WO2021143006A1 - 动性球菌及利用其改善低盐鱼露发酵品质的方法 - Google Patents

动性球菌及利用其改善低盐鱼露发酵品质的方法 Download PDF

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WO2021143006A1
WO2021143006A1 PCT/CN2020/092490 CN2020092490W WO2021143006A1 WO 2021143006 A1 WO2021143006 A1 WO 2021143006A1 CN 2020092490 W CN2020092490 W CN 2020092490W WO 2021143006 A1 WO2021143006 A1 WO 2021143006A1
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fish sauce
planococcus
fish
minutes
sauce
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French (fr)
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高瑞昌
周晶
袁丽
周越
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Jiangsu University
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Jiangsu University
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Priority claimed from CN202010025134.4A external-priority patent/CN111534451B/zh
Priority claimed from CN202010076088.0A external-priority patent/CN111227216B/zh
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Priority to EP20841648.7A priority Critical patent/EP3878945B1/en
Priority to US17/256,008 priority patent/US11337447B2/en
Priority to JP2021513414A priority patent/JP7060902B2/ja
Publication of WO2021143006A1 publication Critical patent/WO2021143006A1/zh
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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 OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • 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 OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L17/00—Food-from-the-sea products; Fish products; Fish meal; Fish-egg substitutes; Preparation or treatment thereof
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L17/00—Food-from-the-sea products; Fish products; Fish meal; Fish-egg substitutes; Preparation or treatment thereof
    • A23L17/20—Fish extracts
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • 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
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/07—Bacillus

Definitions

  • the invention relates to the application field of food microbiological technology, in particular to kinesococcus and a method for improving the fermentation quality of low-salt fish sauce by using it.
  • Fish sauce is a traditional fermented aquatic condiment, also known as fish sauce. It is one of the most popular condiments in Southeast Asia. It has a clear brown-red color and a unique flavor and aroma. It has become a daily diet for people. a part of.
  • Traditional fish sauce production usually uses low-value fish and shrimp or aquatic product processing waste (fish heads, offal, etc.) as raw materials, and uses its own enzymes or microorganisms to naturally ferment.
  • Fish sauce is rich in nutrients, contains a variety of essential amino acids, as well as calcium, magnesium, zinc, iron and other mineral elements that are beneficial to the human body. It is a low-fat and high-protein fermented condiment.
  • fish sauce is also a product of high-value utilization of low-value fish, which has important practical significance to promote the development of the freshwater fish industry.
  • protease is mainly used to catalyze protein hydrolysis.
  • Low-temperature protease is a type of cold-adapted protease produced by low-temperature bacteria under low temperature conditions.
  • Low-temperature protease has high catalytic efficiency under low temperature conditions, and the most suitable reaction temperature is generally 20-40 °C.
  • Low-temperature protease can react at low or room temperature without heating and cooling, which can reduce costs. Therefore, it has the superiority that mesophilic protease cannot be replaced in industrial production, and has broad application prospects in the washing industry, food processing, biopharmaceutical, environmental bioremediation and other fields.
  • Kincococcus has been identified as a cold-adaptive bacteria producing low-temperature protease, and its safety is relatively high.
  • the present invention will utilize kinococcus to ferment fish sauce under low-temperature and low-salt conditions, thereby obtaining fish sauce products with short fermentation period, low salinity, and high flavor and nutritional value, which is the application of kinococcus in food microbial fermentation Provide theoretical basis and methods.
  • the present invention screens out kinesiococcus which can improve the fermentation quality of low-salt fish sauce from traditional shrimp paste, and combines the growth characteristics and enzymatic characteristics of kinesiococcus to ferment fish sauce. This method can effectively shorten the fermentation cycle, reduce the salt content and improve the flavor and nutritional value of fish sauce, so it can provide a more stable, safer and more suitable starter for fish sauce fermentation.
  • the present invention provides the use of the above-mentioned kinesococcus strain to improve the fermented flavor and quality of fish sauce.
  • a method for fermenting fish sauce using motility cocci is carried out according to the following steps:
  • Processing of raw materials use a meat chopper to grind the minced fish meat (waste produced during fish meat processing), add 5-15% (w/w) of pickled sea salt and mix well, and set aside.
  • step (3) Fermentation of fish sauce: the mixture obtained in step (3) is kept and fermented at a temperature of 15-30°C for 5-30 days.
  • step (5) Sterilization of fish sauce: the fish sauce obtained in step (5) is subjected to secondary sterilization at 100° C. for 15-30 minutes, and filled and sealed under aseptic conditions to obtain a finished fish sauce.
  • Evaluation of product physical and chemical indicators amino acid nitrogen content evaluation, the method refers to GB5009.235-2016 colorimetric method; volatile base nitrogen content evaluation, the method refers to GB5009.228-2016 trace diffusion method; determination of histamine content , The method refers to GB5009.208-2016 spectrophotometry.
  • said kinesococcus is marine kinococcus (Planococcus maritimus XJ2) or prakortidis (XJ10) or Dechang kinococcus (Planococcus dechangensis XJ11) or Lebitus kinococcus (Planococcus rifietoensis XJ12) ).
  • a method for improving the fermentation quality of low-salt fish sauce by using mixed strains of kinococcus is carried out according to the following steps:
  • Processing of raw materials use a meat chopper to grind the minced fish meat (waste produced during fish meat processing), add 5-15% (w/w) of pickled sea salt and mix well, and set aside.
  • step (3) Fermentation of fish sauce: the mixture obtained in step (3) is kept and fermented at a temperature of 15-30°C for 5-30 days.
  • step (5) Sterilization of fish sauce: the fish sauce obtained in step (5) is subjected to secondary sterilization at 100° C. for 15-30 minutes, and filled and sealed under aseptic conditions to obtain a finished fish sauce.
  • Evaluation of product physical and chemical indicators amino acid nitrogen content evaluation, the method refers to GB5009.235-2016 colorimetric method; volatile base nitrogen content evaluation, the method refers to GB5009.228-2016 trace diffusion method; determination of histamine content , The method refers to GB5009.208-2016 spectrophotometry.
  • Plant motility cocci Plantococcus maritimus XJ2
  • this strain has been deposited on January 2, 2019 at the Chinese Academy of Microbiology, the Chinese Academy of Sciences, located at No. 3, Beichen West Road, Chaoyang District, Beijing. Microbiology Center (CGMCC), the number is CGMCCNO.17057, and the suggested classification name: Planococcus maritimus.
  • CGMCC Microbiology Center
  • Planococcus plakortidis XJ10 this strain has been deposited on January 02, 2019 at the Chinese Academy of Microbiology Institute of Microbiology, Chinese Academy of Sciences No. 3 Beichen West Road, Chaoyang District, Beijing The General Microbiology Center (CGMCC), numbered CGMCCNO.17058, and the suggested classification name: Planococcus plkortidis.
  • CGMCC General Microbiology Center
  • Planococcus dechangensis XJ11 this strain has been deposited on January 2, 2019 at the Chinese Academy of Microbiology, the Chinese Academy of Sciences' Institute of Microbiology, No. 3, Beichen West Road, Chaoyang District, Beijing The General Microbiology Center (CGMCC), numbered CGMCCNO.17059, and the suggested classification name: Planococcus dechangensis.
  • CGMCC General Microbiology Center
  • Planococcus rifietoensis XJ12 this strain has been deposited on January 2, 2019 at the Chinese Academy of Microbiological Strain Collection Management at the Chinese Academy of Sciences Institute of Microbiology, No. 3 Beichen West Road, Chaoyang District, Beijing Committee General Microbiology Center (CGMCC), the number is CGMCCNO.17060, the proposed classification name: Planococcus rifietoensis.
  • the fermentation period of the fish sauce fermented by the above-mentioned kinesococcus strain is greatly shortened, and the fermentation temperature is lowered.
  • the fish sauce obtained by this method is transparent brown-red, has a unique aroma of aquatic products, and has no suspended or flocculent impurities.
  • the salt content of the finished fish sauce is low, the amino acid nitrogen content is high (according to the Chinese fish sauce industry standard, the fish sauce is classified as first-class fish sauce), the content of volatile basic nitrogen is low, and the content of histamine is lower than the national standard ,
  • the prepared fish sauce tastes delicious and has rich nutritional value, so the starter can be used to produce green and safe fermented fish sauce products.
  • the present invention has the uniqueness of invention patents, has achieved outstanding substantive features, and has achieved remarkable progress in innovation and practicability, and has produced beneficial effects.
  • the colonies are orange-yellow, with smooth surfaces, neat edges, opaque, and dense colonies; Gram staining is positive; the bacteria are spherical and arranged in single or piles.
  • the screened and purified motility cocci were sequenced with 16S rRNA and identified as Planococcus maritimus, Planococcus plakortidis, Planococcus dechangensis and Planococcus rifietoensis, and constructed a biological evolutionary relationship tree.
  • the 4 strains of kinesococci have been deposited on January 2, 2019 at the General Microbiology Center (CGMCC) of the Chinese Academy of Sciences Institute of Microbiology, No. 3 Beichen West Road, Chaoyang District, Beijing.
  • CGMCC General Microbiology Center
  • Plant motility cocci Plantococcus maritimus XJ2
  • this strain has been deposited on January 2, 2019 at the Chinese Academy of Microbiology, the Chinese Academy of Sciences, located at No. 3, Beichen West Road, Chaoyang District, Beijing. Microbiology Center (CGMCC), the number is CGMCCNO.17057, and the suggested classification name: Planococcus maritimus.
  • CGMCC Microbiology Center
  • Planococcus plakortidis XJ10 this strain has been deposited on January 02, 2019 at the Chinese Academy of Microbiology Institute of Microbiology, Chinese Academy of Sciences No. 3 Beichen West Road, Chaoyang District, Beijing The General Microbiology Center (CGMCC), numbered CGMCCNO.17058, and the suggested classification name: Planococcus plkortidis.
  • CGMCC General Microbiology Center
  • Planococcus dechangensis XJ11 this strain has been deposited on January 2, 2019 at the Chinese Academy of Microbiology, the Chinese Academy of Sciences' Institute of Microbiology, No. 3, Beichen West Road, Chaoyang District, Beijing The General Microbiology Center (CGMCC), numbered CGMCCNO.17059, and the suggested classification name: Planococcus dechangensis.
  • CGMCC General Microbiology Center
  • Planococcus rifietoensis XJ12 this strain has been deposited on January 2, 2019 at the Chinese Academy of Microbiological Strain Collection Management at the Chinese Academy of Sciences Institute of Microbiology, No. 3 Beichen West Road, Chaoyang District, Beijing Committee General Microbiology Center (CGMCC), the number is CGMCCNO.17060, the proposed classification name: Planococcus rifietoensis.
  • a method for fermenting fish sauce using the marine kinococcus Planococcus maritimus XJ2 is carried out according to the following steps:
  • step (3) Fermentation of fish sauce: The mixture obtained in step (3) is kept and fermented at a temperature of 15° C. for 30 days.
  • step (5) Sterilization of fish sauce: the fish sauce obtained in step (5) is subjected to secondary sterilization at 100° C. for 30 minutes, and filled and sealed under aseptic conditions to obtain a finished fish sauce.
  • Evaluation of product physical and chemical indicators amino acid nitrogen content evaluation, the method refers to GB5009.235-2016 colorimetric method; volatile base nitrogen content evaluation, the method refers to GB5009.228-2016 trace diffusion method; determination of histamine content , The method refers to GB5009.208-2016 spectrophotometry.
  • Results of physical and chemical indicators of fish sauce The amino acid content of fish sauce fermented by this strain of marine kinococcus can reach 1.284g/100mL.
  • the fish sauce is classified as first-class fish sauce; volatile base The nitrogen content is 107mg/100mL; the histamine content is 19mg/100mL, which is lower than the EU standard of 40mg/100mL.
  • the fish sauce inherent in the raw material of the fish sauce is greatly reduced, the umami taste is obvious, and the flavor is significantly improved compared with the fish sauce fermented with no strain.
  • a method for fermenting fish sauce using the marine kinococcus Planococcus maritimus XJ2 is carried out according to the following steps:
  • step (3) Fermentation of fish sauce: the mixture obtained in step (3) is kept and fermented at a temperature of 20° C. for 15 days.
  • step (5) Sterilization of fish sauce: the fish sauce obtained in step (5) is subjected to secondary sterilization at 100° C. for 20 minutes, and filled and sealed under aseptic conditions to obtain a finished fish sauce.
  • Evaluation of product physical and chemical indicators amino acid nitrogen content evaluation, the method refers to GB5009.235-2016 colorimetric method; volatile base nitrogen content evaluation, the method refers to GB5009.228-2016 trace diffusion method; determination of histamine content , The method refers to GB5009.208-2016 spectrophotometry.
  • Results of the physical and chemical indicators of fish sauce The amino acid content of the fish sauce fermented by this strain of marine kinococcus can reach 1.023g/100mL.
  • the fish sauce is classified as first-class fish sauce; volatile base The nitrogen content is 94mg/100mL; the histamine content is 17mg/100mL, which is lower than the EU standard of 40mg/100mL.
  • the fish sauce inherent in the raw material of the fish sauce is greatly reduced, the umami taste is obvious, and the flavor is significantly improved compared with the fish sauce fermented with no strain.
  • a method for fermenting fish sauce using the marine kinococcus Planococcus maritimus XJ2 is carried out according to the following steps:
  • Processing of raw materials use a meat chopper to grind the minced fish meat (waste produced during fish meat processing), add 15% (w/w) salted sea salt and mix well, and set aside.
  • step (3) Fermentation of fish sauce: the mixture obtained in step (3) is kept and fermented at a temperature of 30° C. for 5 days.
  • step (5) Sterilization of fish sauce: the fish sauce obtained in step (5) is subjected to secondary sterilization at 100° C. for 15 minutes, and filled and sealed under aseptic conditions to obtain a finished fish sauce.
  • Evaluation of product physical and chemical indicators amino acid nitrogen content evaluation, the method refers to GB5009.235-2016 colorimetric method; volatile base nitrogen content evaluation, the method refers to GB5009.228-2016 trace diffusion method; determination of histamine content , The method refers to GB5009.208-2016 spectrophotometry.
  • Results of the physical and chemical indicators of fish sauce The amino acid content of the fish sauce fermented by this strain of marine kinococcus can reach 0.97g/100mL.
  • the fish sauce is classified as first-class fish sauce; volatile base The nitrogen content is 85mg/100mL; the histamine content is 11mg/100mL, which is lower than the EU standard of 40mg/100mL.
  • the fish sauce inherent in the raw material of the fish sauce is greatly reduced, the umami taste is obvious, and the flavor is significantly improved compared with the fish sauce fermented with no strain.
  • a method for fermenting fish sauce using Planococcus plakortidis XJ10 is carried out according to the following steps:
  • step (3) Fermentation of fish sauce: The mixture obtained in step (3) is kept and fermented at a temperature of 15° C. for 30 days.
  • step (5) Sterilization of fish sauce: the fish sauce obtained in step (5) is subjected to secondary sterilization at 100° C. for 30 minutes, and filled and sealed under aseptic conditions to obtain a finished fish sauce.
  • Evaluation of product physical and chemical indicators amino acid nitrogen content evaluation, the method refers to GB5009.235-2016 colorimetric method; volatile base nitrogen content evaluation, the method refers to GB5009.228-2016 trace diffusion method; determination of histamine content , The method refers to GB5009.208-2016 spectrophotometry.
  • the results of the physical and chemical indicators of fish sauce the amino acid content of the fish sauce fermented by this strain of K. praziococcus can reach 1.117g/100mL.
  • the fish sauce is classified as first-class fish sauce; volatile salt The base nitrogen content is 112mg/100mL; the histamine content is 20mg/100mL, which is lower than the EU standard 40mg/100mL.
  • the fish sauce inherent in the raw material of the fish sauce is greatly reduced, the umami taste is obvious, and the flavor is significantly improved compared with the fish sauce fermented with no strain.
  • a method for fermenting fish sauce using Planococcus plakortidis XJ10 is carried out according to the following steps:
  • step (3) Fermentation of fish sauce: the mixture obtained in step (3) is kept and fermented at a temperature of 20° C. for 15 days.
  • step (5) Sterilization of fish sauce: the fish sauce obtained in step (5) is subjected to secondary sterilization at 100° C. for 20 minutes, and filled and sealed under aseptic conditions to obtain a finished fish sauce.
  • Evaluation of product physical and chemical indicators amino acid nitrogen content evaluation, the method refers to GB5009.235-2016 colorimetric method; volatile base nitrogen content evaluation, the method refers to GB5009.228-2016 trace diffusion method; determination of histamine content , The method refers to GB5009.208-2016 spectrophotometry.
  • Results of the physical and chemical indicators of fish sauce The amino acid content of the fish sauce fermented by this strain of K. pragnaticus can reach 1.103g/100mL.
  • the fish sauce is classified as first-class fish sauce; volatile salt The base nitrogen content is 99mg/100mL; the histamine content is 18mg/100mL, which is lower than the EU standard 40mg/100mL.
  • the fish sauce inherent in the raw material of the fish sauce is greatly reduced, the umami taste is obvious, and the flavor is significantly improved compared with the fish sauce fermented with no strain.
  • a method for fermenting fish sauce using Planococcus plakortidis XJ10 is carried out according to the following steps:
  • Processing of raw materials use a meat chopper to grind the minced fish meat (waste produced during fish meat processing), add 15% (w/w) salted sea salt and mix well, and set aside.
  • step (3) Fermentation of fish sauce: the mixture obtained in step (3) is kept and fermented at a temperature of 30° C. for 5 days.
  • step (5) Sterilization of fish sauce: the fish sauce obtained in step (5) is subjected to secondary sterilization at 100° C. for 15 minutes, and filled and sealed under aseptic conditions to obtain a finished fish sauce.
  • Evaluation of product physical and chemical indicators amino acid nitrogen content evaluation, the method refers to GB5009.235-2016 colorimetric method; volatile base nitrogen content evaluation, the method refers to GB5009.228-2016 trace diffusion method; determination of histamine content , The method refers to GB5009.208-2016 spectrophotometry.
  • the results of the physical and chemical indicators of fish sauce the amino acid content of the fish sauce fermented by this strain of K. praziococcus can reach 1.008g/100mL.
  • the fish sauce is classified as first-class fish sauce; volatile salt The base nitrogen content is 92mg/100mL; the histamine content is 17mg/100mL, which is lower than the EU standard 40mg/100mL.
  • the fish sauce inherent in the raw material of the fish sauce is greatly reduced, the umami taste is obvious, and the flavor is significantly improved compared with the fish sauce fermented with no strain.
  • a method for fermenting fish sauce using Dechang kinococcus Planococcus dechangensis XJ11 is carried out according to the following steps:
  • step (3) Fermentation of fish sauce: The mixture obtained in step (3) is kept and fermented at a temperature of 15° C. for 30 days.
  • step (5) Sterilization of fish sauce: the fish sauce obtained in step (5) is subjected to secondary sterilization at 100° C. for 30 minutes, and filled and sealed under aseptic conditions to obtain a finished fish sauce.
  • Evaluation of product physical and chemical indicators amino acid nitrogen content evaluation, the method refers to GB5009.235-2016 colorimetric method; volatile base nitrogen content evaluation, the method refers to GB5009.228-2016 trace diffusion method; determination of histamine content , The method refers to GB5009.208-2016 spectrophotometry.
  • the results of the physical and chemical indicators of fish sauce the amino acid content of fermented fish sauce with this strain of Dechang kinococcus can reach 1.216g/100mL.
  • the fish sauce is classified as first-class fish sauce; volatile salt The base nitrogen content is 109mg/100mL; the histamine content is 19mg/100mL, which is lower than the EU standard 40mg/100mL.
  • the fish sauce inherent in the raw material of the fish sauce is greatly reduced, the umami taste is obvious, and the flavor is significantly improved compared with the fish sauce fermented with no strain.
  • a method for fermenting fish sauce using Dechang kinococcus Planococcus dechangensis XJ11 is carried out according to the following steps:
  • Processing of raw materials use a meat chopper to grind the minced fish meat (waste generated during fish meat processing), add 10% (w/w) pickled sea salt and mix well, and set aside.
  • step (3) Fermentation of fish sauce: the mixture obtained in step (3) is kept and fermented at a temperature of 20° C. for 15 days.
  • step (5) Sterilization of fish sauce: the fish sauce obtained in step (5) is subjected to secondary sterilization at 100° C. for 20 minutes, and filled and sealed under aseptic conditions to obtain a finished fish sauce.
  • Evaluation of product physical and chemical indicators amino acid nitrogen content evaluation, the method refers to GB5009.235-2016 colorimetric method; volatile base nitrogen content evaluation, the method refers to GB5009.228-2016 trace diffusion method; determination of histamine content , The method refers to GB5009.208-2016 spectrophotometry.
  • Results of physical and chemical indicators of fish sauce the amino acid content of fermented fish sauce with this strain of Dechang kinococcus can reach 1.174g/100mL.
  • the fish sauce is classified as first-class fish sauce; volatile salt The base nitrogen content is 98mg/100mL; the histamine content is 20mg/100mL, which is lower than the EU standard 40mg/100mL.
  • the fish sauce inherent in the raw material of the fish sauce is greatly reduced, the umami taste is obvious, and the flavor is significantly improved compared with the fish sauce fermented with no strain.
  • a method for fermenting fish sauce using Dechang kinococcus Planococcus dechangensis XJ11 is carried out according to the following steps:
  • Processing of raw materials use a meat chopper to grind the minced fish meat (waste produced during fish meat processing), add 15% (w/w) salted sea salt and mix well, and set aside.
  • step (3) Fermentation of fish sauce: the mixture obtained in step (3) is kept and fermented at a temperature of 30° C. for 5 days.
  • step (5) Sterilization of fish sauce: the fish sauce obtained in step (5) is subjected to secondary sterilization at 100° C. for 15 minutes, and filled and sealed under aseptic conditions to obtain a finished fish sauce.
  • Evaluation of product physical and chemical indicators amino acid nitrogen content evaluation, the method refers to GB5009.235-2016 colorimetric method; volatile base nitrogen content evaluation, the method refers to GB5009.228-2016 trace diffusion method; determination of histamine content , The method refers to GB5009.208-2016 spectrophotometry.
  • Results of physical and chemical indicators of fish sauce The amino acid content of fermented fish sauce with this strain of Dechang kinococcus can reach 1.089g/100mL.
  • the fish sauce is classified as first-class fish sauce; volatile salt The base nitrogen content is 92mg/100mL; the histamine content is 14mg/100mL, which is lower than the EU standard 40mg/100mL.
  • the fish sauce inherent in the raw material of the fish sauce is greatly reduced, the umami taste is obvious, and the flavor is significantly improved compared with the fish sauce fermented with no strain.
  • a method for fermenting fish sauce by using Lebito Zoococcus Planococcus rifietoensis XJ12 according to the following steps:
  • step (3) Fermentation of fish sauce: The mixture obtained in step (3) is kept and fermented at a temperature of 15° C. for 30 days.
  • step (5) Sterilization of fish sauce: the fish sauce obtained in step (5) is subjected to secondary sterilization at 100° C. for 30 minutes, and filled and sealed under aseptic conditions to obtain a finished fish sauce.
  • Evaluation of product physical and chemical indicators amino acid nitrogen content evaluation, the method refers to GB5009.235-2016 colorimetric method; volatile base nitrogen content evaluation, the method refers to GB5009.228-2016 trace diffusion method; determination of histamine content , The method refers to GB5009.208-2016 spectrophotometry.
  • the amino acid content of the fish sauce fermented by this strain of Lebitococcus Lepitococcus can reach 1.159g/100mL.
  • the fish sauce is classified as first-class fish sauce;
  • the basic nitrogen content is 104mg/100mL;
  • the histamine content is 16mg/100mL, which is lower than the EU standard 40mg/100mL.
  • the fish sauce inherent in the raw material of the fish sauce is greatly reduced, the umami taste is obvious, and the flavor is significantly improved compared with the fish sauce fermented with no strain.
  • a method for fermenting fish sauce by using Lebito Zoococcus Planococcus rifietoensis XJ12 according to the following steps:
  • step (3) Fermentation of fish sauce: the mixture obtained in step (3) is kept and fermented at a temperature of 20° C. for 15 days.
  • step (5) Sterilization of fish sauce: the fish sauce obtained in step (5) is subjected to secondary sterilization at 100° C. for 20 minutes, and filled and sealed under aseptic conditions to obtain a finished fish sauce.
  • Evaluation of product physical and chemical indicators amino acid nitrogen content evaluation, the method refers to GB5009.235-2016 colorimetric method; volatile base nitrogen content evaluation, the method refers to GB5009.228-2016 trace diffusion method; determination of histamine content , The method refers to GB5009.208-2016 spectrophotometry.
  • the results of the physical and chemical indicators of fish sauce the amino acid content of the fish sauce fermented by the strain of Lepitococcus can reach 1.207g/100mL.
  • the fish sauce is classified as first-class fish sauce; volatility The basic nitrogen content is 109mg/100mL; the histamine content is 21mg/100mL, which is lower than the EU standard of 40mg/100mL.
  • the fish sauce inherent in the raw material of the fish sauce is greatly reduced, the umami taste is obvious, and the flavor is significantly improved compared with the fish sauce fermented with no strain.
  • a method for fermenting fish sauce by using Lebito Zoococcus Planococcus rifietoensis XJ12 according to the following steps:
  • Processing of raw materials use a meat chopper to grind the minced fish meat (waste produced during fish meat processing), add 15% (w/w) salted sea salt and mix well, and set aside.
  • step (3) Fermentation of fish sauce: the mixture obtained in step (3) is kept and fermented at a temperature of 30° C. for 5 days.
  • step (5) Sterilization of fish sauce: the fish sauce obtained in step (5) is subjected to secondary sterilization at 100° C. for 15 minutes, and filled and sealed under aseptic conditions to obtain a finished fish sauce.
  • Evaluation of product physical and chemical indicators amino acid nitrogen content evaluation, the method refers to GB5009.235-2016 colorimetric method; volatile base nitrogen content evaluation, the method refers to GB5009.228-2016 trace diffusion method; determination of histamine content , The method refers to GB5009.208-2016 spectrophotometry.
  • the results of the physical and chemical indicators of fish sauce the amino acid content of the fish sauce fermented by this strain of Lebitococcus Lepitococcus can reach 1.012g/100mL.
  • the fish sauce is classified as first-class fish sauce; volatility The basic nitrogen content is 98mg/100mL; the histamine content is 14mg/100mL, which is lower than the EU standard 40mg/100mL.
  • the fish sauce inherent in the raw material of the fish sauce is greatly reduced, the umami taste is obvious, and the flavor is significantly improved compared with the fish sauce fermented with no strain.
  • a method for fermenting fish sauce by using mixed strains of motility cocci is carried out according to the following steps:
  • step (3) Fermentation of fish sauce: The mixture obtained in step (3) is kept and fermented at a temperature of 15° C. for 5 days.
  • step (5) Sterilization of fish sauce: the fish sauce obtained in step (5) is subjected to secondary sterilization at 100° C. for 30 minutes, and filled and sealed under aseptic conditions to obtain a finished fish sauce.
  • Evaluation of product physical and chemical indicators amino acid nitrogen content evaluation, the method refers to GB5009.235-2016 colorimetric method; volatile base nitrogen content evaluation, the method refers to GB5009.228-2016 trace diffusion method; determination of histamine content , The method refers to GB5009.208-2016 spectrophotometry.
  • Results of the physical and chemical indicators of fish sauce The amino acid content of the fish sauce fermented by this strain of marine kinococcus can reach 0.974g/100mL.
  • the fish sauce is classified as first-class fish sauce; volatile base The nitrogen content is 113mg/100mL; the histamine content is 17mg/100mL, which is lower than the EU standard of 40mg/100mL.
  • the fish sauce inherent in the raw material of the fish sauce is greatly reduced, the umami taste is obvious, and the flavor is significantly improved compared with the fish sauce fermented with no strain.
  • a method for fermenting fish sauce by using mixed strains of motility cocci is carried out according to the following steps:
  • step (3) Fermentation of fish sauce: The mixture obtained in step (3) is kept and fermented at a temperature of 15° C. for 30 days.
  • step (5) Sterilization of fish sauce: the fish sauce obtained in step (5) is subjected to secondary sterilization at 100° C. for 30 minutes, and filled and sealed under aseptic conditions to obtain a finished fish sauce.
  • Evaluation of product physical and chemical indicators amino acid nitrogen content evaluation, the method refers to GB5009.235-2016 colorimetric method; volatile base nitrogen content evaluation, the method refers to GB5009.228-2016 trace diffusion method; determination of histamine content , The method refers to GB5009.208-2016 spectrophotometry.
  • results of the physical and chemical indicators of fish sauce The amino acid content of the fish sauce fermented by this strain of marine kinococcus can reach 1.327g/100mL.
  • the fish sauce is classified as first-class fish sauce; volatile base The nitrogen content is 125mg/100mL; the histamine content is 23mg/100mL, which is lower than the EU standard of 40mg/100mL.
  • the fish sauce inherent in the raw material of the fish sauce is greatly reduced, the umami taste is obvious, and the flavor is significantly improved compared with the fish sauce fermented with no strain.
  • a method for fermenting fish sauce by using mixed strains of motility cocci is carried out according to the following steps:
  • step (3) Fermentation of fish sauce: The mixture material obtained in step (3) is incubated and fermented at a temperature of 20° C. for 10 days.
  • step (5) Sterilization of fish sauce: the fish sauce obtained in step (5) is subjected to secondary sterilization at 100° C. for 15 minutes, and filled and sealed under aseptic conditions to obtain a finished fish sauce.
  • Evaluation of product physical and chemical indicators amino acid nitrogen content evaluation, the method refers to GB5009.235-2016 colorimetric method; volatile base nitrogen content evaluation, the method refers to GB5009.228-2016 trace diffusion method; determination of histamine content , The method refers to GB5009.208-2016 spectrophotometry.
  • the results of the physical and chemical indicators of fish sauce the amino acid content of the fish sauce fermented by this strain of marine kinesococcus can reach 1.034g/100mL.
  • the fish sauce is classified as first-class fish sauce; volatile base The nitrogen content is 119mg/100mL; the histamine content is 20mg/100mL, which is lower than the EU standard of 40mg/100mL.
  • the fish sauce inherent in the raw material of the fish sauce is greatly reduced, the umami taste is obvious, and the flavor is significantly improved compared with the fish sauce fermented with no strain.
  • a method for fermenting fish sauce by using mixed strains of motility cocci is carried out according to the following steps:
  • step (3) Fermentation of fish sauce: the mixture obtained in step (3) is kept and fermented at a temperature of 20° C. for 15 days.
  • step (5) Sterilization of fish sauce: the fish sauce obtained in step (5) is subjected to secondary sterilization at 100° C. for 15 minutes, and filled and sealed under aseptic conditions to obtain a finished fish sauce.
  • Evaluation of product physical and chemical indicators amino acid nitrogen content evaluation, the method refers to GB5009.235-2016 colorimetric method; volatile base nitrogen content evaluation, the method refers to GB5009.228-2016 trace diffusion method; determination of histamine content , The method refers to GB5009.208-2016 spectrophotometry.
  • results of physical and chemical indicators of fish sauce the amino acid content of fish sauce fermented by this strain of marine kinesococcus can reach 1.116g/100mL.
  • the fish sauce is classified as first-class fish sauce; volatile base The nitrogen content is 108mg/100mL; the histamine content is 19mg/100mL, which is lower than the EU standard of 40mg/100mL.
  • the fish sauce inherent in the raw material of the fish sauce is greatly reduced, the umami taste is obvious, and the flavor is significantly improved compared with the fish sauce fermented with no strain.
  • a method for fermenting fish sauce by using mixed strains of motility cocci is carried out according to the following steps:
  • step (3) Fermentation of fish sauce: The mixture obtained in step (3) is kept and fermented at a temperature of 25° C. for 20 days.
  • step (5) Sterilization of fish sauce: the fish sauce obtained in step (5) is subjected to secondary sterilization at 100° C. for 15 minutes, and filled and sealed under aseptic conditions to obtain a finished fish sauce.
  • Evaluation of product physical and chemical indicators amino acid nitrogen content evaluation, the method refers to GB5009.235-2016 colorimetric method; volatile base nitrogen content evaluation, the method refers to GB5009.228-2016 trace diffusion method; determination of histamine content , The method refers to GB5009.208-2016 spectrophotometry.
  • results of the physical and chemical indicators of fish sauce the amino acid content of the fish sauce fermented by this strain of marine kinesococcus can reach 1.124g/100mL.
  • the fish sauce is classified as first-class fish sauce; volatile base The nitrogen content is 110mg/100mL; the histamine content is 21mg/100mL, which is lower than the EU standard of 40mg/100mL.
  • the fish sauce inherent in the raw material of the fish sauce is greatly reduced, the umami taste is obvious, and the flavor is significantly improved compared with the fish sauce fermented with no strain.
  • a method for fermenting fish sauce by using mixed strains of motility cocci is carried out according to the following steps:
  • step (3) Fermentation of fish sauce: the mixture obtained in step (3) is incubated and fermented at a temperature of 25° C. for 25 days.
  • step (5) Sterilization of fish sauce: The fish sauce obtained in step (5) is subjected to secondary sterilization at 100°C for 15 minutes, and filled and sealed under aseptic conditions to obtain the finished fish sauce.
  • Evaluation of product physical and chemical indicators amino acid nitrogen content evaluation, the method refers to GB5009.235-2016 colorimetric method; volatile base nitrogen content evaluation, the method refers to GB5009.228-2016 trace diffusion method; determination of histamine content , The method refers to GB5009.208-2016 spectrophotometry.
  • the results of the physical and chemical indicators of fish sauce the amino acid content of the fish sauce fermented by this strain of marine kinesococcus can reach 1.009g/100mL.
  • the fish sauce is classified as first-class fish sauce; volatile base The nitrogen content is 112mg/100mL; the histamine content is 20mg/100mL, which is lower than the EU standard of 40mg/100mL.
  • the fish sauce inherent in the raw material of the fish sauce is greatly reduced, the umami taste is obvious, and the flavor is significantly improved compared with the fish sauce fermented with no strain.
  • a method for fermenting fish sauce by using mixed strains of motility cocci is carried out according to the following steps:
  • Processing of raw materials use a meat chopper to grind the minced fish meat (waste produced during fish meat processing), add 15% (w/w) salted sea salt and mix well, and set aside.
  • step (3) Fermentation of fish sauce: the mixture obtained in step (3) is kept and fermented at a temperature of 30° C. for 5 days.
  • step (5) Sterilization of fish sauce: the fish sauce obtained in step (5) is subjected to secondary sterilization at 100° C. for 25 minutes, and filled and sealed under aseptic conditions to obtain a finished fish sauce.
  • Evaluation of product physical and chemical indicators amino acid nitrogen content evaluation, the method refers to GB5009.235-2016 colorimetric method; volatile base nitrogen content evaluation, the method refers to GB5009.228-2016 trace diffusion method; determination of histamine content , The method refers to GB5009.208-2016 spectrophotometry.
  • the amino acid content of the fish sauce fermented by this strain of marine kinococcus can reach 0.873g/100mL.
  • the fish sauce is classified as first-class fish sauce; volatile base The nitrogen content is 97mg/100mL; the histamine content is 16mg/100mL, which is lower than the EU standard of 40mg/100mL.
  • the fish sauce inherent in the raw material of the fish sauce is greatly reduced, the umami taste is obvious, and the flavor is significantly improved compared with the fish sauce fermented with no strain.
  • a method for fermenting fish sauce by using mixed strains of motility cocci is carried out according to the following steps:
  • Processing of raw materials use a meat chopper to grind the minced fish meat (waste produced during fish meat processing), add 15% (w/w) salted sea salt and mix well, and set aside.
  • step (3) Fermentation of fish sauce: the mixture obtained in step (3) is kept and fermented at a temperature of 30° C. for 30 days.
  • step (5) Sterilization of fish sauce: the fish sauce obtained in step (5) is subjected to secondary sterilization at 100° C. for 25 minutes, and filled and sealed under aseptic conditions to obtain a finished fish sauce.
  • Evaluation of product physical and chemical indicators amino acid nitrogen content evaluation, the method refers to GB5009.235-2016 colorimetric method; volatile base nitrogen content evaluation, the method refers to GB5009.228-2016 trace diffusion method; determination of histamine content , The method refers to GB5009.208-2016 spectrophotometry.
  • the amino acid content of the fish sauce fermented by this strain of marine kinococcus can reach 0.981g/100mL.
  • the fish sauce is classified as first-class fish sauce; volatile base The nitrogen content is 104mg/100mL; the histamine content is 18mg/100mL, which is lower than the EU standard of 40mg/100mL.
  • the fish sauce inherent in the raw material of the fish sauce is greatly reduced, the umami taste is obvious, and the flavor is significantly improved compared with the fish sauce fermented with no strain.

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Abstract

动性球菌及利用其改善低盐鱼露发酵品质的方法。动性球菌已保藏在中国微生物菌种保藏管理委员会普通微生物中心(CGMCC)，编号分别为CGMCCNO.17057、CGMCCNO.17058、CGMCCNO.17059、CGMCCNO.17060。所述利用动性球菌发酵鱼露的方法包括原料处理、发酵剂的制备、发酵剂的添加、发酵、过滤及灭菌步骤。

Description

动性球菌及利用其改善低盐鱼露发酵品质的方法 技术领域
本发明涉及食品微生物技术应用领域，尤其涉及动性球菌及利用其改善低盐鱼露发酵品质的方法。
背景技术
我国是渔业大国，淡水养殖已经成为我国渔业的重要产业支柱。2017年我国淡水养殖总产量约为2905万吨，但是加工利用率却不足15％，从而引起资源浪费、环境污染等问题。因此，如何解决淡水鱼的综合利用问题是我国水产品加工业面临的一大挑战。鱼露(fish sauce)是一种传统水产发酵调味品，又称鱼酱油，是东南亚地区最受欢迎的调味品之一，色泽呈清澈棕红色，具有独特的风味和香气，已经成为人们日常饮食的一部分。传统的鱼露生产通常以低值鱼虾或水产加工废弃物(鱼头、内脏等)为原料，利用自身的酶或微生物自然发酵而成。鱼露营养丰富，含多种必需氨基酸，以及钙、镁、锌、铁等对人体有益的矿物质元素，是一种低脂高蛋白发酵调味品。同时鱼露也是低值鱼高值化利用的产物，对推动淡水鱼产业发展具有重要的现实意义。鱼露发酵过程中主要利用蛋白酶催化蛋白质水解，根据温度对蛋白酶的活性及稳定性的影响可分为低温蛋白酶、中温蛋白酶和高温蛋白酶。低温蛋白酶是由低温菌在低温条件下所产生的一类冷适应蛋白酶。低温蛋白酶在低温条件下有较高的催化效率，一般最适反应温度为20～40℃。低温蛋白酶可在低温或室温下进行反应，无须加热和冷却，可以降低成本。因此在工业生产中有着中温蛋白酶无法取代的优越性，在洗涤业、食品加工、生物制药、环境生物修复等领域有着广阔的应用前景。动性球菌属已被鉴定为产低温蛋白酶的适冷菌，并且安全性较高。本发明将利用动性球菌，在低温低盐的条件下发酵鱼露，从而得到发酵周期短，盐度低以及风味和营养价值较高的鱼露产品，为动性球菌在食品微生物发酵应用上提供理论基础和方法。
发明内容
本发明从传统虾酱中筛选出能够改善低盐鱼露发酵品质的动性球菌，结合动性球菌的生长特性及酶学特性来发酵鱼露。该法可有效的缩短发酵周期，降低盐含量以及提高鱼露的风味和营养价值，因此能为鱼露发酵提供更稳定、更安全、更合适的发酵剂。
本发明提供上述动性球菌菌株的用途，用于鱼露发酵风味和品质的提升。
一种利用动性球菌发酵鱼露的方法，按照下述步骤进行：
(1)原料的处理：将碎鱼肉(鱼肉加工时所产生的废弃物)用碎肉机绞碎，加入腌制海盐5～15％(w/w)混匀，备用。
(2)发酵剂的制备：将动性球菌菌种活化培养三次。将活化好的菌液于4℃、10000r/min离心10min，然后用无菌生理盐水洗涤两次，再悬浮于少量无菌生理盐水中，最后将菌液浓度调整到105～107CFU/mL，备用。
(3)发酵剂的添加：将菌株按最终添加量(105～109CFU/g)混入到预处理过的原料碎鱼肉中，使最终菌数达到添加要求即可。
(4)鱼露的发酵：把步骤(3)得到的混合物料在温度15～30℃下保温发酵5～30d。
(5)鱼露的过滤：将步骤(4)得到的鱼露样品于120℃灭菌10～30min，冷却至室温后于10000r/min离心20min，取上清液用多层纱布过滤以除去固形物和杂质。
(6)鱼露的灭菌：将步骤(5)得到的鱼露在100℃下进行二次灭菌15～30min，在无菌条件下灌装密封，即得鱼露成品。
(7)产品理化指标的评价：氨基酸态氮含量评价，方法参照GB5009.235-2016比色法；挥发性盐基氮含量评价，方法参照GB5009.228-2016微量扩散法；组胺含量的测定，方法参照GB5009.208-2016分光光度法。
其中所述的动性球菌为海洋动性球菌(Planococcus maritimus XJ2)或普拉扣动球菌(Planococcus plakortidis XJ10)或德昌动性球菌(Planococcus dechangensis XJ11)或莱比托游动球菌(Planococcus rifietoensis XJ12)。
一种利用动性球菌混合菌株改善低盐鱼露发酵品质的方法，按照下述步骤进行：
(1)原料的处理：将碎鱼肉(鱼肉加工时所产生的废弃物)用碎肉机绞碎，加入腌制海盐5～15％(w/w)混匀，备用。
(2)混合发酵剂的制备：将海洋动性球菌(Planococcus maritimus XJ2)、普拉扣动球菌(Planococcus plakortidis XJ10)、德昌动性球菌(Planococcus dechangensis XJ11)和莱比托游动球菌(Planococcus rifietoensis XJ12)分别活化培养三次。将活化好的菌液于4℃、10000r/min离心10min，然后用无菌生理盐水洗涤两次，再悬浮于少量无菌生理盐水中，最后将菌液浓度调整到10 5～10 7CFU/mL，备用。
(3)混合发酵剂的添加：将海洋动性球菌(Planococcus maritimus XJ2)、普拉扣动球菌(Planococcus plakortidis XJ10)、德昌动性球菌(Planococcus dechangensis XJ11)和莱比托游动球菌(Planococcus rifietoensis XJ12)按菌数比列(1～3)：(1～3)：(1～3)：(1～3)进行混合并制备成混合发酵剂，按最终添加量(10 5～10 9CFU/g)混入到预处理过的原料碎鱼肉中，使最终菌数达到添加要求即可。
(4)鱼露的发酵：把步骤(3)得到的混合物料在温度15～30℃下保温发酵5～30d。
(5)鱼露的过滤：将步骤(4)得到的鱼露样品于120℃灭菌10～30min，冷却至室温后 于10000r/min离心20min，取上清液用多层纱布过滤以除去固形物和杂质。
(6)鱼露的灭菌：将步骤(5)得到的鱼露在100℃下进行二次灭菌15～30min，在无菌条件下灌装密封，即得鱼露成品。
(7)产品理化指标的评价：氨基酸态氮含量评价，方法参照GB5009.235-2016比色法；挥发性盐基氮含量评价，方法参照GB5009.228-2016微量扩散法；组胺含量的测定，方法参照GB5009.208-2016分光光度法。
海洋动性球菌(Planococcus maritimus XJ2)，该菌株已经于2019年01月02日保藏在位于北京市朝阳区北辰西路1号院3号的中国科学院微生物研究所的中国微生物菌种保藏管理委员会普通微生物中心(CGMCC)，编号为CGMCCNO.17057，建议的分类命名：Planococcus maritimus。
普拉扣动球菌(Planococcus plakortidis XJ10)，该菌株已经于2019年01月02日保藏在位于北京市朝阳区北辰西路1号院3号的中国科学院微生物研究所的中国微生物菌种保藏管理委员会普通微生物中心(CGMCC)，编号为CGMCCNO.17058，建议的分类命名：Planococcus plakortidis。
德昌动性球菌(Planococcus dechangensis XJ11)，该菌株已经于2019年01月02日保藏在位于北京市朝阳区北辰西路1号院3号的中国科学院微生物研究所的中国微生物菌种保藏管理委员会普通微生物中心(CGMCC)，编号为CGMCCNO.17059，建议的分类命名：Planococcus dechangensis。
莱比托游动球菌(Planococcus rifietoensis XJ12)，该菌株已经于2019年01月02日保藏在位于北京市朝阳区北辰西路1号院3号的中国科学院微生物研究所的中国微生物菌种保藏管理委员会普通微生物中心(CGMCC)，编号为CGMCCNO.17060，建议的分类命名：Planococcus rifietoensis。
本发明的有益效果
利用上述动性球菌菌株发酵鱼露的发酵周期大大缩短，发酵温度降低。此法获得的鱼露呈透明棕红色，具有水产品特有的香气，无悬浮或絮状等杂质。成品鱼露的盐含量低，氨基酸态氮含量较高(根据中国鱼露行业标准该鱼露划分为一级鱼露)，挥发性盐基氮的含量较低，组胺的含量低于国家标准，制得的鱼露滋味鲜美且营养价值丰富，因此该发酵剂可用于生产绿色安全的发酵鱼露产品。
综上所述，本发明已经具备了发明专利的独特性、取得突出的实质性特点和显著进步的创新性和实用性，产生了有益的效果。
具体实施方式
1.菌株的筛选纯化：
以传统低温发酵的新鲜虾酱(来源于山东省威海市某一农家餐馆自制的虾酱)为原材料，从虾酱中分离筛选出产低温蛋白酶的动性球菌。将新鲜虾酱稀释涂布于固体培养基上，15℃培养48h，挑取具有典型特征的菌落，进行多次划线分离纯化，最后获得产低温蛋白酶的动性球菌单菌落。将分离纯化后的单菌落接种到斜面上，25℃培养24h，保藏备用。
2.菌株的鉴定
2.1形态学鉴定
菌落呈橘黄色、表面光滑、边缘整齐、不透明、菌落致密；革兰氏染色为阳性；菌体呈球状，单个或成堆排列。
2.2生理特征
4株动性球菌在NaCl浓度为0～15％时均可进行生长；在pH为7～9时，生长较快；在温度范围为15℃～35℃都能进行良好生长；
2.3分子生物学鉴定
对筛选纯化后的动性球菌进行16SrRNA测序，确定为Planococcus maritimus、Planococcus plakortidis、Planococcus dechangensis以及Planococcus rifietoensis，并构建生物进化关系树。
3.菌株的保藏
4株动性球菌均已经于2019年01月02日保藏在位于北京市朝阳区北辰西路1号院3号的中国科学院微生物研究所的中国微生物菌种保藏管理委员会普通微生物中心(CGMCC)，
海洋动性球菌(Planococcus maritimus XJ2)，该菌株已经于2019年01月02日保藏在位于北京市朝阳区北辰西路1号院3号的中国科学院微生物研究所的中国微生物菌种保藏管理委员会普通微生物中心(CGMCC)，编号为CGMCCNO.17057，建议的分类命名：Planococcus maritimus。
普拉扣动球菌(Planococcus plakortidis XJ10)，该菌株已经于2019年01月02日保藏在位于北京市朝阳区北辰西路1号院3号的中国科学院微生物研究所的中国微生物菌种保藏管理委员会普通微生物中心(CGMCC)，编号为CGMCCNO.17058，建议的分类命名：Planococcus plakortidis。
德昌动性球菌(Planococcus dechangensis XJ11)，该菌株已经于2019年01月02日保藏在位于北京市朝阳区北辰西路1号院3号的中国科学院微生物研究所的中国微生物菌种保藏管理委员会普通微生物中心(CGMCC)，编号为CGMCCNO.17059，建议的分类命名：Planococcus dechangensis。
莱比托游动球菌(Planococcus rifietoensis XJ12)，该菌株已经于2019年01月02日保藏 在位于北京市朝阳区北辰西路1号院3号的中国科学院微生物研究所的中国微生物菌种保藏管理委员会普通微生物中心(CGMCC)，编号为CGMCCNO.17060，建议的分类命名：Planococcus rifietoensis。
实施例1：
一种利用海洋动性球菌Planococcus maritimus XJ2发酵鱼露的方法，按照下述步骤进行：
(1)原料的处理：将碎鱼肉(鱼肉加工时所产生的废弃物)用碎肉机绞碎，加入腌制海盐5％(w/w)混匀，备用。
(2)发酵剂的制备：将海洋动性球菌Planococcus maritimusXJ2(CGMCCNO.17057)活化培养三次。将活化好的菌液于4℃、10000r/min离心10min，然后用无菌生理盐水洗涤两次，再悬浮于少量无菌生理盐水中，最后将菌液浓度调整到10 5～10 7CFU/mL，备用。
(3)发酵剂的添加：将海洋动性球菌Planococcus maritimusXJ2(CGMCCNO.17057)按最终添加量(10 5CFU/g)混入到预处理过的原料碎鱼肉中，使最终菌数达到添加要求即可。
(4)鱼露的发酵：把步骤(3)得到的混合物料在温度15℃下保温发酵30d。
(5)鱼露的过滤：将步骤(4)得到的鱼露样品于120℃灭菌10min，冷却至室温后于10000r/min离心20min，取上清液用多层纱布过滤以除去固形物和杂质。
(6)鱼露的灭菌：将步骤(5)得到的鱼露在100℃下进行二次灭菌30min，在无菌条件下灌装密封，即得鱼露成品。
(7)产品理化指标的评价：氨基酸态氮含量评价，方法参照GB5009.235-2016比色法；挥发性盐基氮含量评价，方法参照GB5009.228-2016微量扩散法；组胺含量的测定，方法参照GB5009.208-2016分光光度法。
(8)鱼露理化指标结果：利用该株海洋动性球菌发酵鱼露的氨基酸态含量可达到1.284g/100mL，根据中国鱼露行业标准该鱼露划分为一级鱼露；挥发性盐基氮含量为107mg/100mL；组胺含量为19mg/100mL，低于欧盟标准40mg/100mL。本鱼露中原料中固有的土腥味大大减少，鲜味明显，与未添加菌株发酵的鱼露相比，风味有显著提升。
实施例2：
一种利用海洋动性球菌Planococcus maritimus XJ2发酵鱼露的方法，按照下述步骤进行：
(1)原料的处理：将碎鱼肉(鱼肉加工时所产生的废弃物)用碎肉机绞碎，加入腌制海盐10％(w/w)混匀，备用。
(2)发酵剂的制备：将海洋动性球菌Planococcus maritimusXJ2(CGMCCNO.17057)活化培养三次。将活化好的菌液于4℃、10000r/min离心10min，然后用无菌生理盐水洗涤两次，再悬浮于少量无菌生理盐水中，最后将菌液浓度调整到10 5～10 7CFU/mL，备用。
(3)发酵剂的添加：将海洋动性球菌Planococcus maritimusXJ2(CGMCCNO.17057)按最终添加量(10 7CFU/g)混入到预处理过的原料碎鱼肉中，使最终菌数达到添加要求即可。
(4)鱼露的发酵：把步骤(3)得到的混合物料在温度20℃下保温发酵15d。
(5)鱼露的过滤：将步骤(4)得到的鱼露样品于120℃灭菌20min，冷却至室温后于10000r/min离心20min，取上清液用多层纱布过滤以除去固形物和杂质。
(6)鱼露的灭菌：将步骤(5)得到的鱼露在100℃下进行二次灭菌20min，在无菌条件下灌装密封，即得鱼露成品。
(7)产品理化指标的评价：氨基酸态氮含量评价，方法参照GB5009.235-2016比色法；挥发性盐基氮含量评价，方法参照GB5009.228-2016微量扩散法；组胺含量的测定，方法参照GB5009.208-2016分光光度法。
(8)鱼露理化指标结果：利用该株海洋动性球菌发酵鱼露的氨基酸态含量可达到1.023g/100mL，根据中国鱼露行业标准该鱼露划分为一级鱼露；挥发性盐基氮含量为94mg/100mL；组胺含量为17mg/100mL，低于欧盟标准40mg/100mL。本鱼露中原料中固有的土腥味大大减少，鲜味明显，与未添加菌株发酵的鱼露相比，风味有显著提升。
实施例3：
一种利用海洋动性球菌Planococcus maritimus XJ2发酵鱼露的方法，按照下述步骤进行：
(1)原料的处理：将碎鱼肉(鱼肉加工时所产生的废弃物)用碎肉机绞碎，加入腌制海盐15％(w/w)混匀，备用。
(2)发酵剂的制备：将海洋动性球菌Planococcus maritimusXJ2(CGMCCNO.17057)活化培养三次。将活化好的菌液于4℃、10000r/min离心10min，然后用无菌生理盐水洗涤两次，再悬浮于少量无菌生理盐水中，最后将菌液浓度调整到10 5～10 7CFU/mL，备用。
(3)发酵剂的添加：将海洋动性球菌Planococcus maritimusXJ2(CGMCCNO.17057)按最终添加量(10 9CFU/g)混入到预处理过的原料碎鱼肉中，使最终菌数达到添加要求即可。
(4)鱼露的发酵：把步骤(3)得到的混合物料在温度30℃下保温发酵5d。
(5)鱼露的过滤：将步骤(4)得到的鱼露样品于120℃灭菌30min，冷却至室温后于10000r/min离心20min，取上清液用多层纱布过滤以除去固形物和杂质。
(6)鱼露的灭菌：将步骤(5)得到的鱼露在100℃下进行二次灭菌15min，在无菌条件下灌装密封，即得鱼露成品。
(7)产品理化指标的评价：氨基酸态氮含量评价，方法参照GB5009.235-2016比色法；挥发性盐基氮含量评价，方法参照GB5009.228-2016微量扩散法；组胺含量的测定，方法参照GB5009.208-2016分光光度法。
(8)鱼露理化指标结果：利用该株海洋动性球菌发酵鱼露的氨基酸态含量可达到0.97g/100mL，根据中国鱼露行业标准该鱼露划分为一级鱼露；挥发性盐基氮含量为85mg/100mL；组胺含量为11mg/100mL，低于欧盟标准40mg/100mL。本鱼露中原料中固有的土腥味大大减少，鲜味明显，与未添加菌株发酵的鱼露相比，风味有显著提升。
实施例4：
一种利用普拉扣动球菌Planococcus plakortidis XJ10发酵鱼露的方法，按照下述步骤进行：
(1)原料的处理：将碎鱼肉(鱼肉加工时所产生的废弃物)用碎肉机绞碎，加入腌制海盐5％(w/w)混匀，备用。
(2)发酵剂的制备：将普拉扣动球菌Planococcus plakortidis XJ10(CGMCCNO.17058)活化培养三次。将活化好的菌液于4℃、10000r/min离心10min，然后用无菌生理盐水洗涤两次，再悬浮于少量无菌生理盐水中，最后将菌液浓度调整到10 5～10 7CFU/mL，备用。
(3)发酵剂的添加：将普拉扣动球菌Planococcus plakortidis XJ10(CGMCCNO.17058)按最终添加量(10 5CFU/g)混入到预处理过的原料碎鱼肉中，使最终菌数达到添加要求即可。
(4)鱼露的发酵：把步骤(3)得到的混合物料在温度15℃下保温发酵30d。
(5)鱼露的过滤：将步骤(4)得到的鱼露样品于120℃灭菌10min，冷却至室温后于10000r/min离心20min，取上清液用多层纱布过滤以除去固形物和杂质。
(6)鱼露的灭菌：将步骤(5)得到的鱼露在100℃下进行二次灭菌30min，在无菌条件下灌装密封，即得鱼露成品。
(7)产品理化指标的评价：氨基酸态氮含量评价，方法参照GB5009.235-2016比色法；挥发性盐基氮含量评价，方法参照GB5009.228-2016微量扩散法；组胺含量的测定，方法参照GB5009.208-2016分光光度法。
(8)鱼露理化指标结果：利用该株普拉扣动球菌发酵鱼露的氨基酸态含量可达到1.117g/100mL，根据中国鱼露行业标准该鱼露划分为一级鱼露；挥发性盐基氮含量为112mg/100mL；组胺含量为20mg/100mL，低于欧盟标准40mg/100mL。本鱼露中原料中固有的土腥味大大减少，鲜味明显，与未添加菌株发酵的鱼露相比，风味有显著提升。
实施例5：
一种利用普拉扣动球菌Planococcus plakortidis XJ10发酵鱼露的方法，按照下述步骤进行：
(1)原料的处理：将碎鱼肉(鱼肉加工时所产生的废弃物)用碎肉机绞碎，加入腌制海盐10％(w/w)混匀，备用。
(2)发酵剂的制备：将普拉扣动球菌Planococcus plakortidis XJ10(CGMCCNO.17058)活化培养三次。将活化好的菌液于4℃、10000r/min离心10min，然后用无菌生理盐水洗涤 两次，再悬浮于少量无菌生理盐水中，最后将菌液浓度调整到10 5～10 7CFU/mL，备用。
(3)发酵剂的添加：将普拉扣动球菌Planococcus plakortidis XJ10(CGMCCNO.17058)按最终添加量(10 7CFU/g)混入到预处理过的原料碎鱼肉中，使最终菌数达到添加要求即可。
(4)鱼露的发酵：把步骤(3)得到的混合物料在温度20℃下保温发酵15d。
(5)鱼露的过滤：将步骤(4)得到的鱼露样品于120℃灭菌20min，冷却至室温后于10000r/min离心20min，取上清液用多层纱布过滤以除去固形物和杂质。
(6)鱼露的灭菌：将步骤(5)得到的鱼露在100℃下进行二次灭菌20min，在无菌条件下灌装密封，即得鱼露成品。
(7)产品理化指标的评价：氨基酸态氮含量评价，方法参照GB5009.235-2016比色法；挥发性盐基氮含量评价，方法参照GB5009.228-2016微量扩散法；组胺含量的测定，方法参照GB5009.208-2016分光光度法。
(8)鱼露理化指标结果：利用该株普拉扣动球菌发酵鱼露的氨基酸态含量可达到1.103g/100mL，根据中国鱼露行业标准该鱼露划分为一级鱼露；挥发性盐基氮含量为99mg/100mL；组胺含量为18mg/100mL，低于欧盟标准40mg/100mL。本鱼露中原料中固有的土腥味大大减少，鲜味明显，与未添加菌株发酵的鱼露相比，风味有显著提升。
实施例6：
一种利用普拉扣动球菌Planococcus plakortidis XJ10发酵鱼露的方法，按照下述步骤进行：
(1)原料的处理：将碎鱼肉(鱼肉加工时所产生的废弃物)用碎肉机绞碎，加入腌制海盐15％(w/w)混匀，备用。
(2)发酵剂的制备：将普拉扣动球菌Planococcus plakortidis XJ10(CGMCCNO.17058)活化培养三次。将活化好的菌液于4℃、10000r/min离心10min，然后用无菌生理盐水洗涤两次，再悬浮于少量无菌生理盐水中，最后将菌液浓度调整到10 5～10 7CFU/mL，备用。
(3)发酵剂的添加：将普拉扣动球菌Planococcus plakortidis XJ10(CGMCCNO.17058)按最终添加量(10 9CFU/g)混入到预处理过的原料碎鱼肉中，使最终菌数达到添加要求即可。
(4)鱼露的发酵：把步骤(3)得到的混合物料在温度30℃下保温发酵5d。
(5)鱼露的过滤：将步骤(4)得到的鱼露样品于120℃灭菌30min，冷却至室温后于10000r/min离心20min，取上清液用多层纱布过滤以除去固形物和杂质。
(6)鱼露的灭菌：将步骤(5)得到的鱼露在100℃下进行二次灭菌15min，在无菌条件下灌装密封，即得鱼露成品。
(7)产品理化指标的评价：氨基酸态氮含量评价，方法参照GB5009.235-2016比色法；挥发性盐基氮含量评价，方法参照GB5009.228-2016微量扩散法；组胺含量的测定，方法参 照GB5009.208-2016分光光度法。
(8)鱼露理化指标结果：利用该株普拉扣动球菌发酵鱼露的氨基酸态含量可达到1.008g/100mL，根据中国鱼露行业标准该鱼露划分为一级鱼露；挥发性盐基氮含量为92mg/100mL；组胺含量为17mg/100mL，低于欧盟标准40mg/100mL。本鱼露中原料中固有的土腥味大大减少，鲜味明显，与未添加菌株发酵的鱼露相比，风味有显著提升。
实施例7：
一种利用德昌动性球菌Planococcus dechangensis XJ11发酵鱼露的方法，按照下述步骤进行：
(1)原料的处理：将碎鱼肉(鱼肉加工时所产生的废弃物)用碎肉机绞碎，加入腌制海盐5％(w/w)混匀，备用。
(2)发酵剂的制备：将德昌动性球菌Planococcus dechangensis XJ11(CGMCCNO.17059)活化培养三次。将活化好的菌液于4℃、10000r/min离心10min，然后用无菌生理盐水洗涤两次，再悬浮于少量无菌生理盐水中，最后将菌液浓度调整到10 5～10 7CFU/mL，备用。
(3)发酵剂的添加：将德昌动性球菌Planococcus dechangensis XJ11(CGMCCNO.17059)按最终添加量(10 5CFU/g)混入到预处理过的原料碎鱼肉中，使最终菌数达到添加要求即可。
(4)鱼露的发酵：把步骤(3)得到的混合物料在温度15℃下保温发酵30d。
(5)鱼露的过滤：将步骤(4)得到的鱼露样品于120℃灭菌10min，冷却至室温后于10000r/min离心20min，取上清液用多层纱布过滤以除去固形物和杂质。
(6)鱼露的灭菌：将步骤(5)得到的鱼露在100℃下进行二次灭菌30min，在无菌条件下灌装密封，即得鱼露成品。
(7)产品理化指标的评价：氨基酸态氮含量评价，方法参照GB5009.235-2016比色法；挥发性盐基氮含量评价，方法参照GB5009.228-2016微量扩散法；组胺含量的测定，方法参照GB5009.208-2016分光光度法。
(8)鱼露理化指标结果：利用该株德昌动性球菌发酵鱼露的氨基酸态含量可达到1.216g/100mL，根据中国鱼露行业标准该鱼露划分为一级鱼露；挥发性盐基氮含量为109mg/100mL；组胺含量为19mg/100mL，低于欧盟标准40mg/100mL。本鱼露中原料中固有的土腥味大大减少，鲜味明显，与未添加菌株发酵的鱼露相比，风味有显著提升。
实施例8：
一种利用德昌动性球菌Planococcus dechangensis XJ11发酵鱼露的方法，按照下述步骤进行：
(1)原料的处理：将碎鱼肉(鱼肉加工时所产生的废弃物)用碎肉机绞碎，加入腌制海 盐10％(w/w)混匀，备用。
(2)发酵剂的制备：将德昌动性球菌Planococcus dechangensis XJ11(CGMCCNO.17059)活化培养三次。将活化好的菌液于4℃、10000r/min离心10min，然后用无菌生理盐水洗涤两次，再悬浮于少量无菌生理盐水中，最后将菌液浓度调整到10 5～10 7CFU/mL，备用。
(3)发酵剂的添加：将德昌动性球菌Planococcus dechangensis XJ11(CGMCCNO.17059)按最终添加量(10 7CFU/g)混入到预处理过的原料碎鱼肉中，使最终菌数达到添加要求即可。
(4)鱼露的发酵：把步骤(3)得到的混合物料在温度20℃下保温发酵15d。
(5)鱼露的过滤：将步骤(4)得到的鱼露样品于120℃灭菌20min，冷却至室温后于10000r/min离心20min，取上清液用多层纱布过滤以除去固形物和杂质。
(6)鱼露的灭菌：将步骤(5)得到的鱼露在100℃下进行二次灭菌20min，在无菌条件下灌装密封，即得鱼露成品。
(7)产品理化指标的评价：氨基酸态氮含量评价，方法参照GB5009.235-2016比色法；挥发性盐基氮含量评价，方法参照GB5009.228-2016微量扩散法；组胺含量的测定，方法参照GB5009.208-2016分光光度法。
(8)鱼露理化指标结果：利用该株德昌动性球菌发酵鱼露的氨基酸态含量可达到1.174g/100mL，根据中国鱼露行业标准该鱼露划分为一级鱼露；挥发性盐基氮含量为98mg/100mL；组胺含量为20mg/100mL，低于欧盟标准40mg/100mL。本鱼露中原料中固有的土腥味大大减少，鲜味明显，与未添加菌株发酵的鱼露相比，风味有显著提升。
实施例9：
一种利用德昌动性球菌Planococcus dechangensis XJ11发酵鱼露的方法，按照下述步骤进行：
(1)原料的处理：将碎鱼肉(鱼肉加工时所产生的废弃物)用碎肉机绞碎，加入腌制海盐15％(w/w)混匀，备用。
(2)发酵剂的制备：将德昌动性球菌Planococcus dechangensis XJ11(CGMCCNO.17059)活化培养三次。将活化好的菌液于4℃、10000r/min离心10min，然后用无菌生理盐水洗涤两次，再悬浮于少量无菌生理盐水中，最后将菌液浓度调整到10 5～10 7CFU/mL，备用。
(3)发酵剂的添加：将德昌动性球菌Planococcus dechangensis XJ11(CGMCCNO.17059)按最终添加量(10 9CFU/g)混入到预处理过的原料碎鱼肉中，使最终菌数达到添加要求即可。
(4)鱼露的发酵：把步骤(3)得到的混合物料在温度30℃下保温发酵5d。
(5)鱼露的过滤：将步骤(4)得到的鱼露样品于120℃灭菌30min，冷却至室温后于10000r/min离心20min，取上清液用多层纱布过滤以除去固形物和杂质。
(6)鱼露的灭菌：将步骤(5)得到的鱼露在100℃下进行二次灭菌15min，在无菌条件下灌装密封，即得鱼露成品。
(7)产品理化指标的评价：氨基酸态氮含量评价，方法参照GB5009.235-2016比色法；挥发性盐基氮含量评价，方法参照GB5009.228-2016微量扩散法；组胺含量的测定，方法参照GB5009.208-2016分光光度法。
(8)鱼露理化指标结果：利用该株德昌动性球菌发酵鱼露的氨基酸态含量可达到1.089g/100mL，根据中国鱼露行业标准该鱼露划分为一级鱼露；挥发性盐基氮含量为92mg/100mL；组胺含量为14mg/100mL，低于欧盟标准40mg/100mL。本鱼露中原料中固有的土腥味大大减少，鲜味明显，与未添加菌株发酵的鱼露相比，风味有显著提升。
实施例10：
一种利用莱比托游动球菌Planococcus rifietoensis XJ12发酵鱼露的方法，按照下述步骤进行：
(1)原料的处理：将碎鱼肉(鱼肉加工时所产生的废弃物)用碎肉机绞碎，加入腌制海盐5％(w/w)混匀，备用。
(2)发酵剂的制备：将莱比托游动球菌Planococcus rifietoensis XJ12(CGMCCNO.17060)活化培养三次。将活化好的菌液于4℃、10000r/min离心10min，然后用无菌生理盐水洗涤两次，再悬浮于少量无菌生理盐水中，最后将菌液浓度调整到10 5～10 7CFU/mL，备用。
(3)发酵剂的添加：将莱比托游动球菌Planococcus rifietoensis XJ12(CGMCCNO.17060)按最终添加量(10 5CFU/g)混入到预处理过的原料碎鱼肉中，使最终菌数达到添加要求即可。
(4)鱼露的发酵：把步骤(3)得到的混合物料在温度15℃下保温发酵30d。
(5)鱼露的过滤：将步骤(4)得到的鱼露样品于120℃灭菌10min，冷却至室温后于10000r/min离心20min，取上清液用多层纱布过滤以除去固形物和杂质。
(6)鱼露的灭菌：将步骤(5)得到的鱼露在100℃下进行二次灭菌30min，在无菌条件下灌装密封，即得鱼露成品。
(7)产品理化指标的评价：氨基酸态氮含量评价，方法参照GB5009.235-2016比色法；挥发性盐基氮含量评价，方法参照GB5009.228-2016微量扩散法；组胺含量的测定，方法参照GB5009.208-2016分光光度法。
(8)鱼露理化指标结果：利用该株莱比托游动球菌发酵鱼露的氨基酸态含量可达到1.159g/100mL，根据中国鱼露行业标准该鱼露划分为一级鱼露；挥发性盐基氮含量为104mg/100mL；组胺含量为16mg/100mL，低于欧盟标准40mg/100mL。本鱼露中原料中固有的土腥味大大减少，鲜味明显，与未添加菌株发酵的鱼露相比，风味有显著提升。
实施例11：
一种利用莱比托游动球菌Planococcus rifietoensis XJ12发酵鱼露的方法，按照下述步骤进行：
(1)原料的处理：将碎鱼肉(鱼肉加工时所产生的废弃物)用碎肉机绞碎，加入腌制海盐10％(w/w)混匀，备用。
(2)发酵剂的制备：将莱比托游动球菌Planococcus rifietoensis XJ12(CGMCCNO.17060)活化培养三次。将活化好的菌液于4℃、10000r/min离心10min，然后用无菌生理盐水洗涤两次，再悬浮于少量无菌生理盐水中，最后将菌液浓度调整到10 5～10 7CFU/mL，备用。
(3)发酵剂的添加：将莱比托游动球菌Planococcus rifietoensis XJ12(CGMCCNO.17060)按最终添加量(10 7CFU/g)混入到预处理过的原料碎鱼肉中，使最终菌数达到添加要求即可。
(4)鱼露的发酵：把步骤(3)得到的混合物料在温度20℃下保温发酵15d。
(5)鱼露的过滤：将步骤(4)得到的鱼露样品于120℃灭菌20min，冷却至室温后于10000r/min离心20min，取上清液用多层纱布过滤以除去固形物和杂质。
(6)鱼露的灭菌：将步骤(5)得到的鱼露在100℃下进行二次灭菌20min，在无菌条件下灌装密封，即得鱼露成品。
(7)产品理化指标的评价：氨基酸态氮含量评价，方法参照GB5009.235-2016比色法；挥发性盐基氮含量评价，方法参照GB5009.228-2016微量扩散法；组胺含量的测定，方法参照GB5009.208-2016分光光度法。
(8)鱼露理化指标结果：利用该株莱比托游动球菌发酵鱼露的氨基酸态含量可达到1.207g/100mL，根据中国鱼露行业标准该鱼露划分为一级鱼露；挥发性盐基氮含量为109mg/100mL；组胺含量为21mg/100mL，低于欧盟标准40mg/100mL。本鱼露中原料中固有的土腥味大大减少，鲜味明显，与未添加菌株发酵的鱼露相比，风味有显著提升。
实施例12：
一种利用莱比托游动球菌Planococcus rifietoensis XJ12发酵鱼露的方法，按照下述步骤进行：
(1)原料的处理：将碎鱼肉(鱼肉加工时所产生的废弃物)用碎肉机绞碎，加入腌制海盐15％(w/w)混匀，备用。
(2)发酵剂的制备：将莱比托游动球菌Planococcus rifietoensis XJ12(CGMCCNO.17060)活化培养三次。将活化好的菌液于4℃、10000r/min离心10min，然后用无菌生理盐水洗涤两次，再悬浮于少量无菌生理盐水中，最后将菌液浓度调整到10 5～10 7CFU/mL，备用。
(3)发酵剂的添加：将莱比托游动球菌Planococcus rifietoensis XJ12(CGMCCNO.17060) 按最终添加量(10 9CFU/g)混入到预处理过的原料碎鱼肉中，使最终菌数达到添加要求即可。
(4)鱼露的发酵：把步骤(3)得到的混合物料在温度30℃下保温发酵5d。
(5)鱼露的过滤：将步骤(4)得到的鱼露样品于120℃灭菌30min，冷却至室温后于10000r/min离心20min，取上清液用多层纱布过滤以除去固形物和杂质。
(6)鱼露的灭菌：将步骤(5)得到的鱼露在100℃下进行二次灭菌15min，在无菌条件下灌装密封，即得鱼露成品。
(7)产品理化指标的评价：氨基酸态氮含量评价，方法参照GB5009.235-2016比色法；挥发性盐基氮含量评价，方法参照GB5009.228-2016微量扩散法；组胺含量的测定，方法参照GB5009.208-2016分光光度法。
(8)鱼露理化指标结果：利用该株莱比托游动球菌发酵鱼露的氨基酸态含量可达到1.012g/100mL，根据中国鱼露行业标准该鱼露划分为一级鱼露；挥发性盐基氮含量为98mg/100mL；组胺含量为14mg/100mL，低于欧盟标准40mg/100mL。本鱼露中原料中固有的土腥味大大减少，鲜味明显，与未添加菌株发酵的鱼露相比，风味有显著提升。
实施例13：
一种利用动性球菌混合菌株发酵鱼露的方法，按照下述步骤进行：
(1)原料的处理：将碎鱼肉(鱼肉加工时所产生的废弃物)用碎肉机绞碎，加入腌制海盐5％(w/w)混匀，备用。
(2)混合发酵剂的制备：将海洋动性球菌(Planococcus maritimus XJ2)、普拉扣动球菌(Planococcus plakortidis XJ10)、德昌动性球菌(Planococcus dechangensis XJ11)和莱比托游动球菌(Planococcus rifietoensis XJ12)分别活化培养三次。将活化好的菌液于4℃、10000r/min离心10min，然后用无菌生理盐水洗涤两次，再悬浮于少量无菌生理盐水中，最后将菌液浓度调整到10 5～10 7CFU/mL，备用。
(3)混合发酵剂的添加：将海洋动性球菌(Planococcus maritimus XJ2)、普拉扣动球菌(Planococcus plakortidis XJ10)、德昌动性球菌(Planococcus dechangensis XJ11)和莱比托游动球菌(Planococcus rifietoensis XJ12)按菌数比列1:1:1:1进行混合并制备成混合发酵剂，按最终添加量10 5CFU/g混入到预处理过的原料碎鱼肉中，使最终菌数达到添加要求即可。
(4)鱼露的发酵：把步骤(3)得到的混合物料在温度15℃下保温发酵5d。
(5)鱼露的过滤：将步骤(4)得到的鱼露样品于120℃灭菌10min，冷却至室温后于10000r/min离心20min，取上清液用多层纱布过滤以除去固形物和杂质。
(6)鱼露的灭菌：将步骤(5)得到的鱼露在100℃下进行二次灭菌30min，在无菌条件下灌装密封，即得鱼露成品。
(7)产品理化指标的评价：氨基酸态氮含量评价，方法参照GB5009.235-2016比色法；挥发性盐基氮含量评价，方法参照GB5009.228-2016微量扩散法；组胺含量的测定，方法参照GB5009.208-2016分光光度法。
(8)鱼露理化指标结果：利用该株海洋动性球菌发酵鱼露的氨基酸态含量可达到0.974g/100mL，根据中国鱼露行业标准该鱼露划分为一级鱼露；挥发性盐基氮含量为113mg/100mL；组胺含量为17mg/100mL，低于欧盟标准40mg/100mL。本鱼露中原料中固有的土腥味大大减少，鲜味明显，与未添加菌株发酵的鱼露相比，风味有显著提升。
实施例14：
一种利用动性球菌混合菌株发酵鱼露的方法，按照下述步骤进行：
(1)原料的处理：将碎鱼肉(鱼肉加工时所产生的废弃物)用碎肉机绞碎，加入腌制海盐5％(w/w)混匀，备用。
(2)混合发酵剂的制备：将海洋动性球菌(Planococcus maritimus XJ2)、普拉扣动球菌(Planococcus plakortidis XJ10)、德昌动性球菌(Planococcus dechangensis XJ11)和莱比托游动球菌(Planococcus rifietoensis XJ12)分别活化培养三次。将活化好的菌液于4℃、10000r/min离心10min，然后用无菌生理盐水洗涤两次，再悬浮于少量无菌生理盐水中，最后将菌液浓度调整到10 5～10 7CFU/mL，备用。
(3)混合发酵剂的添加：将海洋动性球菌(Planococcus maritimus XJ2)、普拉扣动球菌(Planococcus plakortidis XJ10)、德昌动性球菌(Planococcus dechangensis XJ11)和莱比托游动球菌(Planococcus rifietoensis XJ12)按菌数比列1:2:2:1进行混合并制备成混合发酵剂，按最终添加量10 5CFU/g混入到预处理过的原料碎鱼肉中，使最终菌数达到添加要求即可。
(4)鱼露的发酵：把步骤(3)得到的混合物料在温度15℃下保温发酵30d。
(5)鱼露的过滤：将步骤(4)得到的鱼露样品于120℃灭菌10min，冷却至室温后于10000r/min离心20min，取上清液用多层纱布过滤以除去固形物和杂质。
(6)鱼露的灭菌：将步骤(5)得到的鱼露在100℃下进行二次灭菌30min，在无菌条件下灌装密封，即得鱼露成品。
(7)产品理化指标的评价：氨基酸态氮含量评价，方法参照GB5009.235-2016比色法；挥发性盐基氮含量评价，方法参照GB5009.228-2016微量扩散法；组胺含量的测定，方法参照GB5009.208-2016分光光度法。
(8)鱼露理化指标结果：利用该株海洋动性球菌发酵鱼露的氨基酸态含量可达到1.327g/100mL，根据中国鱼露行业标准该鱼露划分为一级鱼露；挥发性盐基氮含量为125mg/100mL；组胺含量为23mg/100mL，低于欧盟标准40mg/100mL。本鱼露中原料中固有的土腥味 大大减少，鲜味明显，与未添加菌株发酵的鱼露相比，风味有显著提升。
实施例15：
一种利用动性球菌混合菌株发酵鱼露的方法，按照下述步骤进行：
(1)原料的处理：将碎鱼肉(鱼肉加工时所产生的废弃物)用碎肉机绞碎，加入腌制海盐8％(w/w)混匀，备用。
(2)混合发酵剂的制备：将海洋动性球菌(Planococcus maritimus XJ2)、普拉扣动球菌(Planococcus plakortidis XJ10)、德昌动性球菌(Planococcus dechangensis XJ11)和莱比托游动球菌(Planococcus rifietoensis XJ12)分别活化培养三次。将活化好的菌液于4℃、10000r/min离心10min，然后用无菌生理盐水洗涤两次，再悬浮于少量无菌生理盐水中，最后将菌液浓度调整到10 5～10 7CFU/mL，备用。
(3)混合发酵剂的添加：将海洋动性球菌(Planococcus maritimus XJ2)、普拉扣动球菌(Planococcus plakortidis XJ10)、德昌动性球菌(Planococcus dechangensis XJ11)和莱比托游动球菌(Planococcus rifietoensis XJ12)按菌数比列1:2:1:1进行混合并制备成混合发酵剂，按最终添加量10 6CFU/g混入到预处理过的原料碎鱼肉中，使最终菌数达到添加要求即可。
(4)鱼露的发酵：把步骤(3)得到的混合物料在温度20℃下保温发酵10d。
(5)鱼露的过滤：将步骤(4)得到的鱼露样品于120℃灭菌15min，冷却至室温后于10000r/min离心20min，取上清液用多层纱布过滤以除去固形物和杂质。
(6)鱼露的灭菌：将步骤(5)得到的鱼露在100℃下进行二次灭菌15min，在无菌条件下灌装密封，即得鱼露成品。
(7)产品理化指标的评价：氨基酸态氮含量评价，方法参照GB5009.235-2016比色法；挥发性盐基氮含量评价，方法参照GB5009.228-2016微量扩散法；组胺含量的测定，方法参照GB5009.208-2016分光光度法。
(8)鱼露理化指标结果：利用该株海洋动性球菌发酵鱼露的氨基酸态含量可达到1.034g/100mL，根据中国鱼露行业标准该鱼露划分为一级鱼露；挥发性盐基氮含量为119mg/100mL；组胺含量为20mg/100mL，低于欧盟标准40mg/100mL。本鱼露中原料中固有的土腥味大大减少，鲜味明显，与未添加菌株发酵的鱼露相比，风味有显著提升。
实施例16：
一种利用动性球菌混合菌株发酵鱼露的方法，按照下述步骤进行：
(1)原料的处理：将碎鱼肉(鱼肉加工时所产生的废弃物)用碎肉机绞碎，加入腌制海盐10％(w/w)混匀，备用。
(2)混合发酵剂的制备：将海洋动性球菌(Planococcus maritimus XJ2)、普拉扣动球菌 (Planococcus plakortidis XJ10)、德昌动性球菌(Planococcus dechangensis XJ11)和莱比托游动球菌(Planococcus rifietoensis XJ12)分别活化培养三次。将活化好的菌液于4℃、10000r/min离心10min，然后用无菌生理盐水洗涤两次，再悬浮于少量无菌生理盐水中，最后将菌液浓度调整到10 5～10 7CFU/mL，备用。
(3)混合发酵剂的添加：将海洋动性球菌(Planococcus maritimus XJ2)、普拉扣动球菌(Planococcus plakortidis XJ10)、德昌动性球菌(Planococcus dechangensis XJ11)和莱比托游动球菌(Planococcus rifietoensis XJ12)按菌数比列1:2:1:2进行混合并制备成混合发酵剂，按最终添加量10 6CFU/g混入到预处理过的原料碎鱼肉中，使最终菌数达到添加要求即可。
(4)鱼露的发酵：把步骤(3)得到的混合物料在温度20℃下保温发酵15d。
(5)鱼露的过滤：将步骤(4)得到的鱼露样品于120℃灭菌15min，冷却至室温后于10000r/min离心20min，取上清液用多层纱布过滤以除去固形物和杂质。
(6)鱼露的灭菌：将步骤(5)得到的鱼露在100℃下进行二次灭菌15min，在无菌条件下灌装密封，即得鱼露成品。
(7)产品理化指标的评价：氨基酸态氮含量评价，方法参照GB5009.235-2016比色法；挥发性盐基氮含量评价，方法参照GB5009.228-2016微量扩散法；组胺含量的测定，方法参照GB5009.208-2016分光光度法。
(8)鱼露理化指标结果：利用该株海洋动性球菌发酵鱼露的氨基酸态含量可达到1.116g/100mL，根据中国鱼露行业标准该鱼露划分为一级鱼露；挥发性盐基氮含量为108mg/100mL；组胺含量为19mg/100mL，低于欧盟标准40mg/100mL。本鱼露中原料中固有的土腥味大大减少，鲜味明显，与未添加菌株发酵的鱼露相比，风味有显著提升。
实施例17：
一种利用动性球菌混合菌株发酵鱼露的方法，按照下述步骤进行：
(1)原料的处理：将碎鱼肉(鱼肉加工时所产生的废弃物)用碎肉机绞碎，加入腌制海盐10％(w/w)混匀，备用。
(2)混合发酵剂的制备：将海洋动性球菌(Planococcus maritimus XJ2)、普拉扣动球菌(Planococcus plakortidis XJ10)、德昌动性球菌(Planococcus dechangensis XJ11)和莱比托游动球菌(Planococcus rifietoensis XJ12)分别活化培养三次。将活化好的菌液于4℃、10000r/min离心10min，然后用无菌生理盐水洗涤两次，再悬浮于少量无菌生理盐水中，最后将菌液浓度调整到10 5～10 7CFU/mL，备用。
(3)混合发酵剂的添加：将海洋动性球菌(Planococcus maritimus XJ2)、普拉扣动球菌(Planococcus plakortidis XJ10)、德昌动性球菌(Planococcus dechangensis XJ11)和莱比托游 动球菌(Planococcus rifietoensis XJ12)按菌数比列3:1:1:2进行混合并制备成混合发酵剂，按最终添加量10 7CFU/g混入到预处理过的原料碎鱼肉中，使最终菌数达到添加要求即可。
(4)鱼露的发酵：把步骤(3)得到的混合物料在温度25℃下保温发酵20d。
(5)鱼露的过滤：将步骤(4)得到的鱼露样品于120℃灭菌30min，冷却至室温后于10000r/min离心20min，取上清液用多层纱布过滤以除去固形物和杂质。
(6)鱼露的灭菌：将步骤(5)得到的鱼露在100℃下进行二次灭菌15min，在无菌条件下灌装密封，即得鱼露成品。
(7)产品理化指标的评价：氨基酸态氮含量评价，方法参照GB5009.235-2016比色法；挥发性盐基氮含量评价，方法参照GB5009.228-2016微量扩散法；组胺含量的测定，方法参照GB5009.208-2016分光光度法。
(8)鱼露理化指标结果：利用该株海洋动性球菌发酵鱼露的氨基酸态含量可达到1.124g/100mL，根据中国鱼露行业标准该鱼露划分为一级鱼露；挥发性盐基氮含量为110mg/100mL；组胺含量为21mg/100mL，低于欧盟标准40mg/100mL。本鱼露中原料中固有的土腥味大大减少，鲜味明显，与未添加菌株发酵的鱼露相比，风味有显著提升。
实施例18：
一种利用动性球菌混合菌株发酵鱼露的方法，按照下述步骤进行：
(1)原料的处理：将碎鱼肉(鱼肉加工时所产生的废弃物)用碎肉机绞碎，加入腌制海盐12％(w/w)混匀，备用。
(2)混合发酵剂的制备：将海洋动性球菌(Planococcus maritimus XJ2)、普拉扣动球菌(Planococcus plakortidis XJ10)、德昌动性球菌(Planococcus dechangensis XJ11)和莱比托游动球菌(Planococcus rifietoensis XJ12)分别活化培养三次。将活化好的菌液于4℃、10000r/min离心10min，然后用无菌生理盐水洗涤两次，再悬浮于少量无菌生理盐水中，最后将菌液浓度调整到10 5～10 7CFU/mL，备用。
(3)混合发酵剂的添加：将海洋动性球菌(Planococcus maritimus XJ2)、普拉扣动球菌(Planococcus plakortidis XJ10)、德昌动性球菌(Planococcus dechangensis XJ11)和莱比托游动球菌(Planococcus rifietoensis XJ12)按菌数比列1:1:2:1进行混合并制备成混合发酵剂，按最终添加量10 8CFU/g混入到预处理过的原料碎鱼肉中，使最终菌数达到添加要求即可。
(4)鱼露的发酵：把步骤(3)得到的混合物料在温度25℃下保温发酵25d。
(5)鱼露的过滤：将步骤(4)得到的鱼露样品于120℃灭菌30min，冷却至室温后于10000r/min离心20min，取上清液用多层纱布过滤以除去固形物和杂质。
(6)鱼露的灭菌：将步骤(5)得到的鱼露在100℃下进行二次灭菌15min，在无菌条 件下灌装密封，即得鱼露成品。
(7)产品理化指标的评价：氨基酸态氮含量评价，方法参照GB5009.235-2016比色法；挥发性盐基氮含量评价，方法参照GB5009.228-2016微量扩散法；组胺含量的测定，方法参照GB5009.208-2016分光光度法。
(8)鱼露理化指标结果：利用该株海洋动性球菌发酵鱼露的氨基酸态含量可达到1.009g/100mL，根据中国鱼露行业标准该鱼露划分为一级鱼露；挥发性盐基氮含量为112mg/100mL；组胺含量为20mg/100mL，低于欧盟标准40mg/100mL。本鱼露中原料中固有的土腥味大大减少，鲜味明显，与未添加菌株发酵的鱼露相比，风味有显著提升。
实施例19：
一种利用动性球菌混合菌株发酵鱼露的方法，按照下述步骤进行：
(1)原料的处理：将碎鱼肉(鱼肉加工时所产生的废弃物)用碎肉机绞碎，加入腌制海盐15％(w/w)混匀，备用。
(2)混合发酵剂的制备：将海洋动性球菌(Planococcus maritimus XJ2)、普拉扣动球菌(Planococcus plakortidis XJ10)、德昌动性球菌(Planococcus dechangensis XJ11)和莱比托游动球菌(Planococcus rifietoensis XJ12)分别活化培养三次。将活化好的菌液于4℃、10000r/min离心10min，然后用无菌生理盐水洗涤两次，再悬浮于少量无菌生理盐水中，最后将菌液浓度调整到10 5～10 7CFU/mL，备用。
(3)混合发酵剂的添加：将海洋动性球菌(Planococcus maritimus XJ2)、普拉扣动球菌(Planococcus plakortidis XJ10)、德昌动性球菌(Planococcus dechangensis XJ11)和莱比托游动球菌(Planococcus rifietoensis XJ12)按菌数比列1:1:3:1进行混合并制备成混合发酵剂，按最终添加量10 9CFU/g混入到预处理过的原料碎鱼肉中，使最终菌数达到添加要求即可。
(4)鱼露的发酵：把步骤(3)得到的混合物料在温度30℃下保温发酵5d。
(5)鱼露的过滤：将步骤(4)得到的鱼露样品于120℃灭菌20min，冷却至室温后于10000r/min离心20min，取上清液用多层纱布过滤以除去固形物和杂质。
(6)鱼露的灭菌：将步骤(5)得到的鱼露在100℃下进行二次灭菌25min，在无菌条件下灌装密封，即得鱼露成品。
(7)产品理化指标的评价：氨基酸态氮含量评价，方法参照GB5009.235-2016比色法；挥发性盐基氮含量评价，方法参照GB5009.228-2016微量扩散法；组胺含量的测定，方法参照GB5009.208-2016分光光度法。
(8)鱼露理化指标结果：利用该株海洋动性球菌发酵鱼露的氨基酸态含量可达到0.873g/100mL，根据中国鱼露行业标准该鱼露划分为一级鱼露；挥发性盐基氮含量为97mg/100mL； 组胺含量为16mg/100mL，低于欧盟标准40mg/100mL。本鱼露中原料中固有的土腥味大大减少，鲜味明显，与未添加菌株发酵的鱼露相比，风味有显著提升。
实施例20：
一种利用动性球菌混合菌株发酵鱼露的方法，按照下述步骤进行：
(1)原料的处理：将碎鱼肉(鱼肉加工时所产生的废弃物)用碎肉机绞碎，加入腌制海盐15％(w/w)混匀，备用。
(2)混合发酵剂的制备：将海洋动性球菌(Planococcus maritimus XJ2)、普拉扣动球菌(Planococcus plakortidis XJ10)、德昌动性球菌(Planococcus dechangensis XJ11)和莱比托游动球菌(Planococcus rifietoensis XJ12)分别活化培养三次。将活化好的菌液于4℃、10000r/min离心10min，然后用无菌生理盐水洗涤两次，再悬浮于少量无菌生理盐水中，最后将菌液浓度调整到10 5～10 7CFU/mL，备用。
(3)混合发酵剂的添加：将海洋动性球菌(Planococcus maritimus XJ2)、普拉扣动球菌(Planococcus plakortidis XJ10)、德昌动性球菌(Planococcus dechangensis XJ11)和莱比托游动球菌(Planococcus rifietoensis XJ12)按菌数比列3:3:3:3进行混合并制备成混合发酵剂，按最终添加量10 9CFU/g混入到预处理过的原料碎鱼肉中，使最终菌数达到添加要求即可。
(4)鱼露的发酵：把步骤(3)得到的混合物料在温度30℃下保温发酵30d。
(5)鱼露的过滤：将步骤(4)得到的鱼露样品于120℃灭菌20min，冷却至室温后于10000r/min离心20min，取上清液用多层纱布过滤以除去固形物和杂质。
(6)鱼露的灭菌：将步骤(5)得到的鱼露在100℃下进行二次灭菌25min，在无菌条件下灌装密封，即得鱼露成品。
(7)产品理化指标的评价：氨基酸态氮含量评价，方法参照GB5009.235-2016比色法；挥发性盐基氮含量评价，方法参照GB5009.228-2016微量扩散法；组胺含量的测定，方法参照GB5009.208-2016分光光度法。
(8)鱼露理化指标结果：利用该株海洋动性球菌发酵鱼露的氨基酸态含量可达到0.981g/100mL，根据中国鱼露行业标准该鱼露划分为一级鱼露；挥发性盐基氮含量为104mg/100mL；组胺含量为18mg/100mL，低于欧盟标准40mg/100mL。本鱼露中原料中固有的土腥味大大减少，鲜味明显，与未添加菌株发酵的鱼露相比，风味有显著提升。

Claims (6)

  1. 海洋动性球菌(Planococcus maritimus XJ2)，编号为CGMCC NO.17057。
  2. 普拉扣动球菌(Planococcus plakortidis XJ10)，编号为CGMCC NO.17058。
  3. 德昌动性球菌(Planococcus dechangensis XJ11)，编号为CGMCC NO.17059。
  4. 莱比托游动球菌(Planococcus rifietoensis XJ12)，编号为CGMCC NO.17060。
  5. 利用权利要求1-4所述的动性球菌发酵鱼露的方法，其特征在于按照下述步骤进行：
    (1)原料的处理：将碎鱼肉(鱼肉加工时所产生的废弃物)用碎肉机绞碎，加入腌制海盐5～15％(w/w)混匀，备用；
    (2)发酵剂的制备：将4种动性球菌菌种活化培养三次；将活化好的菌液于4℃、10000r/min离心10min，然后用无菌生理盐水洗涤两次，再悬浮于少量无菌生理盐水中，最后将菌液浓度调整到10 5～10 7CFU/mL，备用；
    (3)发酵剂的添加：将菌株按最终添加量(10 5～10 9CFU/g)混入到预处理过的原料碎鱼肉中，使最终菌数达到添加要求即可；
    (4)鱼露的发酵：把步骤(3)得到的混合物料在温度15～30℃下保温发酵5～30d；
    (5)鱼露的过滤：将步骤(4)得到的鱼露样品于120℃灭菌10～30min，冷却至室温后于10000r/min离心20min，取上清液用多层纱布过滤以除去固形物和杂质；
    (6)鱼露的灭菌：将步骤(5)得到的鱼露在100℃下进行二次灭菌15～30min，在无菌条件下灌装密封，即得鱼露成品。
  6. 一种利用动性球菌混合菌株改善低盐鱼露发酵品质的方法，按照下述步骤进行：
    (1)原料的处理：将碎鱼肉(鱼肉加工时所产生的废弃物)用碎肉机绞碎，加入腌制海盐5～15％(w/w)混匀，备用；
    (2)混合发酵剂的制备：将海洋动性球菌(Planococcus maritimus XJ2)、普拉扣动球菌(Planococcus plakortidis XJ10)、德昌动性球菌(Planococcus dechangensis XJ11)和莱比托游动球菌(Planococcus rifietoensis XJ12)分别活化 培养三次；将活化好的菌液于4℃、10000r/min离心10min，然后用无菌生理盐水洗涤两次，再悬浮于少量无菌生理盐水中，最后将菌液浓度调整到10 5～10 7CFU/mL，备用；
    (3)混合发酵剂的添加：将海洋动性球菌(Planococcus maritimus XJ2)、普拉扣动球菌(Planococcus plakortidis XJ10)、德昌动性球菌(Planococcus dechangensis XJ11)和莱比托游动球菌(Planococcus rifietoensis XJ12)按菌数比列(1～3)：(1～3)：(1～3)：(1～3)进行混合并制备成混合发酵剂，按最终添加量(10 5～10 9CFU/g)混入到预处理过的原料碎鱼肉中，使最终菌数达到添加要求即可；
    (4)鱼露的发酵：把步骤(3)得到的混合物料在温度15～30℃下保温发酵5～30d；
    (5)鱼露的过滤：将步骤(4)得到的鱼露样品于120℃灭菌10～30min，冷却至室温后于10000r/min离心20min，取上清液用多层纱布过滤以除去固形物和杂质；
    (6)鱼露的灭菌：将步骤(5)得到的鱼露在100℃下进行二次灭菌15～30min，在无菌条件下灌装密封，即得鱼露成品。
PCT/CN2020/092490 2020-01-13 2020-05-27 动性球菌及利用其改善低盐鱼露发酵品质的方法 Ceased WO2021143006A1 (zh)

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EP20841648.7A EP3878945B1 (en) 2020-01-13 2020-05-27 Planococcus and method for improving fermentation quality of low-salt fish sauce using same
US17/256,008 US11337447B2 (en) 2020-01-13 2020-05-27 Planococcus and method for improving fermentation quality of low-salt fish sauce by Planococcus
JP2021513414A JP7060902B2 (ja) 2020-01-13 2020-05-27 プラノコッカス及びそれによる減塩魚醤の発酵品質の向上方法

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