CN115944053A - Method for effectively reducing nitrosamine in pickled fish by using leavening agent for pickled fish - Google Patents
Method for effectively reducing nitrosamine in pickled fish by using leavening agent for pickled fish Download PDFInfo
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Abstract
The invention relates to a method for effectively reducing nitrosamine in pickled fish by using a mixed starter, wherein the mixed starter comprises pediococcus pentosaceus and staphylococcus xylosus, and the specific treatment method comprises the following steps: (1) processing raw material fish; (2) domesticating strains; (3) soaking in salt and adding auxiliary materials; (4) inoculating and fermenting: activating and expanding each fermentation strain, centrifuging the culture solution, preparing the mixed starter of claim 1 or 2 with the bacteria content of 800-900cfu/mL by using sterile water, inoculating and fermenting the fish body treated in the step (3), and fermenting the inoculated fish body in a thermostat at 33-35 ℃ for 13-15h; and (5) drying the fish body. The method can greatly shorten the fermentation time, effectively reduce nitrosamine substances, ensure the safety of the salted fish, ensure that the salted fish is not easy to deteriorate due to proper drying temperature and time, and have unique flavor, moderate sour and hot taste and pure and fragrant mouthfeel.
Description
Technical Field
The invention relates to the technical field of microbial technology and food technology, in particular to a method for effectively reducing nitrosamine in pickled fish by using a leavening agent for pickled fish.
Background
The pickled fish is a processed product of traditional pickled and fermented fish in China, and the raw material fish is treated by using auxiliary materials such as salt, sugar, vinegar, vinasse and the like and then is subjected to microbial fermentation, so that the fish has special flavor, color and texture. The traditional salted fish has a long processing period, is easy to generate a large amount of nitrosamine, and has a difficult control process.
CN106962741A discloses a compound microbial preparation for efficiently degrading nitrite, which is composed of three lactic acid bacteria, namely lactobacillus plantarum, pediococcus pentosaceus and lactobacillus brevis, and has a low nitrate degradation characteristic. Although the nitrite content can be reduced under certain conditions, the fermentation time is long, the shelf life cannot be well prolonged, and the quality of the pickled fish is ensured, so that how to shorten the production period and control the product quality is researched.
Disclosure of Invention
In view of the above technical problems, the present invention provides a method for effectively reducing nitrite in salted fish by using a leavening agent for salted fish, which can greatly shorten the fermentation time of the salted fish and effectively reduce nitrosamine substances.
In order to achieve the purpose, the invention adopts the following technical scheme:
first, the present invention provides a mixed starter, which includes pediococcus pentosaceus and staphylococcus xylosus.
Further, the pediococcus pentosaceus and the staphylococcus xylosus are mixed according to a volume ratio of 1:1-3.
Further, the pediococcus pentosaceus and the staphylococcus xylosus are mixed according to a volume ratio of 1:1.
secondly, the invention also provides a method for effectively reducing nitrosamine in the pickled fish by using the mixed leavening agent, and the specific treatment method comprises the following steps:
(1) Processing raw material fish;
(2) Domesticating strains: gradient culture is adopted, and the content of fish meat in the culture medium is gradually increased in an MRS culture medium, so that pediococcus pentosaceus and staphylococcus xylosus are gradually adapted to the growth environment of the flavor salted fish. Adding the marinated fish homogenate into an MRS culture medium according to the mass ratio of 5%, 10%, 20%, 40% and 80%, respectively, placing into a test tube, sterilizing at 120-130 deg.C for 15-20min, and cooling for use; under the aseptic operation condition, inoculating pediococcus pentosaceus and staphylococcus xylosus into an MRS culture medium test tube containing 5% of salted fish homogenate according to the inoculation amount of 3% -4%, culturing in a constant-temperature incubator at 35-40 ℃, and finishing domestication when the content of pediococcus pentosaceus and staphylococcus xylosus reaches 107-120 cfu/g; repeating the domestication for 4-6 times every time, and then inoculating pediococcus pentosaceus and staphylococcus xylosus in the raw materials containing the salted fish into a test tube with a high first-level concentration for domestication in the next gradient until the domestication is finished;
(3) Salt leaching and auxiliary material addition: uniformly wiping fish bodies with fine salt at a fish salt ratio of 10: 1.5, uniformly filling the fish bodies with auxiliary materials, then flatly spreading the fish bodies layer by layer in a container, spreading a layer of cover salt on the uppermost surface, covering gauze on the mouth of the container, and preventing the fish bodies from deteriorating due to spawning;
(4) Inoculating and fermenting: centrifuging culture solution obtained by activating and expanding each fermentation strain, preparing mixed leaven with bacteria content of 800-900cfu/mL according to claim 1 or 2 by using sterile water, inoculating the fish body treated in the step (3) for fermentation, wherein the use amount of the mixed leaven is 6-8% of the total mass of the fish body, soaking the fish body in the leaven for 5-8min, taking out and draining; placing the inoculated fish body in a thermostat at 33-35 ℃ for fermentation for 13-15h;
(5) Drying the fish body: drying the fermented fish in a blower at 23-27 deg.C for 11-15h.
Further, the raw material fish is one of procypris merus, carps, oryza sativa and field fishes.
Compared with the prior art, the invention has the following beneficial effects:
by adopting the method, the fermentation time can be greatly shortened, and nitrosamine substances can be effectively reduced; the traditional method for fermenting the salted fish has longer fermentation time, so that the salt content of the salted fish is increased, the content of nitrosamines is increased, and the salted fish is harmful to the health of human bodies; the mixed leaven containing staphylococcus xylosus and pediococcus pentosaceus greatly reduces the generation of nitrosamine substances, ensures the safety of the salted fish, ensures that the salted fish is not easy to deteriorate due to proper drying temperature and time, and has unique flavor, moderate sour and hot taste and pure and fragrant mouthfeel.
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FIG. 1 is a graph showing the effect of different treatments on the nitrosamine content of salted fish.
In the figure: NDMA: n-nitrosodimethylamine; NPYR: n-nitrosopyrrolidine; NDEA is N-nitrosodiethylamine; NDBA: n-nitrosodibutylamine.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are some embodiments of the present invention, but not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The staphylococcus xylosus and pediococcus pentosaceus used in the following examples and other materials are commercially available from conventional sources, and the experimental methods not specifically described in the examples are all conventional experimental methods.
Example 1
A mixed starter culture comprises Pediococcus pentosaceus and Staphylococcus xylosus.
Pediococcus pentosaceus and staphylococcus xylosus in a volume ratio of 1.
A method for effectively reducing nitrosamine in pickled fish by using a mixed starter comprises the following specific treatment methods:
(1) Processing raw material fish, namely removing scales and internal organs of the raw material fish, cleaning, draining water, and sterilizing for later use; sterilizing with ultraviolet rays at wavelength of 254nm for 30min; the bacteria are afraid of salt, but salmonella which causes inflammation of human intestines and stomachs can survive for several months in meat with the salt content of 10% -15%, so the salmonella can be killed well by using the ultraviolet condition;
(2) Domesticating strains: adopts gradient culture to gradually increase the content of fish meat in a culture medium in an MRS culture medium, so that pediococcus pentosaceus and staphylococcus xylosus are gradually adapted to the growth environment of the flavor salted fish. Adding the marinated fish homogenate into an MRS culture medium according to the mass ratio of 5%, 10%, 20%, 40% and 80%, respectively, placing into a test tube, sterilizing at 120 deg.C for 15min, and cooling for use; under the aseptic operation condition, inoculating pediococcus pentosaceus and staphylococcus xylosus into an MRS culture medium test tube containing 5% of salted fish homogenate according to the inoculation amount of 3%, culturing in a constant-temperature incubator at 35 ℃, and finishing domestication when the content of pediococcus pentosaceus and staphylococcus xylosus reaches 107 cfu/g; repeating the domestication for 6 times every time, and then inoculating pediococcus pentosaceus and staphylococcus xylosus in the raw materials containing the salted fish into a test tube with a higher first-level concentration for domestication in the next gradient until the domestication is finished;
(3) Salt leaching and auxiliary material addition: uniformly wiping fish bodies with fine salt, wherein the ratio of fish salt is 10: 1.5, uniformly filling auxiliary materials into the fish bodies, then flatly spreading the fish bodies layer by layer in a container, scattering a layer of cover salt on the uppermost surface, covering gauze on the mouth of the container, and preventing the fish bodies from going bad due to spawning; the auxiliary materials comprise the following raw materials in percentage by mass: 0.008% of sodium nitrite, 0.7% of white sugar, 0.8% of glucose, 0.3% of monosodium glutamate, 0.7% of hot pepper, 0.1% of black pepper and 0.1% of pepper;
(4) Inoculating and fermenting: centrifuging culture solution obtained by activating and expanding each fermentation strain, preparing the culture solution into the mixed starter of claim 1 or 2 with the bacterium content of 800cfu/mL by using sterile water, inoculating the fish body treated in the step (3) for fermentation, wherein the using amount of the mixed starter is 8% of the total mass of the fish body, soaking the fish body into the starter for 5min, taking out and draining; placing the inoculated fish body in a thermostat at 33 ℃ for fermentation for 15h;
(5) Drying the fish body: drying the fermented fish in a blower at 23 deg.C for 15h.
The raw material fish is the Hehuayu.
Example 2
A mixed starter culture comprises Pediococcus pentosaceus and Staphylococcus xylosus.
Pediococcus pentosaceus and staphylococcus xylosus in a volume ratio of 1:2.
a method for effectively reducing nitrosamine in salted fish by using a mixed starter comprises the following specific treatment methods:
(1) Processing raw material fish, removing scales and internal organs of the raw material fish, cleaning, draining water, and sterilizing for later use; sterilizing with ultraviolet rays at wavelength of 260nm for 35min;
(2) Domesticating strains: adopts gradient culture to gradually increase the content of fish meat in a culture medium in an MRS culture medium, so that pediococcus pentosaceus and staphylococcus xylosus are gradually adapted to the growth environment of the flavor salted fish. Adding the marinated fish homogenate into an MRS culture medium according to the mass ratio of 5%, 10%, 20%, 40% and 80%, respectively, placing into a test tube, sterilizing at 125 deg.C for 18min, and cooling for use; under the aseptic operation condition, inoculating pediococcus pentosaceus and staphylococcus xylosus into an MRS culture medium test tube containing 5% of salted fish homogenate according to the inoculation amount of 3.5%, culturing in a constant-temperature incubator at 37 ℃, and finishing domestication when the content of pediococcus pentosaceus and staphylococcus xylosus reaches 115 cfu/g; repeating the domestication for 5 times every time, and then inoculating pediococcus pentosaceus and staphylococcus xylosus in the raw materials containing the salted fish into a test tube with a higher first-level concentration for domestication in the next gradient until the domestication is finished;
(3) Salt leaching and auxiliary material addition: uniformly wiping fish bodies with fine salt, wherein the ratio of fish salt is 10: 1.5, uniformly filling auxiliary materials into the fish bodies, then flatly spreading the fish bodies layer by layer in a container, scattering a layer of cover salt on the uppermost surface, covering gauze on the mouth of the container, and preventing the fish bodies from going bad due to spawning; the auxiliary materials comprise the following raw materials in percentage by mass: 0.008% of sodium nitrite, 0.7% of white sugar, 0.8% of glucose, 0.3% of monosodium glutamate, 0.7% of hot pepper, 0.1% of black pepper and 0.1% of pepper;
(4) Inoculating and fermenting: centrifuging culture solution obtained by activating and expanding culture of each fermentation strain, preparing into mixed leaven with bacteria content of 850cfu/mL according to claim 1 or 2 with sterile water, inoculating the fish body treated in the step (3) for fermentation, soaking the fish body in the leaven for 6min, taking out, and draining; placing the inoculated fish body in a thermostat at 34 ℃ for fermentation for 14h;
(5) Drying the fish body: drying the fermented fish in an air blower at 25 deg.C for 13 hr.
The fish is fish with rice flower.
Example 3
A mixed starter culture comprises Pediococcus pentosaceus and Staphylococcus xylosus.
Pediococcus pentosaceus and staphylococcus xylosus in a volume ratio of 1:3.
a method for effectively reducing nitrite in salted fish by using a mixed starter comprises the following specific treatment methods:
(1) Processing raw material fish, namely removing scales and internal organs of the raw material fish, cleaning, draining water, and sterilizing for later use; sterilizing with ultraviolet rays at wavelength of 265nm, and irradiating for 40min;
(2) Domesticating strains: adopts gradient culture to gradually increase the content of fish meat in a culture medium in an MRS culture medium, so that pediococcus pentosaceus and staphylococcus xylosus are gradually adapted to the growth environment of the flavor salted fish. Adding the marinated fish homogenate into an MRS culture medium according to the mass ratio of 5%, 10%, 20%, 40% and 80%, respectively, placing into a test tube, sterilizing at 120-130 deg.C for 15-20min, and cooling for use; under the aseptic operation condition, inoculating pediococcus pentosaceus and staphylococcus xylosus into an MRS culture medium test tube containing 5% of salted fish homogenate according to the inoculation amount of 4%, culturing in a constant-temperature incubator at 35-40 ℃, and finishing domestication when the content of pediococcus pentosaceus and staphylococcus xylosus reaches 107 cfu/g; repeating the domestication for 4-6 times every time, and then inoculating pediococcus pentosaceus and staphylococcus xylosus in the raw materials containing the salted fish into a test tube with a high first-level concentration for domestication in the next gradient until the domestication is finished;
(3) Salt leaching and auxiliary material addition: uniformly wiping fish bodies with fine salt at a fish salt ratio of 10: 1.5, uniformly filling the fish bodies with auxiliary materials, then flatly spreading the fish bodies layer by layer in a container, spreading a layer of cover salt on the uppermost surface, covering gauze on the mouth of the container, and preventing the fish bodies from deteriorating due to spawning; the auxiliary materials comprise the following raw materials in percentage by mass: 0.008% of sodium nitrite, 0.7% of white sugar, 0.8% of glucose, 0.3% of monosodium glutamate, 0.7% of hot pepper, 0.1% of black pepper and 0.1% of pepper;
(4) Inoculating and fermenting: centrifuging culture solution obtained by activating and expanding each fermentation strain, preparing a mixed starter of claim 1 or 2 with a bacterium content of 900cfu/mL by using sterile water, inoculating the fish body treated in the step (3) for fermentation, soaking the fish body in the starter for 8min, taking out and draining; placing the inoculated fish body in a thermostat at 35 ℃ for fermentation for 13h;
(5) Drying the fish body: drying the fermented fish in an air blower at 27 deg.C for 11h.
Further, the raw material fish is field fish.
Comparative example 1 is different from example 1 in that the fermentation process is pediococcus pentosaceus fermentation, and the rest of the process is the same as example 1.
Comparative example 2 differs from example 1 in that the fermentation process is a Staphylococcus xylosus fermentation, and the rest of the process is the same as example 1.
Comparative example 3 is different from example 1 in that the fermentation process is a natural fermentation, and the rest of the process is the same as example 1.
Comparative example 4 the same test method was used, and the salted fish obtained by fermentation with the microbial composite preparation of CN106962741A was used as a sampling object, and the specific method was as follows: (1) Selecting single colony of Lactobacillus plantarum, pediococcus pentosaceus and Lactobacillus brevis, transferring to MRS liquid culture medium, culturing at 37 deg.C for 18h in thermostat, centrifuging at 4 deg.C and 5000rpm for 15min, resuspending with sterile normal saline, centrifuging again for three times, counting thallus concentration by viable count method, diluting with normal saline to 10 × 10 10 cfu/mL, stored in a refrigerator at 4 ℃ for future use. (2) Removing scales, viscera and skin of fresh grass carp, cutting into blocks with length, width and thickness of about 4cm and 1cm, draining water under natural condition, and soaking in 60mg/L sodium nitrite solution for 1 hr. Taking out the fish, draining the solvent, accurately weighing 10% of salt, uniformly coating the salt on the surface of the fish, and placing the fish in a sealed tank for pickling for 6 hours at 15 ℃. Soaking the salted dried salt water into the prepared fermentation liquor, fermenting for a certain time at different fermentation temperatures, taking out and drying to obtain the final product.
Each treatment was repeated 10 times.
1. Effect of different treatments on nitrosamine content in salted Fish
TABLE 1 Effect of different treatments on nitrosamine content in salted fish
Note: nd indicates no detection.
The above 9 kinds of nitrosamines (N-Nitrosodiethylamine (NDEA); N-Nitrosopiperidine (NPIP); N-Nitrosodipropylamine (NDPA); N-Nitrosodibutylamine (NDBA); N-nitrosodiphenylamine (NDPhA); N-Nitromethylethylamine (NMEA); N-Nitrosodimethylamine (NDMA); N-Nitrosomorpholine (NMOR); N-Nitrosopyrrolidine (NPYR)) were detected in food, but the nitrosamines frequently detected in the species used for salting fish were NDMA, NPYR, NDEA, NDBA, among which NDMA is the most common and representative. Compared with comparative examples 1-4, the total nitrosamine content of examples 1-3 is the lowest, and the contents of NDMA, NPYR, NDEA and NDBA are also the lowest, which shows that the method of the invention has better effect of reducing nitrosamine content; comparative example 4 was a process according to CN106962741A, with a lower nitrosamine content than in naturally fermented comparative example 3, a slightly lower content than in comparative example 1 with pediococcus pentosaceus fermentation, but a higher content than in comparative example 2 with staphylococcus xylosus fermentation.
The present invention focuses on the effect of comparative examples 1-3 and example 1 on the nitrosamine content of the marinated fish. As can be seen from FIG. 1, 4 nitrosamines, NDMA, NDEA, NPYR and NDBA, were detected in Table 1, and their contents were varied. Wherein the content of 4 nitrosamines in the naturally fermented product is higher, the detected amount of the comparative example 3 of the natural fermentation is 100%, the content of 4 nitrosamines in other 3 treatments is calculated, and the influence of different treatments on the nitrosamine content in the fermentation process of the salted fish product is compared, and the result is shown in figure 1. Comparative example 3 the naturally fermented salted fish products have the highest content of 4 nitrosamines, and the 4 nitrosamines can be effectively reduced in the comparative example 1 of pediococcus pentosaceus fermentation, the comparative example 2 of staphylococcus xylosus fermentation and the example 1; for NDMA, the content of the fermented mixed starter is the lowest and is 70% of that of natural fermentation; for NPYR, the content of pediococcus pentosaceus fermented relative to natural fermentation is slightly reduced to 90% of the content after natural fermentation, while the content of pediococcus xylosus fermented and mixed bacteria fermented is only 56% and 62% of the content after natural fermentation, so that the production amount can be obviously reduced; for NDEA, the content of the pediococcus pentosaceus can be effectively reduced by fermentation, pediococcus xylosus and mixed fermentation, the effect is obvious, and the yield of NDEA of the pickled fish product fermented by the pediococcus pentosaceus is only 50% of that of natural fermentation; for NDBA, the production amount of pediococcus pentosaceus, staphylococcus xylosus and mixed fermentation can be effectively reduced, the content is 67%, 61% and 56% of the natural fermentation production amount respectively, and the production amount of NDBA can be obviously reduced.
In summary, the above description is only a preferred embodiment of the present invention, and all equivalent changes and modifications made in the claims of the present invention should be covered by the claims of the present invention.
Claims (5)
1. The mixed leaven is characterized by comprising pediococcus pentosaceus and staphylococcus xylosus.
2. The mixed starter culture of claim 1, wherein the pediococcus pentosaceus and the staphylococcus xylosus are mixed in a volume ratio of 1:1-3.
3. The mixed starter according to claim 1 or 2, wherein the pediococcus pentosaceus and the staphylococcus xylosus are mixed in a volume ratio of 1:1.
4. a method for effectively reducing nitrosamine in salted fish by using a mixed starter is characterized by comprising the following specific treatment methods:
(1) Processing raw material fish;
(2) Domesticating strains: gradient culture is adopted, the content of fish meat in a culture medium is gradually increased in an MRS culture medium, and pediococcus pentosaceus and staphylococcus xylosus are gradually adapted to the growth environment of the flavor salted fish; adding the marinated fish homogenate into an MRS culture medium according to the mass ratio of 5%, 10%, 20%, 40% and 80%, respectively, placing into a test tube, sterilizing at 120-130 deg.C for 15-20min, and cooling for use; under the aseptic operation condition, inoculating pediococcus pentosaceus and staphylococcus xylosus into an MRS culture medium test tube containing 5% of salted fish homogenate according to the inoculation amount of 3% -4%, culturing in a constant-temperature incubator at 35-40 ℃, and finishing domestication when the content of the pediococcus pentosaceus and the staphylococcus xylosus reaches 107-120 cfu/g; repeating the domestication for 4-6 times, and inoculating Pediococcus pentosaceus and Staphylococcus xylosus in the raw material containing the salted fish into a test tube with a high first-level concentration for domestication in the next gradient until finishing;
(3) Salt leaching and auxiliary material addition: uniformly wiping fish bodies with fine salt, wherein the ratio of fish salt is 10: 1.5, uniformly filling auxiliary materials into the fish bodies, then flatly spreading the fish bodies layer by layer in a container, scattering a layer of cover salt on the uppermost surface, covering gauze on the mouth of the container, and preventing the fish bodies from going bad due to spawning;
(4) Inoculating and fermenting: centrifuging culture solution obtained by activating and expanding each fermentation strain, preparing mixed leaven with bacteria content of 800-900cfu/mL according to claim 1 or 2 by using sterile water, inoculating the fish body treated in the step (3) for fermentation, wherein the use amount of the mixed leaven is 6-8% of the total mass of the fish body, soaking the fish body in the leaven for 5-8min, taking out and draining; placing the inoculated fish body in a thermostat at 33-35 ℃ for fermentation for 13-15h;
(5) Drying the fish body: drying the fermented fish in a blower at 23-27 deg.C for 11-15h.
5. The method of claim 4, wherein the raw fish is one of procypris merus, carpio merus, oryza sativa, and pisces vulgaris.
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