Genetically modified bacillus subtilis strain and use as a live delivery and production system
US-2024390433-A1 · Nov 28, 2024 · US
US11337447B2 · US · B2
| Field | Value |
|---|---|
| Publication number | US-11337447-B2 |
| Application number | US-202017256008-A |
| Country | US |
| Kind code | B2 |
| Filing date | May 27, 2020 |
| Priority date | Jan 13, 2020 |
| Publication date | May 24, 2022 |
| Grant date | May 24, 2022 |
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The present invention discloses Planococcus and a method for improving fermentation quality of a low-salt fish sauce by Planococcus, and relates to the application field of food microbiology technology. The Planococcus strains have been deposited in China General Microbiological Culture Collection Center (CGMCC) under CGMCC NO. 17057, CGMCC NO. 17058, CGMCC NO. 17059, and CGMCC NO. 17060. The production cycle of fermented fish sauce using the Planococcus strains is greatly shortened, and the fermentation temperature is reduced. The fish sauce obtained by this method is transparent brown red, and has the unique aroma of aquatic products without impurities such as suspension or flocculent. The finished fish sauce has low salt content and high amino acid nitrogen content (according to the Chinese fish sauce industry standard, the fish sauce is classified as first-grade fish sauce). The contents of total volatile basic nitrogen and histamine are lower than the national standard.
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What is claimed is: 1. A method for fermenting a fish sauce by using Planococcus maritimus XJ2, designated as CGMCC NO. 17057, the method comprises the following steps: (1) treatment of raw materials: mincing fish trimmings, which are wastes from fish flesh processing with a mincer, and adding 5 to 15% (w/w) of a pickling sea salt thereto, followed by mixing well; (2) preparation of starter cultures: activating and culturing Planococcus maritimus XJ2 three times to obtain an activated bacterial suspension; centrifuging the activated bacterial suspension at 4° C. and 10000 r/min for 10 min to obtain cell pellets, and washing the cell pellets twice with sterile physiological saline, resuspending the cell pellets in a small amount of the sterile physiological saline to obtain a bacterial suspension, and finally adjusting a concentration of the bacterial suspension to 10 5 -10 7 CFU/mL; (3) addition of the starter cultures: mixing the strain into the pretreated raw minced fish flesh to obtain a mixture, so that the final bacterial count meets a final addition amount of 10 5 -10 9 CFU/g; (4) fermentation of fish sauce: incubating and fermenting the mixture obtained in step (3) at a temperature of 15-30° C. for 5-30 days; (5) filtration of the fish sauce: sterilizing the fish sauce sample obtained in step (4) at 120° C. for 10 to 30 mins, cooling the sterilized fish sauce sample to room temperature and centrifuging it at 10000 r/min for 20 mins to obtain a supernatant, and filtering the supernatant with multiple layers of gauze to remove solids and impurities; and (6) sterilization of the fish sauce: performing secondary sterilization on the fish sauce obtained in step (5) at 100° C. for 15 to 30 mins, followed by filling and sealing it under aseptic conditions to obtain a finished fish sauce. 2. A method for shortening a fermentation cycle of a low-salt fish sauce by using mixed Planococcus strains, comprising the following steps: (1) treatment of raw materials: mincing fish trimmings, which are wastes from fish flesh processing with a mincer, and adding 5 to 15% (w/w) of a pickling sea salt thereto, followed by mixing well; (2) preparation of mixed starter: activating and culturing each of Planococcus maritimus XJ2 , Planococcus plakortidis XJ10 , Planococcus dechangensis XJ11 and Planococcus rifietoensis XJ12 three times to obtain an activated bacterial suspension; centrifuging the activated bacterial suspension at 4° C. and 10000 r/min for 10 min to obtain cell pellets, and washing the cell pellets twice with sterile physiological saline, resuspending the cell pellets in a small amount of the sterile physiological saline to obtain a bacterial suspension, and finally adjusting a concentration of the bacterial suspension to 10 5 -10 7 CFU/mL; (3) addition of the mixed starter: mixing Planococcus maritimus XJ2 , Planococcus plakortidis XJ10 , Planococcus dechangensis XJ11 and Planococcus rifietoensis XJ12 according to a bacteria count ratio of (1-3):(1-3):(1-3):(1-3) to prepare the mixed starter, and mixing the mixed starter into the pretreated raw minced fish flesh to obtain a mixture, so that the final bacterial count meets a final addition amount of 10 5 -10 9 CFU/g; (4) fermentation of fish sauce: incubating and fermenting the mixture obtained in step (3) at a temperature of 15-30° C. for 5-30 days; (5) filtration of the fish sauce: sterilizing the fish sauce sample obtained in step (4) at 120° C. for 10 to 30 mins, cooling the sterilized fish sauce sample to room temperature and centrifuging it at 10000 r/min for 20 mins to obtain a supernatant, and filtering the supernatant with multiple layers of gauze to remove solids and impurities; and (6) sterilization of the fish sauce: performing secondary sterilization on the fish sauce obtained in step (5) at 100° C. for 15 to 30 mins, followed by filling and sealing it under aseptic conditions to obtain a finished fish sauce. 3. A method for fermenting a fish sauce by using Planococcus plakortidis XJ10, designated as CGMCC NO. 17058, the method comprises the following steps: (1) treatment of raw materials: mincing fish trimmings, which are wastes from fish flesh processing with a mincer, and adding 5 to 15% (w/w) of a pickling sea salt thereto, followed by mixing well; (2) preparation of starter cultures: activating and culturing Planococcus plakortidis XJ10 three times to obtain an activated bacterial suspension; centrifuging the activated bacterial suspension at 4° C. and 10000 r/min for 10 min to obtain cell pellets, and washing the cell pellets twice with sterile physiological saline, resuspending the cell pellets in a small amount of the sterile physiological saline to obtain a bacterial suspension, and finally adjusting a concentration of the bacterial suspension to 10 5 -10 7 CFU/mL; (3) addition of the starter cultures: mixing the strain into the pretreated raw minced fish flesh to obtain a mixture, so that the final bacterial count meets a final addition amount of 10 5 -10 9 CFU/g; (4) fermentation of fish sauce: incubating and fermenting the mixture obtained in step (3) at a temperature of 15-30° C. for 5-30 days; (5) filtration of the fish sauce: sterilizing the fish sauce sample obtained in step (4) at 120° C. for 10 to 30 mins, cooling the sterilized fish sauce sample to room temperature and centrifuging it at 10000 r/min for 20 mins to obtain a supernatant, and filtering the supernatant with multiple layers of gauze to remove solids and impurities; and (6) sterilization of the fish sauce: performing secondary sterilization on the fish sauce obtained in step (5) at 100° C. for 15 to 30 mins, followed by filling and sealing it under aseptic conditions to obtain a finished fish sauce. 4. A method for fermenting a fish sauce by using Planococcus dechangensis XJ11, designated as CGMCC NO. 17059, the method comprises the following steps: (1) treatment of raw materials: mincing fish trimmings, which are wastes from fish flesh processing with a mincer, and adding 5 to 15% (w/w) of a pickling sea salt thereto, followed by mixing well; (2) preparation of starter cultures: activating and culturing Planococcus dechangensis XJ11 three times to obtain an activated bacterial suspension; centrifuging the activated bacterial suspension at 4° C. and 10000 r/min for 10 min to obtain cell pellets, and washing the cell pellets twice with sterile physiological saline, resuspending the cell pellets in a small amount of the sterile physiological saline to obtain a bacterial suspension, and finally adjusting a concentration of the bacterial suspension to 10 5 -10 7 CFU/mL; (3) addition of the starter cultures: mixing the strain into the pretreated raw minced fish flesh to obtain a mixture, so that the final bacterial count meets a final addition amount of 10 5 -10 9 CFU/g; (4) fermentation of fish sauce: incubating and fermenting the mixture obtained in step (3) at a temperature of 15-30° C. for 5-30 days; (5) filtration of the fish sauce: sterilizing the fish sauce sample obtained in step (4) at 120° C. for 10 to 30 mins, cooling the sterilized fish sauce sample to room temperature and centrifuging it at 10000 r/min for 20 mins to obtain a supernatant, and filtering the supernatant with multiple layers of gauze to remove solids and impurities; and (6) sterilization of the fish sauce: performing secondary sterilization on the fish sauce obtained in step (5) at 100° C. for 15 to 30 mins, followed by filling and sealing it under aseptic conditions to obtain a finished fish sauce. 5. A method for fermenting a fish sauce by using the Planococcus rifietoensis XJ12, designated as CGMCC NO. 17060, the method comprises the following steps: (1) treatment of raw materials: mincing fish trimmings, which are wastes from fish flesh processing with
Fish extracts · CPC title
Bacteria or Actinomycetales {; using bacteria or Actinomycetales} · CPC title
Bacteria; Culture media therefor · CPC title
Bacterial isolates · CPC title
prepared by fermentation · CPC title
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