CN105707473A - Method for preparing green feed-use antibacterial growth promoting agent through high temperature fermentation by means of aquatic protein and Chinese herbal medicine - Google Patents
Method for preparing green feed-use antibacterial growth promoting agent through high temperature fermentation by means of aquatic protein and Chinese herbal medicine Download PDFInfo
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- Fodder In General (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a method for preparing a green feed-use antibacterial growth promoting agent through high temperature fermentation by means of aquatic protein and Chinese herbal medicine. The method comprises the following main steps that fresh low-value aquatic protein is minced for use or aquatic product processing waste water is steamed and concentrated till the protein content ranges from 15% to 20% for use; the Chinese herbal medicine is smashed and sieved through a 60-mesh sieve; the minced low-value aquatic protein or concentrated aquatic product processing waste water is fixed with the smashed Chinese herbal medicine, and nutrients and water are added into the mixture, the materials are mixed to be uniform and serve as a fermentation medium; high-temperature-resistant Bacillus licheniformis HL-3 is inoculated on the fermentation medium for solid aerobic fermentation; a fermented product is subjected to drying, smashing and packaging, and a feed additive finished product is prepared. By means of the method for preparing the green feed-use antibacterial growth promoting agent through high temperature fermentation by means of the aquatic protein and Chinese herbal medicine, the green feed-use antibacterial growth promoting agent which is rich in ingredients of small peptide, Chinese herbal medicine effective ingredients, high-temperature-resistant protease, high-temperature-resistant non-starch polysaccharase and probiotics can be prepared, the good effects of promoting growth and resisting diseases are achieved, the growth promoting agent can completely replace feed-use antibiotics and has a good application prospect and social significance.
Description
Technical field
The present invention relates to a kind of method utilizing aquatic product protein and Chinese herbal medicine hot fermentation to prepare green feeding antivirus somatotropic agent, belong to technical field prepared by feed additive.
Background technology
Since the 1950's, in animal feeding and feedstuff, the growth promotion health-care agents such as antibiotic, chemical synthetic drug, hormone, beta-stimulants, heavy metal, tranquilizer is widely used.The growth promotion health-care agents such as antibiotic applicating history in feedstuff is long-standing, and it creates the revolution of aquaculture and feed industry so that breeding scale is developed rapidly in worldwide, and production quantity and the feed conversion rate of animal are greatly improved.But, as the achievement of industrial civilization has its negative effect as in all, use the non-green feed additives such as antibiotic for a long time, result in following problem: poultry and Aquatic product product drug residue are serious, are directly detrimental to health;Poultry and Aquatic product deterioration in quality, lack the market competitiveness and sense of trust;Bacterial drug resistance strengthens, Resistant strain increases, and threatens human security;Destroy ecological environment etc..Along with feed industry and the fast development of animal husbandry and fishery industry, livestock products and the aquatic products of China have been changed into interim superfluous from shortage at present, whether people are more concerned with food safety, health, thus green consumption has become development trend from now on, in addition China joins WTO, the agricultural product of China's export are required increasingly stricter by developed country, one of major obstacle that current drug residue problem becomes China's livestock products and aquatic products are gone abroad.Hence speed up exploitation, promotion and application novel green feed additive imperative.
Green feed additive, most basic requirement be have no side effect, noresidue, and also want can promote growth of animal.Current internationally recognized growth promotion green feed additive mainly has following several: little peptide, enzyme preparation, plant source product and extract thereof, microbial ecological agent, oligosaccharide and yeast series products etc..
Enzyme preparation has noresidue, pollution-free as a kind of feed additive, improves the value of feedstuff, improves the effect such as breeding performonce fo animals and product quality.Fodder enzyme preparation substantially can be divided into digestive enzyme and non-digestive enzyme two class, generally plays a role in nutrition aspect.Such as improve livestock and poultry alimentary tract endogenous enzyme activity, supplement the deficiency of endogenous enzymes;Destroy plant cell wall, improve the utilization ratio of feedstuff;Eliminate the antinutritional factor in feedstuff, promote digesting and assimilating of nutrient substance.At present, enzyme preparation has been widely used for feed industry, mainly has the enzyme preparations such as protease, non-starch polysaccharide enzyme and phytase.But, the many non-refractories of existing fodder enzyme preparation on market, in feedstuff high temperature granulation process, loss of activity is the biggest.
In recent years, little peptide receives much concern at feed industry as a kind of green feed additive for promoting growth.The preparation method of current little peptide has enzymatic isolation method and microbe fermentation method.It is with protein raw materials as substrate that enzymatic isolation method produces bioactive peptide, uses the zymolysis of protease, produces small-molecular peptides.Enzymatic isolation method is easily controllable, and action condition gentleness is suitable for the production of bioactive peptide.But enzymatic isolation method needs substantial amounts of protease, production cost is high.Therefore, enzymatic isolation method is prepared bioactive peptide and is used for food and pharmaceuticals industry, the method that the feeding bioactive peptide product of low side seldom uses enzymolysis.Microbe fermentation method is with macro-molecular protein as raw material, accesses the microorganism producing protease, utilizes the zymolysis of the protease that fermentation process produces, generation small-molecular peptides.Fermentation method organically combines fermenting and producing and the enzymolysis process of protease, reduces the manufacturing cost of bioactive peptide, is more suitable for the production of bioactive peptide.Patent CNZL200410029787.0 " a kind of activated peptide forage additive and preparation method and purposes " discloses a kind of fermentation and sends out the method preparing small peptide feed additive, it is the mixture of one or more with vegetable protein and animal proteinum, through fermentative degradations such as bacillus cereuss, its degree of hydrolysis is made to control, 20~35%, to be then passed through post processing and can get small peptide feed additive.Little peptide has good application prospect as a kind of novel green feed additive for promoting growth at feed industry, but current little peptide is not widely used at feed industry.The little peptide that either enzymatic isolation method produces, or the little peptide of fermentative Production, high production cost constrains the extensive application at feed industry of the little peptide.It addition, its growth promotion composition of little peptide product prepared by prior art only has little peptide one composition, the most single, growth promotion performance is the most prominent.
Chinese herbal medicine is the rarity in Chinese medicine treasure-house, and Chinese herbal medicine is as feed additive, has begun in Ancient Times in China.Chinese herbal medicine have have no side effect, be difficult to cause the characteristic such as resistance, noresidue, the green succedaneum as a kind of feeding antibiotic has been widely recognized at feed industry.From the eighties in 20th century, Chinese herbal medicine has been used for making feeding antibiotic succedaneum, and achieves certain application effect.After Chinese herbal medicine undergoes microbial fermentation, its effective ingredient is fully discharged so that it is more biological activity and curative effect study hotspot the most especially.China's feedstuff (2010,419(15): 13-17) document discloses the aspergillus oryzae of isolated from different herbal raw materials and ferments Chinese herbal medicine, and fermentation material cooking liquor the most relatively compares raising 36.03%, 60.83%, 27.64%, 41.32% to the bacteriostatic activity of escherichia coli, staphylococcus aureus, proteus vulgaris and Salmonella typhimurium.At present, the main strain used of Chinese herbal medicine fermentation has the strain such as Bacillus licheniformis and aspergillus oryzae.The cell wall main component of Chinese herbal medicine is degraded, so that Chinese herbal medicine effective ingredients discharges by the non-starch polysaccharide enzyme mainly utilizing these microorganisms to produce.But, existing Chinese herbal medicine fermented bacterium is all the strain of room temperature, and the activity of its non-starch polysaccharide enzyme produced during the fermentation can be lost totally in the high temperature granulation process of product drying and feedstuff.Thus, fermentation of Chinese herbal medicine prepared by existing Chinese herbal medicine fermentation technique can only play effect of Chinese herbal medicine in animal body, can not play effect of enzyme preparation.
Summary of the invention
For problem present in the technologies of preparing such as current enzyme preparation, little peptide and fermentation of Chinese herbal medicine, the invention provides a kind of method utilizing aquatic product protein and Chinese herbal medicine hot fermentation to prepare green feeding antivirus somatotropic agent.The inventive method can prepare a kind of feeding antivirus somatotropic agent of green rich in little peptide, Chinese herbal medicine effective ingredients, thermostable proteinase, high temperature resistant non-starch polysaccharide enzyme and these several compositions of probiotic bacteria.
In order to solve the problems referred to above, the technical solution adopted in the present invention is as follows: a kind of method utilizing aquatic product protein and Chinese herbal medicine hot fermentation to prepare green feeding antivirus somatotropic agent, comprises the steps:
(1) by the high temperature resistant Bacillus licheniformis of slant preservation (Bacillus licheniformis) HL-3 is linked into after sterilizing and cools down
LB fluid medium in, in 55 DEG C, 150r/min cultivate 12h, be prepared as Bacillus licheniformis (Bacillus licheniformis) HL-3 seed liquor is standby;
(2) after fresh low-value aquatic product protein being rubbed standby or be 15%~20% standby by aquatic products processing wastewater evaporation and concentration to protein content;
(3) 60 mesh sieves are crossed after being pulverized by Chinese herbal medicine standby;
(4) Chinese herbal medicine that low-value aquatic product protein step (2) rubbed or the aquatic products processing wastewater of concentration and step (3) are pulverized is the ratio mixing of 0.5 ~ 1.5:1 in mass ratio;
(5) mixture obtained toward step (4) adds nutrient and water, and mix, it is thus achieved that final moisture content controls the fermentation medium in the range of 40%~80%;
(6) fermentation culture obtained toward step (5) accesses Bacillus licheniformis prepared by step (1) (Bacillus licheniformis) HL-3 seed liquor, and mix;
(7) fermentation of raw material: the fermentation medium adding bacterium step (6) obtained ferments in following manner: fermentation medium is divided in the fermentation tray sterilized, charging thickness is 4cm~8cm, in tray, fermentation culture primary surface covers the wet gauze of sterilizing, tray is placed in the shelf top fermentation in sterilized solid fermentation room again, control ambient temperature is at 55 DEG C ~ 60 DEG C, and fermentation time is 48h~60h;
(8) product handling system: fermentation ends, will fermentation after obtain product drying, pulverize and pack after, i.e. prepared a kind of of the present invention compound green feed additive for promoting growth finished product.
High temperature resistant Bacillus licheniformis used in described step (1) (Bacillus licheniformis) HL-3 is preserved in Guangdong Province's Culture Collection on January 18th, 2016, deposit number is GDMCC No:60003, and preservation address is Guangdong Microbes Inst, 5th floors, No. 59 building of compound, Xianlie Middle Road, Guangzhou City 100.
One during low-value aquatic product protein is leftovers of tilapia, Antarctic krill, prawn head, scallop body, morrhua leftover bits and pieces, squid leftover bits and pieces, Harengulazunasi (Bleeker) leftover bits and pieces, Mylopharyngodon piceus leftover bits and pieces, Hypophthalmichthys molitrix leftover bits and pieces, Ctenopharyngodon idellus leftover bits and pieces and Skipjack leftover bits and pieces in described step (2).Aquatic products processing wastewater in described step (2) is the one in fish flour processing waste water, the alkali-wash waste water of the chitin course of processing, prawn waste water from cooking and minced fish processing waste water.
Chinese herbal medicine in described step (3) is the mixture of more than a kind of or two of which in Radix Glycyrrhizae, Fructus Forsythiae, Herba Plantaginis, Fructus Perillae, Fructus Schisandrae Chinensis, Herba Verbenae, the Radix Astragali, Flos Lonicerae, Pericarpium Granati, Radix Sanguisorbae, Rhizoma Coptidis, Herba Houttuyniae, Radix Sophorae Flavescentis and Rhizoma Smilacis Glabrae.
The mixture that nutrient in described step (5) is wheat bran, calcium chloride, ferrous chloride and dipotassium hydrogen phosphate are formed.The quality of the addition of the nutrient in described step (5) nutrient to add accounts for fermentation medium gross mass percentages, is respectively as follows: wheat bran 0.01%~15%, calcium chloride 0.01%~5%, ferrous chloride 0.01%~5%, dipotassium hydrogen phosphate 0.01%~5%.
Bacillus licheniformis in described step (6) (Bacillus licheniformis) HL-3 inoculum concentration with access seed liquor quality account for fermentation medium gross mass percentages, be 1%~9%.
The present invention compared with prior art, has the following advantages.
(1) fermented product can play effect of enzyme preparation in letting animals feed body.What the fermentation preparation of existing Chinese herbal medicine zymocyte and little peptide used is all the strain of room temperature, dry run that the activity of its non-starch polysaccharide enzyme produced during the fermentation and protease is prepared at product and losing totally in the high temperature granulation process of feedstuff.Thus, fermentation of Chinese herbal medicine prepared by prior art and little peptide product can not play effect of enzyme preparation in animal body for feedstuff.And this technology resistant to elevated temperatures Bacillus licheniformis of employing (Bacillus licheniformis) ferment, sweat produces resistant to elevated temperatures protease and non-starch polysaccharide enzyme, the activity of these resistant to elevated temperatures protease and non-starch polysaccharide enzyme is dried prepared by product and does not the most lose in the high temperature granulation process of feedstuff, thus the feed additive prepared of this technology can play effect of enzyme preparation in animal body.
(2) ferment effect is good.With existing Chinese herbal medicine fermentation and compared with fermentation method prepares little peptide, the present invention uses hot fermentation, and enzyme activity under conditions of high temperature is obviously enhanced, and this also can be greatly improved Chinese herbal medicine fermentation and fermentation method and prepare the ferment effect of little peptide;It addition, in the technology of the present invention, raw material does not has sterilizing, can give full play to the activity of aquatic product protein endogenous proteinase, this can improve the content of little peptide further.
(3) low cost.Compared with preparing little peptide with existing fermentation method, in the technical scheme that the present invention provides, raw material is without sterilizing, it is possible to decrease energy consumption and cost.It addition, the fermenting and producing of little peptide and the fermenting and producing of fodder enzyme preparation are united two into one by the present invention, the production of fodder enzyme preparation can share part producing cost, thus reduces the production cost of product further.
(4) functional factor of thermal sensitivity in aquatic product protein can effectively be protected.In technical scheme, owing to raw material can not be sterilized, thus, in aquatic product protein, the activity of the functional factor of thermal sensitivity is effectively retained.Such as, the astaxanthin in prawn head and Antarctic krill.
The invention provides a kind of method utilizing aquatic product protein and Chinese herbal medicine hot fermentation to prepare green feeding antivirus somatotropic agent.The inventive method can be prepared a kind of rich in little peptide, Chinese herbal medicine effective ingredients, thermostable proteinase, high temperature resistant non-starch polysaccharide enzyme and the feeding antivirus somatotropic agent of these several compositions of probiotic bacteria.The feeding antivirus somatotropic agent of green that the present invention the obtains pig in South China, chicken, the culture experiment of prawn show, it can promote letting animals feed growth and strengthen the premunition of animal, hence it is evident that reduces aquaculture cost, obtains significant economic benefit.
Detailed description of the invention
In order to be more fully understood that the present invention, with regard to the selection of kind, the kind of Chinese herbal medicine, fermentation parameter and fermentation mode of aquatic product protein, the invention will be further described in conjunction with the embodiments below, but protection scope of the present invention is not limited to this.
Embodiment 1: by the high temperature resistant Bacillus licheniformis of slant preservation (Bacillus licheniformis) HL-3 is linked into after sterilizing in the LB fluid medium of cooling, in 55 DEG C, 150r/min cultivate 12h, be prepared as Bacillus licheniformis (Bacillus licheniformis) HL-3 seed liquor is standby.By standby after fresh leftovers of tilapia rubbing, cross 60 mesh sieves after being pulverized by Radix Glycyrrhizae standby;By 100 kilograms rub leftovers of tilapia and 100 kilograms pulverize Radix Glycyrrhizaes mixing, and add 4900 grams of wheat brans, 455 grams of calcium chloride, 356 grams of ferrous chlorides, 555 grams of dipotassium hydrogen phosphates mix together with 12 kg of water, it is thus achieved that the fermentation medium after mixing;Add in fermentation medium 9 kilograms of Bacillus licheniformis Bacillus licheniformis (Bacillus licheniformis) HL-3 seed liquor, stirring mix homogeneously;Postvaccinal fermentation medium is divided in in the fermentation tray of the ethanol surface sterilization of 75%, the thickness that feeds in tray is 4 centimetres, in tray, material surface covers the wet gauze of sterilization treatment, tray is placed in the shelf top fermentation in the solid fermentation room of sterilizing again, controlling ambient temperature is 55 DEG C, and fermentation time is 48h;After fermentation ends, will fermentation after obtain product drying, pulverize and pack after, i.e. prepared a kind of green feeding antivirus somatotropic agent finished product.
Embodiment 2: by the high temperature resistant Bacillus licheniformis of slant preservation (Bacillus licheniformis) HL-3 is linked into after sterilizing in the LB fluid medium of cooling, in 55 DEG C, 150r/min cultivate 12h, be prepared as Bacillus licheniformis (Bacillus licheniformis) HL-3 seed liquor is standby.By standby after fresh prawn head rubbing, cross 60 mesh sieves after being pulverized by Herba Verbenae standby;By 110 kilograms rub prawn head and 100 kilograms pulverize Herba Verbenae mixing, and add 2350 grams of wheat brans, 459 grams of calcium chloride, 387 grams of ferrous chlorides, 589 grams of dipotassium hydrogen phosphates mix together with 13 kg of water, it is thus achieved that the fermentation medium after mixing;Add in fermentation medium 9 kilograms of Bacillus licheniformis (Bacillus licheniformis) HL-3 seed liquor, stirring mix homogeneously;Postvaccinal fermentation medium is divided in in the fermentation tray of the ethanol surface sterilization of 75%, the thickness that feeds in tray is 8 centimetres, in tray, material surface covers the wet gauze of sterilization treatment, tray is placed in the shelf top fermentation in the solid fermentation room of sterilizing again, controlling ambient temperature is 60 DEG C, and fermentation time is 60h;After fermentation ends, will fermentation after obtain product drying, pulverize and pack after, i.e. prepared a kind of green feeding antivirus somatotropic agent finished product.
Embodiment 3: by the high temperature resistant Bacillus licheniformis of slant preservation (Bacillus licheniformis) HL-3 is linked into after sterilizing in the LB fluid medium of cooling, in 55 DEG C, 150r/min cultivate 12h, be prepared as Bacillus licheniformis (Bacillus licheniformis) HL-3 seed liquor is standby.It is 18% standby by fish flour processing waste water evaporation and concentration to protein content, crosses 60 mesh sieves after Radix Glycyrrhizae and the Radix Astragali being pulverized standby;Chinese herbal medicine (+30 kilograms of Radixs Astragali of the 70 kilograms of Radix Glycyrrhizaes) mixing that 80 kilograms of fish flour processing waste waters concentrated and 100 kilograms are pulverized, and add 3980 grams of wheat brans, 499 grams of calcium chloride, 398 grams of ferrous chlorides, 590 grams of dipotassium hydrogen phosphates mix together with 16 kg of water, it is thus achieved that the fermentation medium after mixing;Add in fermentation medium 9 kilograms of Bacillus licheniformis (Bacillus licheniformis) HL-3 seed liquor, stirring mix homogeneously;Postvaccinal fermentation medium is divided in in the fermentation tray of the ethanol surface sterilization of 75%, the thickness that feeds in tray is 6 centimetres, in tray, material surface covers the wet gauze of sterilization treatment, tray is placed in the shelf top fermentation in the solid fermentation room of sterilizing again, controlling ambient temperature is 57 DEG C, and fermentation time is 56h;After fermentation ends, will fermentation after obtain product drying, pulverize and pack after, i.e. prepared a kind of green feeding antivirus somatotropic agent finished product.
Claims (8)
1. one kind utilizes the method that aquatic product protein and Chinese herbal medicine hot fermentation prepare green feeding antivirus somatotropic agent, it is characterised in that: the method comprises the steps:
(1) by the high temperature resistant Bacillus licheniformis of slant preservation (Bacillus
licheniformis) HL-3 is linked into after sterilizing and cools down
LB fluid medium in, in 55 DEG C, 150r/min cultivate 12h, be prepared as Bacillus licheniformis (Bacillus licheniformis) HL-3 seed liquor is standby;
(2) after fresh low-value aquatic product protein being rubbed standby or be 15%~20% standby by aquatic products processing wastewater evaporation and concentration to protein content;
(3) 60 mesh sieves are crossed after being pulverized by Chinese herbal medicine standby;
(4) Chinese herbal medicine that low-value aquatic product protein step (2) rubbed or the aquatic products processing wastewater of concentration and step (3) are pulverized is the ratio mixing of 0.5 ~ 1.5:1 in mass ratio;
(5) mixture obtained toward step (4) adds nutrient and water, and mix, it is thus achieved that final moisture content controls the fermentation medium in the range of 40%~80%;
(6) fermentation culture obtained toward step (5) accesses Bacillus licheniformis prepared by step (1) (Bacillus licheniformis) HL-3 seed liquor, and mix;
(7) fermentation of raw material: the fermentation medium adding bacterium step (6) obtained ferments in following manner: fermentation medium is divided in the fermentation tray sterilized, charging thickness is 4cm~8cm, in tray, fermentation culture primary surface covers the wet gauze of sterilizing, tray is placed in the shelf top fermentation in sterilized solid fermentation room again, control ambient temperature is at 55 DEG C ~ 60 DEG C, and fermentation time is 48h~60h;
(8) product handling system: fermentation ends, will fermentation after obtain product drying, pulverize and pack after, i.e. prepared a kind of of the present invention compound green feed additive for promoting growth finished product.
A kind of method utilizing aquatic product protein and Chinese herbal medicine hot fermentation to prepare green feeding antivirus somatotropic agent the most according to claim 1, it is characterised in that: high temperature resistant Bacillus licheniformis used in described step (1) (Bacillus licheniformis) HL-3 is GDMCC No:60003 at the deposit number of Guangdong Province's Culture Collection.
A kind of method utilizing aquatic product protein and Chinese herbal medicine hot fermentation to prepare green feeding antivirus somatotropic agent the most according to claim 1, it is characterised in that: the one during low-value aquatic product protein is leftovers of tilapia, Antarctic krill, prawn head, scallop body, morrhua leftover bits and pieces, squid leftover bits and pieces, Harengulazunasi (Bleeker) leftover bits and pieces, Mylopharyngodon piceus leftover bits and pieces, Hypophthalmichthys molitrix leftover bits and pieces, Ctenopharyngodon idellus leftover bits and pieces and Skipjack leftover bits and pieces in described step (2).
A kind of method utilizing aquatic product protein and Chinese herbal medicine hot fermentation to prepare green feeding antivirus somatotropic agent the most according to claim 1, it is characterised in that: the aquatic products processing wastewater in described step (2) is the one in fish flour processing waste water, the alkali-wash waste water of the chitin course of processing, prawn waste water from cooking and minced fish processing waste water.
A kind of method utilizing aquatic product protein and Chinese herbal medicine hot fermentation to prepare green feeding antivirus somatotropic agent the most according to claim 1, it is characterised in that: the Chinese herbal medicine in described step (3) is the mixture of more than a kind of or two of which in Radix Glycyrrhizae, Fructus Forsythiae, Herba Plantaginis, Fructus Perillae, Fructus Schisandrae Chinensis, Herba Verbenae, the Radix Astragali, Flos Lonicerae, Pericarpium Granati, Radix Sanguisorbae, Rhizoma Coptidis, Herba Houttuyniae, Radix Sophorae Flavescentis and Rhizoma Smilacis Glabrae.
A kind of method utilizing aquatic product protein and Chinese herbal medicine hot fermentation to prepare green feeding antivirus somatotropic agent the most according to claim 1, it is characterised in that: the mixture that the nutrient in described step (5) is wheat bran, calcium chloride, ferrous chloride and dipotassium hydrogen phosphate are formed.
A kind of method utilizing aquatic product protein and Chinese herbal medicine hot fermentation to prepare green feeding antivirus somatotropic agent the most according to claim 1, it is characterized in that: the quality of the addition of the nutrient in described step (5) nutrient to add accounts for fermentation medium gross mass percentages, it is respectively as follows: wheat bran 0.01%~15%, calcium chloride 0.01%~5%, ferrous chloride 0.01%~5%, dipotassium hydrogen phosphate 0.01%~5%.
A kind of method utilizing aquatic product protein and Chinese herbal medicine hot fermentation to prepare green feeding antivirus somatotropic agent the most according to claim 1, it is characterised in that: Bacillus licheniformis in described step (6) (Bacillus
licheniformis) HL-3 inoculum concentration with access seed liquor quality account for fermentation medium gross mass percentages, be 1%~9%.
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