CN105166410A - Feed additive with good thermal stability - Google Patents
Feed additive with good thermal stability Download PDFInfo
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- CN105166410A CN105166410A CN201510595387.4A CN201510595387A CN105166410A CN 105166410 A CN105166410 A CN 105166410A CN 201510595387 A CN201510595387 A CN 201510595387A CN 105166410 A CN105166410 A CN 105166410A
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Abstract
The invention relates to a feed additive with good thermal stability and a preparation method thereof, belonging to the field of feed enzyme preparations. The feed additive with good thermal stability is prepared from the following components in parts by weight: 5-10 parts of antibacterial peptide, 10-20 parts of propolis, 15-30 parts of glossy privet fruit extract, 15-30 parts of rhodotorula culture, 25-45 parts of ganoderma lucidum solid fermentation culture, 20-30 parts of toadstool enzymolysis powder and 10-15 parts of folium cortex eucommiae extract, wherein the rhodotorula is Rhodotorula sp. WYN1 with the collection number of CCTCC No: M2014592. Because the rhodotorula used in the invention is capable of producing carotin with high yield and the inorganic copper in a culture medium is converted to chelated copper to be enriched, the growth and development of animals are better promoted, the feed utilization ratio is increased, and the thermal stability of the feed additive is improved.
Description
Technical field:
The invention belongs to field of fodder, particularly relate to feed addictive.
Background technology:
In modern livestock and poultry cultivation process, in order to prevention and therapy Animal diseases, the excessive antibiotic product of normal use guarantees animal health, but excessive antibiotic use often causes the quality of animal meat product and product thereof to decline and antibiotic resistance strengthens, and causes human health to be also subject to having a strong impact on of antibiotic problem by the transmission of food chain.
Be derived from more natural natural materials and not only there is good nutritive peculiarity, also there is the effects such as antiviral, anti-inflammatory, anti-oxidant, conditioner body immunity function simultaneously, have broad application prospects.
The main component of propolis is flavone compound, phenols, lactone, Coumarins, aldehyde ketone, the trace element such as a small amount of iron, calcium, silicon, manganese, lead, nickel, aluminium, copper, zinc and vitamin B complex, vitamin A, and several amino acids, enzyme, polysaccharide and abundant arsanilic acid etc." natural antibiotics " that propolis or both at home and abroad expert generally acknowledge, because it not only has the effect of the external invader of defence and infection pathogen diffusion, also has the effects such as antiviral, anti-inflammatory, anti-oxidant, conditioner body immunity function.
The fruit of glossy privet is also known as glossy privet reality, purpleflower holly fruit, Chinese wax tree, mouse Chinese catalpa.Fruit of glossy privet fruit is containing oleanolic acid (Oleanolicacid), sweet mellow wine, glucose, palmitic acid, stearic acid, oleic acid and leukotrienes.Pericarp is containing ursolic acid (Ursolicacid), oleanolic acid, acetyloleanolic acid (Acetyloleanolicacid).Seed fatty oily 14.9%, in oil, palmitic acid and stearic acid are 19.5%, oleic acid and leukotrienes etc. is 80.5%.The still trace element such as cupric, zinc, iron, manganese in the fruit of glossy privet.
Fruit of glossy privet tonifying liver kidney yin, crow must improving eyesight, and modern medicine study thinks that the fruit of glossy privet can suppress the effect of helicobacter pylori to treat stomach trouble, also has the treatment suppressing purine abnormal metabolism for gout and hyperuricemia.For the deficiency of liver-yin and kidney-yin, soreness of waist tinnitus, poliosis; Unseeing, poor vision; Fever due to yin deficiency, stomach trouble and gout and and hyperuricemia.The fruit of glossy privet is clinical conventional Chinese herbal medicine simply, has begun one's study it in recent years as the effect of feed addictive in Production of Livestock and Poultry.
Antibacterial peptide is the biological small peptides that a class has antibacterial activity, is the important component part of natural animal immune defense system.The stable in physicochemical property of antibacterial peptide, relative molecular mass is little, has a broad antifungal spectrum, and antibacterial mechanisms is unique.Compared with conventional antibiotic, have antibacterial activity strong, have no drug resistance, noresidue and the little advantage of toxicity.In recent years, antibacterial peptide is the focus of research as antibiotic ideal substitute always.It is reported, antibacterial peptide, as additive feeding animals, has and improves immune performance, the generation of minimizing disease and the effect of raising survival rate.Meanwhile, antibacterial peptide has good growth promoting function to animal, can improve the growth performance of animal, reduces feedstuff-meat ratio.Therefore, antibacterial peptide has the potentiality becoming green safety feed addictive.
Glossy ganoderma [Ganoderemalucidum (LeyssexFr.) karst], also known as polyporus lucidus, belongs to Basidiomycotina, Aphyllophorales, Ganodermataceae, Ganoderma.Doctor's allusion quotation in successive dynasties playbacks glossy ganoderma in " medicine-feeding ", and its ranking is before ginseng.Shennong's Herbal, Compendium of Material Medica are recorded, glossy ganoderma materials for improving vision, tonifying liver gas, calm the nerves, beneficial essence, the beneficial motive, beneficial temper, beneficial lung qi, kidney-nourishing gas, logical nine orifices, ear acute hearing, bright, the sharp joint of order, diuresis, skin maintenance, be first-class excellent tonic product.GL-B can promote the synthesis of protein, nucleic acid, has facilitation to the renewal of serum, liver and bone marrow cell protein or nucleic acid, synthesis, and it is one of glossy ganoderma principle active component of strengthening the body resistance to consolidate the constitution.GL-B is mainly present in lucid ganoderma fungus fructification, sclerotium, mycelium or zymotic fluid; The ganoderic acid be present in glossy ganoderma has pain relieving, calmness, suppression histamine releasing, detoxifies, protects the liver, malicious tumoricidal function, is one of principle active component of glossy ganoderma.
The bark of eucommia is the distinctive nonwood forest trees of China, the skin of the bark of eucommia, leaf, really, flower containing many medicinal and nutritional labelings useful to animal health, be important Chinese medicine material.Modern fine abrasive achievement shows: using the bark of eucommia as the additive of animal feed, and to enhancing animal immunizing power, improving nutrients biological transformation function in animal body has positive meaning.
Measure according to Institute of Animal Husbandry, China Academy of Agriculture Scinces's research, Eucommia Leaf Powder gross protein value average out to 11.60% (its contents level of difference according to the folium cortex eucommiae place of production and plucking time is otherness), higher than corn, Chinese sorghum, paddy seed (being generally 7 ~ 9%); Its crude fat average content reaches 7.96%, all higher than the content (being generally 2 ~ 4%) of all common cereal seeds; Folium cortex eucommiae is also containing the abundant nutriment such as calcium (average content is 1.73%), vitamin, amino acid.In addition, the bark of eucommia also has increase hepatic and renal function, removes vivotoxin, thus strengthens the immunologic function of human body, reduces the cholesterol in blood, reduces neutral fat (subcutaneous fat), and promotes the synthesis of collagen and effect of metabolism.Therefore, be very suitable for as feed with folium cortex eucommiae.
In addition, in field of animal feed, mainly with the source of the Inorganic Coppers such as copper sulphate as copper, but it is low to there is absorptivity in Inorganic Copper, there is the problems such as antagonism with other nutriments, chelated copper then can well solve the problem, it can alleviate the antagonism with other nutriments, promote the utilization of copper and other mineral matter elements, promote growing of animal, improve efficiency of feed utilization, improve carcass quality, improve reproductive performance, also there is antibiooxidation and a series of function such as immunity and good stability simultaneously, preferably economic benefit can be brought.
Carotenoid is a class in Polyenes that is yellow, orange or red, and it is the most ubiquity of occurring in nature is also the most stable natural colouring matter, is extensively present in many animals and plants.Carotenoid as a kind of excellent additive and the nutritional supplement improving nutrition, its fine quality and effect, for a long time just generally acknowledge by countries in the world, be described as most promising antioxidant.At present, carotenoid is widely used in feed additive industry, the application prospect good due to it and the function of brilliance, for a long time extremely people's favor.It is significant to feed additive industry development how effective exploitation obtains green feed additive product with natural plants and fungi.
Summary of the invention:
The invention provides a kind of feed addictive of Heat stability is good;
Feed addictive weight fraction is composed as follows:
Antibacterial peptide 5-10 part, propolis 10-20 part, fruit of glossy privet extract 15-30 part, rhodotorula culture 15-30 part, Ganoderma Lucidum solid fermentation culture 25-45 part, hickory chick enzymolysis powder 20-30 part, eucommia leaf extract 10-15 part;
Rhodotorula used in the present invention, be the rich copper ocean rhodotorula WYN1 of a strain High Yield of Carotenoid, described bacterial strain on November 25th, 2014 be preserved in China typical culture collection center (address: China. Wuhan. Wuhan University, postcode 430072), deposit number is: CCTCCNo:M2014592, Classification And Nomenclature: ocean rhodotorula WYN1 (Rhodotorulasp.WYN1).
Described antibacterial peptide, propolis are commercially available prod;
Described fruit of glossy privet extract preparation method is as follows:
The fruit of glossy privet is pulverized, and after crossing 40-50 mesh sieve, add fruit of glossy privet 3-6 times of weight absolute ethyl alcohol Soakage extraction, adjusting temperature after control temperature 30-45 DEG C, 2-4 hour is 55-60 DEG C of 1-2 hour, and extract is concentrated, drying obtains ethanol extract; Add 75-85 DEG C of hot water in fruit of glossy privet residue after alcohol extract, hot water addition is 2-4 times of fruit of glossy privet residue weight, and processing time 30-50 minute, extracts 2-3 time continuously, by spraying dry after extract Vacuum Concentration, obtain hot water extract; Above-mentioned ethanol extract and hot water extract are merged pulverizing, crosses 60 mesh sieves, obtain glossy privet fruit extract.
The preparation method of described eucommia leaf extract is as follows:
Folium cortex eucommiae adds 5-8 times of soft water after pulverizing, with corase grind grinding, and add Radix Isatidis, addition is with the 1-2% of bark of eucommia weight, grind through colloid mill after mixing, the gap of adjustment colloid mill stator and rotor is 50 ~ 100 microns, colloid mill flow-control is 0.1 ~ 0.5 ton/hour, rear slurry add mixed enzyme and carry out enzymolysis, pH5.2-5.5, temperature 45-50 DEG C, enzymolysis time 3-4hr, described mixed enzyme consumption is the 0.05-0.2% of slurry weight, described mixed enzyme parts by weight consist of: pectase 4 parts, 1,4 beta-glucanase 1 part, 2 parts, protease and cellulase 2 parts.Enzymolysis liquid goes out enzyme, filtration, pulverize after freeze concentration, freeze drying both eucommia ulmoides extracts.
The preparation method of described hickory chick enzymolysis powder is as follows:
(1) Morchella esculenta (L.) Pers sporophore drying and crushing;
(2) water-soluble homogeneous: the Morchella esculenta (L.) Pers sporophore after pulverizing is added in stainless steel cylinder, add fructification quality 3-6 water doubly, soak 3-5 hour, then this Morchella esculenta (L.) Pers sporophore liquid is passed through colloid mill, colloid mill operation condition is: adjustment colloid mill stator and the gap of rotor are 0.5-1 micron, colloid mill flow be 0.4-1 ton/hour;
(3) intensification enzymolysis: the Morchella esculenta (L.) Pers sporophore liquid through milling treatment of colloid is transferred in stainless steel enzymatic vessel and is heated to 50-60 DEG C, adjustment pH to 4.5-6.0, add the cellulase of Morchella esculenta (L.) Pers sporophore weight 0.05-0.1%, the 1,4 beta-glucanase of 0.01-0.1%, the protease of 0.01-0.1%, insulation enzymolysis 0.5-1.5 hour, constantly stirs in enzymolysis process;
(4) dry: dry acquisition hickory chick enzymolysis powder after the mash filtrations after enzymolysis.
Described Ganoderma Lucidum solid fermentation culture preparation method is as follows:
Slant strains activation culture: be transferred on slant medium by the glossy ganoderma slant strains of preservation, cultivates 96-120 hours, covers with inclined-plane to mycelia for 22-28 DEG C; Optimum culturing temperature 25 DEG C;
1. liquid first order seed is cultivated: the one piece of access of above-mentioned glossy ganoderma slant strains picking be equipped with in 500 ml shake flasks of 100 milliliters of culture mediums and carry out first order seed cultivation, condition of culture: cultivate 96-130 hours for rotary shaker 80-180 rev/min, 22-28 DEG C; Rotary shaker 150 revs/min is advisable, and cultivation temperature is advisable with 25 DEG C;
2. liquid two stage seed culture: one-level shake-flask seed is equipped with in 500 ml shake flasks of 100 milliliters of culture mediums with the access of 5-20% inoculum concentration and carries out secondary seed cultivation, condition of culture: cultivate 96-130 hours for rotary shaker 80-180 rev/min, 22-28 DEG C; Suitable inoculum concentration is 10%, and rotary shaker 150 revs/min is advisable, and cultivation temperature is advisable with 25 DEG C;
3. solid fermentation is cultivated: be equipped with in the 500L solid-state fermentation tank of solid fermentation culture medium after sterilizing by second-level shake flask seed with the access of 5-10% inoculum concentration, condition of culture: charge 50%, cultivation temperature 20-27 DEG C, humidity 75-90%, ventilation 0.3-1.0 (V/V), incubation time 15-40 days.Stirring 10 minutes is opened every 3-10 days.Adopt SGF solid-state fermentation tank;
4. fermented and cultured terminates, and selects the multiple drying means such as fluid bed, by dry materials to moisture below 7%, pulverizing solid fermentation material both must Ganoderma Lucidum solid fermentation culture.
The carbon source of solid fermentation culture medium of the present invention mainly selects the conventional raw materials such as wheat bran, glucose, sucrose, starch, corn flour, corncob (after pulverizing), and nitrogenous source can select groundnut meal, peptone, De-fatted soya protein Powder, beancake powder, dusty yeast etc.; The inorganic salts added are needed to have potassium dihydrogen phosphate, magnesium sulfate.
Preferably, in above-mentioned solid fermentation culture medium, be added with mass ratio is after 2-6% pulverizes, and after crossing the pulverizing of 40-50 object Radix Angelicae Sinensis, 3-8%, crosses 40-50 object Radix Isatidis, after 2-5% pulverizes, crosses 40-50 object thyme.
The preparation method of described rhodotorula culture is as follows:
Seed culture is cultivated by yeast conventional culture methods;
Fermented and cultured: initial pH is 8, and seed culture fluid is inoculated in fermentation medium by inoculum concentration 3-8%, and shaking speed is 50-80r/min, cultivate for 24-28 DEG C and to adjust temperature after 10-15 hour and be 16-20 DEG C and carry out low temperature static gas wave refrigerator, stop stirring, pH is adjusted to 3-5, keeps 2-4 hour; Stairstepping is warmed up to 24-30 DEG C afterwards, and pH is adjusted to 4.6-5.5, and the addition according to 2.5% adds peptone, and the amount according to 2% adds dusty yeast; Shaking speed is 50-100r/min, continues fermentation 10-15h.
After fermentation ends, 10-1000 μm, fermentation liquor aperture coarse filtration, then concentratedly with 10-20 DEG C of loop ultrafiltration removes moisture and obtains solid content 20-40% concentrate, and concentrate obtains rhodotorula culture through vacuum freeze drying, ultramicro grinding.
Fermentation medium mass volume ratio consists of: glucose 1%, enzymolysis corn flour 1-3%, peptone 2.5%, magnesium sulfate 0.02%, potassium dihydrogen phosphate 0.15%, copper content 150-200mg/L, sodium chloride 20%, and all the other are water;
The preparation method of described enzymolysis corn flour is: by corn flour material-compound tank, be that 2:1 adds pure water with V:m, adjust ph 3-7, add middle temperature amylase, enzyme is lived as 30u/g corn flour, lives as the protease of 30u/g corn flour with enzyme, and warming while stirring is to 45-55 DEG C of insulation 15-30min, then be slowly warming up to 60-65 DEG C of insulation 15-30min, freeze drying is for subsequent use.
Described feed addictive is obtained by following methods:
After pulverizing, Ganoderma Lucidum culture, antibacterial peptide, propolis, fruit of glossy privet extract, eucommia leaf extract, hickory chick enzymolysis powder, rhodotorula culture are proportionally mixed to get feed addictive.
Beneficial effect:
1, the present invention utilizes the rich copper ocean rhodotorula WYN1 being separated from marine environment and obtaining a plant height product carrotene, effectively raises carrotene and copper output in yeast by static gas wave refrigerator method.Adopt said method fermented and cultured ocean rhodotorula WYN1, in its biomass, carotenoid output and born of the same parents, copper content is respectively more than 27g/L, the dry bacterium of 35-40mg/L and 7240-8000 μ g/g.In research and development, inventor surprisingly finds that low temperature static gas wave refrigerator effectively can improve carrot and copper content, is important achievement of the present invention.
2, because rhodotorula used in the present invention can high yield carrotene, and the Inorganic Copper in culture medium is converted into chelated copper and carries out enrichment, better facilitate growing of animal, improve efficiency of feed utilization, can 59.3% be improved for milking sow weight of weaning litter, piglet head daily gain improves 43.3%, diarrhea rate reduces 54.1%, improves 49.1% for weanling pig daily gain, and feedstuff-meat ratio improves 14.7%, diarrhea rate reduces 10%, and death rate improves 3.7%.
3, because chelated copper has good heat endurance, feed addictive provided by the present invention is had and preserves active and heat endurance preferably, make the preservation activity of additive improve 35%, heat endurance improves more than 25%
4, the fruit of glossy privet of the present invention carries out pulverize and break cellular wall process, function factor in the fruit of glossy privet is made to realize full price stripping and high efficiency extraction utilization, the effective extraction, particularly control temperature stage extraction that adopt organic solvent and hot water to realize wherein different efficacies composition respectively effectively improve extraction efficiency and the quality of glossy privet fruit extract especially.
5, the present invention is composite by science, rhodotorula, Ganoderma Lucidum are achieved organic assembling, particularly Ganoderma Lucidum with the addition of traditional Chinese medicine ingredients in cultivating, product of the present invention effectively can reduce the use amount of antibiotics in letting animals feed, improve the security of animal meat product, improve the food utilization efficiency of animal, strengthen the immunity of animal, improve raise benefit.
Detailed description of the invention:
Embodiment 1: the acquisition of ocean rhodotorula bacterium WYN1 and the domestication of resistance to copper ability
(1) by gathering the fresh biological specimen such as extra large shrimp, starfish, marine alga from Huanghai Sea seashore, be placed in aseptic YEPD culture medium, picking redness or pink circular colonies, be further purified cultivation.According to the accumulation of each bacterial classification carotenoid, the bacterial classification that screening carotenoid output is high.The ocean rhodotorula WYN1 that wherein carotenoid output is higher, cell is oval, does not have pseudohypha; The YEPD culture medium of seawater configuration forms red colonies, smooth surface, neat in edge; Product spore culture medium does not produce ascospore, and polygon budding, without ballistopore; Azymic sugar, does not form kind of starch compound, does not assimilate inositol.According to These characteristics, tentatively determine to belong to Rhodotorula (Rhodotorula).Inclined-plane is stored in refrigerator.
(2) fermented and cultured of ocean rhodotorula WYN1
Activate on strain transfer to slant medium, after 28 DEG C of cultivation 24h, be inoculated in liquid seed culture medium, 28 DEG C of shaken cultivation 36h obtain seed liquor, be equipped with in the 250mL triangular flask of 60mL fermentation medium by 10% inoculum concentration access, 28 DEG C, shaken cultivation 48h (shaking speed is 180r/min).Fermentation medium consists of glucose 2%, peptone 1%, yeast extract 1%, magnesium sulfate 0.02%, potassium dihydrogen phosphate 0.15%, salinity 20.48h cultivated by 28 DEG C of shaking tables, and rotating speed is 180r/min.
(3) resistance to copper ability domestication
Ocean rhodotorula bacterium WYN1 being inoculated in content of copper ion is in 50mg/L (copper sulphate configuration) culture medium, is placed in 28 DEG C, and on 180r/min constant-temperature table, 24h is cultivated in concussion.Then from this culture, get 3ml fermentate is inoculated in the malt extract medium of not cupric, cultivates 24h.Repeatedly cultivate several generations, yeast is grown in the culture medium of cupric stable, and when quantity no longer increases, more progressively improve the copper concentration of culture medium.So saccharomycete repeatedly just can be made can to adapt to the environment that content of copper ion is 300mg/l, obtain the characteristic of resistance to copper.The present invention, after ocean rhodotorula being carried out to the domestication of resistance to copper ability, can improve cellular biomass, the output of carotenoid and the rich copper ability of yeast effectively.
(4) mensuration of cellular biomass
By centrifugal for zymotic fluid 3000r/min 10min, abandon supernatant, thalline is again centrifugal after aseptic washing 2 ~ 3 times, and gained thalline is placed in 60 DEG C of baking ovens and is dried to constant weight, correct amount.
(5) extraction of carotenoid:
1g dry mycelium is put into 100mL triangular flask, and add the hydrochloric acid soaking at room temperature 1h of 5mL3mol/L, boiling water bath 4min, cools rapidly, and 3000r/min is centrifugal, and 15min is precipitated as cell residue, is settled to 20mL extraction, obtains carotenoid leaching liquor with acetone.
(6) mensuration of carotenoid
After suitably being diluted by extract, under 475nm condition, measure absorbance value with 722 type spectrophotometers.Be calculated as follows carotenoid content:
In formula: A λ max is the absorbance at 475nm wavelength place; D is extension rate when measuring sample; V is acetone consumption (mL); 0.16 is the molar extinction coefficient of carotenoid; W is rhodotorula dry cell weight (g).
(7) yeast copper content measures
Use aas determination.
(8) culture presevation
The above-mentioned rich copper ocean rhodotorula WYN1 through domestication is preserved in China typical culture collection center, and deposit number is: CCTCCNo:M2014592.
In carotenoid production and born of the same parents, chelated copper content is respectively 27.52g/L, 25.12mg/L and the dry bacterium of 6240 μ g/g.
The feed addictive of embodiment 2 one kinds of Heat stability is goods
Composite feed additive weight fraction is composed as follows:
Antibacterial peptide 8 parts, propolis 15 parts, fruit of glossy privet extract 20 parts, rhodotorula culture 25 parts, Ganoderma Lucidum solid fermentation culture 30 parts, 25 parts, hickory chick enzymolysis powder, eucommia leaf extract 12 parts; Rhodotorula is CCTCCNo:M2014592;
Antibacterial peptide is Chu Tai bio tech ltd, Shanghai sell goods;
Propolis is commercially available;
Described fruit of glossy privet extract preparation method is as follows:
The fruit of glossy privet is pulverized, and crosses after 45 mesh sieves, adds the fruit of glossy privet 5 times of weight absolute ethyl alcohol Soakage extraction, control temperature 40 DEG C, adjusts temperature and be 58 DEG C and maintain 1.5 hours after 3 hours, and concentrated, the drying of extract afterwards obtains ethanol extract; Add 80 DEG C of hot water in fruit of glossy privet residue after alcohol extract, hot water addition is 3 times of fruit of glossy privet residue weight, 40 minutes processing times, extracts 2 times continuously, by spraying dry after extract Vacuum Concentration, obtains hot water extract; Above-mentioned ethanol extract and hot water extract are merged pulverizing, crosses 60 mesh sieves, obtain glossy privet fruit extract.
The preparation method of described eucommia leaf extract is as follows:
The bark of eucommia adds 6 times of soft water after pulverizing, with corase grind grinding, and add Radix Isatidis, addition is with 1.5% of bark of eucommia weight, grind through colloid mill after mixing, the gap of adjustment colloid mill stator and rotor is 75 microns, colloid mill flow-control is 0.25 ton/hour, rear slurry add mixed enzyme and carry out enzymolysis, pH5.3, temperature 48 DEG C, enzymolysis time 3.5hr, described mixed enzyme consumption is 0.1% of slurry weight, and described mixed enzyme parts by weight consist of: pectase 4 parts, 1,4 beta-glucanase 1 part, 2 parts, protease and cellulase 2 parts.Enzymolysis liquid goes out enzyme, filtration, pulverize after freeze concentration, freeze drying both eucommia ulmoides extracts.
The preparation method of described hickory chick enzymolysis powder is as follows:
(1) Morchella esculenta (L.) Pers sporophore drying and crushing is to particle diameter 1mm;
(2) water-soluble homogeneous: the Morchella esculenta (L.) Pers sporophore after pulverizing is added in stainless steel cylinder, add the water of fructification quality 5 times, soak 4 hours, then this Morchella esculenta (L.) Pers sporophore liquid is passed through colloid mill, colloid mill operation condition is: the gap of adjustment colloid mill stator and rotor is 0.8 micron, and colloid mill flow is 0.8 ton/hour;
(3) intensification enzymolysis: the Morchella esculenta (L.) Pers sporophore liquid through milling treatment of colloid is transferred in stainless steel enzymatic vessel and is heated to 55 DEG C, pH is to 5.0 in adjustment, add the cellulase of Morchella esculenta (L.) Pers sporophore weight 0.08%, the 1,4 beta-glucanase of 0.05%, the protease of 0.05%, insulation enzymolysis 1 hour, constantly stirs in enzymolysis process;
(4) dry: dry acquisition hickory chick enzymolysis powder after the mash filtrations after enzymolysis.
Preparation method is as follows for Ganoderma Lucidum solid fermentation culture:
Slant strains activation culture: be transferred on slant medium by the glossy ganoderma slant strains of preservation, cultivates 110 hours, covers with inclined-plane to mycelia for 25 DEG C;
1. liquid first order seed is cultivated: the one piece of access of above-mentioned glossy ganoderma slant strains picking be equipped with in 500 ml shake flasks of 100 milliliters of culture mediums and carry out first order seed cultivation, condition of culture: rotary shaker 150 revs/min, cultivates 110 hours for 25 DEG C;
2. liquid two stage seed culture: one-level shake-flask seed is equipped with in 500 ml shake flasks of 100 milliliters of culture mediums with 10% inoculum concentration access and carries out secondary seed cultivation, condition of culture: rotary shaker 150 revs/min, cultivate 110 hours for 25 DEG C;
3. solid fermentation is cultivated: be equipped with in the 500L solid-state fermentation tank of solid fermentation culture medium after sterilizing by second-level shake flask seed with 8% inoculum concentration access, condition of culture: charge 50%, cultivation temperature 25 DEG C, humidity 80%, ventilation 0.5 (V/V), incubation time 25 days.Open stirring 10 minutes every 5 days, adopt SGF solid-state fermentation tank;
4. fermented and cultured terminates, and selects fluidized bed drying, by dry materials to moisture 5%, pulverizing solid fermentation material both must Ganoderma Lucidum solid fermentation culture;
Lucidum strain used is purchased from Chinese industrial Culture Collection, bacterium CICC14080;
Slant medium composition can be PDA slant medium;
Solid fermentation culture medium quality fraction set becomes as follows: 60% wheat bran, 1.4% sucrose, 10% corn flour, 16% corncob (after pulverizing), 0.1% peptone, 2% De-fatted soya protein Powder, 0.5% dusty yeast; 3% pulverize after cross 50 object Radix Angelicae Sinensis, 4% pulverizing rear mistake 40 object Radix Isatidis, 3% pulverize after cross 40 object thymes; Medium pH nature.Sterilising conditions: 121 DEG C, 2 hours.
Liquid fermentation seed culture medium forms: starch 3%, sucrose 4%, glucose 2%, peptone 0.5%, De-fatted soya protein Powder 2%, potassium dihydrogen phosphate 0.15%, magnesium sulfate 0.12%, etiocobalamin Pm, PH6.5.
Described rhodotorula culture preparation method is as follows:
Rhodozyma culture method carries out seed culture routinely;
Fermented and cultured: initial pH is 8, and seed culture fluid is inoculated in fermentation medium by inoculum concentration 5%, and shaking speed is 65r/min, cultivate for 26 DEG C and to adjust temperature after 12 hours and be 18 DEG C and carry out low temperature static gas wave refrigerator, stop stirring, pH is adjusted to 4, keeps 3 hours; Do not have a hour intensification 2 DEG C to 26 DEG C afterwards, pH is adjusted to 5, and the addition according to 2.5% adds peptone, and the amount according to 2% adds dusty yeast; Shaking speed is 80r/min, continues fermentation 12h;
After fermentation ends, 500 μm, fermentation liquor aperture coarse filtration, then concentratedly with 15 DEG C of loop ultrafiltrations removes moisture and obtains solid content 30% concentrate, and concentrate obtains rhodotorula culture through vacuum freeze drying, ultramicro grinding.
Fermentation medium mass volume ratio consists of: glucose 1%, enzymolysis corn flour 2%, peptone 2.5%, magnesium sulfate 0.02%, potassium dihydrogen phosphate 0.15%, copper content 180mg/L, sodium chloride 20%, and all the other are water;
The preparation method of described enzymolysis corn flour is: by corn flour material-compound tank, be that 2:1 adds pure water with V:m, adjust ph 5, add middle temperature amylase, enzyme is lived as 30u/g corn flour, lives as the protease of 30u/g corn flour with enzyme, warming while stirring to 50 DEG C insulation 25min, then be slowly warming up to 62 DEG C of insulation 25min, freeze drying is for subsequent use.
Adopt said method fermented and cultured ocean rhodotorula WYN1, in zymotic fluid, in biomass, carotenoid output and born of the same parents, chelated copper content is respectively 32.52g/L, 40mg/L and the dry bacterium of 8000 μ g/g.
Described feed addictive is obtained by following methods:
After pulverizing, Ganoderma Lucidum culture, antibacterial peptide, propolis, fruit of glossy privet extract, eucommia leaf extract, hickory chick enzymolysis powder, rhodotorula culture are proportionally mixed to get feed addictive.
The feed addictive of embodiment 3 one kinds of Heat stability is goods
Feed addictive parts by weight are composed as follows:
Antibacterial peptide 5 parts, propolis 10 parts, fruit of glossy privet extract 15 parts, rhodotorula culture 15 parts, Ganoderma Lucidum solid fermentation culture 45 parts, 20 parts, hickory chick enzymolysis powder, eucommia leaf extract 10 parts; Rhodotorula is CCTCCNo:M2014592;
Antibacterial peptide is Chu Tai bio tech ltd, Shanghai sell goods;
Propolis is commercially available;
Described fruit of glossy privet extract preparation method is as follows:
The fruit of glossy privet is pulverized, and crosses after 40 mesh sieves, adds the fruit of glossy privet 3 times of weight absolute ethyl alcohol Soakage extraction, control temperature 30 DEG C, adjusts temperature and be 55 DEG C and maintain 1 hour after 2 hours, and concentrated, the drying of extract afterwards obtains ethanol extract; Add 75 DEG C of hot water in fruit of glossy privet residue after alcohol extract, hot water addition is 2 times of fruit of glossy privet residue weight, 30 minutes processing times, extracts 2 times continuously, by spraying dry after extract Vacuum Concentration, obtains hot water extract; Above-mentioned ethanol extract and hot water extract are merged pulverizing, crosses 60 mesh sieves, obtain glossy privet fruit extract.
The preparation method of described eucommia leaf extract is as follows:
The bark of eucommia adds 5 times of soft water after pulverizing, with corase grind grinding, and add Radix Isatidis, addition is with 1% of bark of eucommia weight, grind through colloid mill after mixing, the gap of adjustment colloid mill stator and rotor is 50 microns, colloid mill flow-control is 0.1 ton/hour, rear slurry add mixed enzyme and carry out enzymolysis, pH5.2, temperature 45 C, enzymolysis time 3hr, described mixed enzyme consumption is 0.05% of slurry weight, and described mixed enzyme parts by weight consist of: pectase 4 parts, 1,4 beta-glucanase 1 part, 2 parts, protease and cellulase 2 parts.Enzymolysis liquid goes out enzyme, filtration, pulverize after freeze concentration, freeze drying both eucommia ulmoides extracts.
The preparation method of described hickory chick enzymolysis powder is as follows:
(1) Morchella esculenta (L.) Pers sporophore drying and crushing is to particle diameter 2mm;
(2) water-soluble homogeneous: the Morchella esculenta (L.) Pers sporophore after pulverizing is added in stainless steel cylinder, add the water of fructification quality 3 times, soak 3 hours, then this Morchella esculenta (L.) Pers sporophore liquid is passed through colloid mill, colloid mill operation condition is: the gap of adjustment colloid mill stator and rotor is 0.5 micron, and colloid mill flow is 0.4 ton/hour;
(3) intensification enzymolysis: the Morchella esculenta (L.) Pers sporophore liquid through milling treatment of colloid is transferred in stainless steel enzymatic vessel and is heated to 50 DEG C, pH is to 4.5 in adjustment, add the cellulase of Morchella esculenta (L.) Pers sporophore weight 0.05%, the 1,4 beta-glucanase of 0.01%, the protease of 0.01%, insulation enzymolysis 0.5 hour, constantly stirs in enzymolysis process;
(4) dry: dry acquisition hickory chick enzymolysis powder after the mash filtrations after enzymolysis.
Preparation method is as follows for Ganoderma Lucidum solid fermentation culture:
Slant strains activation culture: be transferred on slant medium by the glossy ganoderma slant strains of preservation, cultivates 96 hours for 22 DEG C;
1. liquid first order seed is cultivated: the one piece of access of above-mentioned glossy ganoderma slant strains picking be equipped with in 500 ml shake flasks of 100 milliliters of culture mediums and carry out first order seed cultivation, condition of culture: rotary shaker 80 revs/min, cultivates 96 hours for 22 DEG C;
2. liquid two stage seed culture: one-level shake-flask seed is equipped with in 500 ml shake flasks of 100 milliliters of culture mediums with 5% inoculum concentration access and carries out secondary seed cultivation, condition of culture: rotary shaker 80 revs/min, cultivate 96 hours for 22 DEG C;
3. solid fermentation is cultivated: be equipped with in the 500L solid-state fermentation tank of solid fermentation culture medium after sterilizing by second-level shake flask seed with 5% inoculum concentration access, condition of culture: charge 50%, cultivation temperature 20 DEG C, humidity 75%, ventilation 0.3 (V/V), incubation time 15 days, opened stirring 10 minutes every 3 days, adopted SGF solid-state fermentation tank;
4. fermented and cultured terminates, and selects fluidized bed drying, by dry materials to moisture 4%, pulverizing solid fermentation material both must Ganoderma Lucidum solid fermentation culture;
Lucidum strain used is purchased from Chinese industrial Culture Collection, bacterium CICC14080;
Slant medium consists of PDA slant medium;
Solid fermentation culture medium is composed as follows: 60% wheat bran, 1.4% glucose, 10% corn flour, 16% corncob (after pulverizing), 0.1% peanut muffin, 2% De-fatted soya protein Powder, 0.5% dusty yeast; 3% pulverize after cross 50 object Radix Angelicae Sinensis, 4% pulverizing rear mistake 40 object Radix Isatidis, 3% pulverize after cross 40 object thymes; Medium pH nature, sterilising conditions: 121 DEG C, 2 hours;
Liquid fermentation seed culture medium forms: starch 3%, sucrose 4%, glucose 2%, peptone 0.5%, De-fatted soya protein Powder 2%, potassium dihydrogen phosphate 0.15%, magnesium sulfate 0.12%, etiocobalamin Pm, PH6;
Described rhodotorula culture preparation method is as follows:
Seed culture is cultivated by yeast conventional culture methods;
Fermented and cultured: initial pH is 8, and seed culture fluid is inoculated in fermentation medium by inoculum concentration 3%, and shaking speed is 50r/min, cultivate for 24 DEG C and to adjust temperature after 10 hours and be 16 DEG C and carry out low temperature static gas wave refrigerator, stop stirring, pH is adjusted to 3, keeps 2 hours; Intensification per hour 1.5 DEG C is afterwards warmed up to 24-30 DEG C, and pH is adjusted to 4.6, and the addition according to 2.5% adds peptone, and the amount according to 2% adds dusty yeast; Shaking speed is 50r/min, continues fermentation 10h.
After fermentation ends, 100 μm, fermentation liquor aperture coarse filtration, then concentratedly with 10 DEG C of loop ultrafiltrations removes moisture and obtains solid content 20% concentrate, and concentrate obtains rhodotorula culture through vacuum freeze drying, ultramicro grinding.
Fermentation medium mass volume ratio consists of: glucose 1%, enzymolysis corn flour 2%, peptone 2.5%, magnesium sulfate 0.02%, potassium dihydrogen phosphate 0.15%, copper content 150mg/L, sodium chloride 20%, and all the other are water;
The preparation method of described enzymolysis corn flour is: by corn flour material-compound tank, be that 2:1 adds pure water with V:m, adjust ph 3, add middle temperature amylase, enzyme is lived as 30u/g corn flour, lives as the protease of 30u/g corn flour with enzyme, warming while stirring to 45 DEG C insulation 15min, then be slowly warming up to 60 DEG C of insulation 15min, freeze drying is for subsequent use.
Adopt said method fermented and cultured ocean rhodotorula WYN1, in zymotic fluid, in biomass, carotenoid output and born of the same parents, chelated copper content is respectively 29.38g/L, 35mg/L and the dry bacterium of 7240 μ g/g.
Described feed addictive is obtained by following methods:
After pulverizing, Ganoderma Lucidum culture, antibacterial peptide, propolis, fruit of glossy privet extract, eucommia leaf extract, hickory chick enzymolysis powder, rhodotorula culture are proportionally mixed to get feed addictive.
The feed addictive of embodiment 4 one kinds of Heat stability is goods
Feed addictive parts by weight are composed as follows:
Antibacterial peptide 10 parts, propolis 20 parts, fruit of glossy privet extract 30 parts, rhodotorula culture 30 parts, Ganoderma Lucidum solid fermentation culture 25 parts, 30 parts, hickory chick enzymolysis powder, eucommia leaf extract 15 parts; Rhodotorula is CCTCCNo:M2014592;
Antibacterial peptide is Chu Tai bio tech ltd, Shanghai sell goods;
Propolis is commercially available;
Described fruit of glossy privet extract preparation method is as follows:
The fruit of glossy privet is pulverized, and after crossing 50 mesh sieves, adds the fruit of glossy privet 6 times of weight absolute ethyl alcohol Soakage extraction, control temperature 45 DEG C, and adjusting temperature after 4 hours is 60 DEG C of maintenances 2 hours, and extract is concentrated, drying obtains ethanol extract; Add 85 DEG C of hot water in fruit of glossy privet residue after alcohol extract, hot water addition is 4 times of fruit of glossy privet residue weight, 50 minutes processing times, extracts 3 times continuously, by spraying dry after extract Vacuum Concentration, obtains hot water extract; Above-mentioned ethanol extract and hot water extract are merged pulverizing, crosses 60 mesh sieves, obtain glossy privet fruit extract.
The preparation method of described eucommia leaf extract is as follows:
The bark of eucommia adds 8 times of soft water after pulverizing, with corase grind grinding, and add Radix Isatidis, addition is with 2% of bark of eucommia weight, grind through colloid mill after mixing, the gap of adjustment colloid mill stator and rotor is 100 microns, colloid mill flow-control is 0.5 ton/hour, rear slurry add mixed enzyme and carry out enzymolysis, pH5.5, temperature 50 C, enzymolysis time 4hr, described mixed enzyme consumption is 0.2% of slurry weight, and described mixed enzyme parts by weight consist of: pectase 4 parts, 1,4 beta-glucanase 1 part, 2 parts, protease and cellulase 2 parts.Enzymolysis liquid goes out enzyme, filtration, pulverize after freeze concentration, freeze drying both eucommia ulmoides extracts.
The preparation method of described hickory chick enzymolysis powder is as follows:
(1) Morchella esculenta (L.) Pers sporophore drying and crushing is to particle diameter 2mm;
(2) water-soluble homogeneous: the Morchella esculenta (L.) Pers sporophore after pulverizing is added in stainless steel cylinder, add the water of fructification quality 6 times, soak 5 hours, then this Morchella esculenta (L.) Pers sporophore liquid is passed through colloid mill, colloid mill operation condition is: the gap of adjustment colloid mill stator and rotor is 1 micron, and colloid mill flow is 1 ton/hour;
(3) intensification enzymolysis: the Morchella esculenta (L.) Pers sporophore liquid through milling treatment of colloid is transferred in stainless steel enzymatic vessel and is heated to 60 DEG C, pH is to 6.0 in adjustment, add the cellulase of Morchella esculenta (L.) Pers sporophore weight 0.1%, the 1,4 beta-glucanase of 0.1%, the protease of 0.1%, insulation enzymolysis 1.5 hours, constantly stirs in enzymolysis process;
(4) dry: dry acquisition hickory chick enzymolysis powder after the mash filtrations after enzymolysis.Preparation method is as follows for Ganoderma Lucidum solid fermentation culture:
Slant strains activation culture: be transferred on slant medium by the glossy ganoderma slant strains of preservation, cultivates 120 hours for 28 DEG C;
1. liquid first order seed is cultivated: the one piece of access of above-mentioned glossy ganoderma slant strains picking be equipped with in 500 ml shake flasks of 100 milliliters of culture mediums and carry out first order seed cultivation, condition of culture: rotary shaker 180 revs/min, cultivates 130 hours for 28 DEG C;
2. liquid two stage seed culture: one-level shake-flask seed is equipped with in 500 ml shake flasks of 100 milliliters of culture mediums with 20% inoculum concentration access and carries out secondary seed cultivation, condition of culture: rotary shaker 180 revs/min, cultivate 130 hours for 28 DEG C;
3. solid fermentation is cultivated: be equipped with in the 500L solid-state fermentation tank of solid fermentation culture medium after sterilizing by second-level shake flask seed with 10% inoculum concentration access, condition of culture: charge 50%, cultivation temperature 27 DEG C, humidity 90%, ventilation 1.0 (V/V), incubation time 40 days.Open stirring 10 minutes every 10 days, adopt SGF solid-state fermentation tank;
4. fermented and cultured terminates, and selects the multiple drying means such as fluid bed, by dry materials to moisture 6%, pulverizing solid fermentation material both must Ganoderma Lucidum solid fermentation culture;
Lucidum strain used is purchased from Chinese industrial Culture Collection, bacterium CICC14080;
Slant medium composition can be PDA slant medium.
Solid fermentation culture medium mass ratio is composed as follows: 60% wheat bran, 1.4% sucrose, 10% corn flour, 16% corncob (after pulverizing), 0.1% peanut muffin, 2% De-fatted soya protein Powder, 0.5% dusty yeast; 3% pulverize after cross 50 object Radix Angelicae Sinensis, 4% pulverizing rear mistake 40 object Radix Isatidis, 3% pulverize after cross 40 object thymes; Medium pH nature, sterilising conditions: 121 DEG C, 2 hours.
Liquid fermentation seed culture medium forms: starch 3%, sucrose 4%, glucose 2%, peptone 0.5%, De-fatted soya protein Powder 2
Described rhodotorula culture preparation method is as follows:
Seed culture is cultivated by yeast conventional culture methods;
Fermented and cultured: initial pH is 8, and seed culture fluid is inoculated in fermentation medium by inoculum concentration 8%, and shaking speed is 80r/min, cultivate for 28 DEG C and to adjust temperature after 15 hours and be 20 DEG C and carry out low temperature static gas wave refrigerator, stop stirring, pH is adjusted to 5, keeps 4 hours; 3 DEG C per hour are warmed up to 30 DEG C afterwards, and pH is adjusted to 5.5, and the addition according to 2.5% adds peptone, and the amount according to 2% adds dusty yeast; Shaking speed is 100r/min, continues fermentation 15h.
After fermentation ends, 1000 μm, fermentation liquor aperture coarse filtration, then concentratedly with 20 DEG C of loop ultrafiltrations removes moisture and obtains solid content 40% concentrate, and concentrate obtains rhodotorula culture through vacuum freeze drying, ultramicro grinding.
Fermentation medium mass volume ratio consists of: glucose 1%, enzymolysis corn flour 3%, peptone 2.5%, magnesium sulfate 0.02%, potassium dihydrogen phosphate 0.15%, copper content 200mg/L, sodium chloride 20%, and all the other are water;
The preparation method of described enzymolysis corn flour is: by corn flour material-compound tank, be that 2:1 adds pure water with V:m, adjust ph 7, add middle temperature amylase, enzyme is lived as 30u/g corn flour, lives as the protease of 30u/g corn flour with enzyme, warming while stirring to 55 DEG C insulation 30min, then be slowly warming up to 65 DEG C of insulation 30min, freeze drying is for subsequent use.
Adopt said method fermented and cultured ocean rhodotorula WYN1, in zymotic fluid, in biomass, carotenoid output and born of the same parents, chelated copper content is respectively 28.76g/L, 38mg/L and the dry bacterium of 7580 μ g/g.
Described feed addictive is obtained by following methods:
After pulverizing, Ganoderma Lucidum culture, antibacterial peptide, propolis, fruit of glossy privet extract, eucommia leaf extract, hickory chick enzymolysis powder, rhodotorula culture are proportionally mixed to get feed addictive.
The result of use test of embodiment 5 embodiment 2 gained feed addictive in weanling pig
Test method:
Choose 120 weanling pigs, be divided into experimental group and control group by body weight, blood lineage and no sex difference, often organize 3 repetitions, each repetition 20 pigs.Experiment periods is 28 days, and experimental group feed per ton adds additive 120g described in the present embodiment 2, and control group does not add additive, weighs on an empty stomach, calculate feed conversion rate, daily gain, feedstuff-meat ratio, diarrhea rate and death rate morning day at the whole story of experimental period.Experimental result is as shown in the table:
| Daily gain | Feedstuff-meat ratio | Diarrhea rate | Death rate | |
| Control group | 226.31g | 1.49 | 15％ | 5％ |
| Experimental group | 338.12g | 1.27 | 5％ | 1.7％ |
From experimental result, the feed being added with additive of the present invention can make weanling pig daily gain improve 49.41%, and feedstuff-meat ratio improves 14.7%, and diarrhea rate reduces 10%, and death rate improves 3.7%
The result of use test of embodiment 6 example 2 of the present invention feed addictive in milking sow
Test adopts single factor test contrast design, random selecting 30 healthy, farrowing head number is close with birth counterpoise, parity is the milking sow of 2 or 3 tires, be divided at random 2 groups (i.e. test group and control groups), often organize 15 repetitions, carry out the feeding experiment of 21 days by a definite date.Wherein: test group daily ration adds the additive be prepared into by embodiment 2, and control group does not add.Record weaned piglet weight, diarrhea rate, the death rate.
| Project | Test group | Control group |
| Number born alive (head number) | 12 | 10.9 |
| Birth counterpoise (kg) | 1.52 | 1.46 |
| 21 days small weaning pigs (head) | 10 | 8.30 |
| 21 days weight of weaning litters (kg) | 68.8 | 43.2 |
| Wean counterpoise (kg) in 21 days | 6.88 | 5.20 |
| Head net gain (kg) | 5.36 | 3.74 |
| Head daily gain (g) | 255.23 | 178.1 |
| Grice diarrhoea rate (%) | 6.1 | 13.3 |
Test shows: test group can improve 59.3% than control group weight of weaning litter, and piglet head daily gain improves 43.3%, and diarrhea rate reduces 54.1%.
The above results shows that product of the present invention can improve sow and piglet body immunity, reduces antibiotic dosage, increases cultivation quality and benefits.
Embodiment 7 rhodotorula preserves active impact to additive
Rhodotorula in embodiment 2, for sample 1, is replaced with commercially available fodder yeast by the additive obtained with the preparation method described in embodiment 2, and the preservation of comparing two kinds of samples is active.At room temperature preserve above-mentioned sample 6 months, repeat the experiment of embodiment 8 afterwards, characterized the change of additive activity by the change of feedstuff-meat ratio, result shows: the residual activity of sample 1 is more than 85%, and the residual activity of sample 2 remains on about 60%.
Embodiment 8 rhodotorula is on the impact of additive heat endurance
Rhodotorula in embodiment 2, for sample 1, is replaced with commercially available fodder yeast, compares the heat endurance of two kinds of samples by the additive obtained with the preparation method described in embodiment 2.At 45 DEG C, preserve above-mentioned sample 24h, repeat the experiment of embodiment 8 afterwards, characterized the change of additive activity by the change of feedstuff-meat ratio, result shows: the residual activity of sample 1 is more than 75%, and the residual activity of sample 2 remains on about 40%.
Claims (7)
1. the feed addictive of a Heat stability is good, it is characterized in that, be made up of the component of following weight fraction: antibacterial peptide 5-10 part, propolis 10-20 part, fruit of glossy privet extract 15-30 part, rhodotorula culture 15-30 part, Ganoderma Lucidum solid fermentation culture 25-45 part, hickory chick enzymolysis powder 20-30 part, eucommia leaf extract 10-15 part;
Described rhodotorula is specially ocean rhodotorula WYN1 (Rhodotorulasp.WYN1), and deposit number is: CCTCCNo:M2014592.
2. the feed addictive of a kind of Heat stability is good as claimed in claim 1, is characterized in that, described rhodotorula culture preparation method is as follows:
Seed culture is cultivated by yeast conventional culture methods;
Fermented and cultured: initial pH is 8, and seed culture fluid is inoculated in fermentation medium by inoculum concentration 3-8%, and shaking speed is 50-80r/min, cultivate for 24-28 DEG C and to adjust temperature after 10-15 hour and be 16-20 DEG C and carry out low temperature static gas wave refrigerator, stop stirring, pH is adjusted to 3-5, keeps 2-4 hour; Stairstepping is warmed up to 24-30 DEG C afterwards, and pH is adjusted to 4.6-5.5, and the addition according to 2.5% adds peptone, and the amount according to 2% adds dusty yeast; Shaking speed is 50-100r/min, continues fermentation 10-15h.
After fermentation ends, 10-1000 μm, fermentation liquor aperture coarse filtration, then concentratedly with 10-20 DEG C of loop ultrafiltration removes moisture and obtains solid content 20-40% concentrate, and concentrate obtains rhodotorula culture through vacuum freeze drying, ultramicro grinding.
3. the feed addictive of a kind of Heat stability is good as claimed in claim 2, it is characterized in that, described fermentation medium mass volume ratio consists of: glucose 1%, enzymolysis corn flour 1-3%, peptone 2.5%, magnesium sulfate 0.02%, potassium dihydrogen phosphate 0.15%, copper content 150-200mg/L, sodium chloride 20%, all the other are water.
4. the feed addictive of a kind of Heat stability is good as claimed in claim 2, it is characterized in that, the preparation method of described enzymolysis corn flour is: corn flour is placed in material-compound tank, is that 2:1 adds pure water, adjust ph 3-7 with V:m, add middle temperature amylase, enzyme is lived as 30u/g corn flour, lives as the protease of 30u/g corn flour with enzyme, and warming while stirring is to 45-55 DEG C of insulation 15-30min, then be slowly warming up to 60-65 DEG C of insulation 15-30min, freeze drying is for subsequent use.
5. prepare a method for the feed addictive of a kind of Heat stability is good as claimed in claim 1, comprise the steps:
(1) described antibacterial peptide and propolis are commercially available prod;
(2) described fruit of glossy privet extract preparation method is as follows:
The fruit of glossy privet is pulverized, and after crossing 40-50 mesh sieve, add fruit of glossy privet 3-6 times of weight absolute ethyl alcohol Soakage extraction, adjusting temperature after control temperature 30-45 DEG C, 2-4 hour is 55-60 DEG C of 1-2 hour, and extract is concentrated, drying obtains ethanol extract; Add 75-85 DEG C of hot water in fruit of glossy privet residue after alcohol extract, hot water addition is 2-4 times of fruit of glossy privet residue weight, and processing time 30-50 minute, extracts 2-3 time continuously, by spraying dry after extract Vacuum Concentration, obtain hot water extract; Above-mentioned ethanol extract and hot water extract are merged pulverizing, crosses 60 mesh sieves, obtain glossy privet fruit extract;
(3) described Ganoderma Lucidum solid fermentation culture preparation method is as follows:
Slant strains activation culture: be transferred on slant medium by the glossy ganoderma slant strains of preservation, cultivates 96-120 hours, covers with inclined-plane to mycelia for 22-28 DEG C; Optimum culturing temperature 25 DEG C;
1. liquid first order seed is cultivated: the one piece of access of above-mentioned glossy ganoderma slant strains picking be equipped with in 500 ml shake flasks of 100 milliliters of culture mediums and carry out first order seed cultivation, condition of culture: cultivate 96-130 hour for rotary shaker 80-180 rev/min, 22-28 DEG C;
2. liquid two stage seed culture: one-level shake-flask seed is equipped with in 500 ml shake flasks of 100 milliliters of culture mediums with the access of 5-20% inoculum concentration and carries out secondary seed cultivation, condition of culture: cultivate 96-130 hours for rotary shaker 80-180 rev/min, 22-28 DEG C;
3. solid fermentation is cultivated: be equipped with in the 500L solid-state fermentation tank of solid fermentation culture medium after sterilizing by second-level shake flask seed with the access of 5-10% inoculum concentration, condition of culture: charge 50%, cultivation temperature 20-27 DEG C, humidity 75-90%, ventilation 0.3-1.0 (V/V), incubation time 15-40 days, opened stirring 10 minutes every 3-10 days, adopted SGF solid-state fermentation tank;
4. fermented and cultured terminates, and selects fluidized bed drying, by dry materials to moisture below 7%, pulverizing solid fermentation material both must Ganoderma Lucidum solid fermentation culture;
(4) preparation method of described eucommia leaf extract is as follows:
Folium cortex eucommiae adds 5-8 times of soft water after pulverizing, with corase grind grinding, and add Radix Isatidis, addition is with the 1-2% of bark of eucommia weight, grind through colloid mill after mixing, the gap of adjustment colloid mill stator and rotor is 50 ~ 100 microns, colloid mill flow-control is 0.1 ~ 0.5 ton/hour, rear slurry add mixed enzyme and carry out enzymolysis, pH5.2-5.5, temperature 45-50 DEG C, enzymolysis time 3-4hr, described mixed enzyme consumption is the 0.05-0.2% of slurry weight, described mixed enzyme parts by weight consist of: pectase 4 parts, 1,4 beta-glucanase 1 part, 2 parts, protease and cellulase 2 parts.Enzymolysis liquid goes out enzyme, filtration, pulverize after freeze concentration, freeze drying both eucommia ulmoides extracts;
(5) preparation method of described hickory chick enzymolysis powder is as follows:
1. Morchella esculenta (L.) Pers sporophore drying and crushing;
2. water-soluble homogeneous: the Morchella esculenta (L.) Pers sporophore after pulverizing is added in stainless steel cylinder, add fructification quality 3-6 water doubly, soak 3-5 hour, then this Morchella esculenta (L.) Pers sporophore liquid is passed through colloid mill, colloid mill operation condition is: adjustment colloid mill stator and the gap of rotor are 0.5-1 micron, colloid mill flow be 0.4-1 ton/hour;
3. heat up enzymolysis: transferred in stainless steel enzymatic vessel by the Morchella esculenta (L.) Pers sporophore liquid through milling treatment of colloid and be heated to 50-60 DEG C, adjustment pH to 4.5-6.0, add the cellulase of Morchella esculenta (L.) Pers sporophore weight 0.05-0.1%, the 1,4 beta-glucanase of 0.01-0.1%, the protease of 0.01-0.1%, insulation enzymolysis 0.5-1.5 hour, constantly stirs in enzymolysis process;
4. dry: dry acquisition hickory chick enzymolysis powder after the mash filtrations after enzymolysis;
(6) described rhodotorula culture preparation method is as follows:
Seed culture is cultivated by yeast conventional culture methods;
Fermented and cultured: initial pH is 8, and seed culture fluid is inoculated in fermentation medium by inoculum concentration 3-8%, and shaking speed is 50-80r/min, cultivate for 24-28 DEG C and to adjust temperature after 10-15 hour and be 16-20 DEG C and carry out low temperature static gas wave refrigerator, stop stirring, pH is adjusted to 3-5, keeps 2-4 hour; Stairstepping is warmed up to 24-30 DEG C afterwards, and pH is adjusted to 4.6-5.5, and the addition according to 2.5% adds peptone, and the amount according to 2% adds dusty yeast; Shaking speed is 50-100r/min, continues fermentation 10-15h.
After fermentation ends, 10-1000 μm, fermentation liquor aperture coarse filtration, then concentratedly with 10-20 DEG C of loop ultrafiltration removes moisture and obtains solid content 20-40% concentrate, and concentrate obtains rhodotorula culture through vacuum freeze drying, ultramicro grinding;
Fermentation medium mass volume ratio consists of: glucose 1%, enzymolysis corn flour 1-3%, peptone 2.5%, magnesium sulfate 0.02%, potassium dihydrogen phosphate 0.15%, copper content 150-200mg/L, sodium chloride 20%, and all the other are water;
The preparation method of described enzymolysis corn flour is: corn flour is placed in material-compound tank, be that 2:1 adds pure water with V:m, adjust ph 3-7, add middle temperature amylase, enzyme is lived as 30u/g corn flour, lives as the protease of 30u/g corn flour with enzyme, and warming while stirring is to 45-55 DEG C of insulation 15-30min, then be slowly warming up to 60-65 DEG C of insulation 15-30min, freeze drying is for subsequent use.
(6) described composite feed additive is obtained by following methods:
Antibacterial peptide, propolis, fruit of glossy privet extract, rhodotorula culture, Ganoderma Lucidum solid fermentation culture, hickory chick enzymolysis powder, eucommia leaf extract is proportionally mixed to get composite feed additive.
6. the feed addictive of an a kind of Heat stability is good according to claim 1, it is characterized in that, be made up of the component of following weight fraction: antibacterial peptide 8 parts, propolis 15 parts, fruit of glossy privet extract 20 parts, rhodotorula culture 25 parts, Ganoderma Lucidum solid fermentation culture 30 parts, 25 parts, hickory chick enzymolysis powder, eucommia leaf extract 12 parts; Rhodotorula is CCTCCNo:M2014592.
7. the application of feed addictive in animal and fowl fodder of a kind of Heat stability is good according to claim 1.
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Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1415740A (en) * | 2002-10-15 | 2003-05-07 | 湖北安琪酵母股份有限公司 | Drying production technique by bran adsorption for drying oceanic rhodotorula |
| CN102657287A (en) * | 2012-06-05 | 2012-09-12 | 荆州双胞胎饲料有限公司 | Compound feed addictive with bacteria and oxidation resisting functions |
| CN102669435A (en) * | 2012-06-05 | 2012-09-19 | 邵素英 | Feed additive product |
| CN102696884A (en) * | 2012-06-05 | 2012-10-03 | 邵素英 | Composite feed additive product |
| CN102763772A (en) * | 2012-08-02 | 2012-11-07 | 邵素英 | Functional feed additive |
| CN104621356A (en) * | 2014-12-04 | 2015-05-20 | 邵素英 | Micro-ecological complex enzyme preparation and preparation method thereof |
| CN104799016A (en) * | 2014-12-04 | 2015-07-29 | 邵素英 | Composite biological enzyme and preparation method thereof |
-
2015
- 2015-09-17 CN CN201510595387.4A patent/CN105166410A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1415740A (en) * | 2002-10-15 | 2003-05-07 | 湖北安琪酵母股份有限公司 | Drying production technique by bran adsorption for drying oceanic rhodotorula |
| CN102657287A (en) * | 2012-06-05 | 2012-09-12 | 荆州双胞胎饲料有限公司 | Compound feed addictive with bacteria and oxidation resisting functions |
| CN102669435A (en) * | 2012-06-05 | 2012-09-19 | 邵素英 | Feed additive product |
| CN102696884A (en) * | 2012-06-05 | 2012-10-03 | 邵素英 | Composite feed additive product |
| CN102763772A (en) * | 2012-08-02 | 2012-11-07 | 邵素英 | Functional feed additive |
| CN104621356A (en) * | 2014-12-04 | 2015-05-20 | 邵素英 | Micro-ecological complex enzyme preparation and preparation method thereof |
| CN104799016A (en) * | 2014-12-04 | 2015-07-29 | 邵素英 | Composite biological enzyme and preparation method thereof |
Non-Patent Citations (4)
| Title |
|---|
| VERONICA IRAZUSTA ET AL.: "Relationship among carotenoid production，copper bioremediation and oxidative stress in Rhodotorula mucilaginosa RCL-11", 《PROCESS BIOCHEMISTRY》 * |
| 农业部畜牧业司，全国饲料工作办公室编: "《饲料 法规 文件.2014》", 31 May 2014 * |
| 史馨怡，等: "逆境对红酵母发酵生产类胡萝卜素的促进作用", 《生物技术通报》 * |
| 梁宗锁主编: "《杜仲丰产栽培实用技术》", 31 March 2011 * |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109874909A (en) * | 2019-03-28 | 2019-06-14 | 陈欣 | A kind of feed addictive and preparation process based on antibacterial peptide |
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