CN105724764B - A method of antibacterial green feed additive for promoting growth is prepared using aquatic product protein fermentation - Google Patents
A method of antibacterial green feed additive for promoting growth is prepared using aquatic product protein fermentation Download PDFInfo
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- CN105724764B CN105724764B CN201610129706.7A CN201610129706A CN105724764B CN 105724764 B CN105724764 B CN 105724764B CN 201610129706 A CN201610129706 A CN 201610129706A CN 105724764 B CN105724764 B CN 105724764B
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- 238000000855 fermentation Methods 0.000 title claims abstract description 102
- 230000004151 fermentation Effects 0.000 title claims abstract description 102
- 238000000034 method Methods 0.000 title claims abstract description 35
- 102000004169 proteins and genes Human genes 0.000 title claims abstract description 32
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 32
- 239000003674 animal food additive Substances 0.000 title claims abstract description 24
- 230000000844 anti-bacterial effect Effects 0.000 title claims abstract description 20
- 230000001737 promoting effect Effects 0.000 title claims abstract description 20
- 241000194108 Bacillus licheniformis Species 0.000 claims abstract description 36
- 241000411851 herbal medicine Species 0.000 claims abstract description 25
- 108091005804 Peptidases Proteins 0.000 claims abstract description 21
- 239000007788 liquid Substances 0.000 claims abstract description 19
- 102000035195 Peptidases Human genes 0.000 claims abstract description 10
- 235000019833 protease Nutrition 0.000 claims abstract description 10
- 238000010563 solid-state fermentation Methods 0.000 claims abstract description 10
- 238000001035 drying Methods 0.000 claims abstract description 8
- 235000015097 nutrients Nutrition 0.000 claims abstract description 7
- 238000002156 mixing Methods 0.000 claims abstract description 6
- 238000001914 filtration Methods 0.000 claims abstract description 5
- 238000004806 packaging method and process Methods 0.000 claims abstract description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 5
- 239000000047 product Substances 0.000 claims description 63
- 235000021190 leftovers Nutrition 0.000 claims description 17
- 239000002609 medium Substances 0.000 claims description 14
- 230000001954 sterilising effect Effects 0.000 claims description 13
- 239000001963 growth medium Substances 0.000 claims description 8
- 239000007787 solid Substances 0.000 claims description 7
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 6
- 241000238557 Decapoda Species 0.000 claims description 6
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 6
- 239000001110 calcium chloride Substances 0.000 claims description 6
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 6
- 239000008103 glucose Substances 0.000 claims description 6
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- 238000004321 preservation Methods 0.000 claims description 5
- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 claims description 4
- 241000276707 Tilapia Species 0.000 claims description 4
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 4
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- 238000001816 cooling Methods 0.000 claims description 2
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- 238000010564 aerobic fermentation Methods 0.000 abstract 2
- 208000035240 Disease Resistance Diseases 0.000 abstract 1
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- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 150000001805 chlorine compounds Chemical class 0.000 description 2
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- 108010011619 6-Phytase Proteins 0.000 description 1
- 240000006439 Aspergillus oryzae Species 0.000 description 1
- 235000002247 Aspergillus oryzae Nutrition 0.000 description 1
- JEBFVOLFMLUKLF-IFPLVEIFSA-N Astaxanthin Natural products CC(=C/C=C/C(=C/C=C/C1=C(C)C(=O)C(O)CC1(C)C)/C)C=CC=C(/C)C=CC=C(/C)C=CC2=C(C)C(=O)C(O)CC2(C)C JEBFVOLFMLUKLF-IFPLVEIFSA-N 0.000 description 1
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- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 241000588767 Proteus vulgaris Species 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 241000293869 Salmonella enterica subsp. enterica serovar Typhimurium Species 0.000 description 1
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- MBLBDJOUHNCFQT-LXGUWJNJSA-N aldehydo-N-acetyl-D-glucosamine Chemical compound CC(=O)N[C@@H](C=O)[C@@H](O)[C@H](O)[C@H](O)CO MBLBDJOUHNCFQT-LXGUWJNJSA-N 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
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- 229940022405 astaxanthin Drugs 0.000 description 1
- 235000013793 astaxanthin Nutrition 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
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Classifications
-
- 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
Landscapes
- Fodder In General (AREA)
Abstract
The invention discloses a kind of methods for preparing antibacterial green feed additive for promoting growth using aquatic product protein fermentation, including following key step: fresh aquatic product protein is rubbed;Aquatic product protein after rubbing is added in fermentor, and adds nutrients and water mixing as fermentation medium;In fermentor access produce thermostable proteinase bacillus licheniformis (Bacillus licheniformis) HL-3 progress liquid aerobic fermentation;After liquid state fermentation, fermentation liquid continues solid-state aerobic fermentation 46h~56h at 55 DEG C~60 DEG C after mixing after filtering removal precipitating with Chinese herbal medicine;Product after solid state fermentation is prepared into a kind of antibacterial green feed additive for promoting growth finished product after drying, crushing and packaging.The feed addictive of the method for the present invention preparation is rich in small peptide, thermostable proteinase, high temperature resistant non-starch polysaccharide enzyme, Chinese herbal medicine effective ingredients and probiotics, it has good growth promotion performance and disease resistance, there is good application prospect and social effect.
Description
Technical field
The present invention relates to a kind of methods for preparing antibacterial green feed additive for promoting growth using aquatic product protein fermentation, belong to
The technical field of feed addictive preparation.
Background technique
Since the 1950s, antibiotic is widely used in animal feeding and feed, chemical synthetic drug, has swashed
The growth promotions health-care agent such as element, beta-stimulants, heavy metal, sedative.Application of the growth promotions health-care agent such as antibiotic in feed is gone through
History is long-standing, it creates the revolution of aquaculture and feed industry so that breeding scale has obtained fast in worldwide
Speed development, the production quantity and feed conversion rate of animal are greatly improved.But the achievement of industrial civilization is all as in all
As having its negative effect, for a long time using non-green feed addictives such as antibiotic, following problem is resulted in: poultry
Fowl and aquatic products product drug residue are serious, are directly detrimental to health;It is competing to lack market for livestock and poultry and aquatic products deterioration in quality
Strive power and trust;Bacterial drug resistance enhancing, antibody-resistant bacterium increase, and threaten human security;Destroy ecological environment etc..With feed
The fast development of industry and animal husbandry and fishery industry, the livestock products and aquatic products in China are changed into interim surplus from shortage at present,
People's whether safe, health more concerned with food, thus green consumption has become development trend from now on, W has been added in China in addition
TO, developed country require to be increasingly stringenter to the agricultural product of China's export, and medicament residue problem becomes China's livestock products at present
One of the major obstacle gone abroad with aquatic products.Therefore exploitation, promotion and application novel green feed addictive gesture is accelerated to exist
It must go.
Green feed additive, most basic requirement be have no toxic side effect, noresidue, and also want that growth of animal can be promoted.
Current internationally recognized growth promotion green feed additive is mainly include the following types: small peptide, enzyme preparation, plant product-derived and its mention
Take object, probiotics, oligosaccharides (chemical probiotic) and yeast class product (natural prebiotics) etc..
Enzyme preparation has noresidue, pollution-free as a kind of feed addictive, improves the utility value of feed, improves animal
The effects of production performance and product quality.Fodder enzyme preparation can substantially be divided into two class of digestive ferment and non-digestive ferment, usually seek
Feeding level plays a role.For example livestock and poultry alimentary tract endogenous enzyme activity is improved, supplement the deficiency of endogenous enzymes;Plant cell wall is destroyed,
Improve the utilization efficiency of feed;The anti-nutritional factors in feed is eliminated, the digestion and absorption of nutriment is promoted.Currently, enzyme preparation
Feed industry is had been widely used for, mainly there are the enzyme preparations such as protease, non-starch polysaccharide enzyme and phytase.However, in the market
The existing more non-refractories of fodder enzyme preparation, loss of activity is very big in feed high temperature granulation process.
Chinese herbal medicine is the rarity in Chinese medicine treasure-house, and Chinese herbal medicine has been opened as feed addictive in Ancient Times in China
Begin.Chinese herbal medicine, which has, to have no toxic side effect, is not easy to cause the characteristics such as resistance, noresidue, as a kind of feeding antibiotic
Green substitute has been widely recognized in feed industry.From the 1980s, Chinese herbal medicine has been used for making feeding anti-
Raw element substitute, and achieve certain application effect.After Chinese herbal medicine undergoes microbial fermentation, effective component is sufficiently released
It puts, so that it is had more bioactivity and curative effect is even more research hotspot in recent years.Chinese feed (2010,419(15): 13-17) document
It discloses isolated aspergillus oryzae from different herbal raw materials to ferment to Chinese herbal medicine, fermentation material cooking liquor is to large intestine bar
Bacterium, staphylococcus aureus, proteus vulgaris and salmonella typhimurium bacteriostatic activity respectively relatively control improve 36.03%,
60.83%、27.64%、41.32%。
Result of study both domestic and external shows that certain small peptide is added in daily ration can improve body to protein in daily ration
With the utilization rate of amino acid, the growth performance of animal is improved.In recent years, small peptide exists as a kind of green feed additive for promoting growth
Feed industry is concerned.The preparation method of small peptide has enzymatic isolation method and microbe fermentation method at present.Enzymatic isolation method produces active peptide
Using protein raw materials as substrate, with the enzymolysis of protease, small-molecular peptides are produced.Enzymatic isolation method is easily controllable, action condition temperature
With the production of suitable active peptide.But enzymatic isolation method needs a large amount of protease, high production cost.Therefore, enzymatic isolation method prepares active peptide
It is chiefly used in food and pharmaceuticals industry, the feeding active peptide product of low side is seldom using the method for enzymatic hydrolysis.Microbe fermentation method be with
Macro-molecular protein is raw material, and access produces the microorganism of protease, utilizes the enzymatic hydrolysis for the protease that fermentation process generates
Effect generates small-molecular peptides.Fermentation method organically combines the fermenting and producing and enzymolysis process of protease, reduces activity
The manufacturing cost of peptide is more suitable for the production of active peptide.Patent CNZL200410029787.0 " a kind of activated peptide forage additive and
Preparation method and purposes " discloses a kind of method that fermentation hair prepares small peptide feed additive, it is with vegetable protein and animal
One or more of mixtures of albumen make the control of its degree of hydrolysis 20~35%, then pass through through fermentative degradations such as bacillus
Later small peptide feed additive can be obtained in processing.
Small peptide has good application prospect in feed industry as a kind of novel green feed additive for promoting growth, however
There is no be widely used small peptide in feed industry at present.The either small peptide of enzymatic isolation method production or fermentation method production
Small peptide, high production cost constrains small peptide in the extensive use of feed industry.In addition, the small peptide of prior art preparation produces
Product functional components are excessively single, are especially a lack of anti-microbial property.Therefore, the production cost that small peptide is greatly lowered is needed, with
Promote small peptide in the extensive use of feed industry;In addition, the anti-microbial property of small peptide product need to be enhanced, to further enhance its growth-promoting
Long performance.
Summary of the invention
For prior art preparation small peptide due at high cost, be difficult to be widely applied this problem in feed industry, this
The purpose of invention is to provide a kind of method can produce small peptide at low cost.In addition, it is another object of the present invention to by using
High temperature resistant the lichen bacillus ferments prepare small peptide, so that the small peptide product being prepared also is rich in the enzymes systems such as thermostable proteinase
Agent.This not only can be further improved the growth-promoting effect of small peptide product, and can also be by producing the feeding enzyme system of high temperature resistant
Agent and further decrease its production cost.In addition, a further object of the present invention is the mixed fermentation by fermentation liquid and Chinese herbal medicine,
So that the small peptide product of preparation is also equipped with anti-microbial property.
To solve the above-mentioned problems, the technical solution adopted in the present invention is as follows: a kind of to utilize aquatic product protein fermentation preparation
The method of antibacterial green feed additive for promoting growth, includes the following steps:
(1) by slant preservation can produce thermostable proteinase bacillus licheniformis (Bacillus licheniformis)
HL-3 is linked into the LB liquid medium cooled down after sterilizing, is cultivated 12h in 55 DEG C, 150r/min, is prepared into lichens gemma bar
Bacterium (Bacillus licheniformis) HL-3 seed liquor is spare;
(2) spare after rubbing fresh aquatic product protein;
(3) step (2) aquatic product protein obtained rubbed is put into fermentor, and water and battalion is added into fermentor
Object is supported, liquid state fermentation culture medium is obtained after mixing;
(4) bacillus licheniformis prepared by step (1) is accessed in the liquid state fermentation culture obtained toward step (3)
(Bacillus licheniformis) HL-3 seed liquor;
(5) control of fermentation process: fermentation temperature be 55 DEG C~60 DEG C, filtrated air intake be 0.5vvm~
2.5vvm, the revolving speed of fermentor are 150rpm~350rpm, and fermentation time is 36h~48h;
(6) after filtering fermentation liquor removal precipitates after fermentation and the Chinese herbal medicine of 60 meshes is smashed it through by 0.5~1.5:1
Ratio mixing, and stir evenly as the solid-state fermentation culture medium for being connected to strain;
(7) solid state fermentation: the solid-state fermentation culture medium for being connected to strain that step (6) obtain is sent out in following manner
Ferment: fermentation medium is divided in the fermentation tray sterilized, and charging is with a thickness of 4cm~8cm, fermented and cultured primary surface in tray
Sterilized wet gauze is covered, then tray is placed in the shelf top fermentation in sterilized solid fermentation room, controls environment temperature
At 55 DEG C ~ 60 DEG C, fermentation time is 46h~56h;
(8) product handling system: solid state fermentation terminates, and by the product obtained after fermentation after drying, crushing and packaging, obtains
A kind of antibacterial green feed additive for promoting growth finished product.
Can be produced used in the step (1) thermostable proteinase bacillus licheniformis (Bacillus licheniformis) HL-3 is preserved in Guangdong Province's Culture Collection on January 18th, 2016, deposit number is
GDMCC No:60003, preservation address are the compound the 59th of Xianlie Middle Road, Guangzhou City 100 5 building, building Guangdong Microbes Inst.
Aquatic product protein in the step (2) is leftovers of tilapia, krill, shrimp head, scallop body, gadus leftover bits and pieces
One in material, 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 leftover bits and pieces
Kind.
It is total with the quality of the aquatic product protein added to account for liquid state fermentation culture medium for the additional amount of aquatic product protein in the step (3)
Mass percent meter is 10%~40%.Nutrients is glucose, calcium chloride, frerrous chloride and phosphoric acid hydrogen two in the step (3)
The mixture of potassium composition.The additional amount of nutrients in the step (3) accounts for liquid state fermentation training with the quality of the nutrients added
Base gross mass percentages are supported, are respectively as follows: glucose 0.01%~5%, calcium chloride 0.01%~5%, frerrous chloride 0.01%~5%,
Dipotassium hydrogen phosphate 0.01%~5%.
Bacillus licheniformis in the step (4) (Bacillus licheniformis) HL-3 seed liquor inoculum concentration with
The quality of the seed liquor of access accounts for fermentation medium gross mass percentages, is 1%~9%.
Chinese herbal medicine in the step (6) is Radix Glycyrrhizae, Fructus Forsythiae, Asiatic plantain, perillaseed, Schisandra chinensis, Verbena officinalis, Radix Astragali, gold and silver
One of flower, granatum, garden burnet, the coptis, cordate houttuynia, kuh-seng and smilax or in which two or more mixtures.
The present invention has the following advantages compared with the technology of existing fermentation method preparation small peptide.
(1) fermented product can raise the effect of playing protease in animal body.Prepared by the fermentation of existing small peptide uses
Be all room temperature strain, the protease generated during the fermentation activity product preparation drying process and for raising
It can be lost totally in the high temperature granulation process of material.Thus, the small peptide product of existing fermentation technique preparation is for feed in animal body
Interior the effect of can not playing protease.And this technology using bacillus licheniformis resistant to high temperature (Bacillus licheniformis) ferment, fermentation process generates protease resistant to high temperature, and the activity of these thermostable proteinases is producing
Product preparation drying and for not lost substantially in the high temperature granulation process of feed, thus this technology preparation feed addictive
The effect of protease can be played in animal body.
(2) at low cost.Compared with existing fermentation method prepares small peptide, in technical solution provided by the invention, raw material does not have to
Sterilizing, can reduce energy consumption and cost.In addition, the fermenting and producing of small peptide and the fermenting and producing of fodder enzyme preparation conjunction two are by the present invention
One, the production of fodder enzyme preparation can share part producing cost, to further decrease the production cost of small peptide.In addition, this hair
The bright solid state fermentation by the post processor of small peptide product and Chinese herbal medicine organically combines, and the fermenting and producing of Chinese herbal medicine can divide
Part producing cost is spread out, this further reduces the production cost of small peptide.
(3) ferment effect is good, and small peptide content is high.Compared with existing fermentation method prepares small peptide, technology provided by the invention
In scheme, raw material does not have to sterilizing, so that aquatic product protein endogenous enzymes abundant are retained;In addition, the most suitable enzyme of endogenous enzymes
It is consistent with microbial fermentation temperature to solve temperature.Therefore, under the synergistic effect of endogenous enzymes and fermentation institute's producing enzyme, what is be prepared is small
Peptide content can be higher;In the activity that can improve enzyme plus high temperature, small peptide content can be further increased.
(4) product is rich in Chinese herbal medicine effective ingredients, can enhance its premunition, and further increase its growth promotion performance.This
Fermentation liquid containing small peptide is directly carried out mixed fermentation with Chinese herbal medicine by product, by the fermentation of microorganism by Chinese herbal medicine
Effective component releases, so that the finished product finally prepared has good anti-microbial property.
(5) it can effectively protect the function factor of thermal sensitivity in aquatic product protein.In technical solution of the present invention, since raw material can
With not sterilized, thus, the activity of the function factor of thermal sensitivity is effectively retained in aquatic product protein.For example, shrimp head and the South Pole
Astaxanthin in krill.
The present invention has the following advantages that compared with existing Chinese herbal medicine fermentation technique.
(1) compared with existing Chinese herbal medicine fermentation technique, in technical solution provided by the invention, raw material does not have to sterilizing, can
Reduce energy consumption and cost.In addition, the present invention is by the fermenting and producing of Chinese herbal medicine and fodder enzyme preparation (predominantly cellulase and non-shallow lake
Powder polysaccharase) fermenting and producing be combined into one, the production of fodder enzyme preparation can share part producing cost, to further decrease
The cost of Chinese herbal medicine fermentation.
(2) fermented product can raise the effect of playing non-starch polysaccharide enzyme in animal body.The fermentation of existing Chinese herbal medicine
What is used is all the strain of room temperature, and the activity of the non-starch polysaccharide enzyme generated during the fermentation is in the dried of product preparation
Journey and for that can be lost totally in the high temperature granulation process of feed.Thus, the fermentation of Chinese herbal medicine product of prior art preparation is used for
Feed can not play the effect of non-starch polysaccharide enzyme in animal body.And this technology uses bacillus licheniformis resistant to high temperature
(Bacillus licheniformis) ferment, fermentation process generates non-starch polysaccharide enzyme resistant to high temperature, these high temperature resistants
The activity of non-starch polysaccharide enzyme is not lost substantially in the drying of product preparation and the high temperature granulation process for feed, thus
The effect of feed addictive of this technology preparation can play non-starch polysaccharide enzyme in animal body.
The present invention provides a kind of methods for preparing antibacterial green feed additive for promoting growth using aquatic product protein fermentation.This
Obtained antibacterial feed additive for promoting growth is invented to show in the culture experiment of the pig of South China, chicken, prawn with present invention life
The feed of the antibacterial green feed additive for promoting growth cooperation of production, not only can be improved the growth result of raising animal, but also can
The immunity and premunition for improving raising animal can be used directly to prepare green feed, have a good application prospect and social
Meaning.
Specific embodiment
For a better understanding of the present invention, below with regard to the type of aquatic product protein, the type of Chinese herbal medicine, fermentation parameter and fermentation
The invention will be further described in conjunction with the embodiments for the selection of mode, and but the scope of the present invention is not limited thereto.
Embodiment 1: by slant preservation can produce thermostable proteinase bacillus licheniformis (Bacillus licheniformis) HL-3 is linked into cooling LB liquid medium after sterilizing, 12h, system are cultivated in 55 DEG C, 150r/min
It is standby at bacillus licheniformis (Bacillus licheniformis) HL-3 seed liquor is spare.Fresh shrimp head is rubbed into standby
With.By 20 kilograms rub shrimp head and 100 kg of water be added in fermentor, and into fermentor be added 200 grams of glucose,
550 grams of calcium chloride, 580 grams of frerrous chlorides and 750 grams of dipotassium hydrogen phosphates obtain fermentation medium;Fermentation training into fermentor
Support be added in base 9 kilograms of bacillus licheniformis (Bacillus licheniformis) HL-3 seed liquor;Fermentation temperature is 60 DEG C,
Filtrated air intake is 1.5vvm~2.5vvm, and the revolving speed of fermentor is 250rpm~350rpm, fermentation time 48h;Hair
Fermentation liquid is mixed with Radix Glycyrrhizae in the ratio of 1:1 after filtering removes shrimp shell after ferment, and is stirred evenly as the solid-state that strain is added
Culture medium;The solid medium that strain is added is divided in the fermentation tray with 75% alcohol surface sterilization, is charged in tray
With a thickness of 4 centimetres, the wet gauze of fermented and cultured primary surface covering sterilization treatment in tray, then the solid that tray is placed in sterilizing is sent out
Shelf top fermentation in ferment room, control environment temperature are 60 DEG C, fermentation time 53h;After fermentation, it will obtain after fermentation
Product has obtained a kind of antibacterial green growth-promoting feed addition finished product of the present invention after drying, crushing and packaging.
Embodiment 2: by the bacillus licheniformis (Bacillus that can produce thermostable proteinase of slant preservation
Licheniformis) HL-3 is linked into the LB liquid medium cooled down after sterilizing, cultivates 12h, system in 55 DEG C, 150r/min
It is standby at bacillus licheniformis (Bacillus licheniformis) HL-3 seed liquor is spare.Fresh leftovers of tilapia is twisted
It is spare after broken.Leftovers of tilapia that 18 kilograms rub and 100 kg of water are added in fermentor, and are added into fermentor
100 grams of glucose, 520 grams of calcium chloride, 550 grams of frerrous chlorides and 660 grams of dipotassium hydrogen phosphates obtain fermentation medium;Toward fermentation
Be added in fermentation medium in tank 8 kilograms of bacillus licheniformis (Bacillus licheniformis) HL-3 seed liquor;Hair
Ferment temperature is 57 DEG C, and filtrated air intake is 1.5vvm~2.5vvm, and the revolving speed of fermentor is 250rpm~350rpm, fermentation
Time is 45h;Fermentation liquid is mixed with Radix Astragali in the ratio of 0.9:1 after filtering removes fish-bone after fermentation, and stirs evenly conduct
The solid medium of strain is added;The fermentation that the solid medium of addition strain is divided in 75% alcohol surface sterilization is shallow
In disk, charging is in tray with a thickness of 6 centimetres, the wet gauze of fermented and cultured primary surface covering sterilization treatment in tray, then by tray
The shelf top fermentation being placed in the solid fermentation room of sterilizing, control environment temperature are 57 DEG C, fermentation time 50h;Fermentation ends
Afterwards, by the product obtained after fermentation after drying, crushing and packaging, a kind of antibacterial green growth-promoting of the present invention has been obtained
Long feed addition finished product.
Claims (7)
1. a kind of method for preparing antibacterial green feed additive for promoting growth using aquatic product protein fermentation, it is characterised in that: the party
Method includes the following steps:
(1) by slant preservation can produce thermostable proteinase bacillus licheniformis (Bacillus licheniformis) HL-3
It is linked into LB liquid medium cooling after sterilizing, cultivates 12h in 55 DEG C, 150r/min, be prepared into bacillus licheniformis
(Bacillus licheniformis) HL-3 seed liquor is spare;
(2) spare after rubbing fresh aquatic product protein;
(3) step (2) aquatic product protein obtained rubbed is put into fermentor, and water and nutrition is added into fermentor
Object obtains liquid state fermentation culture medium after mixing;
(4) toward step (3) obtain liquid state fermentation culture on access bacillus licheniformis prepared by step (1) (Bacillus licheniformis) HL-3 seed liquor;
(5) control of fermentation process: fermentation temperature is 55 DEG C~60 DEG C, and filtrated air intake is 0.5vvm~2.5vvm, hair
The revolving speed of fermentation tank is 150rpm~350rpm, and fermentation time is 36h~48h;
(6) after filtering fermentation liquor removal precipitates after fermentation and the Chinese herbal medicine of 60 meshes is smashed it through by the ratio of 0.5~1.5:1
Example mixing, and stir evenly as the solid-state fermentation culture medium for being connected to strain;
(7) solid state fermentation: the solid-state fermentation culture medium for being connected to strain that step (6) obtain is fermented in following manner:
Fermentation medium is divided in the fermentation tray sterilized, and charging is with a thickness of 4cm~8cm, and fermented and cultured primary surface covers in tray
Sterilized wet gauze is covered, then tray is placed in the shelf top fermentation in sterilized solid fermentation room, control environment temperature exists
55 DEG C ~ 60 DEG C, fermentation time is 46h~56h;
(8) product handling system: solid state fermentation terminates, and by the product obtained after fermentation after drying, crushing and packaging, has obtained one
Kind antibacterial green feed additive for promoting growth finished product;
Can be produced used in the step (1) thermostable proteinase bacillus licheniformis (Bacillus licheniformis)
HL-3 is GDMCC No:60003 in the deposit number of Guangdong Province's Culture Collection.
2. a kind of side for preparing antibacterial green feed additive for promoting growth using aquatic product protein fermentation according to claim 1
Method, it is characterised in that: the aquatic product protein in the step (2) is leftovers of tilapia, krill, shrimp head, scallop body, cod
Fish pomace, 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 leftover bits and pieces
One of.
3. a kind of side for preparing antibacterial green feed additive for promoting growth using aquatic product protein fermentation according to claim 1
Method, it is characterised in that: the additional amount of aquatic product protein accounts for liquid state fermentation training with the quality of the aquatic product protein added in the step (3)
Base gross mass percentages are supported, are 10%~40%.
4. a kind of side for preparing antibacterial green feed additive for promoting growth using aquatic product protein fermentation according to claim 1
Method, it is characterised in that: nutrients is the mixed of glucose, calcium chloride, frerrous chloride and dipotassium hydrogen phosphate composition in the step (3)
Close object.
5. a kind of side for preparing antibacterial green feed additive for promoting growth using aquatic product protein fermentation according to claim 1
Method, it is characterised in that: the additional amount of the nutrients in the step (3) accounts for liquid state fermentation culture with the quality of the nutrients added
Base gross mass percentages are respectively as follows: glucose 0.01%~5%, calcium chloride 0.01%~5%, frerrous chloride 0.01%~5%, phosphorus
Sour hydrogen dipotassium 0.01%~5%.
6. a kind of side for preparing antibacterial green feed additive for promoting growth using aquatic product protein fermentation according to claim 1
Method, it is characterised in that: bacillus licheniformis in the step (4) (Bacillus licheniformis) HL-3 seed liquor connects
Kind amount accounts for fermentation medium gross mass percentages with the quality of the seed liquor accessed, is 1%~9%.
7. a kind of side for preparing antibacterial green feed additive for promoting growth using aquatic product protein fermentation according to claim 1
Method it is characterized by: Chinese herbal medicine in the step (6) be Radix Glycyrrhizae, Fructus Forsythiae, Asiatic plantain, perillaseed, Schisandra chinensis, Verbena officinalis, Radix Astragali,
One of honeysuckle, granatum, garden burnet, the coptis, cordate houttuynia, kuh-seng and smilax or in which two or more mixtures.
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