CN104686804B - A method of activated peptide forage additive is prepared using aquatic product leftovers - Google Patents
A method of activated peptide forage additive is prepared using aquatic product leftovers Download PDFInfo
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- CN104686804B CN104686804B CN201510092514.9A CN201510092514A CN104686804B CN 104686804 B CN104686804 B CN 104686804B CN 201510092514 A CN201510092514 A CN 201510092514A CN 104686804 B CN104686804 B CN 104686804B
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- bacillus subtilis
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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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- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a kind of methods for preparing activated peptide forage additive using aquatic product leftovers, including following key step: fresh aquatic product leftovers are rubbed；Aquatic product leftovers after rubbing are added in fermentor, and add nutrients and water mixing as fermentation medium；After medium sterilization in fermentor is cooling access can produce subtilin bacillus subtilis (Bacillus subtilis) HL-1 progress liquid aerobic fermentation；Fermentation liquid is warming up to 90 DEG C of enzyme deactivation 5min after fermentation, then obtains filtrate through filtering removal precipitating, filtrate obtains activated peptide forage additive finished product after being concentrated in vacuo and be spray-dried again.For the activated peptide forage additive of the method for the present invention preparation rich in antibacterial peptide and nutrition small peptide, it has good growth promotion performance and disease resistance, has a good application prospect and social effect.
Description
Technical field
The present invention relates to a kind of methods for preparing activated peptide forage additive using aquatic product leftovers, belong to aquatic products and add
Work and the field of utilizing.
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.
A large amount of document shows that the active peptide that separate sources is added in daily ration can improve the production of different raising animals
Performance, the growth promotion performance of active peptide have obtained generally acknowledging for feedstuff industry.Thus, as a kind of novel green feed addictive,
Activated peptide forage additive has good application prospect.Biologically active peptide can be divided into two major classes, i.e. nutrition small peptide and Functional Polypeptides.
Trophism small peptide is only to provide nitrogen frame for protein synthesis, the active peptide without special physiological regulatory function.Functional Polypeptides are tool
There are the peptide of certain specific functions, such as antibacterial peptide, anti-oxidation peptide, immune peptide and Hormone Peptide.
The method of macro-molecular protein chemistry or enzyme is mainly hydrolyzed into the peptide of small molecule by the preparation of active peptide.At present
The preparation method of active peptide has chemical method, enzymatic isolation method and microbe fermentation method.Chemical hydrolysis is existed using acid, alkali chemical reagent
The Fragmentation of protein molecule is promoted to form small-molecular peptides under certain temperature.Although chemical hydrolysis is simple, low at wood,
The disadvantage is that not can be carried out control production regularly, amino acid will receive damage and reduce its nutritive value, and to environment
Pollution is big.Chemical hydrolysis is chiefly used in testing at present, using less in production practices.It is with proteinogen that enzymatic isolation method, which produces active peptide,
Material is substrate, with the enzymolysis of protease, produces small-molecular peptides.Enzymatic isolation method is easily controllable, and action condition is mildly suitble to live
The production of property peptide.But enzymatic isolation method needs a large amount of protease, high production cost.Therefore, enzymatic isolation method prepares active peptide and is chiefly used in eating
Product and pharmaceuticals industry, the feeding active peptide product of low side is seldom using the method for enzymatic hydrolysis.Microbe fermentation method is with macromolecular egg
White matter is raw material, and access produces the microorganism of protease, using the enzymolysis for the protease that fermentation process generates, is generated
Small-molecular peptides.Microbe fermentation method organically combines the fermenting and producing and enzymolysis process of protease, reduces active peptide
Manufacturing cost, be more suitable for the production of active peptide.Patent CNZL200410029787.0 " a kind of activated peptide forage additive and system
Preparation Method and purposes " discloses a kind of method that fermentation hair prepares activated peptide forage 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 activated peptide forage additive can be obtained in processing.But the uncontrollability of the concentration due to fermentation process protease adds enzyme
The randomness of solution, thus the active peptide of above method preparation is mostly nutrition small peptide, and lack Functional Polypeptides, especially feed industry is anxious
The antibacterial peptide needed.
As a kind of substitute of feeding antibiotic, antibacterial peptide usually has heat-resisting, has a broad antifungal spectrum, good water solubility and is not easy
The characteristics such as drug resistance are generated, are concerned in recent years.Antibacterial peptide derives from animal, plant and microorganism, but animal and plant source is anti-
Bacterium peptide is due to limited source, using being restricted；And microbial antibacterial peptide can be mass produced, advantage of lower cost, because
And microbial source antibacterial peptide more has application prospect.At present microbial source antibacterial peptide most study be lactic acid bacteria antibacterial peptide, and
Then seldom to this novel fine bacteriums antibacterial peptide research of the raw subtilin of producing bacillus subtilis in the world, the country is even more few
See.
Summary of the invention
Activated peptide forage additive for the preparation of current fermentation method is mostly nutrition small peptide, and lacks antibacterial peptide, so that its
Effect is not prominent enough.The purpose of the present invention is preparing a kind of activated peptide forage additive rich in antibacterial peptide, further increasing
On the basis of activated peptide forage additive growth promotion performance, its disease resistance is also assigned.Add to significantly improve activated peptide forage
The effect of adding agent, to promote activated peptide forage additive in the extensive use of feed industry.
To solve the above-mentioned problems, the technical solution adopted in the present invention is as follows: a kind of to be prepared using aquatic product leftovers
The method of activated peptide forage additive, includes the following steps:
(1) by slant preservation can produce subtilin bacillus subtilis (Bacillus subtilis) HL-1 access
After to sterilizing in cooling LB liquid medium, 16h is cultivated in 37 DEG C, 150r/min, is prepared into bacillus subtilis
(Bacillus subtilis) HL-1 seed liquor is spare；
(2) spare after rubbing fresh aquatic product leftovers；
(3) step (2) aquatic product leftovers obtained rubbed are put into fermentor, and water is added into fermentor
And nutrients, fermentation medium is obtained after mixing；
(4) fermented and cultured in the fermentor for obtaining step (3) is based on 115 DEG C of sterilizing 20min；
(5) fermentation medium after the sterilizing of step (4) acquisition is cooled to room temperature, it is prepared then accesses step (1)
Bacillus subtilis (Bacillus subtilis) HL-1 seed liquor；
(6) control of fermentation process: fermentation temperature be 28 DEG C~30 DEG C, filtrated air intake be 0.5vvm~
2.5vvm, the revolving speed of fermentor are 150rpm~350rpm, and fermentation time is 35h~45h；
(7) product handling system: fermentation liquid is warming up to 90 DEG C of heat preservation enzyme deactivation 5min after fermentation, and the fermentation liquid after enzyme deactivation is cooling
To room temperature, filtrate is obtained through filtering removal precipitating, filtrate obtains activated peptide forage after vacuum concentration and spray drying again
Additive finished product.
Access in the step (1) Bacillus subtillis (Bacillus subtilis) HL-1 can generate subtilin,
The bacterium is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on January 20th, 2015, and preservation is compiled
Number be CGMCC No.10379, preservation address be Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Chinese Academy of Sciences microbe research
Institute.
Aquatic product leftovers in the step (2) are shrimp head, scallop body, the Rofe fish offal for removing internal organ and fish-skin
Expect, the gadus leftover bits and pieces that removes internal organ and fish-skin, removes the carp leftover bits and pieces of internal organ and fish-skin, removes the black carp of internal organ and fish-skin
Leftover bits and pieces, the silver carp leftover bits and pieces for removing internal organ and fish-skin, the grass carp leftover bits and pieces for removing internal organ and fish-skin and remove internal organ and fish-skin
One of stripped tuna leftover bits and pieces.Aquatic products of the additional amount of aquatic product leftovers to add in fermentor in the step (3)
The quality of leftover bits and pieces accounts for water gross mass percentages, is 10%~20%.
Nutrients in the step (3) is glucose, mixture composed by frerrous chloride and dipotassium hydrogen phosphate；It is described
The additional amount of nutrients in step (3) accounts for water gross mass percentages with the quality of the nutrients added, is respectively as follows: glucose
0.01%~5%, frerrous chloride 0.01%~5%, dipotassium hydrogen phosphate 0.01%~5%.
Bacillus subtilis in the step (5) (Bacillus subtilis) HL-1 seed liquor inoculum concentration to access
The quality of seed liquor account for fermentation medium gross mass percentages, be 0.1%~5%.
The present invention is compared with existing technology, and the present invention carries out purpose using the bacillus subtilis that can produce subtilin
Fermentation, is also obtained this bacteriocin class antibacterial peptide of subtilin, thus the method for the present invention system while obtaining nutrition small peptide
Standby activated peptide forage additive is rich in antibacterial peptide.Rich in antibacterial peptide, so that feed addictive prepared by the present invention is further
On the basis of improving activated peptide forage additive growth promotion performance, its disease resistance is also assigned, activated peptide forage is significantly improved
The effect of additive.
The present invention provides a kind of methods for preparing activated peptide forage additive using aquatic product leftovers.The present invention obtains
Activated peptide forage additive show that active peptide produced by the invention is raised in the culture experiment of the pig of South China, chicken, prawn
The feed of feed additives cooperation, not only can be improved the growth result of raising animal, but also the immunity of raising animal can be improved
And premunition, it can be used directly to prepare green feed, have a good application prospect and social effect.
Specific embodiment
For a better understanding of the present invention, below with regard to the choosing of the type of aquatic product leftovers, fermentation parameter and fermentation method
Select in conjunction with the embodiments that the invention will be further described, but the scope of the present invention is not limited thereto.
Embodiment 1: by slant preservation can produce subtilin bacillus subtilis (Bacillus subtilis) HL-1
It is linked into LB liquid medium cooling after sterilizing, cultivates 16h in 37 DEG C, 150r/min, be prepared into bacillus subtilis
(Bacillus subtilis) HL-1 seed liquor is spare.After the fresh leftovers of tilapia rubbing for removing internal organ and fish-skin
It is spare.Leftovers of tilapia that 10 kilograms rub and 100 kg of water are added in fermentor, and 800 are added into fermentor
Gram glucose, 300 grams of frerrous chlorides and 500 grams of dipotassium hydrogen phosphates obtain fermentation medium；Fermentation cylinder for fermentation culture is based on
115 DEG C of sterilizing 20min；Then be added in fermentation raw material cooling after sterilizing 2 kilograms of bacillus subtilises (Bacillus subtilis) HL-1 seed liquor；Fermentation temperature is 30 DEG C, and filtrated air intake is 1.5vvm~2.5vvm, and fermentor turns
Speed is 250rpm~350rpm, fermentation time 35h；Fermentation liquid is warming up to 90 DEG C of enzyme deactivation 5min after fermentation, after enzyme deactivation
After fermentation liquid is cooled to room temperature, filtrate is obtained through filtering removal precipitating, then filtrate is made after vacuum concentration and spray drying
Obtained a kind of activated peptide forage additive finished product prepared using aquatic product leftovers of the present invention.
Embodiment 2: by slant preservation can produce subtilin bacillus subtilis (Bacillus subtilis) HL-1
It is linked into LB liquid medium cooling after sterilizing, cultivates 16h in 37 DEG C, 150r/min, be prepared into bacillus subtilis
(Bacillus subtilis) HL-1 seed liquor is spare.It is spare after fresh shrimp head is rubbed.By 15 kilograms rub shrimp head and
100 kg of water are added in fermentor, and 900 grams of glucose, 300 grams of frerrous chlorides and 550 grams of dipotassium hydrogen phosphates are added together
It mixes, the fermentation medium after being mixed；Fermentation cylinder for fermentation culture is based on 115 DEG C of sterilizing 20min；Then cold toward after sterilizing
But be added in fermentation raw material 2 kilograms of bacillus subtilises (Bacillus subtilis) HL-1 seed liquor；Fermentation temperature is 28
DEG C, filtrated air intake is 0.5vvm~1.5vvm, and the revolving speed of fermentor is 150rpm~250rpm, fermentation time 45h；
Fermentation liquid is warming up to 90 DEG C of enzyme deactivation 5min after fermentation, after the fermentation liquid after enzyme deactivation is cooled to room temperature, through filtering removal precipitating
Filtrate is obtained, then by filtrate after vacuum concentration and spray drying, obtained of the present invention a kind of using under aquatic products
The activated peptide forage additive finished product of heel preparation.
Embodiment 3: by slant preservation can produce subtilin bacillus subtilis (Bacillus subtilis) HL-1
It is linked into LB liquid medium cooling after sterilizing, cultivates 16h in 37 DEG C, 150r/min, be prepared into bacillus subtilis
(Bacillus subtilis) HL-1 seed liquor is spare.It is spare after fresh scallop body is rubbed.The fan that 20 kilograms are rubbed
Shellfish skirt and 100 kg of water are added in fermentor, and 600 grams of glucose, 300 grams of frerrous chlorides and 550 grams of phosphoric acid hydrogen are added
Dipotassium mixes together, the fermentation medium after being mixed；Fermentation cylinder for fermentation culture is based on 115 DEG C of sterilizing 20min；Then past
Be added in cooling fermentation raw material after sterilizing 2 kilograms of bacillus subtilises (Bacillus subtilis) HL-1 seed liquor；Fermentation
Temperature is 29 DEG C, and filtrated air intake is 1.0vvm~2.0vvm, and the revolving speed of fermentor is 180rpm~280rpm, when fermentation
Between be 40h；Fermentation liquid is warming up to 90 DEG C of enzyme deactivation 5min after fermentation, after the fermentation liquid after enzyme deactivation is cooled to room temperature, through filtering
Removal precipitating obtains filtrate, then by filtrate after vacuum concentration and spray drying, has obtained a kind of utilization of the present invention
The activated peptide forage additive finished product of aquatic product leftovers preparation.
Claims (5)
1. a kind of method for preparing activated peptide forage additive using aquatic product leftovers, it is characterised in that: this method includes such as
Lower step:
(1) by slant preservation can produce subtilin bacillus subtilis (Bacillus subtilis) HL-1 is linked into and goes out
After bacterium in cooling LB liquid medium, cultivate 16h in 37 DEG C, 150r/min, be prepared into bacillus subtilis (Bacillus subtilis) HL-1 seed liquor is spare；
(2) spare after rubbing fresh aquatic product leftovers；
(3) step (2) aquatic product leftovers obtained rubbed are put into fermentor, and water and battalion is added into fermentor
Object is supported, obtains fermentation medium after mixing；
(4) fermented and cultured in the fermentor for obtaining step (3) is based on 115 DEG C of sterilizing 20min；
(5) fermentation medium after the sterilizing of step (4) acquisition is cooled to room temperature, is then accessed prepared by step (1) withered
Careless bacillus (Bacillus subtilis) HL-1 seed liquor；
(6) control of fermentation process: fermentation temperature is 28 DEG C~30 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 35h~45h；
(7) product handling system: fermentation liquid is warming up to 90 DEG C of heat preservation enzyme deactivation 5min after fermentation, and the fermentation liquid after enzyme deactivation is cooled to room
Wen Hou obtains filtrate through filtering removal precipitating；Filtrate obtains activated peptide forage addition after vacuum concentration and spray drying again
Agent finished product；
Bacillus subtilis used in the step (1) (Bacillus subtilis) HL-1 Chinese microorganism strain protect
The deposit number for hiding administration committee's common micro-organisms center is CGMCC No.10379；Aquatic products are got a foothold in the step (3)
The additional amount of material accounts for water gross mass percentages with the quality of the aquatic product leftovers added, is 10%~20%.
2. a kind of method for preparing activated peptide forage additive using aquatic product leftovers according to claim 1, special
Sign is: the aquatic product leftovers in the step (2) are shrimp head, scallop body, the Rofe fish offal for removing internal organ and fish-skin
Expect, the gadus leftover bits and pieces that removes internal organ and fish-skin, removes the carp leftover bits and pieces of internal organ and fish-skin, removes the black carp of internal organ and fish-skin
Leftover bits and pieces, the silver carp leftover bits and pieces for removing internal organ and fish-skin, the grass carp leftover bits and pieces for removing internal organ and fish-skin and remove internal organ and fish-skin
One of stripped tuna leftover bits and pieces.
3. a kind of method for preparing activated peptide forage additive using aquatic product leftovers according to claim 1, special
Sign is: nutrients is glucose, mixture composed by frerrous chloride and dipotassium hydrogen phosphate in the step (3).
4. a kind of method for preparing activated peptide forage additive using aquatic product leftovers according to claim 1, special
Sign is: the additional amount of the nutrients in the step (3) accounts for water gross mass percentages with the quality of the nutrients added, point
Not are as follows: glucose 0.01%~5%, frerrous chloride 0.01%~5%, dipotassium hydrogen phosphate 0.01%~5%.
5. a kind of method for preparing activated peptide forage additive using aquatic product leftovers according to claim 1, special
Sign is: bacillus subtilis in the step (5) (Bacillus subtilis) HL-1 seed liquor inoculum concentration to access
The quality of seed liquor accounts for fermentation medium gross mass percentages, is 0.1%~5%.
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| CN105231000B (en) * | 2015-09-25 | 2020-01-07 | 广东海洋大学 | Method for producing antibacterial nutritional compound feed additive by using aquatic product leftovers |
| CN109953176A (en) * | 2017-12-23 | 2019-07-02 | 青岛麦迪尔生物工程有限公司 | A method of peptone is prepared using aquatic products processing leftover bits and pieces |
| CN109123726A (en) * | 2018-10-08 | 2019-01-04 | 何秋宏 | A kind of preparation method of feed fermentation fish meal |
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| JPH0347051A (en) * | 1989-04-13 | 1991-02-28 | Kanzakiya:Kk | Preparation of raw solution of seasoning |
| CN1114352C (en) * | 2000-09-13 | 2003-07-16 | 山东大学 | Process for preparing nutritive activated peptide as additive |
| CN1112124C (en) * | 2001-05-15 | 2003-06-25 | 山东大学 | Process for prpearing marine active peptide as food additive |
| US6960451B2 (en) * | 2002-02-06 | 2005-11-01 | Green Earth Industries | Proteolytic fermenter |
| CN1262198C (en) * | 2003-08-15 | 2006-07-05 | 中国农业大学 | Active peptide feed additive and its preparation method and use |
| CN101564081B (en) * | 2008-04-25 | 2012-12-05 | 上海创博生态工程有限公司 | Production method for producing fish meal replacement products by utilizing fermentation process |
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Effective date of registration: 20200131 Address after: 524394 Gas Station on 374 Provincial Road, Yangqing Town, Suixi County, Zhanjiang City, Guangdong Province Patentee after: Zhanjiang silver Biotechnology Co., Ltd. Address before: 524088 Guangdong city of Zhanjiang province sea Mazhang District Road No. 1 Patentee before: Guangdong Ocean University |