish sauce fermentation process, and the fish sauce can be divided into low-temperature protease, medium-temperature protease and high-temperature protease according to the influence of temperature on the activity and stability of the protease. Low temperature proteases are a class of cold adapted proteases produced by low temperature bacteria under low temperature conditions. The low-temperature protease has higher catalytic efficiency under the low-temperature condition, and the optimal reaction temperature is 20-40 ℃ generally. The low-temperature protease can be reacted at low temperature or room temperature, heating and cooling are not needed, and the cost can be reduced. Therefore, the method has the advantage that the mesophilic protease can not be substituted in industrial production, and has wide application prospect in the fields of washing industry, food processing, biopharmaceutical industry, environmental bioremediation and the like. Planococcus has been identified as a psychrophile producing low temperature protease and is highly safe. The invention utilizes the planococcus to ferment the fish gravy under the condition of low temperature and low salt, thereby obtaining the fish gravy product with short fermentation period, low salinity and higher flavor and nutritive value, and providing theoretical basis and method for the planococcus in the application of food microbial fermentation.
Disclosure of Invention
The invention screens 4 new strains capable of improving the fermentation quality of the low-salt fish sauce from the traditional shrimp paste, and combines the growth characteristics and the enzymatic characteristics of the 4 strains to produce the fish sauce. The 4 strains of the bacillus subtilis can effectively shorten the fermentation period, reduce the salt content, improve the flavor and the nutritive value of the fish gravy, and provide a more stable, safer and more appropriate leavening agent for fish gravy fermentation.
The technical scheme adopted by the invention is as follows:
the invention provides a Planococcus marinus XJ2 which is a novel strain capable of being applied to improving the fermentation quality of fish gravy. The strain is preserved in China general microbiological culture Collection center (CGMCC) of China academy of sciences microorganism institute No. 3, West Lu No.1, North Chen, Xilu, Beijing, on 2019 at 02.01.3, the serial number is CGMCC NO.17057, and the suggested classification nomenclature is as follows: planococcus maritimus.
The invention provides a puccinia pulcherrima (Planococcus plakortidis XJ10), which is a new strain capable of being applied to improve the fermentation quality of fish gravy. The strain is preserved in China general microbiological culture Collection center (CGMCC) of China academy of sciences microorganism institute No. 3, West Lu No.1, North Chen, Xilu, Beijing, on 2019 at 02.01.3, the serial number is CGMCC NO.17058, and the suggested classification nomenclature is as follows: planococcus platortiis.
The Planococcus decellularis XJ11 provided by the invention is a novel strain which can be applied to improve the fermentation quality of fish gravy. The strain is preserved in China general microbiological culture Collection center (CGMCC) of China academy of sciences microorganism institute No. 3, West Lu No.1, North Chen, Xilu, Beijing, on 2019 at 02.01.3, the serial number is CGMCC NO.17059, and the suggested classification nomenclature is as follows: planococcus decalensis.
The Planococcus rivitoensis XJ12 provided by the invention is a new strain which can be applied to improve the fermentation quality of fish gravy. The strain is preserved in China general microbiological culture Collection center (CGMCC) of China academy of sciences microorganism institute No. 3, West Lu 1, North Chen, Xilu, Beijing, on 2019, at 02.01.3, the serial number is CGMCC NO.17060, and the suggested classification nomenclature is as follows: planococcus rieitoensis.
In one aspect of the present invention, there is provided the use of the aforementioned Planococcus marinus (Planococcus maritimus XJ2), Planococcus pulcherrimus (Planococcus platkortidis XJ10), Planococcus delbrueckii (Planococcus decalensis XJ11) and Planococcus leboensis (Planococcus rieitoensis XJ12) for improving the flavor and quality of fish sauce fermentation.
A method for fermenting fish sauce by utilizing planococcus comprises the following steps:
(1) treatment of raw materials: mincing minced fish (waste generated in fish processing) by using a mincing machine, adding 5-15% (w/w) of pickled sea salt, and uniformly mixing for later use.
(2) Preparing a leavening agent: the planococcus strain is activated and cultured for three times. Centrifuging the activated bacteria liquid at 4 deg.C and 10000r/min for 10min, washing with sterile normal saline twice, suspending in small amount of sterile normal saline, and adjusting the bacteria liquid concentration to 105～107CFU/mL, preparedThe application is as follows.
(3) Addition of a starter: adding the strain according to the final adding amount (10)5～109CFU/g) is mixed into the pretreated raw material minced fish, so that the final bacteria number reaches the addition requirement.
(4) Fermentation of fish sauce: and (4) fermenting the mixed material obtained in the step (3) at the temperature of 15-30 ℃ for 5-30 days in a heat preservation manner.
(5) Filtering the fish sauce: and (4) sterilizing the fish sauce sample obtained in the step (4) at 120 ℃ for 10-30 min, cooling to room temperature, centrifuging at 10000r/min for 20min, and filtering supernate with multiple layers of gauze to remove solid matters and impurities.
(6) Sterilizing the fish sauce: and (5) carrying out secondary sterilization on the fish sauce obtained in the step (5) at 100 ℃ for 15-30 min, and filling and sealing under aseptic conditions to obtain the fish sauce finished product.
(7) And (3) evaluating the physical and chemical indexes of the product: evaluating the content of amino acid nitrogen by reference to a GB5009.235-2016 colorimetric method; evaluating the content of volatile basic nitrogen by reference to a GB5009.228-2016 microdiffusion method; the method for measuring the histamine content refers to GB5009.208-2016 spectrophotometry.
Wherein the Planococcus is Planococcus maritimus (Planococcus maritimus XJ2) or Planococcus pulmonarius (Planococcus plateaus XJ10) or Planococcus derangensis (Planococcus mutandis XJ11) or Planococcus rabilis (Planococcus rieitoensis XJ 12).
The invention has the beneficial effects that:
the fermentation period of the fish sauce fermented by the 4 strains of the Planococcus is greatly shortened, and the fermentation temperature is reduced. The fish sauce obtained by the method is transparent brownish red, has the special fragrance of aquatic products, and has no suspended or flocculent impurities and the like. The salt content of the finished fish sauce is low, the amino acid nitrogen content is high (the fish sauce is divided into first-grade fish sauce according to the Chinese fish sauce industry standard), the content of volatile basic nitrogen is low, the content of histamine is lower than the national standard, and the prepared fish sauce is delicious in taste and rich in nutritional value, so that the 4 planococcus can be used for producing green and safe fermented fish sauce products.
In conclusion, the invention has the uniqueness, outstanding substantive features and obvious innovativeness and practicability of the invention patent, and produces beneficial effects.
Drawings
FIG. 1 shows the colony morphology and the cell morphology of Planococcus maritimus XJ 2;
FIG. 2 is a phylogenetic tree constructed from the 16S rRNA sequence of Planococcus maritimus XJ 2.
FIG. 3 shows the colony morphology and the cell morphology of Planococcus plakortidis XJ 10;
FIG. 4 is a phylogenetic tree constructed from the 16S rRNA sequence of Planococcus plakortidis XJ 10.
FIG. 5 shows the colony morphology and the cell morphology of Planococcus decehangensis XJ 11;
FIG. 6 is a phylogenetic tree constructed from the 16S rRNA sequence of Planococcus decehangensis XJ 11.
FIG. 7 shows the colony morphology and the cell morphology of Planococcus rieitoensis XJ 12;
FIG. 8 is a phylogenetic tree constructed from the 16S rRNA sequence of Planococcus rieitoensis XJ 12;
(Note: XJ-2 is Planococcus maritimus XJ2 of the present invention, XJ-10 is Planococcus maritimus XJ10 of the present invention, XJ-11 is Planococcus replacing XJ11 of the present invention, and XJ-12 is Planococcus rievetoinensis XJ12 of the present invention).
Detailed Description
1. Screening and purifying strains:
fresh shrimp paste (from a homemade shrimp paste in a farmhouse restaurant in Weihai city, Shandong province) which is fermented at low temperature is taken as a raw material, and the low-temperature protease producing planococcus spp is separated and screened from the shrimp paste. Diluting and coating fresh shrimp paste on a solid culture medium, culturing for 48h at 15 ℃, picking a colony with typical characteristics, carrying out streaking separation and purification for many times, and finally obtaining a single colony of the planococcus producing low-temperature protease. And inoculating the separated and purified single colony to a slant, culturing at 25 ℃ for 24h, and preserving for later use.
2. Identification of strains
2.1 morphological characterisation
The bacterial colony is orange yellow, has smooth surface, neat edge, non-transparency and compact bacterial colony; gram staining positive; the cells are spherical, and arranged singly or in a pile (see FIGS. 1, 3, 5 and 7).
2.2 physiological characteristics
4 planococcus versicolor can grow when the NaCl concentration is 0-15%; when the pH value is 7-9, the growth is fast; can grow well at the temperature of 15-35 ℃;
2.3 molecular biological identification
The selected and purified Planococcus were subjected to 16S rRNA sequencing, and identified as Planococcus maritimus, Planococcus plakortidis, Planococcus decalingnsis, and Planococcus riefieensis, and a biological evolutionary relationship tree was constructed (see FIGS. 2, 4, 6, and 8).
3. Preservation of the Strain
4 Planococcus strains have been deposited in the China general microbiological culture Collection center (CGMCC) of the institute of microbiology, China institute of sciences, west Lu No.1, 3, located in the south facing area, Beijing on the year 01 and 02 days in 2019, and the Planococcus maritime (XJ 2), which has been deposited in the China general microbiological culture Collection center (CGMCC) of the institute of microbiology, China institute of sciences, China school of sciences, west Lu No.1, 3, located in the south facing area, Beijing on the year 01 and 02 days in 2019, and the serial number is CGMCC NO.17057, and the suggested classification names are as follows: planococcus maritimus.
The strain is preserved in China general microbiological culture Collection center (CGMCC) of China academy of sciences microbiological research institute, China institute of microbiology 1, No. 3, located in the morning area of the south facing the Yangtze area in Beijing on the 02 th day of 2019, the number is CGMCC NO.17058, and the suggested classification name is as follows: planococcus platortiis.
The trichomonas germanica (Planococcus dechhangensis XJ11), which has been deposited in 2019 on 02.01.3 at China center for culture Collection of microorganisms (CGMCC) of institute of microbiology, China academy of sciences, institute of microbiology, No.1, North Cheng Luo, No. 3, located in the area of the rising of Beijing, with the number of CGMCC NO.17059, proposed classification name: planococcus decalensis.
Planococcus rifietonensis XJ12, which has been deposited in 2019 on 02.01.02 at the China center for culture Collection of microorganisms (CGMCC) of the institute of microbiology, China academy of sciences, institute of microbiology, No.1, North Cheng, Xilu, No. 3, located in the area of the rising of Beijing, under the number CGMCC NO.17060, proposed taxonomic nomenclature: planococcus rieitoensis is a bacterium of Planococcus rieitoensis.
Example 1:
a method for fermenting fish sauce by using Planococcus maritimus XJ2 comprises the following steps:
(1) treatment of raw materials: mincing minced fish (waste generated during fish processing) with a mincing machine, adding 5% (w/w) of pickled sea salt, and mixing.
(2) Preparing a leavening agent: planococcus marinus XJ2(CGMCC NO.17057) was activated and cultured three times. Centrifuging the activated bacteria liquid at 4 deg.C and 10000r/min for 10min, washing with sterile normal saline twice, suspending in small amount of sterile normal saline, and adjusting the bacteria liquid concentration to 105～107CFU/mL, spare.
(3) Addition of a starter: adding Planococcus maritimus XJ2(CGMCC NO.17057) into the mixture at final addition amount of 105CFU/g) is mixed into the pretreated raw material minced fish, so that the final bacteria number reaches the addition requirement.
(4) Fermentation of fish sauce: and (4) fermenting the mixed material obtained in the step (3) at the temperature of 15 ℃ for 30 d.
(5) Filtering the fish sauce: and (4) sterilizing the fish sauce sample obtained in the step (4) at 120 ℃ for 10min, cooling to room temperature, centrifuging at 10000r/min for 20min, and filtering supernate with multilayer gauze to remove solid matters and impurities.
(6) Sterilizing the fish sauce: and (5) performing secondary sterilization on the fish sauce obtained in the step (5) at 100 ℃ for 30min, filling and sealing under aseptic conditions to obtain the fish sauce finished product.
(7) And (3) evaluating the physical and chemical indexes of the product: evaluating the content of amino acid nitrogen by reference to a GB5009.235-2016 colorimetric method; evaluating the content of volatile basic nitrogen by reference to a GB5009.228-2016 microdiffusion method; the method for measuring the histamine content refers to GB5009.208-2016 spectrophotometry.
(8) The fish sauce has the following physical and chemical index results: the amino acid state content of the fish sauce fermented by the strain of the marine planococcus can reach 1.284g/100mL, and the fish sauce is divided into first-grade fish sauce according to the Chinese fish sauce industry standard; the content of volatile basic nitrogen is 107mg/100 mL; the histamine content was 19mg/100mL, which is lower than EU standard 40mg/100 mL. The inherent earthy smell in the raw materials of the fish sauce is greatly reduced, the delicate flavor is obvious, and the flavor is obviously improved compared with the fish sauce which is not fermented by adding bacterial strains.
Example 2:
a method for fermenting fish sauce by using Planococcus maritimus XJ2 comprises the following steps:
(1) treatment of raw materials: mincing minced fish (waste generated during fish processing) with a mincing machine, adding 10% (w/w) of pickled sea salt, and mixing.
(2) Preparing a leavening agent: planococcus marinus XJ2(CGMCC NO.17057) was activated and cultured three times. Centrifuging the activated bacteria liquid at 4 deg.C and 10000r/min for 10min, washing with sterile normal saline twice, suspending in small amount of sterile normal saline, and adjusting the bacteria liquid concentration to 105～107CFU/mL, spare.
(3) Addition of a starter: adding Planococcus maritimus XJ2(CGMCC NO.17057) into the mixture at final addition amount of 107CFU/g) is mixed into the pretreated raw material minced fish, so that the final bacteria number reaches the addition requirement.
(4) Fermentation of fish sauce: and (4) fermenting the mixed material obtained in the step (3) at the temperature of 20 ℃ for 15 days in a heat preservation way.
(5) Filtering the fish sauce: and (4) sterilizing the fish sauce sample obtained in the step (4) at 120 ℃ for 20min, cooling to room temperature, centrifuging at 10000r/min for 20min, and filtering supernate with multilayer gauze to remove solid matters and impurities.
(6) Sterilizing the fish sauce: and (5) performing secondary sterilization on the fish sauce obtained in the step (5) at 100 ℃ for 20min, filling and sealing under aseptic conditions to obtain the finished product of the fish sauce.
(7) And (3) evaluating the physical and chemical indexes of the product: evaluating the content of amino acid nitrogen by reference to a GB5009.235-2016 colorimetric method; evaluating the content of volatile basic nitrogen by reference to a GB5009.228-2016 microdiffusion method; the method for measuring the histamine content refers to GB5009.208-2016 spectrophotometry.
(8) The fish sauce has the following physical and chemical index results: the amino acid state content of the fish sauce fermented by the strain of the marine planococcus can reach 1.023g/100mL, and the fish sauce is divided into first-grade fish sauce according to the Chinese fish sauce industry standard; the content of volatile basic nitrogen is 94mg/100 mL; the histamine content was 17mg/100mL, which is lower than EU standard 40mg/100 mL. The inherent earthy smell in the raw materials of the fish sauce is greatly reduced, the delicate flavor is obvious, and the flavor is obviously improved compared with the fish sauce which is not fermented by adding bacterial strains.
Example 3:
a method for fermenting fish sauce by using Planococcus maritimus XJ2 comprises the following steps:
(1) treatment of raw materials: mincing minced fish (waste generated during fish processing) with a mincing machine, adding 15% (w/w) of pickled sea salt, and mixing.
(2) Preparing a leavening agent: planococcus marinus XJ2(CGMCC NO.17057) was activated and cultured three times. Centrifuging the activated bacteria liquid at 4 deg.C and 10000r/min for 10min, washing with sterile normal saline twice, suspending in small amount of sterile normal saline, and adjusting the bacteria liquid concentration to 105～107CFU/mL, spare.
(3) Addition of a starter: adding Planococcus maritimus XJ2(CGMCC NO.17057) into the mixture at final addition amount of 109CFU/g) is mixed into the pretreated raw material minced fish, so that the final bacteria number reaches the addition requirement.
(4) Fermentation of fish sauce: and (4) fermenting the mixed material obtained in the step (3) at the temperature of 30 ℃ for 5 d.
(5) Filtering the fish sauce: and (4) sterilizing the fish sauce sample obtained in the step (4) at 120 ℃ for 30min, cooling to room temperature, centrifuging at 10000r/min for 20min, and filtering supernate with multilayer gauze to remove solid matters and impurities.
(6) Sterilizing the fish sauce: and (5) carrying out secondary sterilization on the fish sauce obtained in the step (5) at 100 ℃ for 15min, filling and sealing under aseptic conditions to obtain the finished product of the fish sauce.
(7) And (3) evaluating the physical and chemical indexes of the product: evaluating the content of amino acid nitrogen by reference to a GB5009.235-2016 colorimetric method; evaluating the content of volatile basic nitrogen by reference to a GB5009.228-2016 microdiffusion method; the method for measuring the histamine content refers to GB5009.208-2016 spectrophotometry.
(8) The fish sauce has the following physical and chemical index results: the amino acid state content of the fish sauce fermented by the strain of the marine planococcus can reach 0.97g/100mL, and the fish sauce is divided into first-grade fish sauce according to the Chinese fish sauce industry standard; the content of volatile basic nitrogen is 85mg/100 mL; the histamine content was 11mg/100mL, which is lower than EU standard 40mg/100 mL. The inherent earthy smell in the raw materials of the fish sauce is greatly reduced, the delicate flavor is obvious, and the flavor is obviously improved compared with the fish sauce which is not fermented by adding bacterial strains.
Example 4:
a method for fermenting fish sauce by using Micrococcus platensis XJ10 comprises the following steps:
(1) treatment of raw materials: mincing minced fish (waste generated during fish processing) with a mincing machine, adding 5% (w/w) of pickled sea salt, and mixing.
(2) Preparing a leavening agent: activation culture was carried out three times on the P.pulcherrimus platortis XJ10(CGMCC NO. 17058). Centrifuging the activated bacteria liquid at 4 deg.C and 10000r/min for 10min, washing with sterile normal saline twice, suspending in small amount of sterile normal saline, and adjusting the bacteria liquid concentration to 105～107CFU/mL, spare.
(3) Addition of a starter: the Pneumococcus pulcherrimus platortidis XJ10(CGMCC NO.17058) is added according to the final addition amount of 105CFU/g) is added to the pretreated ground fish meat to increase the final bacterial countAdding the requirements.
(4) Fermentation of fish sauce: and (4) fermenting the mixed material obtained in the step (3) at the temperature of 15 ℃ for 30 d.
(5) Filtering the fish sauce: and (4) sterilizing the fish sauce sample obtained in the step (4) at 120 ℃ for 10min, cooling to room temperature, centrifuging at 10000r/min for 20min, and filtering supernate with multilayer gauze to remove solid matters and impurities.
(6) Sterilizing the fish sauce: and (5) performing secondary sterilization on the fish sauce obtained in the step (5) at 100 ℃ for 30min, filling and sealing under aseptic conditions to obtain the fish sauce finished product.
(7) And (3) evaluating the physical and chemical indexes of the product: evaluating the content of amino acid nitrogen by reference to a GB5009.235-2016 colorimetric method; evaluating the content of volatile basic nitrogen by reference to a GB5009.228-2016 microdiffusion method; the method for measuring the histamine content refers to GB5009.208-2016 spectrophotometry.
(8) The fish sauce has the following physical and chemical index results: the amino acid state content of the fish sauce fermented by the strain of the streptococci pulchella can reach 1.117g/100mL, and the fish sauce is divided into first-grade fish sauce according to the Chinese fish sauce industry standard; the content of volatile basic nitrogen is 112mg/100 mL; the histamine content was 20mg/100mL, which is lower than EU standard 40mg/100 mL. The inherent earthy smell in the raw materials of the fish sauce is greatly reduced, the delicate flavor is obvious, and the flavor is obviously improved compared with the fish sauce which is not fermented by adding bacterial strains.
Example 5:
a method for fermenting fish sauce by using Micrococcus platensis XJ10 comprises the following steps:
(1) treatment of raw materials: mincing minced fish (waste generated during fish processing) with a mincing machine, adding 10% (w/w) of pickled sea salt, and mixing.
(2) Preparing a leavening agent: activation culture was carried out three times on the P.pulcherrimus platortis XJ10(CGMCC NO. 17058). Centrifuging the activated bacteria liquid at 4 deg.C and 10000r/min for 10min, washing with sterile normal saline twice, suspending in small amount of sterile normal saline, and adjusting the bacteria liquid concentration to 105～107CFU/mL, spare.
(3) Hair-like deviceAdding a fermentation agent: the Pneumococcus pulcherrimus platortidis XJ10(CGMCC NO.17058) is added according to the final addition amount of 107CFU/g) is mixed into the pretreated raw material minced fish, so that the final bacteria number reaches the addition requirement.
(4) Fermentation of fish sauce: and (4) fermenting the mixed material obtained in the step (3) at the temperature of 20 ℃ for 15 days in a heat preservation way.
(5) Filtering the fish sauce: and (4) sterilizing the fish sauce sample obtained in the step (4) at 120 ℃ for 20min, cooling to room temperature, centrifuging at 10000r/min for 20min, and filtering supernate with multilayer gauze to remove solid matters and impurities.
(6) Sterilizing the fish sauce: and (5) performing secondary sterilization on the fish sauce obtained in the step (5) at 100 ℃ for 20min, filling and sealing under aseptic conditions to obtain the finished product of the fish sauce.
(7) And (3) evaluating the physical and chemical indexes of the product: evaluating the content of amino acid nitrogen by reference to a GB5009.235-2016 colorimetric method; evaluating the content of volatile basic nitrogen by reference to a GB5009.228-2016 microdiffusion method; the method for measuring the histamine content refers to GB5009.208-2016 spectrophotometry.
(8) The fish sauce has the following physical and chemical index results: the amino acid state content of the fish sauce fermented by the strain of the pulenococcus pulveratus can reach 1.103g/100mL, and the fish sauce is divided into first-grade fish sauce according to the Chinese fish sauce industry standard; the content of volatile basic nitrogen is 99mg/100 mL; the histamine content was 18mg/100mL, which is lower than EU standard 40mg/100 mL. The inherent earthy smell in the raw materials of the fish sauce is greatly reduced, the delicate flavor is obvious, and the flavor is obviously improved compared with the fish sauce which is not fermented by adding bacterial strains.
Example 6:
a method for fermenting fish sauce by using Micrococcus platensis XJ10 comprises the following steps:
(1) treatment of raw materials: mincing minced fish (waste generated during fish processing) with a mincing machine, adding 15% (w/w) of pickled sea salt, and mixing.
(2) Preparing a leavening agent: activation culture was carried out three times on the P.pulcherrimus platortis XJ10(CGMCC NO. 17058). Centrifuging the activated bacteria liquid at 4 deg.C and 10000r/min for 10min, and sterilizingWashing twice with normal saline, suspending in small amount of sterile normal saline, and adjusting the concentration of bacterial liquid to 105～107CFU/mL, spare.
(3) Addition of a starter: the Pneumococcus pulcherrimus platortidis XJ10(CGMCC NO.17058) is added according to the final addition amount of 109CFU/g) is mixed into the pretreated raw material minced fish, so that the final bacteria number reaches the addition requirement.
(4) Fermentation of fish sauce: and (4) fermenting the mixed material obtained in the step (3) at the temperature of 30 ℃ for 5 d.
(5) Filtering the fish sauce: and (4) sterilizing the fish sauce sample obtained in the step (4) at 120 ℃ for 30min, cooling to room temperature, centrifuging at 10000r/min for 20min, and filtering supernate with multilayer gauze to remove solid matters and impurities.
(6) Sterilizing the fish sauce: and (5) carrying out secondary sterilization on the fish sauce obtained in the step (5) at 100 ℃ for 15min, filling and sealing under aseptic conditions to obtain the finished product of the fish sauce.
(7) And (3) evaluating the physical and chemical indexes of the product: evaluating the content of amino acid nitrogen by reference to a GB5009.235-2016 colorimetric method; evaluating the content of volatile basic nitrogen by reference to a GB5009.228-2016 microdiffusion method; the method for measuring the histamine content refers to GB5009.208-2016 spectrophotometry.
(8) The fish sauce has the following physical and chemical index results: the amino acid state content of the fish sauce fermented by the strain of the pulenococcus pulveratus can reach 1.008g/100mL, and the fish sauce is divided into first-grade fish sauce according to the Chinese fish sauce industry standard; the content of volatile basic nitrogen is 92mg/100 mL; the histamine content was 17mg/100mL, which is lower than EU standard 40mg/100 mL. The inherent earthy smell in the raw materials of the fish sauce is greatly reduced, the delicate flavor is obvious, and the flavor is obviously improved compared with the fish sauce which is not fermented by adding bacterial strains.
Example 7:
a method for fermenting fish sauce by using Planococcus decalensis XJ11 of Dryobalanops delavayi comprises the following steps:
(1) treatment of raw materials: mincing minced fish (waste generated during fish processing) with a mincing machine, adding 5% (w/w) of pickled sea salt, and mixing.
(2) Preparing a leavening agent: activation culture was carried out three times on Planococcus decalensis XJ11(CGMCC NO. 17059). Centrifuging the activated bacteria liquid at 4 deg.C and 10000r/min for 10min, washing with sterile normal saline twice, suspending in small amount of sterile normal saline, and adjusting the bacteria liquid concentration to 105～107CFU/mL, spare.
(3) Addition of a starter: the Planococcus dechhangensis XJ11(CGMCC NO.17059) is added according to the final adding amount of 105CFU/g) is mixed into the pretreated raw material minced fish, so that the final bacteria number reaches the addition requirement.
(4) Fermentation of fish sauce: and (4) fermenting the mixed material obtained in the step (3) at the temperature of 15 ℃ for 30 d.
(5) Filtering the fish sauce: and (4) sterilizing the fish sauce sample obtained in the step (4) at 120 ℃ for 10min, cooling to room temperature, centrifuging at 10000r/min for 20min, and filtering supernate with multilayer gauze to remove solid matters and impurities.
(6) Sterilizing the fish sauce: and (5) performing secondary sterilization on the fish sauce obtained in the step (5) at 100 ℃ for 30min, filling and sealing under aseptic conditions to obtain the fish sauce finished product.
(7) And (3) evaluating the physical and chemical indexes of the product: evaluating the content of amino acid nitrogen by reference to a GB5009.235-2016 colorimetric method; evaluating the content of volatile basic nitrogen by reference to a GB5009.228-2016 microdiffusion method; the method for measuring the histamine content refers to GB5009.208-2016 spectrophotometry.
(8) The fish sauce has the following physical and chemical index results: the amino acid state content of the fish sauce fermented by the strain of the Trichosporon dachang can reach 1.216g/100mL, and the fish sauce is divided into first-grade fish sauce according to the Chinese fish sauce industry standard; the content of volatile basic nitrogen is 109mg/100 mL; the histamine content was 19mg/100mL, which is lower than EU standard 40mg/100 mL. The inherent earthy smell in the raw materials of the fish sauce is greatly reduced, the delicate flavor is obvious, and the flavor is obviously improved compared with the fish sauce which is not fermented by adding bacterial strains.
Example 8:
a method for fermenting fish sauce by using Planococcus decalensis XJ11 of Dryobalanops delavayi comprises the following steps:
(1) treatment of raw materials: mincing minced fish (waste generated during fish processing) with a mincing machine, adding 10% (w/w) of pickled sea salt, and mixing.
(2) Preparing a leavening agent: activation culture was carried out three times on Planococcus decalensis XJ11(CGMCC NO. 17059). Centrifuging the activated bacteria liquid at 4 deg.C and 10000r/min for 10min, washing with sterile normal saline twice, suspending in small amount of sterile normal saline, and adjusting the bacteria liquid concentration to 105～107CFU/mL, spare.
(3) Addition of a starter: the Planococcus dechhangensis XJ11(CGMCC NO.17059) is added according to the final adding amount of 107CFU/g) is mixed into the pretreated raw material minced fish, so that the final bacteria number reaches the addition requirement.
(4) Fermentation of fish sauce: and (4) fermenting the mixed material obtained in the step (3) at the temperature of 20 ℃ for 15 days in a heat preservation way.
(5) Filtering the fish sauce: and (4) sterilizing the fish sauce sample obtained in the step (4) at 120 ℃ for 20min, cooling to room temperature, centrifuging at 10000r/min for 20min, and filtering supernate with multilayer gauze to remove solid matters and impurities.
(6) Sterilizing the fish sauce: and (5) performing secondary sterilization on the fish sauce obtained in the step (5) at 100 ℃ for 20min, filling and sealing under aseptic conditions to obtain the finished product of the fish sauce.
(7) And (3) evaluating the physical and chemical indexes of the product: evaluating the content of amino acid nitrogen by reference to a GB5009.235-2016 colorimetric method; evaluating the content of volatile basic nitrogen by reference to a GB5009.228-2016 microdiffusion method; the method for measuring the histamine content refers to GB5009.208-2016 spectrophotometry.
(8) The fish sauce has the following physical and chemical index results: the amino acid state content of the fish sauce fermented by the strain of the Trichosporon dachang can reach 1.174g/100mL, and the fish sauce is divided into first-grade fish sauce according to the Chinese fish sauce industry standard; the content of volatile basic nitrogen is 98mg/100 mL; the histamine content was 20mg/100mL, which is lower than EU standard 40mg/100 mL. The inherent earthy smell in the raw materials of the fish sauce is greatly reduced, the delicate flavor is obvious, and the flavor is obviously improved compared with the fish sauce which is not fermented by adding bacterial strains.
Example 9:
a method for fermenting fish sauce by using Planococcus decalensis XJ11 of Dryobalanops delavayi comprises the following steps:
(1) treatment of raw materials: mincing minced fish (waste generated during fish processing) with a mincing machine, adding 15% (w/w) of pickled sea salt, and mixing.
(2) Preparing a leavening agent: activation culture was carried out three times on Planococcus decalensis XJ11(CGMCC NO. 17059). Centrifuging the activated bacteria liquid at 4 deg.C and 10000r/min for 10min, washing with sterile normal saline twice, suspending in small amount of sterile normal saline, and adjusting the bacteria liquid concentration to 105～107CFU/mL, spare.
(3) Addition of a starter: the Planococcus dechhangensis XJ11(CGMCC NO.17059) is added according to the final adding amount of 109CFU/g) is mixed into the pretreated raw material minced fish, so that the final bacteria number reaches the addition requirement.
(4) Fermentation of fish sauce: and (4) fermenting the mixed material obtained in the step (3) at the temperature of 30 ℃ for 5 d.
(5) Filtering the fish sauce: and (4) sterilizing the fish sauce sample obtained in the step (4) at 120 ℃ for 30min, cooling to room temperature, centrifuging at 10000r/min for 20min, and filtering supernate with multilayer gauze to remove solid matters and impurities.
(6) Sterilizing the fish sauce: and (5) carrying out secondary sterilization on the fish sauce obtained in the step (5) at 100 ℃ for 15min, filling and sealing under aseptic conditions to obtain the finished product of the fish sauce.
(7) And (3) evaluating the physical and chemical indexes of the product: evaluating the content of amino acid nitrogen by reference to a GB5009.235-2016 colorimetric method; evaluating the content of volatile basic nitrogen by reference to a GB5009.228-2016 microdiffusion method; the method for measuring the histamine content refers to GB5009.208-2016 spectrophotometry.
(8) The fish sauce has the following physical and chemical index results: the amino acid state content of the fish sauce fermented by the strain of the Trichosporon dachang can reach 1.089g/100mL, and the fish sauce is divided into first-level fish sauce according to the Chinese fish sauce industry standard; the content of volatile basic nitrogen is 92mg/100 mL; the histamine content was 14mg/100mL, which is lower than EU standard 40mg/100 mL. The inherent earthy smell in the raw materials of the fish sauce is greatly reduced, the delicate flavor is obvious, and the flavor is obviously improved compared with the fish sauce which is not fermented by adding bacterial strains.
Example 10:
a method for fermenting fish sauce with Planococcus riefieensis XJ12, which comprises the following steps:
(1) treatment of raw materials: mincing minced fish (waste generated during fish processing) with a mincing machine, adding 5% (w/w) of pickled sea salt, and mixing.
(2) Preparing a leavening agent: activate culture of Planococcus riefietonensis XJ12(CGMCC NO.17060) three times. Centrifuging the activated bacteria liquid at 4 deg.C and 10000r/min for 10min, washing with sterile normal saline twice, suspending in small amount of sterile normal saline, and adjusting the bacteria liquid concentration to 105～107CFU/mL, spare.
(3) Addition of a starter: adding Planococcus riefieiensis XJ12(CGMCC NO.17060) into the mixture at a final addition amount of 105CFU/g) is mixed into the pretreated raw material minced fish, so that the final bacteria number reaches the addition requirement.
(4) Fermentation of fish sauce: and (4) fermenting the mixed material obtained in the step (3) at the temperature of 15 ℃ for 30 d.
(5) Filtering the fish sauce: and (4) sterilizing the fish sauce sample obtained in the step (4) at 120 ℃ for 10min, cooling to room temperature, centrifuging at 10000r/min for 20min, and filtering supernate with multilayer gauze to remove solid matters and impurities.
(6) Sterilizing the fish sauce: and (5) performing secondary sterilization on the fish sauce obtained in the step (5) at 100 ℃ for 30min, filling and sealing under aseptic conditions to obtain the fish sauce finished product.
(7) And (3) evaluating the physical and chemical indexes of the product: evaluating the content of amino acid nitrogen by reference to a GB5009.235-2016 colorimetric method; evaluating the content of volatile basic nitrogen by reference to a GB5009.228-2016 microdiffusion method; the method for measuring the histamine content refers to GB5009.208-2016 spectrophotometry.
(8) The fish sauce has the following physical and chemical index results: the amino acid state content of the fish sauce fermented by the deinococcus lepigone can reach 1.159g/100mL, and the fish sauce is divided into first-grade fish sauce according to the Chinese fish sauce industry standard; the content of volatile basic nitrogen is 104mg/100 mL; the histamine content was 16mg/100mL, which is lower than EU standard 40mg/100 mL. The inherent earthy smell in the raw materials of the fish sauce is greatly reduced, the delicate flavor is obvious, and the flavor is obviously improved compared with the fish sauce which is not fermented by adding bacterial strains.
Example 11:
a method for fermenting fish sauce with Planococcus riefieensis XJ12, which comprises the following steps:
(1) treatment of raw materials: mincing minced fish (waste generated during fish processing) with a mincing machine, adding 10% (w/w) of pickled sea salt, and mixing.
(2) Preparing a leavening agent: activate culture of Planococcus riefietonensis XJ12(CGMCC NO.17060) three times. Centrifuging the activated bacteria liquid at 4 deg.C and 10000r/min for 10min, washing with sterile normal saline twice, suspending in small amount of sterile normal saline, and adjusting the bacteria liquid concentration to 105～107CFU/mL, spare.
(3) Addition of a starter: adding Planococcus riefieiensis XJ12(CGMCC NO.17060) into the mixture at a final addition amount of 107CFU/g) is mixed into the pretreated raw material minced fish, so that the final bacteria number reaches the addition requirement.
(4) Fermentation of fish sauce: and (4) fermenting the mixed material obtained in the step (3) at the temperature of 20 ℃ for 15 days in a heat preservation way.
(5) Filtering the fish sauce: and (4) sterilizing the fish sauce sample obtained in the step (4) at 120 ℃ for 20min, cooling to room temperature, centrifuging at 10000r/min for 20min, and filtering supernate with multilayer gauze to remove solid matters and impurities.
(6) Sterilizing the fish sauce: and (5) performing secondary sterilization on the fish sauce obtained in the step (5) at 100 ℃ for 20min, filling and sealing under aseptic conditions to obtain the finished product of the fish sauce.
(7) And (3) evaluating the physical and chemical indexes of the product: evaluating the content of amino acid nitrogen by reference to a GB5009.235-2016 colorimetric method; evaluating the content of volatile basic nitrogen by reference to a GB5009.228-2016 microdiffusion method; the method for measuring the histamine content refers to GB5009.208-2016 spectrophotometry.
(8) The fish sauce has the following physical and chemical index results: the amino acid state content of the fish sauce fermented by the deinococcus lepigone can reach 1.207g/100mL, and the fish sauce is divided into first-grade fish sauce according to the Chinese fish sauce industry standard; the content of volatile basic nitrogen is 109mg/100 mL; the histamine content was 21mg/100mL, which is lower than EU standard 40mg/100 mL. The inherent earthy smell in the raw materials of the fish sauce is greatly reduced, the delicate flavor is obvious, and the flavor is obviously improved compared with the fish sauce which is not fermented by adding bacterial strains.
Example 12:
a method for fermenting fish sauce with Planococcus riefieensis XJ12, which comprises the following steps:
(1) treatment of raw materials: mincing minced fish (waste generated during fish processing) with a mincing machine, adding 15% (w/w) of pickled sea salt, and mixing.
(2) Preparing a leavening agent: activate culture of Planococcus riefietonensis XJ12(CGMCC NO.17060) three times. Centrifuging the activated bacteria liquid at 4 deg.C and 10000r/min for 10min, washing with sterile normal saline twice, suspending in small amount of sterile normal saline, and adjusting the bacteria liquid concentration to 105～107CFU/mL, spare.
(3) Addition of a starter: adding Planococcus riefieiensis XJ12(CGMCC NO.17060) into the mixture at a final addition amount of 109CFU/g) is mixed into the pretreated raw material minced fish, so that the final bacteria number reaches the addition requirement.
(4) Fermentation of fish sauce: and (4) fermenting the mixed material obtained in the step (3) at the temperature of 30 ℃ for 5 d.
(5) Filtering the fish sauce: and (4) sterilizing the fish sauce sample obtained in the step (4) at 120 ℃ for 30min, cooling to room temperature, centrifuging at 10000r/min for 20min, and filtering supernate with multilayer gauze to remove solid matters and impurities.
(6) Sterilizing the fish sauce: and (5) carrying out secondary sterilization on the fish sauce obtained in the step (5) at 100 ℃ for 15min, filling and sealing under aseptic conditions to obtain the finished product of the fish sauce.
(7) And (3) evaluating the physical and chemical indexes of the product: evaluating the content of amino acid nitrogen by reference to a GB5009.235-2016 colorimetric method; evaluating the content of volatile basic nitrogen by reference to a GB5009.228-2016 microdiffusion method; the method for measuring the histamine content refers to GB5009.208-2016 spectrophotometry.
(8) The fish sauce has the following physical and chemical index results: the amino acid state content of the fish sauce fermented by the deinococcus lepigone can reach 1.012g/100mL, and the fish sauce is divided into first-grade fish sauce according to the Chinese fish sauce industry standard; the content of volatile basic nitrogen is 98mg/100 mL; the histamine content was 14mg/100mL, which is lower than EU standard 40mg/100 mL. The inherent earthy smell in the raw materials of the fish sauce is greatly reduced, the delicate flavor is obvious, and the flavor is obviously improved compared with the fish sauce which is not fermented by adding bacterial strains.

Claims (2)

1. Planococcus marinus (Planococcus maritimus) XJ2 with the preservation number of CGMCC NO.17057, classified name: planococcus maritimus.
2. The method for fermenting fish gravy by using planococcus marinus as claimed in claim 1, which is carried out according to the following steps:
(1) treatment of raw materials: mincing minced fish meat by using a meat mincer, adding 5-15% (w/w) of pickled sea salt, and uniformly mixing for later use;
(2) preparing a leavening agent: activating and culturing the planococcus strain for three times; centrifuging the activated bacteria liquid at 4 deg.C and 10000r/min for 10min, washing with sterile normal saline twice, suspending in small amount of sterile normal saline, and adjusting the bacteria liquid concentration to 105～107CFU/mL for standby;
(3) addition of a starter: adding the strain at a final addition amount of 105～109Mixing CFU/g into the pretreated minced fish meat to make the final bacteria number meet the addition requirement;
(4) fermentation of fish sauce: fermenting the mixed material obtained in the step (3) at the temperature of 15-30 ℃ for 5-30 d in a heat preservation manner;
(5) filtering the fish sauce: sterilizing the fish sauce sample obtained in the step (4) at 120 ℃ for 10-30 min, cooling to room temperature, centrifuging at 10000r/min for 20min, and filtering supernate with multiple layers of gauze to remove solid matters and impurities;
(6) sterilizing the fish sauce: and (5) carrying out secondary sterilization on the fish sauce obtained in the step (5) at 100 ℃ for 15-30 min, and filling and sealing under aseptic conditions to obtain the fish sauce finished product.
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