CN109170130B - Method for preparing fish meal by solid-state fermentation of aquatic product leftovers - Google Patents
Method for preparing fish meal by solid-state fermentation of aquatic product leftovers Download PDFInfo
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- CN109170130B CN109170130B CN201810896870.XA CN201810896870A CN109170130B CN 109170130 B CN109170130 B CN 109170130B CN 201810896870 A CN201810896870 A CN 201810896870A CN 109170130 B CN109170130 B CN 109170130B
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- fermentation
- fish meal
- aquatic
- leftovers
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- 238000010563 solid-state fermentation Methods 0.000 title claims abstract description 16
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- CHHHXKFHOYLYRE-UHFFFAOYSA-M 2,4-Hexadienoic acid, potassium salt (1:1), (2E,4E)- Chemical compound [K+].CC=CC=CC([O-])=O CHHHXKFHOYLYRE-UHFFFAOYSA-M 0.000 claims abstract description 11
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Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/12—Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/20—Animal feeding-stuffs from material of animal origin
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/20—Animal feeding-stuffs from material of animal origin
- A23K10/22—Animal feeding-stuffs from material of animal origin from fish
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/105—Aliphatic or alicyclic compounds
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/111—Aromatic compounds
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/163—Sugars; Polysaccharides
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/174—Vitamins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/20—Inorganic substances, e.g. oligoelements
- A23K20/26—Compounds containing phosphorus
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/20—Inorganic substances, e.g. oligoelements
- A23K20/30—Oligoelements
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- 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, e.g. protozoa; 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
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; 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, e.g. protozoa; 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
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- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Polymers & Plastics (AREA)
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- Tropical Medicine & Parasitology (AREA)
- Mycology (AREA)
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- General Health & Medical Sciences (AREA)
- Inorganic Chemistry (AREA)
- Botany (AREA)
- Marine Sciences & Fisheries (AREA)
- Sustainable Development (AREA)
- Fodder In General (AREA)
- Feed For Specific Animals (AREA)
Abstract
The invention discloses a method for preparing fish meal by solid-state fermentation of aquatic leftovers, which adopts a microbial fermentation method to prepare the fish meal, and potassium sorbate and (S) - (+) -3- (1-naphthoxy) -1-phenyl-1-propanol which are protective agents are added in the preparation process of microbial mixed bacterial powder; the fermentation process comprises the following steps: subpackaging the fermentation medium in a sterilized fermentation tray, covering the surface of the fermentation medium in the tray with sterilized wet gauze, and placing the tray on a shelf in a sterilized solid fermentation room for fermentation, wherein the fermentation medium contains (Z) -3-hexenol and riboflavin. Has the advantages that: the invention uses the aquatic product leftovers as raw materials, ensures lower cost, adopts the microbial fermentation method to produce the fish meal, avoids high-temperature cooking of the traditional processing method, and can effectively avoid loss and damage of nutritional ingredients in the fish meal; the fish meal prepared by the method has the advantages of antibiosis, antivirus and high digestibility.
Description
Technical Field
The invention relates to the technical field of aquatic products, in particular to a method for preparing fish meal by solid-state fermentation of aquatic product leftovers.
Background
The fish meal is the main raw material of high protein feed, and is a powdery or granular product obtained by using aquatic animals such as fish, shrimp, crab and the like or aquatic product processing waste as raw materials and performing deoiling, drying and crushing. With the rapid development of the aquaculture industry and the fishing industry, the demand of aquatic feed protein is increasing. The high-quality fish meal can promote the absorption and utilization of high-nutrient substances by aquatic animals such as fish, shrimps and the like, further promote the growth and reproduction of the fish, the shrimps and the like, and has an advantage position in the current aquatic feeds.
With the rapid development of the aquaculture industry, the demand of feed protein is increasing. In particular to high-quality fish meal, because the fish meal is favorable for promoting aquatic animals such as fish, shrimp and the like to absorb and utilize high nutrient substances and stimulating the growth and reproduction of the fish, the shrimp and the like, the efficacy of the fish meal in aquatic feeds is incomparable with other protein sources at present. The fish meal is processed by taking aquatic animals such as fish, shrimp, crab and the like or aquatic product processing waste as raw materials, a large amount of leftovers such as fish heads, fish skins, fish bones, swim bladders, shrimp shells, crab shells and the like are generated in the processing process of the aquatic products every year, the leftovers account for about 40-55% of the raw materials of the fish, most of the leftovers are roughly processed or directly discarded as feed raw materials, resources are wasted, and the environment is polluted. The leftovers are used as raw materials for producing the fish meal, so that the method saves resources, is favorable for environmental protection and has good application prospect.
The traditional processing mode of the fish meal is a dry method and a wet method, the fish meal in the dry method is brown due to the long-time high-temperature drying, the nutritional ingredients of the fish meal are damaged, and the nutritional value of the fish meal is reduced; the wet method fish meal is easy to cause the loss of nutrient components such as free amino acid, water-soluble protein and the like in the production process, so that the nutrient value of the fish meal is reduced.
In order to overcome the defects of the prior dry and wet processing technology of the traditional fish meal, a brand new method capable of providing high-quality fish meal needs to be found.
The prior art discloses a preparation method of fish meal small peptide feed, such as a Chinese patent with an authorization publication number of CN 104041710B: mixing commercially available inferior fish meal, molasses, brown sugar or glucose, water, microbial protease, lactobacillus strain liquid and yeast strain liquid to prepare a liquid fermentation culture medium, and performing primary liquid fermentation enzymolysis for 2-120h at 20-70 ℃ to prepare fish meal protein small peptide liquid; mixing the fish meal protein small peptide liquid, the corn flour and the alfalfa meal to prepare a solid fermentation culture medium, performing solid state fermentation and enzymolysis for 6-360h at 10-50 ℃ for the second time to prepare the fish meal small peptide feed which can eliminate anti-nutritional factors, kill pathogenic microorganisms, generate rich small peptides, lactic acid, yeast protein and probiotics, eliminate odor, present fishy fragrance and sour taste, improve the feed intake of livestock and poultry, promote the growth of piglets and improve the milk yield of lactating sows. However, this method is high in raw material cost.
Disclosure of Invention
The invention aims to provide a method for preparing fish meal by solid-state fermentation of aquatic leftovers, which utilizes the aquatic leftovers as raw materials, ensures lower cost, adopts a microbial fermentation method to produce the fish meal, avoids high-temperature cooking of the traditional processing method, and can effectively avoid loss and damage of nutritional ingredients in the fish meal; the fish meal prepared by the method has the advantages of antibiosis, antivirus and high digestibility.
Aiming at the problems mentioned in the background technology, the invention adopts the technical scheme that:
the strain used in the invention is purchased from Zhejiang Lukang sourced biotechnology limited.
The method for preparing fish meal by solid-state fermentation of aquatic product leftovers comprises the following steps:
s1, respectively placing the rhodotorula benthica and the bacillus subtilis on a potato sugar agar culture medium for slant culture; inoculating the obtained rhodotorula benthica and bacillus subtilis after slant culture into a rhodotorula benthica and bacillus subtilis seed bottle culture medium for propagation, inoculating 5-7% of inoculum size of rhodotorula benthica and bacillus subtilis seed bottle bacterial liquid according to the volume ratio of 2-4:1-3, performing mixed culture, centrifuging after culture, taking precipitate, drying at 40-50 ℃, and grinding to obtain rhodotorula benthica-bacillus subtilis mixed bacterial powder; the rhodotorula benthica is rich in active substances such as beta-glucan, mannan oligosaccharide, active polypeptide, astaxanthin and the like, can provide abundant nutrient substances for raw material fermentation, and meanwhile, the active polypeptide can inhibit inflammation and resist microbial infection, so that a healthy and harmonious culture medium system can be created in the early fermentation stage, and the fermentation efficiency is improved; the bacillus subtilis can improve the micro-ecological environment, promote the reproduction of beneficial microorganisms and improve the immune function of organisms; the fish meal feed can degrade anti-nutritional molecules in the fish meal, reduce the viscosity of chyme in animal intestinal tracts, reduce the generation of toxic and harmful putrefactive substances in the intestinal tracts, reduce the damage to intestinal walls and the pollution to the environment and improve the digestion and utilization rate of nutritional substances;
s2, mincing fresh aquatic product leftovers, adding nutrients, and uniformly mixing to obtain a fermentation culture medium;
s3, sterilizing the fermentation culture obtained in the step S2 for 20-30min at the temperature of 115-;
s4, cooling the sterilized fermentation medium obtained in the step S3 to room temperature, inoculating the rhodotorula ocellata-bacillus subtilis mixed powder prepared in the step S1, and stirring and uniformly mixing;
s5, fermenting the fermentation medium obtained in the step S4 according to the following method: subpackaging the fermentation medium in a sterilized fermentation tray, wherein the thickness of the material is 4-8cm, covering sterilized wet gauze on the surface of the fermentation medium in the tray, placing the tray on a shelf in a sterilized solid fermentation room for fermentation, and controlling the environmental temperature at 28-30 ℃ for 30-100 h;
s6, after the fermentation is finished, drying and concentrating the fermentation product to obtain the high-quality fish meal product prepared from the aquatic product leftovers.
Preferably, the protective agent is added in the fermentation process of the mixed strain, and the protective agent comprises 8 to 10 percent of skim milk, 5 to 6 percent of xanthan gum, 2 to 4 percent of trehalose, 0.6 to 1.2 percent of potassium sorbate and 0.001 to 0.003 percent of (S) - (+) -3- (1-naphthoxy) -1-phenyl-1-propanol according to mass percentage; one of the special existence of potassium sorbate and (S) - (+) -3- (1-naphthoxy) -1-phenyl-1-propanol can promote the growth and the propagation of fermentation strains, inhibit the infection of harmful bacteria and create a good environment for the fermentation of microbial strains; the microbial bacterial feed is beneficial to promoting microbial bacteria to secrete various active enzymes, the enzymes have the effects of inhibiting inflammation and resisting pathogen infection, can provide a safe and reliable environment for subsequent raw material fermentation, improves the fermentation efficiency, improves the antibacterial and antiviral effects of a fermentation product, and can enhance the immunity of animals when being used as a feed ingredient; the second is beneficial to enhancing the stability of the microorganism mixed bacteria liquid, delaying the deterioration time and improving the enzyme activity retention rate.
Preferably, the nutrient in step S2 is a mixture of glucose, ferrous chloride and dipotassium hydrogen phosphate; the mass of the nutrients in the total mass percentage of the fermentation medium is respectively as follows: 0.01-5% of glucose, 0.01-5% of ferrous chloride and 0.01-5% of dipotassium phosphate.
Preferably, (Z) -3-hexenol in an amount of 0.001-0.003% by weight of the medium and riboflavin in an amount of 0.002-0.003% by weight of the medium are added to the fermentation medium before the fermentation in step S5; the (Z) -3-hexenol and riboflavin can regulate and control the metabolic mechanism of microbial strains in the culture medium, so that the composite flora metabolizes more components such as polysaccharide, amino acid and the like through the proliferation and synergistic effects, the nutrition balance of a fermentation product is improved, and the improvement of the metabolic level of the microbes is favorable for improving the fermentation rate; on the other hand, the active sites of the microbial protease acting on the substrate in the culture medium can be increased, the affinity of the enzyme and the substrate is improved, the substrate is accelerated to be decomposed into small molecular substances, and the obtained fermentation product serving as a feed protein source is favorable for reducing the feed coefficient and improving the digestibility of the feed.
Compared with the prior art, the invention has the advantages that: 1) the invention utilizes the aquatic product leftovers as the raw materials of the fish meal, ensures lower cost, adopts the microbial fermentation method to produce the fish meal, avoids high-temperature cooking of the traditional processing method, can effectively avoid the loss and damage of nutritional ingredients in the fish meal, and improves the quality of the fish meal; 2) potassium sorbate and (S) - (+) -3- (1-naphthoxy) -1-phenyl-1-propanol which are protective agents are added in the mixed strain fermentation to promote the microbial strains to secrete various active enzymes, the enzymes have the effects of inhibiting inflammation and resisting pathogen infection, can provide a safe and reliable environment for subsequent raw material fermentation, improve the fermentation efficiency, simultaneously improve the antibacterial and antiviral effects of a fermentation product, and can enhance the immunity of animals when being used as a feed ingredient; the stability of the microorganism mixed bacteria liquid is enhanced, the time for the microorganism mixed bacteria liquid to deteriorate is delayed, and the enzyme activity retention rate is improved; 3) the (Z) -3-hexenol and riboflavin are added into the fermentation medium, so that the metabolic mechanism of microbial strains in the medium can be regulated and controlled, the composite flora metabolizes more components such as polysaccharide, amino acid and the like through the proliferation effect and the synergistic effect, the nutrition balance of the fermentation product is improved, and the improvement of the metabolic level of the microorganisms is favorable for improving the fermentation rate; on the other hand, the active sites of the microbial protease acting on the substrate in the culture medium can be increased, the affinity of the enzyme and the substrate is improved, the substrate is accelerated to be decomposed into small molecular substances, and the obtained fermentation product serving as a feed protein source is favorable for reducing the feed coefficient and improving the digestibility of the feed.
Detailed Description
The scheme of the invention is further illustrated by the following examples:
example 1:
the method for preparing fish meal by solid-state fermentation of aquatic product leftovers comprises the following steps:
(1) respectively placing the rhodotorula benthica and the bacillus subtilis on a potato sugar agar culture medium for slant culture; inoculating the obtained rhodotorula benthica and bacillus subtilis after slant culture into a rhodotorula benthica and bacillus subtilis seed bottle culture medium for propagation, inoculating 5% of inoculum size of rhodotorula benthica and bacillus subtilis seed bottle bacterial liquid according to the volume ratio of 2:1, performing mixed culture, centrifuging, taking precipitate, drying at 40 ℃, and grinding to obtain rhodotorula benthica-bacillus subtilis mixed bacterial powder; the rhodotorula benthica is rich in active substances such as beta-glucan, mannan oligosaccharide, active polypeptide, astaxanthin and the like, can provide abundant nutrient substances for raw material fermentation, and meanwhile, the active polypeptide can inhibit inflammation and resist microbial infection, so that a healthy and harmonious culture medium system can be created in the early fermentation stage, and the fermentation efficiency is improved; the bacillus subtilis can improve the micro-ecological environment, promote the reproduction of beneficial microorganisms and improve the immune function of organisms; the fish meal feed can degrade anti-nutritional molecules in the fish meal, reduce the viscosity of chyme in animal intestinal tracts, reduce the generation of toxic and harmful putrefactive substances in the intestinal tracts, reduce the damage to intestinal walls and the pollution to the environment and improve the digestion and utilization rate of nutritional substances;
adding a protective agent into the mixed strain fermentation process, wherein the protective agent comprises 8% of skim milk, 5% of xanthan gum, 2% of trehalose, 0.6% of potassium sorbate and 0.001% of (S) - (+) -3- (1-naphthoxy) -1-phenyl-1-propanol by mass percent; one of the special existence of potassium sorbate and (S) - (+) -3- (1-naphthoxy) -1-phenyl-1-propanol can promote the growth and the propagation of fermentation strains, inhibit the infection of harmful bacteria and create a good environment for the fermentation of microbial strains; the microbial bacterial feed is beneficial to promoting microbial bacteria to secrete various active enzymes, the enzymes have the effects of inhibiting inflammation and resisting pathogen infection, can provide a safe and reliable environment for subsequent raw material fermentation, improves the fermentation efficiency, improves the antibacterial and antiviral effects of a fermentation product, and can enhance the immunity of animals when being used as a feed ingredient; the second is beneficial to enhancing the stability of the microorganism mixed bacteria liquid, delaying the deterioration time of the microorganism mixed bacteria liquid and improving the enzyme activity retention rate of the microorganism mixed bacteria liquid;
(2) mincing fresh aquatic product leftovers, adding nutrients, and mixing uniformly to obtain a fermentation culture medium; the nutrient is a mixture of glucose, ferrous chloride and dipotassium hydrogen phosphate; the mass of the nutrients in the total mass percentage of the fermentation medium is respectively as follows: 0.01 percent of glucose, 0.01 percent of ferrous chloride and 0.01 percent of dipotassium phosphate;
(3) sterilizing the fermentation culture medium obtained in the step (2) for 20min at 115 ℃;
(4) cooling the sterilized fermentation medium obtained in the step (3) to room temperature, then inoculating 5% of rhodotorula benthica-bacillus subtilis mixed bacterial powder, and stirring and uniformly mixing;
(5) fermenting the fermentation medium obtained in the step (4) according to the following method: subpackaging the fermentation medium in a sterilized fermentation tray, wherein the material loading thickness is 4cm, covering sterilized wet gauze on the surface of the fermentation medium in the tray, placing the tray on a shelf in a sterilized solid fermentation room for fermentation, and controlling the environmental temperature at 28 ℃ for 30 h;
adding (Z) -3-hexenol accounting for 0.001 percent of the weight of the culture medium and riboflavin accounting for 0.002 percent of the weight of the culture medium into a fermentation culture medium before fermentation; the (Z) -3-hexenol and riboflavin can regulate and control the metabolic mechanism of microbial strains in the culture medium, so that the composite flora metabolizes more components such as polysaccharide, amino acid and the like through the proliferation and synergistic effects, the nutrition balance of a fermentation product is improved, and the improvement of the metabolic level of the microbes is favorable for improving the fermentation rate; on the other hand, the active sites of the microbial protease acting on the substrate in the culture medium can be increased, the affinity of the enzyme with the substrate is improved, the substrate is accelerated to be decomposed into small molecular substances, and the obtained fermentation product serving as a feed protein source is favorable for reducing the feed coefficient and improving the digestibility of the feed;
(6) after the fermentation is finished, drying and concentrating the fermentation product to obtain the fish meal product with excellent quality prepared by the aquatic product leftovers.
Example 2:
the method for preparing fish meal by solid-state fermentation of aquatic product leftovers comprises the following steps:
(1) respectively placing the rhodotorula benthica and the bacillus subtilis on a potato sugar agar culture medium for slant culture; inoculating the obtained rhodotorula benthica and bacillus subtilis after slant culture into a rhodotorula benthica and bacillus subtilis seed bottle culture medium for propagation, inoculating 6% of inoculum size of rhodotorula benthica and bacillus subtilis seed bottle bacterial liquid according to the volume ratio of 3:2, performing mixed culture, centrifuging, taking precipitate, drying at 45 ℃, and grinding to obtain rhodotorula benthica-bacillus subtilis mixed bacterial powder; the rhodotorula benthica is rich in active substances such as beta-glucan, mannan oligosaccharide, active polypeptide, astaxanthin and the like, can provide abundant nutrient substances for raw material fermentation, and meanwhile, the active polypeptide can inhibit inflammation and resist microbial infection, so that a healthy and harmonious culture medium system can be created in the early fermentation stage, and the fermentation efficiency is improved; the bacillus subtilis can improve the micro-ecological environment, promote the reproduction of beneficial microorganisms and improve the immune function of organisms; the fish meal feed can degrade anti-nutritional molecules in the fish meal, reduce the viscosity of chyme in animal intestinal tracts, reduce the generation of toxic and harmful putrefactive substances in the intestinal tracts, reduce the damage to intestinal walls and the pollution to the environment and improve the digestion and utilization rate of nutritional substances;
adding a protective agent into the mixed strain fermentation process, wherein the protective agent comprises 9% of skim milk, 5% of xanthan gum, 3% of trehalose, 1.0% of potassium sorbate and 0.002% of (S) - (+) -3- (1-naphthoxy) -1-phenyl-1-propanol by mass percent;
(2) mincing fresh aquatic product leftovers, adding nutrients, and mixing uniformly to obtain a fermentation culture medium; the nutrient is a mixture of glucose, ferrous chloride and dipotassium hydrogen phosphate; the mass of the nutrients in the total mass percentage of the fermentation medium is respectively as follows: 2.15 percent of glucose, 3.72 percent of ferrous chloride and 1.69 percent of dipotassium phosphate;
(3) sterilizing the fermentation culture medium obtained in the step (2) for 30min at 121 ℃;
(4) cooling the sterilized fermentation medium obtained in the step (3) to room temperature, then inoculating 5% of rhodotorula benthica-bacillus subtilis mixed bacterial powder, and stirring and uniformly mixing;
(5) fermenting the fermentation medium obtained in the step (4) according to the following method: subpackaging the fermentation medium in a sterilized fermentation tray, wherein the material loading thickness is 6cm, covering sterilized wet gauze on the surface of the fermentation medium in the tray, placing the tray on a shelf in a sterilized solid fermentation room for fermentation, and controlling the environmental temperature at 29 ℃ for 85 h; adding (Z) -3-hexenol accounting for 0.002% of the weight of the culture medium and riboflavin accounting for 0.002% of the weight of the culture medium into a fermentation culture medium before fermentation;
(6) after the fermentation is finished, drying and concentrating the fermentation product to obtain the fish meal product with excellent quality prepared by the aquatic product leftovers.
Example 3:
the method for preparing fish meal by solid-state fermentation of aquatic product leftovers comprises the following steps:
(1) respectively placing the rhodotorula benthica and the bacillus subtilis on a potato sugar agar culture medium for slant culture; inoculating the obtained rhodotorula benthica and bacillus subtilis after slant culture into a rhodotorula benthica and bacillus subtilis seed bottle culture medium for propagation, inoculating 7% of inoculum size of rhodotorula benthica and bacillus subtilis seed bottle bacterial liquid according to the volume ratio of 4:3, performing mixed culture, centrifuging, taking precipitate, drying at 50 ℃, and grinding to obtain rhodotorula benthica-bacillus subtilis mixed bacterial powder; the rhodotorula benthica is rich in active substances such as beta-glucan, mannan oligosaccharide, active polypeptide, astaxanthin and the like, can provide abundant nutrient substances for raw material fermentation, and meanwhile, the active polypeptide can inhibit inflammation and resist microbial infection, so that a healthy and harmonious culture medium system can be created in the early fermentation stage, and the fermentation efficiency is improved; the bacillus subtilis can improve the micro-ecological environment, promote the reproduction of beneficial microorganisms and improve the immune function of organisms; the fish meal feed can degrade anti-nutritional molecules in the fish meal, reduce the viscosity of chyme in animal intestinal tracts, reduce the generation of toxic and harmful putrefactive substances in the intestinal tracts, reduce the damage to intestinal walls and the pollution to the environment and improve the digestion and utilization rate of nutritional substances;
adding a protective agent into the mixed strain fermentation process, wherein the protective agent comprises 10% of skim milk, 6% of xanthan gum, 4% of trehalose, 1.2% of potassium sorbate and 0.003% of (S) - (+) -3- (1-naphthoxy) -1-phenyl-1-propanol by mass percent;
(2) mincing fresh aquatic product leftovers, adding nutrients, and mixing uniformly to obtain a fermentation culture medium; the nutrient is a mixture of glucose, ferrous chloride and dipotassium hydrogen phosphate; the mass of the nutrients in the total mass percentage of the fermentation medium is respectively as follows: 5% of glucose, 5% of ferrous chloride and 5% of dipotassium phosphate;
(3) sterilizing the fermentation culture medium obtained in the step (2) for 30min at 130 ℃;
(4) cooling the sterilized fermentation medium obtained in the step (3) to room temperature, then inoculating 5% of rhodotorula benthica-bacillus subtilis mixed bacterial powder, and stirring and uniformly mixing;
(5) fermenting the fermentation medium obtained in the step (4) according to the following method: subpackaging the fermentation medium in a sterilized fermentation tray, wherein the material loading thickness is 8cm, covering sterilized wet gauze on the surface of the fermentation medium in the tray, placing the tray on a shelf in a sterilized solid fermentation room for fermentation, and controlling the environmental temperature at 30 ℃ for 100 hours; adding (Z) -3-hexenol accounting for 0.003 percent of the weight of the culture medium and riboflavin accounting for 0.003 percent of the weight of the culture medium into a fermentation culture medium before fermentation;
(6) after the fermentation is finished, drying and concentrating the fermentation product to obtain the fish meal product with excellent quality prepared by the aquatic product leftovers.
Comparative example 1:
the protective agent added in the mixed strain fermentation does not contain potassium sorbate and (S) - (+) -3- (1-naphthoxy) -1-phenyl-1-propanol, and the rest is completely the same as that in example 2.
Comparative example 2:
no (Z) -3-hexenol and riboflavin were added to the fermentation medium before the fermentation of the starting materials, the remainder being identical to that of example 2.
Determination of main components of fish meal: the fermentation product is frozen and dried in vacuum to prepare fish meal, and the main indexes are measured as follows: crude protein, GB/T6432-1994, Kjeldahl method; crude fat, GB/T6432-1994, Soxhlet extraction; ash, GB/T6432-1994, high temperature firing; moisture, GB/T6432-1994, constant weight method.
The method for measuring the total amount of free amino acid in the fermentation liquor comprises the following steps:
1) preparation of amino acid standard curve:
accurately sucking 0.0, 0.5, 1.0, 1.5, 2.0, 2.5 and 3.0mL of 0.2mg/mL glutamic acid standard solution, respectively placing the solution in a 25mL colorimetric tube with a plug, then adding 1mL each of a 2% ninhydrin solution and a phosphate buffer solution (pH 6.7), uniformly mixing, heating in a water bath for 15min, taking out, rapidly cooling to room temperature, adding water to a marked line, shaking uniformly, standing for 15min, and measuring the light absorption value (A) of each solution at a wavelength of 570nm by taking a reagent blank as a reference. A standard curve was drawn with the number of milligrams of amino acid as the abscissa and the absorbance (A) as the ordinate.
2) And (3) measuring the total amount of free amino acid in the fermentation liquor:
accurately weighing 1.0g of sample, adding water to dilute by 100 times, and uniformly mixing for later use. Absorbing 1.0mL of sample solution, determining the light absorption value (A) under the same conditions according to the standard curve preparation steps, and checking the corresponding amino acid milligrams by using the determined A value on the standard curve.
3) And (4) calculating a result:
in the formula:Cthe number of milligrams of amino acid searched from a standard curve;mthe sample solution for the measurement corresponds to the mass (g) of the sample.
The protein content is a key index for evaluating the quality of the fish meal and is also an important factor influencing the price of the fish meal. The higher the protein content is, the better the quality of the fish meal is, in addition, the crude fat, ash content, water content and the like in the fish meal are also important indexes for evaluating the fish meal, the fish meal mainly containing unsaturated fatty acid is easy to oxidize to cause the quality reduction of the fish meal, the higher the ash content is, the more fishbone, gravel and the like in the fish meal are, and the poorer the quality of the fish meal is; the fish meal component prepared in the embodiment 2 of the invention is measured, and the protein content is 63.28 percent and reaches the first-grade standard; the content of crude fat reaches 8.56 percent and reaches the standard of a special grade product; the ash content is 6.58 percent, and reaches the standard of a superfine product; the water content is 5.32 percent, which meets the standard of fish meal.
The effective viable count of the microbial strains cultured in example 2 and comparative example 1 was respectively detected by using a plate counting method, and the results were 32 hundred million/mL and 25 hundred million/mL respectively, which indicates that the special presence of potassium sorbate and (S) - (+) -3- (1-naphthoxy) -1-phenyl-1-propanol can optimize the effect of the protective agent in microbial strain fermentation, promote the growth and reproduction of the strains, and enhance the stability of the microbial strains.
The total amount of free amino acid in the fermentation liquor of the aquatic leftovers in example 2 and comparative example 2 is measured to be 4.05g/100g and 2.16g/100g respectively, which shows that (Z) -3-hexenol and riboflavin have synergistic effect, and the content of small molecular amino acid in the fermentation product can be improved, so that the prepared fish meal is used as a protein source of aquatic feed, and the digestion utilization rate of the feed can be greatly improved.
The conventional operations in the operation steps of the present invention are well known to those skilled in the art and will not be described herein.
The embodiments described above are intended to illustrate the technical solutions of the present invention in detail, and it should be understood that the above-mentioned embodiments are only specific embodiments of the present invention, and are not intended to limit the present invention, and any modification, supplement or similar substitution made within the scope of the principles of the present invention should be included in the protection scope of the present invention.
Claims (6)
1. A method for preparing fish meal by solid-state fermentation of aquatic leftovers adopts a microbial fermentation method, and is characterized in that: the preparation process of the mixed microbial powder comprises the following steps: respectively placing the rhodotorula benthica and the bacillus subtilis on a potato sugar agar culture medium for slant culture; inoculating the obtained rhodotorula benthica and bacillus subtilis after slant culture into a rhodotorula benthica and bacillus subtilis seed bottle culture medium for propagation, inoculating, fermenting, mixing and culturing, centrifuging, taking precipitate, drying, and grinding to obtain rhodotorula benthica-bacillus subtilis mixed powder;
adding a protective agent into the mixed bacterial powder in the fermentation process, wherein the protective agent comprises 8-10% of skim milk, 5-6% of xanthan gum, 2-4% of trehalose, 0.6-1.2% of potassium sorbate and 0.001-0.003% of (S) - (+) -3- (1-naphthoxy) -1-phenyl-1-propanol by mass percentage;
the preparation process of the fermentation medium comprises the following steps: mincing fresh aquatic product leftovers, adding nutrients, and mixing uniformly to obtain a fermentation culture medium.
2. The method for preparing fish meal by solid state fermentation of aquatic leftovers according to claim 1, wherein the method comprises the following steps: the nutrient is a mixture consisting of glucose, ferrous chloride and dipotassium hydrogen phosphate; the mass of the nutrients in the total mass percentage of the fermentation medium is respectively as follows: 0.01-5% of glucose, 0.01-5% of ferrous chloride and 0.01-5% of dipotassium phosphate.
3. The method for preparing fish meal by solid state fermentation of aquatic leftovers according to claim 1, wherein the method comprises the following steps: the fermentation culture is based on sterilization at the temperature of 115 ℃ and 130 ℃ for 20-30 min.
4. The method for preparing fish meal by solid state fermentation of aquatic leftovers according to claim 1, wherein the method comprises the following steps: the fermentation process comprises the following steps: cooling the sterilized fermentation medium to room temperature, then adding the mixed powder of the rhodotorula benthica and the bacillus subtilis, and uniformly stirring; subpackaging the fermentation medium in a sterilized fermentation tray, wherein the thickness of the material is 4-8cm, covering sterilized wet gauze on the surface of the fermentation medium in the tray, and placing the tray on a shelf in a sterilized solid fermentation room for fermentation.
5. The method for preparing fish meal by solid state fermentation of aquatic leftovers according to claim 4, wherein the method comprises the following steps: the fermentation temperature is controlled at 28-30 ℃, and the fermentation time is 30-100 h.
6. The method for preparing fish meal by solid state fermentation of aquatic leftovers according to claim 4, wherein the method comprises the following steps: adding (Z) -3-hexenol accounting for 0.001-0.003 percent of the weight of the culture medium and riboflavin accounting for 0.002-0.003 percent of the weight of the culture medium into a fermentation culture medium before fermentation.
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