CN105746890A - Method of utilizing fermentation of aquatic product protein and plant feed protein to prepare fish meal replacing protein - Google Patents
Method of utilizing fermentation of aquatic product protein and plant feed protein to prepare fish meal replacing protein Download PDFInfo
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- CN105746890A CN105746890A CN201610129699.0A CN201610129699A CN105746890A CN 105746890 A CN105746890 A CN 105746890A CN 201610129699 A CN201610129699 A CN 201610129699A CN 105746890 A CN105746890 A CN 105746890A
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
- Y02A40/818—Alternative feeds for fish, e.g. in aquacultures
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
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Abstract
The invention discloses a method of utilizing fermentation of aquatic product protein and plant feed protein to prepare fish meal replacing protein.The method mainly includes following steps: culturing high-temperature-resistant facultative anaerobe methyl nutritional bacillus (Bacillus methylotrophicus) HL-4 generating protease to obtain a microbial agent for standby use; mixing minced low-value aquatic product protein or concentrated aquatic product processing wastewater with smashed plant feed protein, adding nutrient and water, and mixing well to form a fermentation medium; adding Bacillus methylotrophicus inoculant and enzyme into the fermentation medium, and mixing well for heap building fermentation in a natural standing manner for 48-72 h; drying, smashing and packaging a product after fermentation to obtain the fish meal replacing protein.The fish meal replacing protein prepared by the method can be used in aquatic product feed to substantially replace imported high-quality fish meal and obviously lower feed cost, and has good application prospect and great realistic significance.
Description
Technical field
The present invention relates to a kind of method utilizing aquatic product protein and vegetable feed proteins fermentation to prepare fish meal replacement albumen, belong to
Technical field in processing of aquatic products Yu utilization.
Background technology
Fish meal is good, nutritious because of palatability, especially amino acid Compositional balance and the rich content of UGF,
It it is indispensable high-quality protein source in aquatic feeds all the time.In aquatic feeds, the consumption of fish meal generally accounts for the 30% of formula
～60%, it is part maximum in its cost.Along with the continuous expansion of aquatic animal raising scale, the whole world in world wide
Demand sustainable growth to fish meal.At present, the annual fish meal supply in the whole world is about 7,000,000 tons.But owing to bad climate affects,
Overfishing and the pollution of marine environment, the many reasons such as destruction of ecological environment, fishery resources are caused by heavy damage entirely
Ball fish meal supply continuous decrease, it is difficult to meet the whole aquatic products industry great demand to fish meal.Therefore fish meal prices one tunnel in recent years
Raise up, and then cause aquaculture cost relatively to raise.But the market competition of fierceness, the market price of aquatic products the most constantly declines,
Directly result in economic benefit to decline, greatly have impact on the enthusiasm for production of culturist.At marine fishery resources growing tension
The protein raw material finding Peru Fish Dietary under situation is the key of the sustainable development maintaining culture fishery.
In recent years, aquatic product protein and vegetable feed proteins mixed fermentation prepare fish meal substitute albumen obtained researcher's
Pay close attention to.Patent application 201210024181.2 " the preparation side of the forage protein of a kind of leftovers of tilapia and dregs of beans mixed fermentation
Method " disclose a kind of with leftovers of tilapia and dregs of beans as raw material, use bacillus, saccharomycete and lactobacillus-fermented to prepare one
Plant the method that fish meal substitutes albumen.Patent application 201010565938.X " a kind of tilapia gruel feed protein and processing method " is public
Open one to starch with vegetable feed proteins as raw material with fish, use bacillus, saccharomycete and lactobacillus-fermented preparation one fish
Powder substitutes the method for albumen.
The existing bacterial classification for fodder protein fermentation is mainly the bacillus subtilis of normal temperature.This bacterium is a kind of aerobic
Bacterium, in this large-scale solid-state submerged fermentation of forage protein, supports wretched insufficiency, needs constantly to be spaced ventilation；And send out
Ferment is a kind of exothermic reaction, and the temperature of charge in sweat constantly raises, and can suppress proceeding of fermentation, thus also to need
Constantly to stir cooling.Thus, prior art prepares fermented feed protein, needs constantly to stir to reach cooling and the mesh of ventilation
's.This mode not only labour intensity is big, and production cost is high.
Summary of the invention
Prepare the problem existing for fermented feed protein for prior art, the invention provides one and utilize aquatic product protein
The method that fish meal substitutes albumen is prepared with vegetable feed proteins fermentation.A kind of preparation that need not stir of the main object of the present invention is sent out
The method of ferment forage protein.
In order to solve the problems referred to above, the technical solution adopted in the present invention is as follows: one utilizes aquatic product protein and plant to raise
Material protein fermentation prepares the method that fish meal substitutes albumen, comprises the steps:
(1) by the high temperature resistant facultative anaerobe Methylotrophic bacillus (Bacillus producing protease of slant preservation
Methylotrophicus) after HL-4 is linked into sterilizing in the LB fluid nutrient medium of cooling, in 55 DEG C, 150r/min cultivation
12h, is prepared as Bacillus methylotrophicus HL-4 seed liquor standby；
(2) by wheat bran and water be in mass ratio 1:1 ratio mixing, and mix be prepared as bran mass, bran mass in
After 121 DEG C of sterilizing 30min standby；
(3) Bacillus methylotrophicus HL-4 seed liquor step (1) prepared is linked into what step (2) obtained
After sterilizing in the bran mass of cooling and mix, then by the bran mass that accesses bacterial classification, to be divided in the fermentation sterilized shallow
In dish, charging thickness is 4cm, and in tray, fermented and cultured primary surface covers the wet gauze of sterilizing, then is placed in by tray sterilized
Shelf top fermentation in solid fermentation room, control environment temperature is at 55 DEG C, and fermentation time is 48h, and the fermentation after fermentation ends is produced
It is prepared as Bacillus methylotrophicus microbial inoculum standby after thing drying and pulverizing；
(4) by low-value aquatic product protein rub after standby or by aquatic products processing wastewater be concentrated by evaporation to protein content be 10%～
20% is standby；
(5) 60 mesh sieves are crossed after being pulverized by vegetable feed proteins standby；
(6) plant that low-value aquatic product protein step (4) rubbed or the aquatic products processing wastewater of concentration and step (5) are pulverized is raised
Material albumen is the ratio mixing of 0.5～1.5:1 in mass ratio, then add nutrients and water be uniformly mixed so as to obtain water content be 40%～
Fermentation medium in the range of 85%；
(7) fermentation medium being prepared into toward step (6) adds enzyme and Bacillus prepared by step (3)
Methylotrophicus microbial inoculum, and mix；
(8) fermentation medium adding enzyme and microbial inoculum step (7) obtained is packed together, and the THICKNESS CONTROL of material is at 50cm
～150cm, and the fermentation materials plastic sheeting building heap is covered good seal, standing for fermentation 48h～72h；
(9) product handling system: by the product drying obtained after fermentation ends, pulverize and pack after, i.e. prepared a kind of fish meal and replaced
For albumen finished product.
The high temperature resistant facultative anaerobe Methylotrophic bacillus producing protease used in described step (1)
(Bacillus methylotrophicus) HL-4 is preserved in Guangdong Province's Microbiological Culture Collection on January 18th, 2016
The heart, deposit number is GDMCC No:60002, and preservation address is 5th floors, No. 59 building of compound, Xianlie Middle Road, Guangzhou City 100 Guangdong Province
Institute of microbiology.
The inoculum concentration of Bacillus methylotrophicus HL-4 seed liquor used in described step (3) is to access
The quality of seed liquor accounts for bran mass gross mass percentages, is 0.1%～5%.
Low-value aquatic product protein used in described step (4) be leftovers of tilapia, krill, shrimp head, scallop body,
Cod leftover bits and pieces, squid leftover bits and pieces, Harengula zunasi leftover bits and pieces, black carp leftover bits and pieces, silver carp leftover bits and pieces, grass carp leftover bits and pieces and stripped tuna are got a foothold
One in material.Aquatic products processing wastewater used in described step (4) is prawn waste water from cooking, minced fillet processing waste water, crust
One in the alkali-wash waste water of element process and fish meal processing waste water.
Vegetable feed proteins used in described step (5) is the one in dregs of beans, peanut meal, the dish dregs of rice and Cottonseed Meal.
Nutrients used in described step (6) is corn flour, calcium chloride, zinc chloride, frerrous chloride and dipotassium hydrogen phosphate
The mixture of composition.The addition of nutrients used in described step (6) accounts for fermentation medium with the quality adding nutrients
The percentages of gross mass, is respectively as follows: corn flour 0.01%～5%, calcium chloride 0.01%～5%, zinc chloride 0.01%～5%, chlorination
Ferrous 0.01%～5%, dipotassium hydrogen phosphate 0.01%～5%.
Enzyme used in described step (7) is cellulase, zytase, 1,4 beta-glucanase, 'beta '-mannase, fat
Enzyme and protease.In described step (7), the quality of the addition of the enzyme enzyme to add accounts for fermentation medium gross mass percentages,
It is respectively as follows: cellulase 0.001%～2%, zytase 0.001%～2%, 1,4 beta-glucanase 0.001%～2%, beta-mannase
Enzyme 0.001%～2%, lipase 0.001%～2%, proteinase-10 .001%～2%.Bacillus in described step (7)
The quality of the addition of the methylotrophicus microbial inoculum microbial inoculum to add accounts for fermentation medium gross mass percentages, for
0.1%～9%.
Compared with prior art, the present invention has the following advantages and feature.
(1) production process is not required to stir, and reduces production cost.The present invention uses high temperature resistant facultative anaerobe
Bacillus methylotrophicus HL-4 has relatively wide fermentation temperature as fermented bacterium, this bacterium, at 10 DEG C～75
All can ferment within DEG C, optimum fermentation temp is 50 DEG C～55 DEG C.This bacterial classification is a kind of facultative anaerobe, thus can carry out big
The submerged fermentation of scale；Additionally this bacterium is high temperature resistant bacterial classification, and its optimum fermentation temp is 50 DEG C～55 DEG C, therefore solid ferment process
Heat production heat up and be conducive to the fermentation of bacterial classification on the contrary, thus sweat need not be lowered the temperature, and takes direct standing for fermentation.To sum up institute
Stating, the present invention is fermented by resistant to elevated temperatures facultative anaerobe, can be directly to take the spontaneous fermentation mode stood, production cost
It is substantially reduced.
(2) ferment effect significantly improves.The fermentation of existing forage protein mainly uses this aerobic bacteria of bacillus subtilis,
And the fermentation of forage protein is that a kind of solid-state submerged fermentation oxygen delivery capacity is very poor, thus, use bacillus subtilis the most aerobic
Bacterium carries out fodder protein fermentation poor effect.And the present invention uses high temperature resistant facultative anaerobe to ferment, very poor in oxygen supply
In the case of also have good ferment effect.Adding the present invention and use hot fermentation, at high temperature the activity of enzyme is notable increases
By force, thus improve ferment effect further, improve the little peptide content of the forage protein of preparation.
The invention provides a kind of method utilizing aquatic product protein and vegetable feed proteins fermentation to prepare fish meal replacement albumen.
The fish meal that the present invention obtains substitutes the culture experiment table of the aquatic livestocks such as albumen prawn, cobio and Tilapia mossambica in South China
Bright, the fish meal prepared by the present invention substitutes albumen and can significantly substitute high-quality imported fish meal, hence it is evident that reduce feed cost, obtains aobvious
The economic benefit write.
Detailed description of the invention
In order to be more fully understood that the present invention, below with regard to aquatic product protein and the kind of vegetable feed proteins, fermentation parameter with send out
The invention will be further described in conjunction with the embodiments for ferment way choice, but protection scope of the present invention is not limited to this.
Embodiment 1: by the product protease high temperature resistant facultative anaerobe Bacillus methylotrophicus of slant preservation
HL-4 is linked into after sterilizing in the LB fluid nutrient medium of cooling, in 55 DEG C, 150r/min cultivate 12h, be prepared as Bacillus
Methylotrophicus HL-4 seed liquor is standby；It is the ratio mixing of 1:1 in mass ratio by wheat bran and water, and mixes preparation
Becoming bran mass, bran mass is standby after 121 DEG C of sterilizing 30min；Bacillus by preparation
Methylotrophicus HL-4 seed liquor be linked into sterilizing after cooling bran mass in and mix, then will access bacterium
The bran mass planted is divided in the fermentation tray sterilized, and charging thickness is 4cm, and in tray, fermented and cultured primary surface covers
The wet gauze of sterilizing, then tray is placed in the shelf top fermentation in sterilized solid fermentation room, controls environment temperature at 55 DEG C,
Fermentation time is 48h, is prepared as Bacillus after the tunning drying after fermentation ends and pulverizing
Methylotrophicus microbial inoculum is standby.By standby after krill rubbing, cross 60 mesh sieves after being pulverized by dregs of beans standby；By 1 ton
The krill rubbed and 1 ton of dregs of beans mixing, and add 30 kilograms of corn flour, 1 kilogram of calcium chloride, 1 kilogram of zinc chloride, 1.8 public affairs
Jin frerrous chloride, 2 kg of phosphoric acid hydrogen dipotassiums and 10 kg of water mix, and obtain fermentation medium；Add in fermentation medium
Enter 1 kilogram of dimension element enzyme, 1 kilogram of zytase, 1 kilogram of 1,4 beta-glucanase, 1 kilogram of 'beta '-mannase, 0.5 kilogram of lipase, 1
Kilogram protease and 5 kilograms of Bacillus methylotrophicus microbial inoculums, and mix；Add bacterial classification and the fermented and cultured of enzyme
Base carries out building heap, and the material thickness building heap is 130cm, and the fermentation materials plastic sheeting building heap is covered good seal, stands
Fermentation 50h；After fermentation ends, substitute albumen finished product by a kind of fish meal can be prepared as after tunning drying and pulverizing.
Embodiment 2: by the high temperature resistant facultative anaerobe Bacillus producing protease of slant preservation
In the LB fluid nutrient medium that methylotrophicus HL-4 cools down after being linked into sterilizing, in 55 DEG C, 150r/min cultivation
12h, is prepared as Bacillus methylotrophicus HL-4 seed liquor standby；It is 1:1's in mass ratio by wheat bran and water
Ratio mixes, and mixing is prepared as bran mass, and bran mass is standby after 121 DEG C of sterilizing 30min；By preparation
Bacillus methylotrophicus HL-4 seed liquor be linked into sterilizing after cooling bran mass in and mix, then
Being divided in the fermentation tray sterilized by the bran mass accessing bacterial classification, charging thickness is 4cm, fermentation medium in tray
Surface covers the wet gauze of sterilizing, then tray is placed in the shelf top fermentation in sterilized solid fermentation room, controls environment temperature
Degree is at 55 DEG C, and fermentation time is 48h, is prepared as Bacillus after the tunning drying after fermentation ends and pulverizing
Methylotrophicus microbial inoculum is standby.It is 15% standby that prawn waste water from cooking heating evaporation is concentrated into protein content, will flower
It is standby that the raw dregs of rice cross 60 mesh sieves after pulverizing；1 ton is concentrated waste water and the mixing of 1 ton of peanut meal, and add 40 kilograms of corn flour, 1 kilogram
Calcium chloride, 1 kilogram of zinc chloride, 1.2 kilograms of frerrous chlorides, 1.5 kg of phosphoric acid hydrogen dipotassiums and 5 kg of water mix, and are sent out
Ferment culture medium；Add in fermentation medium 1.8 kilograms of cellulases, 1 kilogram of zytase, 1.8 kilograms of 1,4 beta-glucanases, 1
Kilogram 'beta '-mannase, 0.1 kilogram of lipase, 1 kilogram of protease and 8 kilograms of Bacillus methylotrophicus bacterium
Agent, and mix；The fermentation medium adding bacterial classification and enzyme carries out building heap, and the material thickness building heap is 130cm, and will build sending out of heap
Ferment material plastic sheeting covers good seal, standing for fermentation 70h；After fermentation ends, by after tunning drying and pulverizing it is
A kind of fish meal can be prepared as and substitute albumen finished product.
Claims (11)
1. one kind utilizes aquatic product protein and vegetable feed proteins fermentation to prepare the method that fish meal substitutes albumen, it is characterised in that: should
Method comprises the steps:
(1) by the high temperature resistant facultative anaerobe Methylotrophic bacillus (Bacillus producing protease of slant preservation
Methylotrophicus) after HL-4 is linked into sterilizing in the LB fluid nutrient medium of cooling, in 55 DEG C, 150r/min cultivation
12h, is prepared as Bacillus methylotrophicus HL-4 seed liquor standby；
(2) by wheat bran and water be in mass ratio 1:1 ratio mixing, and mix be prepared as bran mass, bran mass in
After 121 DEG C of sterilizing 30min standby；
(3) Bacillus methylotrophicus HL-4 seed liquor step (1) prepared is linked into what step (2) obtained
After sterilizing in the bran mass of cooling and mix, then by the bran mass that accesses bacterial classification, to be divided in the fermentation sterilized shallow
In dish, charging thickness is 4cm, and in tray, fermented and cultured primary surface covers the wet gauze of sterilizing, then is placed in by tray sterilized
Shelf top fermentation in solid fermentation room, control environment temperature is at 55 DEG C, and fermentation time is 48h, and the fermentation after fermentation ends is produced
It is prepared as Bacillus methylotrophicus microbial inoculum standby after thing drying and pulverizing；
(4) by low-value aquatic product protein rub after standby or by aquatic products processing wastewater be concentrated by evaporation to protein content be 10%～
20% is standby；
(5) 60 mesh sieves are crossed after being pulverized by vegetable feed proteins standby；
(6) plant that low-value aquatic product protein step (4) rubbed or the aquatic products processing wastewater of concentration and step (5) are pulverized is raised
Material albumen is the ratio mixing of 0.5～1.5:1 in mass ratio, then add nutrients and water be uniformly mixed so as to obtain water content be 40%～
Fermentation medium in the range of 85%；
(7) fermentation medium being prepared into toward step (6) adds enzyme and Bacillus prepared by step (3)
Methylotrophicus microbial inoculum, and mix；
(8) fermentation medium adding enzyme and microbial inoculum step (7) obtained is packed together, and the THICKNESS CONTROL of material is at 50cm
～150cm, and the fermentation materials plastic sheeting building heap is covered good seal, standing for fermentation 48h～72h；
(9) product handling system: by the product drying obtained after fermentation ends, pulverize and pack after, i.e. prepared a kind of fish meal and replaced
For albumen finished product.
The most according to claim 1 a kind of utilize aquatic product protein and vegetable feed proteins fermentation to prepare fish meal to substitute albumen
Method, it is characterised in that: the high temperature resistant facultative anaerobe Bacillus producing protease used in described step (1)
Methylotrophicus HL-4 is GDMCC No:60002 at the deposit number of Guangdong Province's Culture Collection.
The most according to claim 1 a kind of utilize aquatic product protein and vegetable feed proteins fermentation to prepare fish meal to substitute albumen
Method, it is characterised in that: the inoculation of Bacillus methylotrophicus HL-4 seed liquor used in described step (3)
Measure and account for bran mass gross mass percentages with the quality accessing seed liquor, be 0.1%～5%.
The most according to claim 1 a kind of utilize aquatic product protein and vegetable feed proteins fermentation to prepare fish meal to substitute albumen
Method, it is characterised in that: low-value aquatic product protein used in described step (4) be leftovers of tilapia, krill, shrimp head,
Scallop body, cod leftover bits and pieces, squid leftover bits and pieces, Harengula zunasi leftover bits and pieces, black carp leftover bits and pieces, silver carp leftover bits and pieces, grass carp leftover bits and pieces
With the one in stripped tuna leftover bits and pieces.
The most according to claim 1 a kind of utilize aquatic product protein and vegetable feed proteins fermentation to prepare fish meal to substitute albumen
Method, it is characterised in that: aquatic products processing wastewater used in described step (4) be prawn waste water from cooking, minced fillet processing waste water,
One in the alkali-wash waste water of chitin process and fish meal processing waste water.
The most according to claim 1 a kind of utilize aquatic product protein and vegetable feed proteins fermentation to prepare fish meal to substitute albumen
Method, it is characterised in that: vegetable feed proteins used in described step (5) is in dregs of beans, peanut meal, the dish dregs of rice and Cottonseed Meal
A kind of.
The most according to claim 1 a kind of utilize aquatic product protein and vegetable feed proteins fermentation to prepare fish meal to substitute albumen
Method, it is characterised in that: nutrients used in described step (6) is corn flour, calcium chloride, zinc chloride, frerrous chloride and phosphorus
The mixture of acid hydrogen dipotassium composition.
The most according to claim 1 a kind of utilize aquatic product protein and vegetable feed proteins fermentation to prepare fish meal to substitute albumen
Method, it is characterised in that: the addition of nutrients used in described step (6) accounts for fermented and cultured with the quality adding nutrients
The percentages of base gross mass, is respectively as follows: corn flour 0.01%～5%, calcium chloride 0.01%～5%, zinc chloride 0.01%～5%, chlorine
Change ferrous 0.01%～5%, dipotassium hydrogen phosphate 0.01%～5%.
The most according to claim 1 a kind of utilize aquatic product protein and vegetable feed proteins fermentation to prepare fish meal to substitute albumen
Method, it is characterised in that: enzyme used in described step (7) is that cellulase, zytase, 1,4 beta-glucanase, β-sweet dew gather
Carbohydrase, lipase and protease.
One the most according to claim 1 utilizes aquatic product protein and vegetable feed proteins fermentation to prepare fish meal and substitutes albumen
Method, it is characterised in that: in described step (7), the addition of enzyme accounts for fermentation medium gross mass hundred with the quality of enzyme added
Proportion by subtraction meter, is respectively as follows: cellulase 0.001%～2%, zytase 0.001%～2%, 1,4 beta-glucanase 0.001%～2%, β-sweet
Dew dextranase 0.001%～2%, lipase 0.001%～2%, proteinase-10 .001%～2%.
11. one according to claim 1 utilize aquatic product protein and vegetable feed proteins fermentation to prepare fish meal and substitute albumen
Method, it is characterised in that: in described step (7) addition of Bacillus methylotrophicus microbial inoculum with add
The quality of microbial inoculum accounts for fermentation medium gross mass percentages, is 0.1%～9%.
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| CN106689692A (en) * | 2016-11-25 | 2017-05-24 | 彭程 | Protein feed for substituting soybean meal or fish meal |
| CN106721054A (en) * | 2016-12-14 | 2017-05-31 | 新昌县拜瑞珂生物技术有限公司 | The method that forage protein is prepared using alcohol effluent and crop material |
| CN107094984A (en) * | 2017-03-31 | 2017-08-29 | 上海优久生物科技有限公司 | A kind of Tilapia mossambica foreign protein processing method and its application |
| CN108925747A (en) * | 2018-06-01 | 2018-12-04 | 东莞市银华生物科技有限公司 | A kind of functional protein and its method using aquatic product protein and saccharomycete preparation |
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