CN114045227B - Pediococcus praecox starter for improving fermentation quality of low-salt fish sauce - Google Patents

Pediococcus praecox starter for improving fermentation quality of low-salt fish sauce Download PDF

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CN114045227B
CN114045227B CN202110951662.7A CN202110951662A CN114045227B CN 114045227 B CN114045227 B CN 114045227B CN 202110951662 A CN202110951662 A CN 202110951662A CN 114045227 B CN114045227 B CN 114045227B
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fish sauce
fish
planococcus
sauce
fermenting
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CN114045227A (en
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高瑞昌
周晶
袁丽
周越
王柱梅
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Shenzhen Wanzhida Technology Co ltd
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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

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  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Food Science & Technology (AREA)
  • Health & Medical Sciences (AREA)
  • Nutrition Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Polymers & Plastics (AREA)
  • Marine Sciences & Fisheries (AREA)
  • Zoology (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Soy Sauces And Products Related Thereto (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Abstract

The invention discloses a fermentation agent for improving fermentation quality of low-salt fish gravy, and relates to the field of application of food microorganism technology. The enterococcus prasuavis is preserved in China general microbiological culture Collection center (CGMCC), the preservation number is CGMCC NO.17058, and the proposed classification and naming are: planococcus plakortidis. The production period of fermenting fish sauce by utilizing the enterococcus prasugrel (Planococcus plakortidis) XJ10 is greatly shortened, and the fermentation temperature is reduced. The fish sauce obtained by the method is transparent and brownish red, has special fragrance of aquatic products, and has no suspended or flocculent impurities.

Description

Pediococcus praecox starter for improving fermentation quality of low-salt fish sauce
The application is a divisional application of Chinese invention patent, the application date of the original application is 2020, 01 and 13 days, the application number is 202010025134.4, and the invention is named as 'a enterococcus faecalis starter for improving the fermentation quality of low-salt fish sauce'.
Technical Field
The invention relates to the field of application of food microorganism technology, in particular to a rhodococcus mobilis starter for improving the fermentation quality of low-salt fish gravy.
Background
China is a large fishery country, and freshwater aquaculture has become an important industrial prop for fishery in China. In 2017, the total yield of the freshwater aquaculture in China is about 2905 ten thousand tons, but the processing utilization rate is less than 15 percent, so that the problems of resource waste, environmental pollution and the like are caused. Therefore, how to solve the comprehensive utilization problem of freshwater fish is a great challenge facing the aquatic product processing industry in China. Fish sauce is one of the most popular condiments in southeast Asia, and has clear brownish red color, unique flavor and fragrance, and has become part of people's daily diet. The traditional fish sauce production is usually carried out by taking low-value fish and shrimp or aquatic product processing waste (fish heads, viscera and the like) as raw materials and naturally fermenting by utilizing enzymes or microorganisms of the raw materials. The fish sauce is nutritious, contains multiple essential amino acids, and mineral elements such as calcium, magnesium, zinc, iron and the like beneficial to human body, and is a low-fat high-protein fermented condiment. Meanwhile, the fish gravy is also a product of high-valued utilization of low-valued fish, and has important practical significance for promoting the healthy development of the freshwater fish industry. In the fish sauce fermentation process, protease is mainly used for catalyzing protein hydrolysis, and the influence of temperature on the activity and stability of the protease can be divided into low-temperature protease, medium-temperature protease and high-temperature 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 generally 20-40 ℃. The low-temperature protease can react at low temperature or room temperature without heating and cooling, and the cost can be reduced. Therefore, the method has the advantage that the mesophilic protease cannot be replaced in industrial production, and has wide application prospects in the fields of washing industry, food processing, biopharmaceuticals, environmental bioremediation and the like. The genus Planococcus has been identified as a cryoprotectant producing low temperature proteases and has high safety. The invention ferments fish sauce under the condition of low temperature and low salt by utilizing the enterococcus, thereby obtaining fish sauce products with short fermentation period, low salinity and higher flavor and nutrition value, and providing theoretical basis and method for the enterococcus in the food microorganism fermentation application.
Disclosure of Invention
According to the invention, 4 new strains capable of improving the fermentation quality of the low-salt fish sauce are screened from the traditional shrimp sauce, and the fish sauce is produced by combining the growth characteristics and the enzymatic characteristics of the 4 strains. The 4 strains can effectively shorten the fermentation period, reduce the salt content and improve the flavor and the nutritional value of the fish gravy, and provide a more stable, safer and more suitable starter for fish gravy fermentation.
The technical scheme adopted by the invention is as follows:
the marine enterococcus (Planococcus maritimus XJ 2) provided by the invention is a novel strain which can be applied to improving the fermentation quality of fish gravy. The strain has been preserved in 2019, 01 and 02 days, and is named as CGMCC No.17057, suggested classification and naming by China general microbiological culture Collection center (CGMCC) of China national institute of culture Collection of microorganisms, national institute of sciences, no.1 and 3, beijing, kogyo, and the same. Planococcus maritimus.
The enterococcus prasugrel (Planococcus plakortidis XJ) provided by the invention is a novel strain which can be applied to improving the fermentation quality of fish gravy. The strain has been preserved in 2019, 01 and 02 days, and is named as CGMCC No.17058, suggested classification and naming by China general microbiological culture Collection center (CGMCC) of China national institute of culture Collection of microorganisms, national institute of sciences, no.1 and 3, beijing, kogyo, and the same. Planococcus plakortidis.
The Dechang active coccus (Planococcus dechangensis XJ) provided by the invention is a novel strain which can be applied to improving the fermentation quality of fish gravy. The strain has been preserved in 2019, 01 and 02 days, and is named as CGMCC No.17059, suggested classification and naming by China general microbiological culture Collection center (CGMCC) of China national institute of culture Collection of microorganisms, national institute of sciences, no.1 and 3, beijing, kogyo, and the same. Planococcus dechangensis.
The Laibotozoococcus (Planococcus rifietoensis XJ) provided by the invention is a novel strain which can be applied to improving the fermentation quality of fish gravy. The strain has been preserved in 2019, 01 and 02 days, and is named as CGMCC No.17060, suggested classification and naming by China general microbiological culture Collection center (CGMCC) of China national institute of culture Collection of microorganisms, national institute of sciences, no.1 and 3, beijing, kogyo, and the same. Planococcus rifietoensis.
In one aspect of the invention there is provided the use of a marine enterococcus (Planococcus maritimus XJ), a Praeparata (Planococcus plakortidis XJ), a Desmococcus changii (Planococcus dechangensis XJ 11) and a Leibotuberculosis (Planococcus rifietoensis XJ) as described above for improving the fermentation flavour and quality of fish maws.
A method for fermenting fish sauce by utilizing enterococcus faecalis comprises the following steps:
(1) And (3) raw material treatment: mincing minced fish meat (waste generated during fish processing) with a meat mincing machine, adding 5-15% (w/w) of salted sea salt, and mixing well for standby.
(2) Preparation of the starter: the active coccus strain is activated and cultured for three times. Centrifuging activated bacteria solution at 4deg.C and 10000r/min for 10min, washing with sterile physiological saline twice, suspending in small amount of sterile physiological saline, and regulating bacteria solution concentration to 10 5 ～10 7 CFU/mL, ready for use.
(3) Addition of a starter: the strain was added in the final amount (10 5 ～10 9 CFU/g) is mixed into the pretreated raw material minced fish meat to enable the final bacterial count to reach the addition requirement.
(4) Fermenting fish sauce: and (3) preserving heat and fermenting the mixed material obtained in the step (3) for 5-30 d at the temperature of 15-30 ℃.
(5) Filtering 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 the supernatant with multi-layer gauze to remove solids and impurities.
(6) Sterilization of fish sauce: and (3) carrying out secondary sterilization on the fish sauce obtained in the step (5) at 100 ℃ for 15-30 min, and filling and sealing under the aseptic condition to obtain a fish sauce finished product.
(7) Evaluation of physical and chemical indexes of products: amino acid nitrogen content evaluation, wherein the method refers to GB5009.235-2016 colorimetric method; the method refers to GB5009.228-2016 micro-diffusion method for evaluating the content of volatile basic nitrogen; the measurement of histamine content was carried out by the method described in GB5009.208-2016 spectrophotometry.
Wherein the enterococcus is marine enterococcus (Planococcus maritimus XJ) or Prinsepia (Planococcus plakortidis XJ) or Dechang (Planococcus dechangensis XJ 11) or Leibotuber's zoococcus (Planococcus rifietoensis XJ).
The invention has the beneficial effects that:
the fermentation period of the fish sauce is greatly shortened by respectively utilizing the 4 strains of the active cocci, and the fermentation temperature is reduced. The fish sauce obtained by the method is transparent and brownish red, has special fragrance of aquatic products, and has no suspended or flocculent impurities. The finished fish sauce has low salt content, high amino acid nitrogen content (the fish sauce is divided into first-class fish sauce according to the Chinese fish sauce industry standard), low volatile basic nitrogen content and low histamine content, and the prepared fish sauce has delicious taste and rich nutrition value, so that the 4-strain active coccus can be used for producing green and safe fermented fish sauce products.
In summary, the invention has the unique, outstanding and substantial features and significant advances in innovations and utilities of the invention patent, producing beneficial results.
Drawings
FIG. 1 shows colony morphology and cell morphology of Pediococcus celeriacus Planococcus maritimus XJ 2;
FIG. 2 is a phylogenetic tree constructed from the 16S rRNA sequence of P.mobilis Planococcus maritimus XJ 2.
FIG. 3 shows colony morphology and cell morphology of Pediococcus celeriacus Planococcus plakortidis XJ;
FIG. 4 is a phylogenetic tree constructed from the 16S rRNA sequence of P.mobilis Planococcus plakortidis XJ.
FIG. 5 shows colony morphology and cell morphology of Pediococcus celeriacus Planococcus dechangensis XJ 11;
FIG. 6 is a phylogenetic tree constructed from the 16S rRNA sequence of P.mobilis Planococcus dechangensis XJ 11.
FIG. 7 shows colony morphology and cell morphology of Pediococcus celeriacus Planococcus rifietoensis XJ;
FIG. 8 is a phylogenetic tree constructed from the 16S rRNA sequence of P.mobilis Planococcus rifietoensis XJ;
(note: xj-2 is the enterococcus Planococcus maritimus XJ of the present invention, xj-10 is the enterococcus Planococcus plakortidis XJ of the present invention, xj-11 is the enterococcus Planococcus dechangensis XJ of the present invention, xj-12 is the enterococcus Planococcus rifietoensis XJ of the present invention.).
Detailed Description
1. Screening and purifying strains:
taking traditional fresh shrimp paste (shrimp paste from farmhouse restaurant in WiHai of Shandong province) fermented at low temperature as raw material, separating and screening out enterococcus faecalis producing low-temperature protease from shrimp paste. And (3) diluting and coating fresh shrimp paste on a solid culture medium, culturing for 48 hours at 15 ℃, picking out bacterial colonies with typical characteristics, carrying out repeated streak separation and purification, and finally obtaining the single bacterial colony of the enterococcus with low-temperature protease. Inoculating the separated and purified single colony onto an inclined plane, culturing for 24 hours at 25 ℃, and preserving for later use.
2. Identification of strains
2.1 morphological identification
The bacterial colony is orange, smooth in surface, neat in edge, opaque and compact; gram staining was positive; the cells are spherical, and are arranged singly or in a pile (see fig. 1, 3, 5 and 7).
2.2 physiological characteristics
4 active cocci can grow when the NaCl concentration is 0-15%; when the pH is 7-9, the growth is faster; good growth can be carried out at the temperature ranging from 15 ℃ to 35 ℃;
2.3 molecular biological identification
The screened purified enterococci were subjected to 16S rRNA sequencing, identified as Planococcus maritimus, planococcus plakortidis, planococcus dechangensis and Planococcus rifietoensis, and a biological evolutionary relationship tree was constructed (see fig. 2, 4, 6 and 8).
3. Preservation of strains
4 Planococcus mobilis have been preserved in 2019, 01 and 02 in China general microbiological culture collection center (CGMCC) of China national institute of microbiological culture Collection center (CGMCC) located in national institute of sciences, national center for use in North Chen, lu No.1, 3, korea, beijing,
marine pediococcus (Planococcus maritimus XJ), which has been deposited on 2019, 01 and 02 in China center for culture collection (CGMCC), number CGMCC No.17057, proposed classification and nomenclature: planococcus maritimus.
The strain is preserved in China general microbiological culture Collection center (CGMCC) with the number of CGMCC NO.17058 and proposed classification and naming in China national institutes of microbiology institute of China, national academy of sciences, no.1, no. 3, the Korean region of Beijing, at the month 01 and 02 of Prinsepia (Planococcus plakortidis XJ) of Prinsepia). Planococcus plakortidis.
Dechang's active coccus (Planococcus dechangensis XJ), which has been deposited on 2019, 01 and 02 in China center for culture collection (CGMCC), number CGMCC No.17059, proposed classification and nomenclature: planococcus dechangensis.
The Laibotoxin zoococcus (Planococcus rifietoensis XJ) has been preserved in 2019, 01 and 02 in China center for culture collection (CGMCC), number CGMCC No.17060, the proposed classification and nomenclature: leptozoococcus mobilis Planococcus rifietoensis.
Example 1:
a method for fermenting fish sauce by utilizing marine enterococcus Planococcus maritimus XJ, which comprises the following steps:
(1) And (3) raw material treatment: mincing minced fish (waste generated during fish processing) with a meat mincing machine, adding 5% (w/w) of salted sea salt, and mixing.
(2) Preparation of the starter: the marine enterococcus Planococcus maritimus XJ (CGMCC No. 17057) is activated and cultured for three times. Centrifuging activated bacteria solution at 4deg.C and 10000r/min for 10min, washing with sterile physiological saline twice, suspending in small amount of sterile physiological saline, and regulating bacteria solution concentration to 10 5 ～10 7 CFU/mL, ready for use.
(3) Addition of a starter: adding marine coccus Planococcus maritimus XJ (CGMCC No. 17057) in a final amount (10 5 CFU/g) is mixed into the pretreated raw material minced fish meat to enable the final bacterial count to reach the addition requirement.
(4) Fermenting fish sauce: and (3) preserving heat and fermenting the mixed material obtained in the step (3) for 30d at the temperature of 15 ℃.
(5) Filtering fish sauce: 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 the supernatant with a multi-layer gauze to remove solids and impurities.
(6) Sterilization of fish sauce: and (3) carrying out secondary sterilization on the fish sauce obtained in the step (5) at 100 ℃ for 30min, and filling and sealing under the aseptic condition to obtain a fish sauce finished product.
(7) Evaluation of physical and chemical indexes of products: amino acid nitrogen content evaluation, wherein the method refers to GB5009.235-2016 colorimetric method; the method refers to GB5009.228-2016 micro-diffusion method for evaluating the content of volatile basic nitrogen; the measurement of histamine content was carried out by the method described in GB5009.208-2016 spectrophotometry.
(8) Fish sauce physical and chemical index results: the amino acid content of the fish sauce fermented by the marine active coccus can reach 1.284g/100mL, and the fish sauce is divided into first-class fish sauce according to the Chinese fish sauce industry standard; the volatile basic nitrogen content is 107mg/100mL; the histamine content is 19mg/100mL, which is lower than the European Union standard 40mg/100mL. The inherent earthy taste of the raw materials in the fish sauce is greatly reduced, the fresh taste is obvious, and compared with the fish sauce fermented by the strain, the flavor is obviously improved.
Example 2:
a method for fermenting fish sauce by utilizing marine enterococcus Planococcus maritimus XJ, which comprises the following steps:
(1) And (3) raw material treatment: mincing minced fish (waste generated during fish processing) with a meat mincing machine, adding 10% (w/w) of salted sea salt, and mixing.
(2) Preparation of the starter: the marine enterococcus Planococcus maritimus XJ (CGMCC No. 17057) is activated and cultured for three times. Centrifuging activated bacteria solution at 4deg.C and 10000r/min for 10min, washing with sterile physiological saline twice, suspending in small amount of sterile physiological saline, and regulating bacteria solution concentration to 10 5 ～10 7 CFU/mL, ready for use.
(3) Addition of a starter: adding marine coccus Planococcus maritimus XJ (CGMCC No. 17057) in a final amount (10 7 CFU/g) is mixed into the pretreated raw material minced fish meat to enable the final bacterial count to reach the addition requirement.
(4) Fermenting fish sauce: and (3) preserving heat and fermenting the mixed material obtained in the step (3) for 15d at the temperature of 20 ℃.
(5) Filtering fish sauce: 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 the supernatant with a multi-layer gauze to remove solids and impurities.
(6) Sterilization of fish sauce: and (3) carrying out secondary sterilization on the fish sauce obtained in the step (5) at 100 ℃ for 20min, and filling and sealing under the aseptic condition to obtain a fish sauce finished product.
(7) Evaluation of physical and chemical indexes of products: amino acid nitrogen content evaluation, wherein the method refers to GB5009.235-2016 colorimetric method; the method refers to GB5009.228-2016 micro-diffusion method for evaluating the content of volatile basic nitrogen; the measurement of histamine content was carried out by the method described in GB5009.208-2016 spectrophotometry.
(8) Fish sauce physical and chemical index results: the amino acid content of the fish gravy fermented by the marine active coccus can reach 1.023g/100mL, and the fish gravy is divided into first-class fish gravy according to the Chinese fish gravy industry standard; the volatile basic nitrogen content is 94mg/100mL; the histamine content is 17mg/100mL, which is lower than the European Union standard 40mg/100mL. The inherent earthy taste of the raw materials in the fish sauce is greatly reduced, the fresh taste is obvious, and compared with the fish sauce fermented by the strain, the flavor is obviously improved.
Example 3:
a method for fermenting fish sauce by utilizing marine enterococcus Planococcus maritimus XJ, which comprises the following steps:
(1) And (3) raw material treatment: mincing minced fish (waste generated during fish processing) with a meat mincing machine, adding 15% (w/w) of salted sea salt, and mixing.
(2) Preparation of the starter: the marine enterococcus Planococcus maritimus XJ (CGMCC No. 17057) is activated and cultured for three times. Centrifuging activated bacteria solution at 4deg.C and 10000r/min for 10min, washing with sterile physiological saline twice, suspending in small amount of sterile physiological saline, and regulating bacteria solution concentration to 10 5 ～10 7 CFU/mL, ready for use.
(3) Addition of a starter: adding marine coccus Planococcus maritimus XJ (CGMCC No. 17057) in a final amount (10 9 CFU/g) is mixed into the pretreated raw material minced fish meat to enable the final bacterial count to reach the addition requirement.
(4) Fermenting fish sauce: and (3) preserving heat and fermenting the mixed material obtained in the step (3) for 5 days at the temperature of 30 ℃.
(5) Filtering fish sauce: 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 the supernatant with a multi-layer gauze to remove solids and impurities.
(6) Sterilization of fish sauce: and (3) carrying out secondary sterilization on the fish sauce obtained in the step (5) at 100 ℃ for 15min, and filling and sealing under the aseptic condition to obtain a fish sauce finished product.
(7) Evaluation of physical and chemical indexes of products: amino acid nitrogen content evaluation, wherein the method refers to GB5009.235-2016 colorimetric method; the method refers to GB5009.228-2016 micro-diffusion method for evaluating the content of volatile basic nitrogen; the measurement of histamine content was carried out by the method described in GB5009.208-2016 spectrophotometry.
(8) Fish sauce physical and chemical index results: the amino acid content of the fish sauce fermented by the marine active coccus can reach 0.97g/100mL, and the fish sauce is divided into first-class fish sauce according to the Chinese fish sauce industry standard; the volatile basic nitrogen content is 85mg/100mL; the histamine content is 11mg/100mL, which is lower than the European Union standard 40mg/100mL. The inherent earthy taste of the raw materials in the fish sauce is greatly reduced, the fresh taste is obvious, and compared with the fish sauce fermented by the strain, the flavor is obviously improved.
Example 4:
a method for fermenting fish sauce by utilizing the enterococcus prasugrel Planococcus plakortidis XJ comprises the following steps:
(1) And (3) raw material treatment: mincing minced fish (waste generated during fish processing) with a meat mincing machine, adding 5% (w/w) of salted sea salt, and mixing.
(2) Preparation of the starter: the Pediococcus praecox Planococcus plakortidis XJ (CGMCC No. 17058) was subjected to activation culture three times. Centrifuging activated bacteria solution at 4deg.C and 10000r/min for 10min, washing with sterile physiological saline twice, suspending in small amount of sterile physiological saline, and regulating bacteria solution concentration to 10 5 ～10 7 CFU/mL, ready for use.
(3) Addition of a starter: the enterococcus prasugrel Planococcus plakortidis XJ (CGMCC No. 17058) is added according to the final adding amount (10) 5 CFU/g) is mixed into the pretreated raw material minced fish meat to enable the final bacterial count to reach the addition requirement.
(4) Fermenting fish sauce: and (3) preserving heat and fermenting the mixed material obtained in the step (3) for 30d at the temperature of 15 ℃.
(5) Filtering fish sauce: 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 the supernatant with a multi-layer gauze to remove solids and impurities.
(6) Sterilization of fish sauce: and (3) carrying out secondary sterilization on the fish sauce obtained in the step (5) at 100 ℃ for 30min, and filling and sealing under the aseptic condition to obtain a fish sauce finished product.
(7) Evaluation of physical and chemical indexes of products: amino acid nitrogen content evaluation, wherein the method refers to GB5009.235-2016 colorimetric method; the method refers to GB5009.228-2016 micro-diffusion method for evaluating the content of volatile basic nitrogen; the measurement of histamine content was carried out by the method described in GB5009.208-2016 spectrophotometry.
(8) Fish sauce physical and chemical index results: the amino acid content of the fermentation fish gravy of the strain of the enterococcus prasuavis can reach 1.117g/100mL, and the fish gravy is divided into first-class fish gravy according to the industry standard of Chinese fish gravy; the volatile basic nitrogen content is 112mg/100mL; the histamine content is 20mg/100mL, which is lower than the European Union standard 40mg/100mL. The inherent earthy taste of the raw materials in the fish sauce is greatly reduced, the fresh taste is obvious, and compared with the fish sauce fermented by the strain, the flavor is obviously improved.
Example 5:
a method for fermenting fish sauce by utilizing the enterococcus prasugrel Planococcus plakortidis XJ comprises the following steps:
(1) And (3) raw material treatment: mincing minced fish (waste generated during fish processing) with a meat mincing machine, adding 10% (w/w) of salted sea salt, and mixing.
(2) Preparation of the starter: the Pediococcus praecox Planococcus plakortidis XJ (CGMCC No. 17058) was subjected to activation culture three times. Centrifuging activated bacteria solution at 4deg.C and 10000r/min for 10min, washing with sterile physiological saline twice, suspending in small amount of sterile physiological saline, and regulating bacteria solution concentration to 10 5 ～10 7 CFU/mL, ready for use.
(3) Addition of a starter: the enterococcus prasugrel Planococcus plakortidis XJ (CGMCC No. 17058) is added according to the final adding amount (10) 7 CFU/g) is mixed into the pretreated raw material minced fish meat to enable the final bacterial count to reach the addition requirement.
(4) Fermenting fish sauce: and (3) preserving heat and fermenting the mixed material obtained in the step (3) for 15d at the temperature of 20 ℃.
(5) Filtering fish sauce: 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 the supernatant with a multi-layer gauze to remove solids and impurities.
(6) Sterilization of fish sauce: and (3) carrying out secondary sterilization on the fish sauce obtained in the step (5) at 100 ℃ for 20min, and filling and sealing under the aseptic condition to obtain a fish sauce finished product.
(7) Evaluation of physical and chemical indexes of products: amino acid nitrogen content evaluation, wherein the method refers to GB5009.235-2016 colorimetric method; the method refers to GB5009.228-2016 micro-diffusion method for evaluating the content of volatile basic nitrogen; the measurement of histamine content was carried out by the method described in GB5009.208-2016 spectrophotometry.
(8) Fish sauce physical and chemical index results: the amino acid content of the fermentation fish gravy of the strain of the enterococcus prasuavis can reach 1.103g/100mL, and the fish gravy is divided into first-class fish gravy according to the industry standard of Chinese fish gravy; the volatile basic nitrogen content is 99mg/100mL; the histamine content is 18mg/100mL, which is lower than the European Union standard 40mg/100mL. The inherent earthy taste of the raw materials in the fish sauce is greatly reduced, the fresh taste is obvious, and compared with the fish sauce fermented by the strain, the flavor is obviously improved.
Example 6:
a method for fermenting fish sauce by utilizing the enterococcus prasugrel Planococcus plakortidis XJ comprises the following steps:
(1) And (3) raw material treatment: mincing minced fish (waste generated during fish processing) with a meat mincing machine, adding 15% (w/w) of salted sea salt, and mixing.
(2) Preparation of the starter: the Pediococcus praecox Planococcus plakortidis XJ (CGMCC No. 17058) was subjected to activation culture three times. Centrifuging activated bacteria solution at 4deg.C and 10000r/min for 10min, washing with sterile physiological saline twice, suspending in small amount of sterile physiological saline, and regulating bacteria solution concentration to 10 5 ～10 7 CFU/mL, ready for use.
(3) Addition of a starter: the enterococcus prasugrel Planococcus plakortidis XJ (CGMCC No. 17058) is added according to the final adding amount (10) 9 CFU/g) is mixed into the pretreated raw material minced fish meat to enable the final bacterial count to reach the addition requirement.
(4) Fermenting fish sauce: and (3) preserving heat and fermenting the mixed material obtained in the step (3) for 5 days at the temperature of 30 ℃.
(5) Filtering fish sauce: 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 the supernatant with a multi-layer gauze to remove solids and impurities.
(6) Sterilization of fish sauce: and (3) carrying out secondary sterilization on the fish sauce obtained in the step (5) at 100 ℃ for 15min, and filling and sealing under the aseptic condition to obtain a fish sauce finished product.
(7) Evaluation of physical and chemical indexes of products: amino acid nitrogen content evaluation, wherein the method refers to GB5009.235-2016 colorimetric method; the method refers to GB5009.228-2016 micro-diffusion method for evaluating the content of volatile basic nitrogen; the measurement of histamine content was carried out by the method described in GB5009.208-2016 spectrophotometry.
(8) Fish sauce physical and chemical index results: the amino acid content of the fermentation fish gravy of the strain of the enterococcus prasuavis can reach 1.008g/100mL, and the fish gravy is divided into first-class fish gravy according to the industry standard of Chinese fish gravy; the volatile basic nitrogen content is 92mg/100mL; the histamine content is 17mg/100mL, which is lower than the European Union standard 40mg/100mL. The inherent earthy taste of the raw materials in the fish sauce is greatly reduced, the fresh taste is obvious, and compared with the fish sauce fermented by the strain, the flavor is obviously improved.
Example 7:
a method for fermenting fish sauce by using De-Mexico Planococcus dechangensis XJ11 comprises the following steps:
(1) And (3) raw material treatment: mincing minced fish (waste generated during fish processing) with a meat mincing machine, adding 5% (w/w) of salted sea salt, and mixing.
(2) Preparation of the starter: dechang rhodococcus Planococcus dechangensis XJ11 (CGMCC No. 17059) was activated and cultured three times. Centrifuging activated bacteria solution at 4deg.C and 10000r/min for 10min, washing with sterile physiological saline twice, suspending in small amount of sterile physiological saline, and regulating bacteria solution concentration to 10 5 ～10 7 CFU/mL, ready for use.
(3) Addition of a starter: adding Dechang coccus Planococcus dechangensis XJ (CGMCC No. 17059) in a final amount (10 5 CFU/g) is mixed into the pretreated raw material minced fish meat to enable the final bacterial count to reach the addition requirement.
(4) Fermenting fish sauce: and (3) preserving heat and fermenting the mixed material obtained in the step (3) for 30d at the temperature of 15 ℃.
(5) Filtering fish sauce: 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 the supernatant with a multi-layer gauze to remove solids and impurities.
(6) Sterilization of fish sauce: and (3) carrying out secondary sterilization on the fish sauce obtained in the step (5) at 100 ℃ for 30min, and filling and sealing under the aseptic condition to obtain a fish sauce finished product.
(7) Evaluation of physical and chemical indexes of products: amino acid nitrogen content evaluation, wherein the method refers to GB5009.235-2016 colorimetric method; the method refers to GB5009.228-2016 micro-diffusion method for evaluating the content of volatile basic nitrogen; the measurement of histamine content was carried out by the method described in GB5009.208-2016 spectrophotometry.
(8) Fish sauce physical and chemical index results: the amino acid content of the fish sauce fermented by the Dechang active coccus can reach 1.216g/100mL, and the fish sauce is divided into first-class fish sauce according to the Chinese fish sauce industry standard; the volatile basic nitrogen content is 109mg/100mL; the histamine content is 19mg/100mL, which is lower than the European Union standard 40mg/100mL. The inherent earthy taste of the raw materials in the fish sauce is greatly reduced, the fresh taste is obvious, and compared with the fish sauce fermented by the strain, the flavor is obviously improved.
Example 8:
a method for fermenting fish sauce by using De-Mexico Planococcus dechangensis XJ11 comprises the following steps:
(1) And (3) raw material treatment: mincing minced fish (waste generated during fish processing) with a meat mincing machine, adding 10% (w/w) of salted sea salt, and mixing.
(2) Preparation of the starter: dechang rhodococcus Planococcus dechangensis XJ11 (CGMCC No. 17059) was activated and cultured three times. Centrifuging activated bacteria solution at 4deg.C and 10000r/min for 10min, washing with sterile physiological saline twice, suspending in small amount of sterile physiological saline, and regulating bacteria solution concentration to 10 5 ～10 7 CFU/mL, ready for use.
(3) Addition of a starter: adding Dechang coccus Planococcus dechangensis XJ (CGMCC No. 17059) in a final amount (10 7 CFU/g) is mixed into the pretreated raw material minced fish meat to enable the final bacterial count to reach the addition requirement.
(4) Fermenting fish sauce: and (3) preserving heat and fermenting the mixed material obtained in the step (3) for 15d at the temperature of 20 ℃.
(5) Filtering fish sauce: 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 the supernatant with a multi-layer gauze to remove solids and impurities.
(6) Sterilization of fish sauce: and (3) carrying out secondary sterilization on the fish sauce obtained in the step (5) at 100 ℃ for 20min, and filling and sealing under the aseptic condition to obtain a fish sauce finished product.
(7) Evaluation of physical and chemical indexes of products: amino acid nitrogen content evaluation, wherein the method refers to GB5009.235-2016 colorimetric method; the method refers to GB5009.228-2016 micro-diffusion method for evaluating the content of volatile basic nitrogen; the measurement of histamine content was carried out by the method described in GB5009.208-2016 spectrophotometry.
(8) Fish sauce physical and chemical index results: the amino acid content of the fish sauce fermented by the Dechang active coccus can reach 1.174g/100mL, and the fish sauce is divided into first-class fish sauce according to the Chinese fish sauce industry standard; the volatile basic nitrogen content is 98mg/100mL; the histamine content is 20mg/100mL, which is lower than the European Union standard 40mg/100mL. The inherent earthy taste of the raw materials in the fish sauce is greatly reduced, the fresh taste is obvious, and compared with the fish sauce fermented by the strain, the flavor is obviously improved.
Example 9:
a method for fermenting fish sauce by using De-Mexico Planococcus dechangensis XJ11 comprises the following steps:
(1) And (3) raw material treatment: mincing minced fish (waste generated during fish processing) with a meat mincing machine, adding 15% (w/w) of salted sea salt, and mixing.
(2) Preparation of the starter: dechang rhodococcus Planococcus dechangensis XJ11 (CGMCC No. 17059) was activated and cultured three times. Centrifuging activated bacteria solution at 4deg.C and 10000r/min for 10min, washing with sterile physiological saline twice, suspending in small amount of sterile physiological saline, and regulating bacteria solution concentration to 10 5 ～10 7 CFU/mL, ready for use.
(3) Addition of a starter: adding Dechang coccus Planococcus dechangensis XJ (CGMCC No. 17059) in a final amount (10 9 CFU/g) is mixed into the pretreated raw material minced fish to achieve the final bacterial countAnd adding requirements.
(4) Fermenting fish sauce: and (3) preserving heat and fermenting the mixed material obtained in the step (3) for 5 days at the temperature of 30 ℃.
(5) Filtering fish sauce: 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 the supernatant with a multi-layer gauze to remove solids and impurities.
(6) Sterilization of fish sauce: and (3) carrying out secondary sterilization on the fish sauce obtained in the step (5) at 100 ℃ for 15min, and filling and sealing under the aseptic condition to obtain a fish sauce finished product.
(7) Evaluation of physical and chemical indexes of products: amino acid nitrogen content evaluation, wherein the method refers to GB5009.235-2016 colorimetric method; the method refers to GB5009.228-2016 micro-diffusion method for evaluating the content of volatile basic nitrogen; the measurement of histamine content was carried out by the method described in GB5009.208-2016 spectrophotometry.
(8) Fish sauce physical and chemical index results: the amino acid content of the fish sauce fermented by the Dechang active coccus can reach 1.089g/100mL, and the fish sauce is divided into first-class fish sauce according to the Chinese fish sauce industry standard; the volatile basic nitrogen content is 92mg/100mL; the histamine content is 14mg/100mL, which is lower than the European Union standard 40mg/100mL. The inherent earthy taste of the raw materials in the fish sauce is greatly reduced, the fresh taste is obvious, and compared with the fish sauce fermented by the strain, the flavor is obviously improved.
Example 10:
a method for fermenting fish sauce by utilizing Laibotozoococcus Planococcus rifietoensis XJ, which comprises the following steps:
(1) And (3) raw material treatment: mincing minced fish (waste generated during fish processing) with a meat mincing machine, adding 5% (w/w) of salted sea salt, and mixing.
(2) Preparation of the starter: the Laiyitozoococcus bifidus Planococcus rifietoensis XJ (CGMCC No. 17060) is activated and cultured for three times. Centrifuging activated bacteria solution at 4deg.C and 10000r/min for 10min, washing with sterile physiological saline twice, suspending in small amount of sterile physiological saline, and regulating bacteria solution concentration to 10 5 ～10 7 CFU/mL, ready for use.
(3) Addition of a starter: adding Laibotozoococcus Planococcus rifietoensis XJ (CGMCC No. 17060) in a final amount (10 5 CFU/g) is mixed into the pretreated raw material minced fish meat to enable the final bacterial count to reach the addition requirement.
(4) Fermenting fish sauce: and (3) preserving heat and fermenting the mixed material obtained in the step (3) for 30d at the temperature of 15 ℃.
(5) Filtering fish sauce: 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 the supernatant with a multi-layer gauze to remove solids and impurities.
(6) Sterilization of fish sauce: and (3) carrying out secondary sterilization on the fish sauce obtained in the step (5) at 100 ℃ for 30min, and filling and sealing under the aseptic condition to obtain a fish sauce finished product.
(7) Evaluation of physical and chemical indexes of products: amino acid nitrogen content evaluation, wherein the method refers to GB5009.235-2016 colorimetric method; the method refers to GB5009.228-2016 micro-diffusion method for evaluating the content of volatile basic nitrogen; the measurement of histamine content was carried out by the method described in GB5009.208-2016 spectrophotometry.
(8) Fish sauce physical and chemical index results: the amino acid content of the fish sauce fermented by the Laiyitong zoococcus can reach 1.159g/100mL, and the fish sauce is divided into first-class fish sauce according to the Chinese fish sauce industry standard; the volatile basic nitrogen content is 104mg/100mL; the histamine content is 16mg/100mL, which is lower than the European Union standard 40mg/100mL. The inherent earthy taste of the raw materials in the fish sauce is greatly reduced, the fresh taste is obvious, and compared with the fish sauce fermented by the strain, the flavor is obviously improved.
Example 11:
a method for fermenting fish sauce by utilizing Laibotozoococcus Planococcus rifietoensis XJ, which comprises the following steps:
(1) And (3) raw material treatment: mincing minced fish (waste generated during fish processing) with a meat mincing machine, adding 10% (w/w) of salted sea salt, and mixing.
(2) Preparation of the starter: the Laiyitozoococcus bifidus Planococcus rifietoensis XJ (CGMCC No. 17060) is activated and cultured for three times. Isolating the activated bacterial liquid at 4deg.C and 10000r/minHeart for 10min, washing twice with sterile physiological saline, suspending in small amount of sterile physiological saline, and regulating bacterial liquid concentration to 10 5 ～10 7 CFU/mL, ready for use.
(3) Addition of a starter: adding Laibotozoococcus Planococcus rifietoensis XJ (CGMCC No. 17060) in a final amount (10 7 CFU/g) is mixed into the pretreated raw material minced fish meat to enable the final bacterial count to reach the addition requirement.
(4) Fermenting fish sauce: and (3) preserving heat and fermenting the mixed material obtained in the step (3) for 15d at the temperature of 20 ℃.
(5) Filtering fish sauce: 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 the supernatant with a multi-layer gauze to remove solids and impurities.
(6) Sterilization of fish sauce: and (3) carrying out secondary sterilization on the fish sauce obtained in the step (5) at 100 ℃ for 20min, and filling and sealing under the aseptic condition to obtain a fish sauce finished product.
(7) Evaluation of physical and chemical indexes of products: amino acid nitrogen content evaluation, wherein the method refers to GB5009.235-2016 colorimetric method; the method refers to GB5009.228-2016 micro-diffusion method for evaluating the content of volatile basic nitrogen; the measurement of histamine content was carried out by the method described in GB5009.208-2016 spectrophotometry.
(8) Fish sauce physical and chemical index results: the amino acid content of the fish sauce fermented by the Laiyitong zoococcus can reach 1.207g/100mL, and the fish sauce is divided into first-class fish sauce according to the Chinese fish sauce industry standard; the volatile basic nitrogen content is 109mg/100mL; the histamine content is 21mg/100mL, which is lower than the European Union standard 40mg/100mL. The inherent earthy taste of the raw materials in the fish sauce is greatly reduced, the fresh taste is obvious, and compared with the fish sauce fermented by the strain, the flavor is obviously improved.
Example 12:
a method for fermenting fish sauce by utilizing Laibotozoococcus Planococcus rifietoensis XJ, which comprises the following steps:
(1) And (3) raw material treatment: mincing minced fish (waste generated during fish processing) with a meat mincing machine, adding 15% (w/w) of salted sea salt, and mixing.
(2) Preparation of the starter: the Laiyitozoococcus bifidus Planococcus rifietoensis XJ (CGMCC No. 17060) is activated and cultured for three times. Centrifuging activated bacteria solution at 4deg.C and 10000r/min for 10min, washing with sterile physiological saline twice, suspending in small amount of sterile physiological saline, and regulating bacteria solution concentration to 10 5 ～10 7 CFU/mL, ready for use.
(3) Addition of a starter: adding Laibotozoococcus Planococcus rifietoensis XJ (CGMCC No. 17060) in a final amount (10 9 CFU/g) is mixed into the pretreated raw material minced fish meat to enable the final bacterial count to reach the addition requirement.
(4) Fermenting fish sauce: and (3) preserving heat and fermenting the mixed material obtained in the step (3) for 5 days at the temperature of 30 ℃.
(5) Filtering fish sauce: 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 the supernatant with a multi-layer gauze to remove solids and impurities.
(6) Sterilization of fish sauce: and (3) carrying out secondary sterilization on the fish sauce obtained in the step (5) at 100 ℃ for 15min, and filling and sealing under the aseptic condition to obtain a fish sauce finished product.
(7) Evaluation of physical and chemical indexes of products: amino acid nitrogen content evaluation, wherein the method refers to GB5009.235-2016 colorimetric method; the method refers to GB5009.228-2016 micro-diffusion method for evaluating the content of volatile basic nitrogen; the measurement of histamine content was carried out by the method described in GB5009.208-2016 spectrophotometry.
(8) Fish sauce physical and chemical index results: the amino acid content of the fish sauce fermented by the Laiyitong zoococcus can reach 1.012g/100mL, and the fish sauce is divided into first-class fish sauce according to the Chinese fish sauce industry standard; the volatile basic nitrogen content is 98mg/100mL; the histamine content is 14mg/100mL, which is lower than the European Union standard 40mg/100mL. The inherent earthy taste of the raw materials in the fish sauce is greatly reduced, the fresh taste is obvious, and compared with the fish sauce fermented by the strain, the flavor is obviously improved.

Claims (2)

1. The preservation number of the enterococcus prasugrel (Planococcus plakortidis) XJ10 is CGMCC No.17058.
2. A method for fermenting fish gravy using the acinetococcus praecox (Planococcus plakortidis) XJ10 according to claim 1, comprising the steps of:
(1) Pretreatment of raw materials: mincing minced fish with a meat mincing machine, adding 5-15% (w/w) of salted sea salt, and mixing well for later use;
(2) Preparation of the starter: activating and culturing the actinococcus prasuavis strain for three times; centrifuging activated bacteria solution at 4deg.C and 10000r/min for 10min, washing with sterile physiological saline twice, suspending in small amount of sterile physiological saline, and regulating bacteria solution concentration to 10 5 ～10 7 CFU/mL for standby;
(3) Addition of a starter: the strain is added in a final amount of 10 5 ～10 9 Mixing CFU/g into pretreated raw material minced fish;
(4) Fermenting fish sauce: fermenting the mixed material obtained in the step (3) at 15-30 ℃ for 5-30 d;
(5) Filtering 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 the supernatant with multi-layer gauze to remove solids and impurities;
(6) Sterilization of fish sauce: and (3) carrying out secondary sterilization on the fish sauce obtained in the step (5) at 100 ℃ for 15-30 min, and filling and sealing under the aseptic condition to obtain a fish sauce finished product.
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