CN106819632A

CN106819632A - A kind of catfish feed complex enzyme containing yeast culture and preparation method thereof

Info

Publication number: CN106819632A
Application number: CN201611216878.4A
Authority: CN
Inventors: 田岗; 邢晓慧
Original assignee: Individual
Current assignee: Individual
Priority date: 2016-12-26
Filing date: 2016-12-26
Publication date: 2017-06-13

Abstract

The invention discloses a kind of catfish feed complex enzyme containing yeast culture and preparation method thereof, based on catfish nutritional need, water body environment, with reference to its digestion characteristics, optimizing raw material is formulated, science Cultures of S. cerevisiae of the compounding with functional characteristics such as powerful trophism, bioactivity and inoxidizability；Calcium, iron and Soluble Fiber, the calcium berry extract with powerful embedding effect, extension feed complex enzyme shelf-life can effectively be supplemented；Feed complex enzyme cryoprotective effects and stability are remarkably improved, catfish feed water resistance and glossiness is improved, wormwood seed powder of the feed containing powder rate is reduced；Agreeable to the taste dose of feed palatability and digestibility can be greatly improved；The Wheat Sprout Powder of abundant nutrition material, bioactivator and phytoenzyme source can be provided；The final one kind that provides is remarkably improved catfish digestibility and production performance, reduces feed coefficient, prevents water pollution, maintains the catfish feed complex enzyme containing yeast culture of fine water.

Description

A kind of catfish feed complex enzyme containing yeast culture and preparation method thereof

Technical field

The present invention relates to feed complex enzyme, and in particular to a kind of catfish feed complex enzyme and its preparation containing yeast culture Method.

Background technology

Catfish is animal feeding fish, is the improved seeds in fresh-water fishes, and its is nutritious, aqueous in every 100 grams of flesh of fish Point 64.1 grams, 14.4 grams of protein, and containing several mineral materials and trace element, be particularly suitable for it is weak it is deficient, malnutritive it People eats.In addition to the roe of catfish has miscellaneous taste inedibility, whole body is precious, and catfish is rare nutrient excellent product, early in history records In just it is on the books, can be compared favourably with shark's fin, wild soft-shelled turtle, be treasure in fish.Its dietary function and medical value is other Not available for fish, strong able-bodied bone and the effect of lengthening one's life are the characteristics of it are unique.Catfish is the good merchantable brand of lactagogue, its sweet property simultaneously Temperature, and play the role of nourishing yin and nourishing blood, bowl spares gas, appetizing, diuresis, is adapted to common people and eats, especially with old, young, women's postpartum and The not good people of digestive function is applicable the most, with wide market prospects and development potentiality, as the concern of raiser in recent years Focus.

As other fish, catfish is used as a kind of animal feeding habits fingerling for catfish, it is not necessary to maintain constant body temperature, growth Required energy is low, and can ingest natural bait, by the gill, skin absorption inorganic salts, and easily excludes waste, causes artificial feeding Material is relative to save mineral matter；The differentiation of catfish digestive organs is simple and short, and it is more small than terrestrial animal that the ratio between alimentary canal is long with body, The undeveloped ability of glandula digestive, it is impossible to eccrine fiber element enzyme, can not digest crude fibre substantially；Protease, amylase secretion are not enough, It is poor to protein, carbohydrate Utilization ability；Although strong to fat utilization ability, unrighted acid is mainly needed；It is right Protein requirements are high, calcium phosphorus ration 1:1-1.5, vitamin (B family vitamin especially B₁, B₂, vitamin C, vitamin A) requirement Big etc., the compounding relative to animal and fowl fodder complex enzyme is more complicated.

Adding functional feed complex enzyme in catfish feed can：1) degraded the anti-nutrition present in catfish feed because Son.These materials can not be degraded by catfish endogenous enzymes, so as to disturb the eubolism of catfish, cause catfish indigestion, production Hydraulic performance decline.2) utilization rate of starch, protein and mineral matter is improved.These materials or by rich fibrous cell membrane bag Enclose, or can not be by structure types of catfish digestion in the presence of (such as in plant feed raw material, substantial amounts of phosphorus is with phytic acid with some The form presence of phosphorus).3) degrade in the feed some specific chemical bonds.These chemical bonds can not be dropped by the enzyme of catfish itself Solution, more nutrients can be discharged after addition exogenous enzymes.4) it is endogenous because young age catfish own digestive system is also immature Enzyme is not enough, and addition exogenous enzymes can improve feed digestibility, prevent indigestion symptom.Profit except daily ration can be improved Beyond with rate, the enzyme-added difference that can also be reduced between feedstuff improves the accuracy of feed formula, while can also improve The regularity of catfish growth, reduces management cost, increases economic efficiency.Can be with environmental protection using enzyme preparation.Due to feed Utilization rate improve, corresponding excrement discharge capacity have dropped.In the case where effect is obvious, the discharge capacity of excrement can be with Reduce 20% or so.For phytate phosphorus, pollution of the phosphorus to water body environment can be greatly reduced.

The A of Chinese patent CN 106071399 disclose a kind of catfish feed, including：It is soy meal, analysis for soybean powder, black bean powder, small Flour, millet powder, glutinous millet powder, glutinous rice powder, corn flour, bran powder, oatmeal, black sesame powder, linseed meal, eggshell, crab shell, pigeon breast Meat, sunflower seeds, rapeseed, fish oil, carrot, composite trace element, citric acid, lactic acid, ostreae testa pulverata, fish meal, glutathione, phosphorus Fat, vitamin C, vitamin E, vitamin A, calcium gluconae, zinc gluconate, acetic acid, DHA, phytoenzyme and algin.The hair Bright disclosed fish meal is nutritious, rich in several amino acids, protein and trace element, while rich in various organic acids and Enzyme, not only increases the survival rate of fish, and the flesh of fish matter that can be effectively improved.

The B of Chinese patent CN 103202408 disclose a kind of environment-friendlynutritional nutritional type compound feed of mullet, it is desirable to provide Yi Zhongti The growth performance and efficiency of feed utilization of Mullet fishes high, reduce mixed feed of the breeding waste to the pollution of breeding environment；Its technology Scheme is：The feed is made up of following components：Fish meal 3~10%, Soybean Meal 10~25%, the cotton benevolence dregs of rice 5~15%, rapeseed meal 5~ 15%th, flour 15~25%, brewer's yeast 5~15%, cuttlefish cream 1~5%, extra dry white wine vinasse 10~25%, wash rice bran 5~ 15%th, soyabean lecithin oil 2~10%, phagostimulant 0.4~1%, B B-complex 0.2~0.5%, composite mineral matter 1~3%, Calcium dihydrogen phosphate 1~3%, salt 0.05~0.15%, mould inhibitor 0.02~0.06%, Choline Chloride 0.1~0.5%, dimension life Plain C acetates 0.2~0.5%, beta glucan 0.01~0.05%, vitamin E 0.01~0.05%, high temperature resistant phytase 0.01~0.05%；Belong to fish meal technical field.

The A of Chinese patent CN 104543450 disclose a kind of fermentation catfish feed, are made from the following raw materials in parts by weight： Soy noodle 80-90, sunflower meal 20-25, corn protein powder 6-8, red date powder 10-14, oyster meat 10-15, squid chip 5-10, bean curd Newborn 13-17, peach kernel 5-7, corn stem leaf 10-12, lemon 13-16, flower of Panax ginseng 1-2, palma 2-3, leek 2-3, yolk powder 7-9, butter 3-4, saccharomycete 2-3, phagostimulant 3-4, appropriate amount of water；The fermentation catfish feed of the invention, the oyster meat of addition, squid Piece is rich in multiple nutritional components such as the several mineral materials such as protein, calcium.The lemon of addition can play antibacterial, heat-clearing, promote appetite Effect.Obtained catfish feed is rich in various protein, fat, vitamin and mineral matter needed for catfish, and proportioning is suitable, accelerates The growth of catfish, improves immunity of organisms and tissue metabolism's ability.

The enzyme preparation added in fish meal disclosed above is single enzyme, and has compounded saccharomyces cerevisiae, but digestibility and feed Utilization rate is not still highly desirable.

The A of Chinese patent CN 105581024 disclose a kind of fish meal, including following portions by weight raw material composition： Wheat bran 100-200, dregs of beans 50-100, cornstarch 10-50, yeast 3-6, seawood meal 20-30, B B-complex 1-5, mineral unit Plain 1-5, complex enzyme formulation 1-3；Each composition has complementary functions, coordinated, can meet the nutritional need that fry is raised and grows, its In, by the use of wheat bran and dregs of beans as primary raw material, cost is relatively low, and improves forage protein content using seawood meal, can seawood meal Interior Rich in Trace Element, with B B-complex, mineral element, complex enzyme formulation synergy, can supplement greater protein matter, improve Flesh quality, improves the immunity of machine matter and body, and prophylactic generation improves the speed of growth of fish, reduces feeding cost, The complex enzyme formulation is zytase, cellulase, amylase, pectase, in proportion 1:1:1:1 is made.

The complex enzyme list enzyme class added in fish meal disclosed above is not comprehensive, and saccharomycete feature is poor, secretion Enzyme system is single, and feed digestion utilization rate is still undesirable.

Therefore, with reference to factors such as catfish digestion characteristics, nutritional need and water body environments, prepare that a kind of feed coefficient is low, disappear The catfish feed complex enzyme containing yeast culture that rate is high, production performance is good is imperative.

The content of the invention

Technical problem solved by the invention is the defect for overcoming existing catfish feed complex enzyme, with catfish nutritional need, Based on water body environment, with reference to its digestion characteristics, optimizing raw material formula, science compounding have powerful trophism, bioactivity and The Cultures of S. cerevisiae of the functional characteristics such as inoxidizability；Calcium, iron and Soluble Fiber can effectively be supplemented, made with powerful embedding With, extension the feed complex enzyme shelf-life calcium berry extract；Feed complex enzyme cryoprotective effects and stability are remarkably improved, are improved Catfish feed water resistance and glossiness, reduce wormwood seed powder of the feed containing powder rate；Feed palatability and digestibility can be greatly improved Agreeable to the taste dose；The Wheat Sprout Powder of abundant nutrition material, bioactivator and phytoenzyme source can be provided；The final one kind that provides can have ANFs in effect degraded catfish feed, promotes the utilization rate of the nutriments such as starch, protein and mineral matter, improves Nian The protein level of fish, body antioxidation activity and digestibility, significantly improve catfish digestibility and production performance, reduce and raise Material coefficient, prevents water pollution, maintains the catfish feed complex enzyme containing yeast culture of fine water.

In order to achieve the above object, the present invention uses following technical scheme：

A kind of catfish feed complex enzyme containing yeast culture, is mainly prepared by the raw material of following parts by weight：

Yeast culture 45-55 parts, 15-30 parts of calcium berry extract, Wheat Sprout Powder 10-20 parts, acidic xylanase 10-20 Part, acid protease 10-20 parts, phytase 10-20 parts, 1,4 beta-glucanase 6-10 parts, cellulase 5-9 parts, amylase 5-9 Part, 'beta '-mannase 3-7 parts, pectase 3-7 parts, lipase 2-6 parts, phosphate 2-6 parts, agreeable to the taste dose 1-5 parts, wormwood seed Powder 1-3 parts；

The acidic xylanase, acid protease, phytase, 1,4 beta-glucanase, cellulase, amylase, β-sweet dew Dextranase, pectase, lipase, phosphate are food-grade enzyme preparation；

Further, the yeast culture is with saccharomyces cerevisiae (Saccharomyces cerevisiae) tlj2016 To set out, bacterium prepares through spreading cultivation step by step, fermenting；

Preferably, the preparation method of the yeast culture, comprises the following steps：Take the saccharomyces cerevisiae after the activation of 1 ring Tlj2016 slant strains, be seeded in the 500m L triangular flasks equipped with 100m L seed culture mediums cultivate, 28~30 DEG C, shake 25~30h is cultivated under the conditions of 150~200r/min of bed rotating speed, primary seed solution is obtained；Primary seed solution is pressed into 5~10% (v/ V) inoculum concentration, being seeded in the 500m L triangular flasks equipped with 100m L fermentation mediums carries out fermented and cultured, fermentation time 25 ~30h, 28~30 DEG C of temperature, 150~200r/min of shaking speed obtains secondary seed solution；By secondary seed solution with 5~10% (v/v) inoculum concentration is linked into the 5L fermentation tank ferment at constant temperature of dress 3L fermentation mediums, 28~30 DEG C of temperature, 4~6L/ of ventilation Min, 150~200r/min of rotating speed, fermentation full adjustment zymotic fluid pH value are 6.0~6.5, when fermentation is to 25~30h, with every The addition of 25~30mmol of zymotic fluid is risen to disposable addition Cys in zymotic fluid, is continued fermentation 20-30h and is obtained most Whole zymotic fluid；Final zymotic fluid is concentrated under reduced pressure into solid content for 18-26%, and freeze-drying obtains final product yeast culture；

GSH final concentrations reach 3268-3308mg/L in the final zymotic fluid；

Viable bacteria content is in the yeast culture：7×10¹²-9×10¹²cfu/g；

The quality group of the seed culture medium turns into：(NH₄)₂SO₄6g/L, glucose 35g/L, K₂HPO₄·3H₂O 3g/ L, KH₂PO₄0.5g/L, dusty yeast 11g/L, MnSO₄0.1g/L, KCL 0.1g/L, FeSO₄0.1g/L, MgSO₄·7H₂O 0.1g/L, balance of water, pH 6.0；

The quality group of the fermentation medium turns into：(NH₄)₂SO₄10g/L, molasses 150g/L, K₂HPO₄·3H₂O 8g/ L, KH₂PO₄0.5g/L, dusty yeast 5g/L, corn pulp 10g/L, MnSO₄0.1g/L, KCL 0.1g/L, FeSO₄0.1g/L, MgSO₄·7H₂O 0.1g/L, balance of water, pH 6.0.

The saccharomyces cerevisiae is specially saccharomyces cerevisiae tlj2016, and it is micro- that the bacterial strain is preserved in China on July 15th, 2016 Biological inoculum preservation administration committee common micro-organisms center, deposit number is CGMCC No.12789, preservation address：Beijing The institute 3 of Chaoyang District North Star West Road 1, Institute of Microorganism, Academia Sinica, postcode 100101；

The saccharomyces cerevisiae tlj2016 has following characteristic：1) tolerance to glucose reaches 300g/L, beneficial to it GSH is produced under the conditions of high concentration glucose；2) 3308mg/L is reached in 5L fermentation cylinder for fermentation production GSH final concentrations；3) it is resistance to By the ability extra-heavy of Cys, slow growth is remained under the effect of 5mmol/L Cys, in 40mmol/L L- half Remain to keep GSH largely to synthesize under cystine effect；4) salt resistance ability reaches 18%, is conducive to extending its application field.

Further, the calcium berry extract is with calcium, iron and dietary fiber content calcium stem end higher, leaf of GAIGUO and chrysanthemum Powder is raw material, through ultrasonic cleaning, biology enzyme enzymolysis, microwave drying, extrusion, ultramicro grinding and be obtained；Its Soluble Fiber Cellulose content is high, bioactivity is strong, have important, positive role fiber to beneficial flora in catfish body, can particularly supplement calcium The shelf-life of iron, extension feed, compared with full diet fiber, its feature, bioactivity are more powerful；

Preferably, the preparation method of the calcium berry extract, comprises the following steps：Calcium stem end, leaf of GAIGUO and inulin are pressed Mass ratio 5-9:3-6:1-3 uniformly mixes, and is placed in supersonic wave cleaning machine and cleans 3-5min in 200W, 30KHz, drains, and crushes To particle diameter 0.3-0.7mm, the water of 0.5-1.5 times of fragment quality is added, regulation pH value is 5-7, adds mixture quality 0.1- 0.3% biology enzyme, 10-15min is digested in 50-60 DEG C；Enzymolysis liquid is concentrated under reduced pressure into solid content for 40-60%, is put into Microwave dryer carries out intermittent drying in 2000W, 130-150 DEG C, is allowed to moisture and reaches 4-6%, is then crushed to particle diameter 0.4-0.6mm, adds the sodium acid carbonate of crushed material quality 0.2-0.4%, and uniform mixing, adjustment mixture moisture is 12- 14%, room temperature, sealing and standing 2-4h, in screw speed 105-115r/min, 140-160 DEG C of condition extrusion of temperature, then Ultramicro grinding obtains final product calcium berry extract to particle diameter 0.1-0.3mm；

The biology enzyme is cellulase, laccase, dextranase, seminase, pectase 2-4 in mass ratio:1-3:1- 3:0.5-1.5:0.4-1 uniformly mixes.

Further, the preparation method of the Wheat Sprout Powder, comprises the following steps：By wheat be placed on temperature for 10-18 DEG C, To soak 20-40min in the soak of 6-8, soak contains the sodium acid carbonate that mass percent concentration is 0.1-0.3% to pH value With the compound enzyme of 0.2-0.4%, 28-32 DEG C then is warming up to the speed of 3-5 DEG C/min, continues to soak 3-5h, finally with 5-7 DEG C/speed of min is warming up to 50-60 DEG C of immersion 5-8min, whole immersion process is divulged information once every 15-20min, ventilation pressure 0.13-0.15MPa, while in electric-field intensity 3-5kV/cm, burst length 100-300 μ s, pulse frequency 60-80Hz conditions are carried out High-pressure pulse electric treatment, until wheat water content content is 40-45%；Wheat after immersion is drained, and is carried out in growing floor dark Germination, dark germination temperature is kept for 20-24 DEG C, and the dark germination time is 20-24h, is through the wheat drying after germination to moisture 5-8%, low-temperature grinding crosses 60-80 mesh sieves and obtains final product Wheat Sprout Powder；

The compound enzyme is cellulase, 1,4 beta-glucanase, zytase, pentosanase 2-4 in mass ratio:2-4:1-3: 1-2 uniformly mixes.

Further, described agreeable to the taste dose is science compounding sheep blood and earthworm, through high-pressure pulse electric, complex enzyme segmentation enzyme Solution, ultrafiltration, segmentation reaction, the low temperature processing techniques such as freeze-drying and prepare have sheep blood concurrently and the composite flavor of earthworm local flavor is in Gustin, is not only greatly improved the palatability of catfish feed, improves efficiency of feed utilization, and with compared with strong anti-oxidation effect, disappear Except free radical；

Preferably, described agreeable to the taste dose of preparation method, comprises the following steps：1-3 in mass ratio:2-8 weighs fresh sheep blood And earthworm, mix, rub to obtain compound, the solion of 2-5 times of its quality is added, mix, regulation pH value is 6-8, in room temperature Under in electric-field intensity 25-35kV/cm, burst length 300-500 μ s, high-voltage pulse is carried out under the conditions of pulse frequency 200-300Hz Electric field treatment 20-30min；The complex enzyme of mixture quality 0.04-0.08% is subsequently adding, 30-50min is digested in 50-60 DEG C, Then 35-45 DEG C of enzymolysis 40-60min, 90 DEG C of enzyme 10min that go out are cooled to, room temperature is down to, aseptic filtration obtains enzymolysis liquid, enzymolysis liquid By the ultrafiltration membrance filter of 1000Da and 5000Da, the permeate between two films is taken, be concentrated under reduced pressure into solid content for 18- 22% must digest concentrate；The reduced sugar of enzymolysis concentrate quality 5-7% is added, 30-50min is reacted in 112-118 DEG C successively After 135-145 DEG C of reaction 45-65min；By reaction solution by the ultrafiltration membrance filter of 1000Da and 5000Da, take between two films Permeate, is concentrated under reduced pressure into solid content for 26-30% must react concentrate；To reaction concentrate in add its quality 0.2- 0.4% sericin peptide taken, mixes, and freeze-drying obtains final product agreeable to the taste dose.

Preferably, the solion is to contain sodium ion 17-22mg/L, zinc ion 15-20mg/L, potassium ion 12- The aqueous solution of 14mg/L, calcium ion 9-11mg/L and magnesium ion 6-10mg/L；

Preferably, the complex enzyme is earthworm serine protease, alkali protease, flavor protease, lipase by matter Amount compares 4-6:2-4:0.2-0.4:0.1-0.3 uniformly mixes；

Preferably, the reduced sugar is one or more in xylose, ribose, glucose, fructose, galactolipin；

It is highly preferred that the quality group of the reduced sugar turns into：Xylose:Ribose:Glucose:Fructose:Galactolipin=3-5:2- 4:1-3:1-3:1-2。

Further, the wormwood seed powder is with wormwood seed as raw material, through high-pressure electrostatic, high-pressure pulse electric auxiliary bigcatkin willow Crushed after acid soak, refrigerating treatment, freeze and be obtained；

Preferably, the preparation method of the wormwood seed powder, comprises the following steps：By wormwood seed sabot, first in electric-field strength Degree 1-3kV/cm high-pressure electrostatic treatment 5-7min；Then concentration for 3-5mg/L salicylic acid solution in soaking at room temperature 2-5h, together When in electric-field intensity 2-6kV/cm, burst length 50-100 μ s, high-pressure pulse electric is carried out under the conditions of pulse frequency 100-150Hz Treatment；Rinse, drain, in 3-5 DEG C stand 18-24h, then successively 1-3 DEG C refrigerate 2-4d, -3--5 DEG C freezing 1-3d, - 15--18 DEG C of freezing 10-15h, is immediately placed at outdoor natural light according to being crushed immediately after 2-4h, freezes, and adds freeze-dried powder quality The sodium bicarbonate of 3-8%, uniform mixing is crossed 80-100 mesh sieves and obtains final product wormwood seed powder.

Another object of the present invention is to provide the preparation method of the above-mentioned catfish feed complex enzyme containing yeast culture：Including such as Lower step：According to formula, first successively by wormwood seed powder, agreeable to the taste dose, phosphate, lipase, pectase, beta-mannase Enzyme, amylase, cellulase, 1,4 beta-glucanase, phytase, acid protease, acidic xylanase, Wheat Sprout Powder, yeast training Support thing, the 10-30% of calcium berry extract quality adds V-type blending tank uniformly to mix 15-25min, speed of agitator 20-40r/min, It is subsequently adding remaining calcium berry extract and uniformly mixes 20-40min, speed of agitator 40-60r/min, sterile filling, sealing, packaging Obtain final product the catfish feed complex enzyme containing yeast culture.

Beneficial effect：

The present invention containing yeast culture catfish feed complex enzyme based on catfish nutritional need, water body environment, with reference to Its digestion characteristics, optimizing raw material formula, science compounding is with functional characteristics such as powerful trophism, bioactivity and inoxidizability Cultures of S. cerevisiae；Calcium, iron and Soluble Fiber can effectively be supplemented, protected with powerful embedding effect, extension feed complex enzyme The calcium berry extract of matter phase；Feed complex enzyme cryoprotective effects and stability are remarkably improved, catfish feed water resistance and light is improved Pool property, reduces wormwood seed powder of the feed containing powder rate；Agreeable to the taste dose of feed palatability and digestibility can be greatly improved；Can provide abundant The Wheat Sprout Powder of nutriment, bioactivator and phytoenzyme source；During a kind of catfish feed that can effectively degrade finally is provided ANFs, promotes the utilization rate of the nutriments such as starch, protein and mineral matter, improves protein level, the machine of catfish Body antioxidation activity and digestibility, significantly improve catfish digestibility and production performance, reduce feed coefficient, and anti-water-stop body is dirty Dye, the feed complex enzyme for maintaining fine water.1) feeding experiment shows：With same feeding and acclimation method, same is daily Management, feeds catfish, with identical addition when same fish pond condition and same native catfish fingerling and injected volume The feed palatability of the catfish feed complex enzyme containing yeast culture prepared by the addition present invention is strong, and adult fish specification gap is small, whole Together, and then catfish baits, competes that the situation of looking for food is slight, it is only 3% that the death rate is low, than the reduction of control group commercial diet complex enzyme 75%；Disease resistance is strong, and the incidence of disease is only 3%, than control group commercial diet complex enzyme reduction by 66.67%；Digestibility is high, and tail increases Weight is strong, and 55.74% is improved than control group commercial diet complex enzyme；Feed coefficient is low, and feedstuff-meat ratio is low, more multiple than control group commercial diet Synthase reduction by 36.09%；Significantly, output increased 59.58%, greatly reduces and cultivates into culture benefit in the case of same injected volume This, with preferable practicality and advance.2) stability test shows：Stored 12 months under the conditions of -12 DEG C and 40 DEG C, this Compared with commercial diet complex enzyme, the loss of its proteinase activity is reduced catfish feed complex enzyme of the invention containing yeast culture respectively 43% and 18.9%, with excellent temperature storage, enzyme activity is highly stable, while also illustrating that the present invention contains yeast culture Catfish feed complex enzyme in the loss late of other bioactivators that contains it is extremely low, with preferable freeze proof, heat resistanceheat resistant performance, Product quality stabilization, long shelf-life.Particular technique effect is shown in embodiment 7-11；Particular technique principle is as follows：

1. the viable bacteria content in Cultures of S. cerevisiae of the present invention is up to 7 × 10¹²-9×10¹²Cfu/g, final zymotic fluid Middle GSH final concentrations reach 3268-3308mg/L, not only increase the protein level and enzymatic activity of catfish feed, and improve The bioactivity and inoxidizability of catfish feed, has greatly manifested the functional characteristic of feed complex enzyme, saccharomyces cerevisiae Tlj2016 except possess common yeast it is probiotic in addition to also have following functions characteristic：1) tolerance to glucose reaches 300g/L, GSH is produced beneficial to it under the conditions of high concentration glucose；2) reached in 5L fermentation cylinder for fermentation production GSH final concentrations 3308mg/L；3) the ability extra-heavy of Cys is tolerated, slow growth is remained under the effect of 5mmol/L Cys, Remain to keep GSH largely to synthesize under the effect of 40mmol/L Cys；4) salt resistance ability reaches 18%；By its science compounding Can be further in catfish feed：(1) poisonous substances such as free radical, peroxide, heavy metal and aflatoxin are removed；(2) participate in The transhipment of amino acid (glutaminase, cysteine and other neutral amino acids)；(3) absorption beneficial to iron, the absorption of selenium, calcium Absorb, glutathione can also make the peroxidating aliphatic acid in feed revert to normal fat when absorbing or after absorbing；(4) Protection gastrointestinal tract mucosa epithelium, prevents the damage to intestinal mucosa because of inflammation, ischaemic, oxidation material etc.；(5) store and provide Its composition amino acid (especially cysteine)；(6) synthesis of protein and DNA is participated in；(7) as reducing substances, beneficial to dimension Raw element E, ascorbic reduction, maintain sulfydryl enzymatic activity.

2. the wormwood seed powder that prepared by the present invention, with stronger caking property, is fitted very much while abundant nutrition material is provided Close and prepare catfish pellet, be added into catfish feed being greatly improved pellet into graininess and integrality, prepare Catfish feed water-tolerant, glossiness is strong, and rate containing powder is low, and efficiency of feed utilization is high；Wormwood cold resistance is strong simultaneously, (wormwood seed) wormwood artemisia Grass seed is rich in antifreeze protein, through high-pressure electrostatic treatment, high-pressure pulse electric auxiliary Induced by Salicylic Acid, low temperature segmentation Stress treatment Organically combined with natural lighting so that active seed containing freeze proof matrix itself resists under the stress and induction of outer boundary's environment Freeze matrix components and obtained most comprehensive, most abundant synthesis and accumulation, improve the content of antifreeze protein, and then improve feed The cryoprotective effects and stability of complex enzyme, extend the shelf-life of feed complex enzyme.It is former with calcium berry extract, agreeable to the taste dose etc. other Material science is compounded, better.

3. the calcium berry extract that prepared by the present invention is with calcic, iron and dietary fiber content calcium stem end, leaf of GAIGUO higher With inulin be raw material, through ultrasonic cleaning, biology enzyme enzymolysis, microwave drying, extrusion, ultramicro grinding and be obtained；It is soluble Content of cellulose is high, bioactivity is strong, have important, positive role fiber to beneficial flora in catfish body, can particularly mend Calcium iron, the shelf-life of extension feed are filled, compared with full diet fiber, its feature, bioactivity are more powerful；Simultaneously through ultra micro After crushing, add a certain amount of sodium acid carbonate, sealing and standing after adjusted moisture, can significantly increase dietary fiber puffed degree and The content of soluble dietary fiber, improves the feature of dietary fiber, and gained calcium berry extract thickening property is stronger, than single side Modified dietary fiber prepared by method improves 40-50%, and is not influenceed by acid, alkali, salt, significantly improves the water resistance of feed； Soluble Fiber cellulose content is high, and 20-40% is improved than modified dietary fiber prepared by single method, it is easier to by lactic acid bacteria profit With, growth and fertility of the lactic acid bacteria in animal body enteron aisle are improved, increase the type and quantity of probiotics flora, reduce animal Body gut pH, improves animal body intestine microenvironment；High adsorption capacity, after modified, the specific surface area increase of cellulose, Network enrich, absorption affinity enhancing, chelating, absorption cholesterol and bile acids organic molecule ability it is stronger, suppress animal Absorption of the body to them；Ion-exchange capacity strengthens, and to metallic element, particularly heavy metal element adsorption effect is stronger, effectively Prevent the weight of animals metal poisoning；Regulation and the resident time of maintenance gut flora, strengthen digestion and the absorbability of enteron aisle, Improve immunity of organisms；Gastrointestinal peristalsis is effectively facilitated, slows down and eliminate the adverse reactions such as gasteremphraxis, abdominal distension；Powerful embedding effect Environment (oxygen, temperature, the illumination, water activity etc.) influence of factor to feed complex enzyme quality can be prevented, is further stabilized The bioactivity and stability of feed complex enzyme, extend the shelf-life of feed complex enzyme.

4. agreeable to the taste dose of present invention preparation is science compounding sheep blood and earthworm, through high-pressure pulse electric, complex enzyme segmentation enzyme Solution, ultrafiltration, segmentation reaction, the low temperature processing techniques such as freeze-drying and prepare have sheep blood concurrently and the composite flavor of earthworm local flavor is in Gustin, taste compound enriches, and is particularly suitable for catfish taste, can not only improve the palatability of catfish feed, but also can be complete Face promotes the digestion and absorption of catfish feed nutrient, improves efficiency of feed utilization, organic in excreta so as to greatly reduce The discharge rate of the nutriments such as thing, nitrogen phosphorus, reduces pollution of the undigested material to water environment in feed.Whole process uses low temperature process Technology, it is to avoid because high temperature and caused by albuminous degeneration and other bioactivators and fragrance matter lose, using high-tension pulse Electric field technology is rushed, extraction process living contaminants can be not only prevented, and it is significantly loose between sheep blood and earthworm protein and tissue Structure, increased the permeability of tissue, be conducive to the infiltration and contact of follow-up complex enzyme, improve enzymolysis efficiency and flavor peptides Recovery rate, while also can directly increase the dissolution of soluble flavor peptides, pre-processes more efficient, more environmentally friendly relative to existing high temperature； Using subsection enzymolysis and segmentation Maillard reaction, the yield of agreeable to the taste dose of composite flavor is further increased；Solion and compound The science compounding of enzyme further increases the activity of complex enzyme, improves enzymolysis efficiency, reduces the usage amount of enzyme preparation, maximum Improve to limit the acquisition of composite flavor peptide；Using freeze-drying, especially compounded with excellent freeze proof to reaction solution science The sericin peptide taken of effect, the oxidational losses and freezing for greatly reducing agreeable to the taste dose is lost, and improves agreeable to the taste dose of inoxidizability, finally A kind of palatability for being not only greatly improved catfish feed is obtained, efficiency of feed utilization is improved, and with compared with strong anti-oxidation effect, Eliminate agreeable to the taste dose of free radical.

5. the Wheat Sprout Powder that prepared by the present invention carries out high-pressure pulse electric treatment to the wheat of immersion process and effectively prevent Soak pollution microbes, it is to avoid generation stink is penetrated into Wheat Sprout Powder product, meanwhile, can effectively strengthen cereal kind chrotoplast The relative permeability of wall and cell membrane, the water absorption rate for improving full cereal seed, promote seed to sprout in advance, improve superoxide anion certainly By generation speed and atriphos (ATP) content of base, the activation of the various biology enzymes of stratification early stage wheat seed and release Put, endosperm dissolves and functional nutrient composition, bioactive ingredients, the synthesis of antioxidant content, promotes the breathing generation of seed The effect of thanking, accelerates nutrition and functional materials, bioactive ingredients, the enrichment process of antioxidant content, when shortening enrichment Between, nutrition and functional materials, bioactive ingredients, the content of antioxidant content are improve, soaked with segmentation, rapidly heated up, Biological enzymolysis technique is organically combined, and can further degrade kind of hide fiber element and a hemicellulose structure, increase kind of a skin permeability, activation Various bioactivator (endogenous enzymes etc.) vigor are most strong, and enriching quantity is bigger, and Soluble Fiber cellulose content increases therewith, is wheat Catfish feed in bud powder provides trophic factors, better notable；By segmentation immersion, high-voltage pulse electric field technology and biology Enzymolysis is organically combined, and shortens soak time, improves germination percentage and germination uniformity；Wheat Sprout Powder heat can to greatest extent be kept Quick property content of material, especially polyphenoils (glutathione, IP6, vitamin C, polyphenol etc.), to greatest extent Ensure natural colored, the taste and flavor of Wheat Sprout Powder, while may also function as bactericidal action；Particularly moderately killed through high temperature, short time After embryo, in the case where the bioactivity of wheat malt seed is not influenceed, with the absorption of moisture, wheat embryo is suppressed but not shadow The growth and the dissolving of endosperm of items enzyme and bioactivator are rung, bud growth is very short, and respiration is weak, and sprout damage is low, carries The high yield and quality of Wheat Sprout Powder, significantly improves functional characteristic, digestion characteristics and the palatability of feed complex enzyme.

6. the preparation method process is simple of catfish feed complex enzyme of the present invention, low for equipment requirements, will be with powerful embedding The calcium berry extract of effect is added, mixed at twice, can completely be embedded wherein other enzyme preparations and bioactivator, maximum Loss of the feed complex enzyme in catfish feed granulating, storage is reduced to limit, the biology of feed complex enzyme is improve Activity and quality stability, effectively extend the shelf-life of feed complex enzyme.

It should be noted that the technique effect of the catfish feed complex enzyme of the present invention containing yeast culture and preparation method thereof It is result that each component technical characteristic and method characteristic are mutually cooperateed with, interacted, not simple technical characteristic (component work( Can) superposition, the effect that the combination of each component technical characteristic and collaboration are produced is considerably beyond each monotechnics feature functionality With the superposition of effect, with preferable advanced and practicality.

Specific embodiment

The present invention is described below by specific embodiment.Unless stated otherwise, technological means used in the present invention It is method known in those skilled in the art.In addition, embodiment is interpreted as illustrative, it is not intended to limit the present invention Scope, the spirit and scope of the invention are limited only by the claims that follow.To those skilled in the art, without departing substantially from this On the premise of invention spirit and scope, the various changes that are carried out to the material component and consumption in these embodiments or change Belong to protection scope of the present invention.

It is prepared by the raw material of embodiment 1

1. the preparation of yeast culture

The preparation method of the yeast culture, comprises the following steps：The saccharomyces cerevisiae tlj2016 taken after the activation of 1 ring is oblique Face strain, is seeded in the 500m L triangular flasks equipped with 100m L seed culture mediums and cultivates, in 30 DEG C, shaking speed 150r/min Under the conditions of cultivate 28h, obtain primary seed solution；By primary seed solution by the inoculum concentration of 8% (v/v), it is seeded to equipped with 100m L Carry out fermented and cultured in the 500m L triangular flasks of fermentation medium, fermentation time 28h, 30 DEG C of temperature, shaking speed 180r/min, Obtain secondary seed solution；Secondary seed solution is linked into the 5L fermentation tank constant temperature of dress 3L fermentation mediums with 8% (v/v) inoculum concentration Fermentation, 30 DEG C of temperature, ventilation 5L/min, rotating speed 200r/min, fermentation full adjustment zymotic fluid pH value is 6.0~6.5, works as hair When ferment is to 30h, with every liter of addition of zymotic fluid 28mmol to disposable addition Cys in zymotic fluid, continue to ferment 25h obtains final zymotic fluid；It is 22% that final zymotic fluid is concentrated under reduced pressure into solid content, and freeze-drying obtains final product Yeast Cultivation Thing；

GSH final concentrations reach 3308mg/L in the final zymotic fluid；

Viable bacteria content is in the yeast culture：9×10¹²cfu/g；

2. the preparation of Wheat Sprout Powder

The preparation method of the Wheat Sprout Powder, comprises the following steps：Wheat is placed on the leaching that temperature is 15 DEG C, pH value is 7 30min is soaked in bubble liquid, soak contains the sodium acid carbonate and 0.3% compound enzyme that mass percent concentration is 0.2%, then 30 DEG C are warming up to the speed of 4 DEG C/min, continue to soak 4h, being finally warming up to 55 DEG C with the speed of 6 DEG C/min soaks 7min, whole Individual immersion process is divulged information once every 18min, ventilation pressure 0.14MPa, while in electric-field intensity 4kV/cm, the μ of burst length 200 S, pulse frequency 70Hz condition carry out high-pressure pulse electric treatment, until wheat water content content is 42%；Wheat drip after immersion It is dry, dark germination is carried out in growing floor, dark germination temperature is kept for 22 DEG C, and the dark germination time is 22h, through the wheat drying after germination It is 6% to moisture, low-temperature grinding crosses 80 mesh sieves and obtains final product Wheat Sprout Powder；

The compound enzyme is cellulase, 1,4 beta-glucanase, zytase, pentosanase in mass ratio 3:3:2:1.5 is equal Even mixing.

3. the preparation of calcium berry extract

The preparation method of the calcium berry extract, comprises the following steps：By calcium stem end, leaf of GAIGUO and inulin in mass ratio 7: 5:2 uniform mixing, are placed in supersonic wave cleaning machine and clean 4min in 200W, 30KHz, drain, and are crushed to particle diameter 0.5mm, add The water that 1 times of fragment quality, regulation pH value is 6, adds the biology enzyme of mixture quality 0.2%, and 12min is digested in 55 DEG C；Enzyme It is 50% that solution liquid is concentrated under reduced pressure into solid content, and be put into microwave dryer carries out intermittent drying in 2000W, 140 DEG C, is allowed to Moisture reaches 5%, is then crushed to particle diameter 0.5mm, adds the sodium acid carbonate of crushed material quality 0.3%, and uniform mixing, adjustment is mixed Compound moisture is 13%, room temperature, sealing and standing 3h, in screw speed 110r/min, 150 DEG C of condition extrusions of temperature, Then ultramicro grinding obtains final product calcium berry extract to particle diameter 0.2mm；

The biology enzyme is cellulase, laccase, dextranase, seminase, pectase in mass ratio 3:2:2:1:0.7 Uniform mixing.

4. agreeable to the taste dose of preparation

Described agreeable to the taste dose of preparation method, comprises the following steps：In mass ratio 2:5 weigh fresh sheep blood and earthworm, mix, Compound is rubbed to obtain, the solion of 4 times of its quality is added, mixed, regulation pH value is 7, at room temperature in electric-field intensity 30kV/ Cm, the μ s of burst length 400, carry out high-pressure pulse electric treatment 25min under the conditions of pulse frequency 250Hz；It is subsequently adding compound The complex enzyme of quality 0.06%, 40min is digested in 55 DEG C, is then cooled to 40 DEG C of enzymolysis 50min, 90 DEG C of enzyme 10min that go out, and is down to Room temperature, aseptic filtration obtains enzymolysis liquid, and the ultrafiltration membrance filter that enzymolysis liquid passes through 1000Da and 5000Da takes the transmission between two films Liquid, is concentrated under reduced pressure into solid content for 20% must digest concentrate；The reduced sugar of enzymolysis concentrate quality 6% is added, successively In after 40min and 140 DEG C of reaction 55min of 115 DEG C of reactions；The ultrafiltration membrance filter that reaction solution is passed through into 1000Da and 5000Da, takes Permeate between two films, is concentrated under reduced pressure into solid content for 28% must react concentrate；To reaction concentrate in add it The sericin peptide taken of quality 0.3%, mixes, and freeze-drying obtains final product agreeable to the taste dose.

The solion is to contain sodium ion 20mg/L, zinc ion 18mg/L, potassium ion 13mg/L, calcium ion 10mg/L With the aqueous solution of magnesium ion 8mg/L；

The complex enzyme is earthworm serine protease, alkali protease, flavor protease, lipase in mass ratio 5:3: 0.3:0.2 uniform mixing；

The quality group of the reduced sugar turns into：Xylose:Ribose:Glucose:Fructose:Galactolipin=4:3:2:2:1.5.

5. the preparation of wormwood seed powder

The preparation method of the wormwood seed powder, comprises the following steps：By wormwood seed sabot, first in electric-field intensity 2kV/cm High-pressure electrostatic processes 6min；Then concentration for 4mg/L salicylic acid solution in soaking at room temperature 4h, while in electric-field intensity 4kV/ Cm, the μ s of burst length 80, carries out high-pressure pulse electric treatment under the conditions of pulse frequency 120Hz；Rinse, drain, in 4 DEG C of standings 21h, then refrigerates 3d, -4 DEG C of freezing 2d, -16 DEG C of freezing 12h at 2 DEG C successively, be immediately placed at outdoor natural light according to after 3h immediately Crushed, freezed, added the sodium bicarbonate of freeze-dried powder quality 5%, uniform mixing is crossed 80 mesh sieves and obtains final product wormwood seed powder.

Yeast culture that example 2 below -6 is used, Wheat Sprout Powder, calcium berry extract, agreeable to the taste dose, wormwood seed powder it is equal For prepared by embodiment 1, other raw materials are purchased in market.

Embodiment 2

50 parts of yeast culture, 22 parts of calcium berry extract, 15 parts of Wheat Sprout Powder, 15 parts of acidic xylanase, acidic protein 15 parts of enzyme, 15 parts of phytase, 8 parts of 1,4 beta-glucanase, 7 parts of cellulase, 7 parts of amylase, 5 parts of 'beta '-mannase, pectase 5 Part, 4 parts of lipase, 4 parts of phosphate, agreeable to the taste dose 3 parts, 2 parts of wormwood seed powder；

Preparation method：Comprise the following steps：According to formula, first successively by wormwood seed powder, agreeable to the taste dose, phosphate, fat Fat enzyme, pectase, 'beta '-mannase, amylase, cellulase, 1,4 beta-glucanase, phytase, acid protease, acid wood Dextranase, Wheat Sprout Powder, yeast culture, 20% addition V-type blending tank of calcium berry extract quality uniformly mix 20min, stir Mix rotating speed 30r/min, is subsequently adding remaining calcium berry extract and uniformly mixes 30min, speed of agitator 50r/min, sterile filling, close Envelope, packaging obtain final product the catfish feed complex enzyme containing yeast culture.

Embodiment 3

45 parts of yeast culture, 15 parts of calcium berry extract, 10 parts of Wheat Sprout Powder, 10 parts of acidic xylanase, acidic protein 10 parts of enzyme, 10 parts of phytase, 6 parts of 1,4 beta-glucanase, 5 parts of cellulase, 5 parts of amylase, 3 parts of 'beta '-mannase, pectase 3 Part, 2 parts of lipase, 2 parts of phosphate, agreeable to the taste dose 1 part, 1 part of wormwood seed powder；

Preparation method：Comprise the following steps：According to formula, first successively by wormwood seed powder, agreeable to the taste dose, phosphate, fat Fat enzyme, pectase, 'beta '-mannase, amylase, cellulase, 1,4 beta-glucanase, phytase, acid protease, acid wood Dextranase, Wheat Sprout Powder, yeast culture, 10% addition V-type blending tank of calcium berry extract quality uniformly mix 15min, stir Mix rotating speed 20r/min, is subsequently adding remaining calcium berry extract and uniformly mixes 20min, speed of agitator 40r/min, sterile filling, close Envelope, packaging obtain final product the catfish feed complex enzyme containing yeast culture.

Embodiment 4

55 parts of yeast culture, 30 parts of calcium berry extract, 20 parts of Wheat Sprout Powder, 20 parts of acidic xylanase, acidic protein 20 parts of enzyme, 20 parts of phytase, 10 parts of 1,4 beta-glucanase, 9 parts of cellulase, 9 parts of amylase, 7 parts of 'beta '-mannase, pectase 7 parts, 6 parts of lipase, 6 parts of phosphate, agreeable to the taste dose 5 parts, 3 parts of wormwood seed powder；

Preparation method：Comprise the following steps：According to formula, first successively by wormwood seed powder, agreeable to the taste dose, phosphate, fat Fat enzyme, pectase, 'beta '-mannase, amylase, cellulase, 1,4 beta-glucanase, phytase, acid protease, acid wood Dextranase, Wheat Sprout Powder, yeast culture, 30% addition V-type blending tank of calcium berry extract quality uniformly mix 25min, stir Mix rotating speed 40r/min, is subsequently adding remaining calcium berry extract and uniformly mixes 40min, speed of agitator 60r/min, sterile filling, close Envelope, packaging obtain final product the catfish feed complex enzyme containing yeast culture.

Embodiment 5

45 parts of yeast culture, 30 parts of calcium berry extract, 10 parts of Wheat Sprout Powder, 20 parts of acidic xylanase, acidic protein 10 parts of enzyme, 20 parts of phytase, 6 parts of 1,4 beta-glucanase, 9 parts of cellulase, 5 parts of amylase, 7 parts of 'beta '-mannase, pectase 3 Part, 6 parts of lipase, 2 parts of phosphate, agreeable to the taste dose 5 parts, 1 part of wormwood seed powder；

Preparation method：Comprise the following steps：According to formula, first successively by wormwood seed powder, agreeable to the taste dose, phosphate, fat Fat enzyme, pectase, 'beta '-mannase, amylase, cellulase, 1,4 beta-glucanase, phytase, acid protease, acid wood Dextranase, Wheat Sprout Powder, yeast culture, 30% addition V-type blending tank of calcium berry extract quality uniformly mix 15min, stir Mix rotating speed 40r/min, is subsequently adding remaining calcium berry extract and uniformly mixes 20min, speed of agitator 60r/min, sterile filling, close Envelope, packaging obtain final product the catfish feed complex enzyme containing yeast culture.

Embodiment 6

55 parts of yeast culture, 15 parts of calcium berry extract, 20 parts of Wheat Sprout Powder, 10 parts of acidic xylanase, acidic protein 20 parts of enzyme, 10 parts of phytase, 10 parts of 1,4 beta-glucanase, 5 parts of cellulase, 9 parts of amylase, 3 parts of 'beta '-mannase, pectase 7 parts, 2 parts of lipase, 6 parts of phosphate, agreeable to the taste dose 1 part, 3 parts of wormwood seed powder；

Preparation method：Comprise the following steps：According to formula, first successively by wormwood seed powder, agreeable to the taste dose, phosphate, fat Fat enzyme, pectase, 'beta '-mannase, amylase, cellulase, 1,4 beta-glucanase, phytase, acid protease, acid wood Dextranase, Wheat Sprout Powder, yeast culture, 10% addition V-type blending tank of calcium berry extract quality uniformly mix 25min, stir Mix rotating speed 20r/min, is subsequently adding remaining calcium berry extract and uniformly mixes 40min, speed of agitator 40r/min, sterile filling, close Envelope, packaging obtain final product the catfish feed complex enzyme containing yeast culture.

The culture efficiency experiment of catfish feed complex enzyme of the present invention containing yeast culture of embodiment 7

1. fingerling and its source：It is purchased from the native catfish of certain aquaculture base；

2. the selection in fish pond is tested：Two mouthfuls of fish ponds respectively experimental group and control group, fish pond have been selected in certain production of catfish Area is 3 mu, and the cultivation depth of water is 2.0-2.5 meters.The cultivation abundant water in fish pond is tested, water quality is pure and fresh pollution-free.Fish pond sets Independent culvert system, water inlet gate-valve chamber sets three banister grooves, and water jacket installs coarse strainer, and inside groove installs conic net, rhone peace Dress anti-escape network.

3. test period：2015.3.18-2015.11.18

4. cultural method：Test group and control group use following cultural method：

(1) pond preparation

1. pond is leveled：Including the dry pool, desilting and solarization pond.

2. drug disinfection：7 days before fingerling stocking, change water with quick lime 100~130,000 gram/acre and splash sterilization of cleaning up the pond, it is desirable to Splash uniform, pond Bu Liu dead angles.

3. pond water filling：Water filling on the 5th before fingerling stocking, fill level 1.2 meters before fingerling stocking, after fingerling stocking Peak level is gradually filled to again.

(2) fingerling stocking：Experiment cultivation fingerling delivers the date on March 20th, 2015, during selection fingerling, it is desirable to fingerling body Mass color slide, without disability, fingerling embathes sterilization 3~5 minutes before entering pond with 5% saline solution.It is thrown to test group and control at random Group fish pond, input situation is shown in Table 1.

The soil Nian input situations of table 1

Fish pond	Kind	Specification (g/ tails)	Total mantissa	Average mu mantissa	Mu amount (Kg)	Total amount (Kg)
							Test group	Native catfish	50-100	9453	3151	236.33	708.98
Control group	Native catfish	50-100	9450	3150	236.25	708.75

(3) tame：Sat in the north facing the south in fishpond direction, with a distance from 2.5 meters from pond bank, away from 60 centimetres of bottom of pond, taken with asbestos shingle If 3~5 food platforms.The same day that fingerling enters pond starts domestication and feeds, and people sends sound before bait throwing in, makes fish Production conditions anti- Penetrate, when sound pauses, granular bait for fish is fed with wisp near bait table.2 times a day, 30~40 minutes every time, each bait throwing in Interval 10 seconds or so, so adheres to domestication 7~10 days, and fingerling ingests granular bait for fish quantity from less to more, and the intensity of feeding is by weak To strong transformation, until taming successfully.

(4) raise：Being trained containing yeast for the preparation of the embodiment of the present invention 2 is added in the feed of test group and control group fed respectively The catfish feed complex enzyme and commercially available same type, same specification, the catfish feed complex enzyme with the date of manufacture of thing are supported, addition is feeding The 0.4% of material quality, feeds 2 times, with 9 daily:00―10:00、15:00―16:00 respectively feeds once.The bait after fish are tamed Material should be thrown on food platform.The feeding rate of fish be 2%~6%, daily ration of feeding should regard cultivate concrete condition flexibly grasp, every time with It is degree to be eaten up in 1 hour." four determine " principle should be also followed when feeding.

(5) daily management

1. regulating and controlling water quality：The preceding pond water level of fingerling stocking filled new water after fingerling stocking at 1.2 meters or so every 5~7 days Once.Add water 10~15 centimetres every time.The high temperature cultivation season water of 7-September adds to 2.0~2.5 meters, and former commonly using EM Dew improvement water quality, water transparency is 30~40 centimetres.

2. the pool is patrolled：It is daily patrol the pool and should follow early, middle and late patrol pool system.Check food situation, the variation of water of fish Deng pinpointing the problems and take measures in time.Particularly weather is sultry or has the situation of floating head omen to open aerator in time, keeps It is more than the mg/litre of water dissolved oxygen 5.

3. fish disease prevention and cure：Fish disease prevention and cure according to " put prevention first, prevent and treat combination " policy.Accomplish it is disease-free first prevent, cultivation Period is commonly used chlorine dioxide or lime disinfection water body.

5. pilot project and result：

Observation, detection catfish incidence, death condition are simultaneously noted down in the whole test period, statistical experiment group and control Group the fish pond incidence of disease and the death rate；Statistical experiment group and control group yield when going out the pool (mantissa, cabrage, gross weight, tail increase weight)； Result such as table 2：

Table 2：Native catfish feeding experiment result

Above result of the test shows：With same feeding and acclimation method, same daily management, same fish pond condition Catfish is fed with when same native catfish fingerling and injected volume, with the addition present invention preparation of identical addition containing ferment The feed palatability of the catfish feed complex enzyme of female culture is strong, and adult fish specification gap is small, and neatly, and then catfish baits, competes Situation of looking for food is slight, and it is only 3% that the death rate is low, than control group commercial diet complex enzyme reduction by 75%；Disease resistance is strong, and the incidence of disease is only It is 3%, than control group commercial diet complex enzyme reduction by 66.67%；Digestibility is high, and tail increases weight by force, than control group commercial diet Complex enzyme improves 55.74%；Feed coefficient is low, and feedstuff-meat ratio is low, than control group commercial diet complex enzyme reduction by 36.09%；Cultivation Remarkable benefit, output increased 59.58%, greatly reduces aquaculture cost in the case of same injected volume, with preferable practicality And advance.

It should be noted that：Catfish feed complex enzyme containing yeast culture prepared by embodiment of the present invention 3-6 equally has There is above-mentioned experiment effect, it is between each embodiment and little with above-mentioned experiment effect otherness.

Bioactivity (stability) experiment of catfish feed complex enzyme of the present invention containing yeast culture of embodiment 8

Catfish feed complex enzyme containing yeast culture prepared by the embodiment of the present invention 2 and commercially available identical date of manufacture Mash feed complex enzyme is stored 12 months under the conditions of -12 DEG C and 40 DEG C, is used《The GB/T23527-2009 China people are total to With state's national standard protease preparation》Middle detection method determines the enzyme activity of protease, calculates enzyme activity loss late, enzyme activity loss late Refer to that actually detected enzyme activity accounts for the percentage for marking enzyme activity with the difference of product mark enzyme activity, as a result such as table 3

The Storage period enzyme activity loss late of table 3

Result above shows, is stored 12 months under the conditions of -12 DEG C and 40 DEG C that catfish of the present invention containing yeast culture is raised Compared with commercial diet complex enzyme, the loss of its proteinase activity reduces by 43% and 18.9% to material complex enzyme respectively, with excellent Temperature storage, enzyme activity is highly stable, while also illustrating to contain in catfish feed complex enzyme of the present invention containing yeast culture Other bioactivators loss late it is extremely low, with preferable freeze proof, heat resistanceheat resistant performance, product quality stabilization, long shelf-life.

GSH capacity experimentals are produced in saccharomyces cerevisiae tlj2016 fermentations under the conditions of the sugar high of embodiment 9

(1) Shaking culture

The ring of tlj2016 slant strains one is taken, 150rpm, 30 DEG C in the 250mL shaking flasks equipped with 30mL Shake flask mediums is accessed Culture 30h obtains seed liquor；

The Shake flask medium quality group turns into：(NH4)₂SO₄6g/L, glucose 35g/L, K₂HPO4·3H₂O 3g/L, KH₂PO₄0.5g/L, dusty yeast 11g/L, MnSO₄0.1g/L, KCL 0.1g/L, FeSO₄0.1g/L, MgSO₄·7H₂O 0.1g/L, balance of water, pH 6.0；

(2) 5L fermentation tank cultures

Seed liquor is pressed into 10% inoculum concentration, is accessed and is equipped with the fermentation tank of 3L fermentation mediums, 30 DEG C, throughput 6L/ Min, tank pressure 0.03MPa, 500rpm, carry out fermented and cultured under the conditions of permanent pH6.0, ferment during to 30h, disposably add final concentration It is the Cys of 25mmol/L, total fermentation time is 50h；

The fermentation medium quality group turns into：(NH₄)₂SO₄10g/L, glucose 100g/L, K₂HPO₄·3H₂O 8g/ L, KH₂PO₄0.5g/L, dusty yeast 11g/L, MnSO₄0.1g/L, KCL 0.1g/L, FeSO4 0.1g/L, MgSO₄·7H₂O 0.1g/L, balance of water, pH6.0；

After fermentation ends, the content for determining GSH in zymotic fluid is 3308mg/L.

The saccharomyces cerevisiae tlj2016L- cysteines tolerance of embodiment 10 is tested

By starting strain and each ring of tlj2016 slant strains, the 250mL equipped with 30mL Shake flask mediums is respectively connected to 150rpm in shaking flask, 30 DEG C are cultivated, when culture is to 12h, to the Guang ammonia of L- half that different final concentrations are added in shaking flask Acid, is further cultured for 10h, determines dry cell weight, as a result table 4,5；

Shake flask medium (g/L)：(NH₄)₂SO₄6th, glucose 20, K₂HPO₄·3H₂O 3、KH₂PO₄0.5th, dusty yeast 11, MnSO₄ 0.1、KCL 0.1、FeSO4 0.1、MgSO₄·7H₂O 0.1, pH6.0；

Table 4：Starting strain Cys tolerance

Cys concentration mmol/L	0	5	10	15	20	40
							Starting strain dry weight g/L	22.6	15.7	10.2	4.3	2.2	0.8
GSH concentration (mg/L)	35.6	46.7	43.2	40.7	37.9	25.3

The tlj2016L- cysteine tolerances of table 5

Cys concentration mmol/L	0	5	10	15	20	40
							Tlj2016 dry weights g/L	25.7	28.5	23.6	21.2	20.6	18.7
GSH concentration (mg/L)	73.2	98.3	113.5	121.7	127.5	135.8

Be can be seen that for starting strain from the result of table 4,5, Cys are added in culture medium, cell stops life It is long, and start self-dissolving, cause GSH growth rates to be reduced with the rising of Cys concentration；And the Guang ammonia of low concentration L- half Under acid, tlj2016 still is able to slow growth, and with the raising of Cys concentration, the dry cell weight of tlj2016 bacterial strains is slow Decline, and GSH concentration sustainable growths, this result is beneficial in GSH production processes by adding the Guangs of precusor amino acids-L- half Propylhomoserin proposes the production for promoting GSH.

The saccharomyces cerevisiae tlj2016 salt resistance abilities of embodiment 11 are tested

Take tlj2016 bacterium solutions 1mL inoculation strain in containing different NaCl concentrations (concentration gradients be 0%, 2%, 5%, 10%th, 15%, 10mL YPD fluid nutrient mediums (pH=6.5) 18%), 24h is cultivated at being placed in 30 DEG C respectively, each treatment 3 Individual repetition.Respectively take 1ml samples bacterium solution to be mixed in 9ml physiological saline, prepare dilution factor solution, take 0.1ml dilutions solid in YPD It is coated with body flat board, culture 36 hours (each dilution factor do 3 parallel) record is inverted in 30 DEG C of biochemical cultivation cases and calculates flat Bacterium number number on plate.The results are shown in Table 6, it is known that the resistance to salinity of the bacterium is 18%, illustrate that saccharomyces cerevisiae tlj2016 not only can be with Survived in conventional environment, still there is vigor under high salt conditions, can be applied to the high salt food processing such as soy sauce, curing food Sugar is consumed in journey and produces glutathione.

The salt resistance ability of table 6 detection (× 10⁷cfu/ml)

Claims

1. a kind of catfish feed complex enzyme containing yeast culture, is mainly prepared by the raw material of following parts by weight：Yeast Cultivation Thing 45-55 parts, 15-30 parts of calcium berry extract, Wheat Sprout Powder 10-20 parts, acidic xylanase 10-20 parts, acid protease 10- 20 parts, phytase 10-20 parts, 1,4 beta-glucanase 6-10 parts, cellulase 5-9 parts, amylase 5-9 parts, 'beta '-mannase 3-7 Part, pectase 3-7 parts, lipase 2-6 parts, phosphate 2-6 parts, agreeable to the taste dose 1-5 parts, 1-3 parts of wormwood seed powder.

2. the catfish feed complex enzyme of yeast culture is contained as claimed in claim 1, it is characterised in that the yeast culture Preparation method, comprise the following steps：The saccharomyces cerevisiae tlj2016 slant strains after the activation of 1 ring are taken, is seeded to equipped with 100mL In the 500mL triangular flasks of seed culture medium cultivate, under the conditions of 28~30 DEG C, 150~200r/min of shaking speed cultivate 25~ 30h, obtains primary seed solution；By primary seed solution by the inoculum concentration of 5~10% (v/v), it is seeded to equipped with 100mL fermented and cultureds Fermented and cultured, 25~30h of fermentation time, 28~30 DEG C of temperature, 150~200r/ of shaking speed are carried out in the 500mL triangular flasks of base Min, obtains secondary seed solution；Secondary seed solution is sent out with the 5L that 5~10% (v/v) inoculum concentrations are linked into dress 3L fermentation mediums Fermentation tank ferment at constant temperature, 28~30 DEG C of temperature, 4~6L/min of ventilation, 150~200r/min of rotating speed, fermentation full adjustment fermentation Liquid pH value is 6.0~6.5, when fermentation is to 25~30h, with every liter of addition of 25~30mmol of zymotic fluid in zymotic fluid one Secondary property adds Cys, continues fermentation 20-30h and obtains final zymotic fluid；Final zymotic fluid is concentrated under reduced pressure into solid content and contains It is 18-26% to measure, and freeze-drying obtains final product yeast culture；

The saccharomyces cerevisiae tlj2016 deposit numbers are CGMCC No.12789.

3. the catfish feed complex enzyme of yeast culture is contained as claimed in claim 2, it is characterised in that the yeast culture Middle viable bacteria content is：7×10¹²-9×10¹²cfu/g。

4. the catfish feed complex enzyme of yeast culture is contained as claimed in claim 2, it is characterised in that the final zymotic fluid Middle GSH final concentrations reach 3268-3308mg/L.

5. the catfish feed complex enzyme of yeast culture is contained as claimed in claim 2, it is characterised in that the fermentation medium Quality group turn into：(NH₄)₂SO₄10g/L, molasses 150g/L, K₂HPO₄·3H₂O 8g/L, KH₂PO₄0.5g/L, dusty yeast 5g/L, corn pulp 10g/L, MnSO₄0.1g/L, KCL 0.1g/L, FeSO₄0.1g/L, MgSO₄·7H₂O 0.1g/L, surplus It is water, pH 6.0.

6. the catfish feed complex enzyme of yeast culture is contained as claimed in claim 1, it is characterised in that the calcium berry extract Preparation method, comprise the following steps：By calcium stem end, leaf of GAIGUO and inulin 5-9 in mass ratio:3-6:1-3 uniformly mixes, and is placed in 3-5min is cleaned in 200W, 30KHz in supersonic wave cleaning machine, is drained, be crushed to particle diameter 0.3-0.7mm, add fragment quality 0.5-1.5 times of water, regulation pH value is 5-7, adds the biology enzyme of mixture quality 0.1-0.3%, and 10- is digested in 50-60 DEG C 15min；Enzymolysis liquid is concentrated under reduced pressure into solid content for 40-60%, and be put into microwave dryer is carried out in 2000W, 130-150 DEG C Intermittent drying, is allowed to moisture and reaches 4-6%, is then crushed to particle diameter 0.4-0.6mm, adds crushed material quality 0.2-0.4% Sodium acid carbonate, uniform mixing, adjustment mixture moisture is 12-14%, room temperature, sealing and standing 2-4h, in screw speed 105-115r/min, 140-160 DEG C of condition extrusion of temperature, then ultramicro grinding to particle diameter 0.1-0.3mm obtain final product calcium fruit carry Take thing；

The biology enzyme is cellulase, laccase, dextranase, seminase, pectase 2-4 in mass ratio:1-3:1-3: 0.5-1.5:0.4-1 uniformly mixes.

7. the catfish feed complex enzyme of yeast culture is contained as claimed in claim 1, it is characterised in that the Wheat Sprout Powder Preparation method, comprises the following steps：Wheat is placed on temperature for 10-18 DEG C, pH value to soak 20- in the soak of 6-8 40min, soak contain mass percent concentration be 0.1-0.3% sodium acid carbonate and 0.2-0.4% compound enzyme, then with The speed of 3-5 DEG C/min is warming up to 28-32 DEG C, continues to soak 3-5h, is finally warming up to 50-60 DEG C with the speed of 5-7 DEG C/min Immersion 5-8min, whole immersion process is divulged information once every 15-20min, ventilation pressure 0.13-0.15MPa, while in electric-field strength Degree 3-5kV/cm, burst length 100-300 μ s, pulse frequency 60-80Hz conditions carry out high-pressure pulse electric treatment, until wheat Moisture is 40-45%；Wheat after immersion is drained, and dark germination is carried out in growing floor, and dark germination temperature keeps 20-24 DEG C, the dark germination time is 20-24h, is 5-8% through the wheat drying after germination to moisture, and low-temperature grinding crosses 60-80 mesh Sieve obtains final product Wheat Sprout Powder；

The compound enzyme is cellulase, 1,4 beta-glucanase, zytase, pentosanase 2-4 in mass ratio:2-4:1-3:1-2 Uniform mixing.

8. the catfish feed complex enzyme of yeast culture is contained as claimed in claim 1, it is characterised in that described agreeable to the taste dose of system Preparation Method, comprises the following steps：1-3 in mass ratio:2-8 weighs fresh sheep blood and earthworm, mixes, rubs to obtain compound, adds The solion of 2-5 times of its quality, mixes, and regulation pH value is 6-8, at room temperature in electric-field intensity 25-35kV/cm, during pulse Between 300-500 μ s, carried out under the conditions of pulse frequency 200-300Hz high-pressure pulse electric treatment 20-30min；It is subsequently adding mixing The complex enzyme of material quality 0.04-0.08%, 30-50min is digested in 50-60 DEG C, is then cooled to 35-45 DEG C of enzymolysis 40- 60min, 90 DEG C of enzyme 10min that go out, are down to room temperature, and aseptic filtration obtains enzymolysis liquid, the ultrafiltration that enzymolysis liquid passes through 1000Da and 5000Da Membrane filtration, takes the permeate between two films, is concentrated under reduced pressure into solid content for 18-22% must digest concentrate；Add enzymolysis The reduced sugar of concentrate quality 5-7%, after reacting 30-50min and 135-145 DEG C of reaction 45-65min in 112-118 DEG C successively； The ultrafiltration membrance filter that reaction solution is passed through into 1000Da and 5000Da, takes the permeate between two films, is concentrated under reduced pressure into solid content and contains Measure as 26-30% must react concentrate；To the sericin peptide taken that its quality 0.2-0.4% is added in reaction concentrate, mix, freezing is dry It is dry to obtain final product agreeable to the taste dose；

The solion is to contain sodium ion 17-22mg/L, zinc ion 15-20mg/L, potassium ion 12-14mg/L, calcium ion The aqueous solution of 9-11mg/L and magnesium ion 6-10mg/L；

The complex enzyme is earthworm serine protease, alkali protease, flavor protease, lipase 4-6 in mass ratio:2-4: 0.2-0.4:0.1-0.3 uniformly mixes；

The quality group of the reduced sugar turns into：Xylose:Ribose:Glucose:Fructose:Galactolipin=3-5:2-4:1-3:1-3:1-2.

9. the catfish feed complex enzyme of yeast culture is contained as claimed in claim 1, it is characterised in that the wormwood seed powder Preparation method, comprises the following steps：By wormwood seed sabot, 5-7min is processed in electric-field intensity 1-3kV/cm high-pressure electrostatics first； Then concentration for 3-5mg/L salicylic acid solution in soaking at room temperature 2-5h, while in electric-field intensity 2-6kV/cm, burst length 50-100 μ s, carry out high-pressure pulse electric treatment under the conditions of pulse frequency 100-150Hz；Rinse, drain, 18- is stood in 3-5 DEG C 24h, then refrigerates 2-4d, -3--5 DEG C of freezing 1-3d, -15--18 DEG C of freezing 10-15h at 1-3 DEG C successively, is immediately placed at outdoor Crushed immediately after natural lighting 2-4h, freezed, added the sodium bicarbonate of freeze-dried powder quality 3-8%, 80-100 is crossed in uniform mixing Mesh sieve obtains final product wormwood seed powder.

10. the preparation method of the catfish feed complex enzyme of yeast culture is contained as described in claim 1-9 is any, and its feature exists In comprising the following steps：According to formula, first successively by wormwood seed powder, agreeable to the taste dose, phosphate, lipase, pectase, β- Mannonase amylase, cellulase, 1,4 beta-glucanase, phytase, acid protease, acidic xylanase, wheat malt Powder, yeast culture, the 10-30% of calcium berry extract quality add V-type blending tank uniformly to mix 15-25min, speed of agitator 20-40r/min, is subsequently adding remaining calcium berry extract and uniformly mixes 20-40min, speed of agitator 40-60r/min, aseptic filling Dress, sealing, packaging obtain final product the catfish feed complex enzyme containing yeast culture.