CN104996722A - Method of two-step united multi-strain fermented feed - Google Patents
Method of two-step united multi-strain fermented feed Download PDFInfo
- Publication number
- CN104996722A CN104996722A CN201510498368.XA CN201510498368A CN104996722A CN 104996722 A CN104996722 A CN 104996722A CN 201510498368 A CN201510498368 A CN 201510498368A CN 104996722 A CN104996722 A CN 104996722A
- Authority
- CN
- China
- Prior art keywords
- culture
- pure culture
- seed
- parts
- raw material
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Landscapes
- Fodder In General (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a method of a two-step united multi-strain fermented feed, and belongs to the technical field of feeds. The fermented feed disclosed by the invention is obtained through two steps of aerobic fermentation and anaerobic fermentation by using maize flour, palm dregs, soybean meal, bran and middling as raw materials and by using rhizopus nigricans, aspergillus oryzae, bacillus subtilis, lactobacillus plantarum and beer yeast as fermenting strains. The feed fermented through the method disclosed by the invention has the advantages that the protein content is as high as 30-34% which is increased by 45-62% than that of raw materials, the content of alpha-amylase is as high as 90-110U/g, the enzyme activity of acidic protein is as high as 110-130U/g, and the feed is rich in protein, rich in probiotics and high in enzymic activity. The method disclosed by the invention has the characteristics of high maneuverability, being suitable for large-scale industrial production and the like, and has favorable market prospects.
Description
Technical field
The present invention relates to a kind of method of multi-cultur es two step combined ferment feed, belong to field of feed.
Background technology
In recent years, fermented feed becomes study hotspot, and the research of multiple fermented feed obtains certain development.Fermented feed is the effect utilizing microorganism, feedstuff is converted into the biological fermentation feed that microbial bacteria body protein, bioactive micro peptide amino acid, microbial activity probio and complex enzyme formulation are integrated.Fermented feed not only can make up the amino acid easily lacked in conventional feed, and roughage material composition can be made to degrade, improve food conversion ratio, antigen protein in degraded dregs of beans, in addition, fermented feed can also reduce the diarrhea rate of pig and significantly reduce thickness of fat, and lactobacillus-fermented feed also has the effect reducing pig house ammonia content.
Fermented feed divides from fermentation raw material can be divided into ensilage fermentation, concentrated feed fermentation, the fermentation of processing of farm products leftover bits and pieces etc., is divided into solid fermentation, liquid fermentation and semisolid fermentation from the physical state row of fermentation substrate.With corn flour, the palm fibre dregs of rice etc. are that the solid fermentation feed of primary raw material is relative to fruits and vegetables slag, soybean whey liquid, dining room hogwash, ensilage etc., product quality is more stable, more be appropriate to storage and transport, be conducive to realizing commercialization large-scale production, mainly comprise following several production method: (1) only has the fermentation process of anaerobic fermentation stage, after bacterial classification mixes with feed, direct envelope carries out Anaerobic culturel, the method of this directly pack fermentation is because of oxygen deficiency, aerobic bacteria growth is limited, cause hydrolytic enzyme activities in raw material low, can not the macromolecular substances in hydrolysate feed be small-molecule substance, and then sufficient nutrient can not be provided for the probio of anaerobic fermentation.In the feed of final acquisition, protein content improves not remarkable, and hydrolytic enzyme activities is low, and probio total plate count is low.(2) aerobicly anaerobism two-part fermentation process is added, make produced fermented feed while generation organic acid and lactic acid bacteria by the aerobic fermentation process combined with Solid anaerobic two kinds of modes of solid, containing a high proportion of small molecular protein through degraded, and lower content of cellulose.
Some fermented feed only takes bacterium, saccharomycete and lactic acid bacteria to carry out anaerobic fermentation, and this kind of combination needs outer interpolation enzyme preparation or add the early growth that monose is flora outward to provide nutrition.Some only takes yeast and mold or bacillus subtilis etc. to carry out aerobic fermentation and without anaerobic fermentation, this kind of fermentation is significantly improved to feed on raising protein content.
The present invention for raw material, adopts mould, saccharomycete, bacterium and lactic acid bacteria combined ferment with palm kernel meal, dregs of beans, corn etc., and sweat divides supports and two stages of anaerobism carry out.In the aerobic fermentation stage, mould, bacillus subtilis vigorous growth, produce the multiple biology enzymes such as amylase, protease, cellulase, the macromolecular substances such as the starch in hydrolysis material, protein, cellulose are Small molecular nutriment, for the growth of saccharomycete, bacterium and lactic acid bacteria provides nutriment; At anaerobic stages, mould is dissolved, intracellular hydrolase is discharged in feed, macro-nutrients in further hydrolysate feed is Small molecular nutriment, bacillus subtilis, saccharomycete and lactic acid bacteria continued growth, the lactic acid generated and anaerobic condition can suppress the growth of harmful bacteria preferably, ensure the activity of probio, finally obtain the agreeable to the taste feed of fragrance of rich protein, rich probio and enzymatic activity high.
Summary of the invention
The object of this invention is to provide a kind of method of multi-cultur es two step combined ferment feed, object is aerobic, anaerobism two-step fermentation by multiple-microorganism, obtains the agreeable to the taste feed of fragrance of rich protein, rich probio and enzymatic activity high.
Technical solution of the present invention mainly comprises the following steps:
(1) activated spawn, and the pure culture preparing head mold, aspergillus oryzae, bacillus subtilis, Lactobacillus plantarum, brewer's yeast respectively;
(2) fermentation raw material pretreatment, comprises raw material pulverizing, mixes the rear sterilizing of water mixing;
(3) aerobic fermentation: inoculation head mold, aspergillus oryzae, bacillus subtilis, Lactobacillus plantarum, brewer's yeast inoculum are in fermentation raw material, example inoculation in mass ratio, material after every 100 parts of sterilizings, inoculation rhizopus wheat bran seed pure culture 0.2 ~ 0.7 part, aspergillus oryzae wheat bran seed pure culture 0.2 ~ 0.7 part, bacillus subtilis pure culture 0.8 ~ 1.2 part, Lactobacillus plantarum pure culture 0.5 ~ 1.5 part, brewer's yeast pure culture 2 ~ 8 parts; Mix thoroughly after inoculation, be placed in 28 ~ 30 DEG C of aerobic cultivation 36 ~ 48h of moisturizing, temperature raises and is placed on 37 DEG C of constant incubator aerlbic culture 24h, controls relative humidity 85% ~ 95%;
(4) anaerobic fermentation: after aerobic stage terminates, breaks up, mixes fermented feed thoroughly, loads one-way exhaust sealing bag, is placed in Anaerobic culturel 4 ~ 8d under room temperature.
In one embodiment of the invention, described fermentation raw material pretreatment comprises: the pulverizing of (1) fermentation raw material: get corn 10 ~ 15 parts in mass ratio, 10 ~ 15 parts, wheat bran, dregs of beans 15 ~ 25 parts, palm kernel meal 35 ~ 45 parts, 10 ~ 15 parts, secondary powder, 1 ~ 2 part, urea, glucose 0.5 ~ 1 part, mixing, pulverizing, sieve, fineness is 20 ~ 40 orders; (2) fermentation raw material mix water, material-water ratio is 1:0.6 ~ 1.0, mixes thoroughly, sabot; (3) sterilizing of fermentation raw material: carry out high pressure steam sterilization to fermentation raw material, the condition of sterilizing is 0.1Mpa ~ 0.2Mpa, 15 ~ 30min.
In one embodiment of the invention, material after every 100 parts of sterilizings, inoculation rhizopus wheat bran seed pure culture 0.2 ~ 0.7 part, aspergillus oryzae wheat bran seed pure culture 0.2 ~ 0.7 part, the inoculum concentration of bacillus subtilis pure culture is 1 part, the inoculum concentration of Lactobacillus plantarum pure culture is 1 part, and the inoculum concentration of brewer's yeast pure culture is 6 parts.
In one embodiment of the invention, the activation of head mold, that the spore of inoculation rhizopus is on potato glucose slant tube culture medium, 28 ~ 30 DEG C of constant temperature culture 24 ~ 48h, the newborn mycelium of picking, be transferred on potato glucose slant tube culture medium, 28 ~ 30 DEG C of constant temperature culture 24 ~ 48h.
In one embodiment of the invention, the activation of aspergillus oryzae, that the spore of inoculation aspergillus oryzae is on potato glucose slant tube culture medium, 28 ~ 30 DEG C of constant temperature culture 24 ~ 48h, the newborn mycelium of picking, be transferred on potato glucose slant tube culture medium, 28 ~ 30 DEG C of constant temperature culture 24 ~ 48h.
In one embodiment of the invention, the activation of bacillus subtilis is seeded to by bacillus subtilis on LB slant tube culture medium, 37 DEG C of constant temperature culture 20 ~ 24h, then be transferred on LB slant tube culture medium, 37 DEG C of constant temperature culture 20 ~ 24h.
In one embodiment of the invention, the activation of Lactobacillus plantarum is seeded to by Lactobacillus plantarum on MRS fluid nutrient medium, 37 DEG C of constant temperature culture 20 ~ 24h, then transfer once on MRS fluid nutrient medium, 37 DEG C of constant temperature culture 20 ~ 24h.
In one embodiment of the invention, the activation of brewer's yeast is by brewer's yeast, is seeded on YPD fluid nutrient medium, 37 DEG C of constant temperature culture 20 ~ 24h, then transfers once on YPD fluid nutrient medium, 37 DEG C of constant temperature culture 20 ~ 24h.
In one embodiment of the invention, the preparation of head mold seed pure culture: inoculate the rhizopus filament after activation on dregs of beans bran mass solid medium, 28 ~ 30 DEG C of constant temperature culture 3 ~ 5d.
In one embodiment of the invention, the preparation of aspergillus oryzae seed pure culture: inoculate the aspergillus oryzae filament after activation on dregs of beans bran mass solid medium, 28 ~ 30 DEG C of constant temperature culture 3 ~ 5d.
In one embodiment of the invention, the preparation of bacillus subtilis seed pure culture: inoculate the bacillus subtilis after activation to 50ml LB seed culture fluid, 37 DEG C of constant-temperature shaking culture 10 ~ 18h.
In one embodiment of the invention, the preparation of Lactobacillus plantarum seed pure culture: switching 1ml ~ 2ml activation after Lactobacillus plantarum liquid medium to 50ml MRS seed culture fluid, 37 DEG C of quiescent culture 10 ~ 18h.
In one embodiment of the invention, the preparation of brewer's yeast seed pure culture: switching 1ml ~ 2ml activation after brewer's yeast liquid nutrient solution to 50ml YPD seed culture fluid, 28 ~ 30 DEG C of quiescent culture 12 ~ 18h.
The present invention, compared to classical production process, has the following advantages:
1, the activity of amylase and protease in feed has been increased substantially.The aerobic incubation of multi-cultur es two-step method fermented feed, is beneficial to the vigorous growth breeding of mould and bacillus subtilis, produces abundant hydrolase system, especially amylase and protease.Solve when only utilizing saccharomycete and bacterial fermentation and need outer problem of adding enzyme preparation.In sweat, alpha-amylase activity is up to and reaches more than 160U/g, and acid protease activity is up to more than 140U/g; At the end of anaerobic stages, alpha-amylase activity still can remain on 90 ~ 110U/g, and acid protease activity remains on 110 ~ 130U/g.These hydrolases can the macromolecular substances in hydrolysis material be Small molecular nutrients.
2, the content of feed protein has been increased substantially.The dietary protein level utilizing this method to ferment, up to 30% ~ 34%, improves 45% ~ 62% than raw material.Produce multiple hydrolase in aerobic incubation, the macromolecular substances in hydrolysis material is Small molecular nutrients, for the growth of saccharomycete, bacillus subtilis, lactic acid bacteria continuously provides nutrition, obtains a large amount of mycoprotein.Which solve and only cause protein content to increase little problem with in bacterium and yeast anaerobic fermentation feed because hydrolase is not enough.
3, the content of probio in feed has been increased substantially.By the cultivation of aerobic stage and anaerobic stages, the growth and breeding that aspergillus oryzae, head mold, brewer's yeast, bacillus subtilis and lactic acid bacteria are a large amount of, containing abundant beneficial flora in the final feed obtained.After fermentation ends, lactic acid bacteria is 5.0 × 10
10~ 5.0 × 10
11cfu/g, bacillus subtilis is 2.0 × 10
9cfu/g ~ 2.0 × 10
10cfu/g, saccharomycete is 1.5 × 10
4~ 2.0 × 10
3cfu/g.
The inventive method has workable, is applicable to the features such as large-scale industrial production, has good market prospects.
Accompanying drawing explanation
The content of thick protein in Fig. 1 feed
Alpha-amylase activity in Fig. 2 feed
Acid protease activity in Fig. 3 feed
Lactic acid bacteria total plate count logarithm value in Fig. 4 feed
Detailed description of the invention
The content of protein measures according to standard GB/T 5009.5-2010 method.
α-diastatic content measures according to GB/T5521-2008 method.
Lactic acid bacteria colony counting measures according to GB4789.35-2010 method.
Dregs of beans bran mass solid medium consists of bean cake powder 35 ~ 45g, wheat bran 30 ~ 40g, water 40 ~ 50mL, charging thickness 1 ~ 2cm.
Potato dextrose agar is formulated by following method: potato 200 parts, adds 500mL water boil 30min, filters, gets filtrate, boil, add 20 parts of agar strips, be stirred to and dissolve completely, add glucose 20 parts, moisturizing to 1000 parts, 115 ~ 121 DEG C of moist heat sterilization 20min.
LB culture medium is the preparation of reagents pressing row weight portion: beef extract 3 parts, peptone 10 parts, 5 parts, sodium chloride, deionized water 1000 parts, adjusts pH to 7.0 ~ 7.2,115 ~ 121 DEG C of moist heat sterilization 20min after mixing reagent.
YPD culture medium is the preparation of reagents pressing row weight portion: dusty yeast 10 parts, peptone 20 parts, glucose 20 parts, deionized water 1000 parts, mixing, 115 ~ 121 DEG C of moist heat sterilization 20min.
MRS broth bouillon is the preparation of reagents pressing row weight portion: peptone: 10 parts; Beef extract: 10 parts; Yeast extract: 5 parts; Glucose: 20 parts; Dipotassium hydrogen phosphate: 2 parts; Diammonium hydrogen citrate: 2 parts; Anhydrous sodium acetate: 5 parts; Magnesium sulfate: 0.58 part; Manganese sulfate: 0.25 part; Tween 80: 1 part; Deionized water: 1000 parts; PH to 6.2 ~ 6.4 are adjusted, 115 ~ 121 DEG C of moist heat sterilization 20min after mixing reagent.
Embodiment 1 liang of bacterial classification is aerobic, anaerobism two-step fermentation feed
Concrete implementation step is as follows:
A. the cultivation of seed
A. the preparation of bacillus subtilis seed pure culture: switching activation after bacillus subtilis liquid medium to LB seed culture fluid, 37 DEG C of constant-temperature shaking culture 10 ~ 18h.
B. the preparation of Lactobacillus plantarum seed pure culture: the Lactobacillus plantarum liquid medium after switching activation is trained to MRS seed
Nutrient solution, 37 DEG C of quiescent culture 10 ~ 18h.
B. the pulverizing of fermentation raw material
Get corn 10 ~ 15 parts, 10 ~ 15 parts, wheat bran, dregs of beans 15 ~ 25 parts, palm kernel meal 35 ~ 45 parts, 10 ~ 15 parts, secondary powder, 1 ~ 2 part, urea, glucose 0.5 ~ 1 part, mixing, pulverizing, sieve, fineness is 20 ~ 40 orders.
D. fermentation raw material mix water
Material-water ratio is 1:0.6 ~ 1.0, mixes thoroughly, sabot.
E. the sterilizing of fermentation raw material
Carry out high pressure steam sterilization to fermentation raw material, the condition of sterilizing is 0.1Mpa ~ 0.2Mpa, 15 ~ 30min.
F. aerobic fermentation
Inoculation bacillus subtilis inoculum is in fermentation raw material, inoculative proportion bacillus subtilis: the mass ratio of raw material is 1.5 ~ 2.5:100, mix thoroughly after inoculation, be placed in 28 ~ 30 DEG C of aerobic cultivations of incubator moisturizing, temperature raises and is placed on 37 DEG C of constant incubator aerlbic cultures, controls relative humidity 85% ~ 95%.
After aerobic fermentation 2d terminates, inoculation Lactobacillus plantarum inoculum, inoculative proportion is Lactobacillus plantarum: the mass ratio of raw material is 4 ~ 6:100, mixes thoroughly, dress one-way exhaust sealing bag, is placed in Anaerobic culturel 6d under room temperature.Measure the content of protein in feed, α-diastatic content, the content of acid protease and the viable count of lactic acid bacteria.
Embodiment 2 multi-cultur es anaerobic fermentation feed
Concrete implementation step is as follows:
A. the preparation of seed liquor
A. the preparation of bacillus subtilis seed pure culture: switching 1ml ~ 2ml activation after bacillus subtilis liquid medium to 50mlLB seed culture fluid, 37 DEG C of constant-temperature shaking culture 10 ~ 18h.
B. the preparation of Lactobacillus plantarum seed pure culture: switching 1ml ~ 2ml activation after Lactobacillus plantarum liquid medium to 50mlMRS seed culture fluid, 37 DEG C of quiescent culture 10 ~ 18h.
C. the preparation of brewer's yeast seed pure culture: switching 1ml ~ 2ml activation after brewer's yeast liquid nutrient solution to 50mlYPD seed culture fluid, 28 ~ 30 DEG C of quiescent culture 12 ~ 18h.
B. the pulverizing of fermentation raw material
Get corn 10 ~ 15 parts, 10 ~ 15 parts, wheat bran, dregs of beans 15 ~ 25 parts, palm kernel meal 35 ~ 45 parts, 10 ~ 15 parts, secondary powder, 1 ~ 2 part, urea, glucose 0.5 ~ 1 part, mixing, pulverizing, sieve, fineness is 20 ~ 40 orders.
C. fermentation raw material mix water
Material-water ratio is 1:0.6 ~ 1.0, mixes thoroughly, sabot.
D. the sterilizing of fermentation raw material
Carry out high pressure steam sterilization to fermentation raw material, the condition of sterilizing is 0.1Mpa ~ 0.2Mpa, 15 ~ 30min.
E. anaerobic fermentation
Inoculation bacillus subtilis, Lactobacillus plantarum, brewer's yeast inoculum are in fermentation raw material, inoculation quality ratio is as follows, bacillus subtilis: raw material 0.5 ~ 2:100, Lactobacillus plantarum: raw material 0.5 ~ 2:100: brewer's yeast: raw material 2 ~ 8:100.Mix thoroughly after inoculation, dress one-way exhaust sealing bag, anaerobic fermentation.The content of protein in feed, α-diastatic content, the content of acid protease and the viable count of lactic acid bacteria is measured after fermentation 6d.
The method of embodiment 3 Fodder making of the present invention
Concrete implementation step is as follows:
A. the activation of bacterial classification
A. the activation of head mold: the spore of picking head mold from the test tube slant being stored in 0-4 DEG C of refrigerator, be seeded on potato glucose slant tube culture medium, 28 ~ 30 DEG C of constant temperature culture 24 ~ 48h, the newborn mycelium of picking, be transferred on potato glucose slant tube culture medium, 28 ~ 30 DEG C of constant temperature culture 24 ~ 48h.
B. the activation of aspergillus oryzae: the spore of picking aspergillus oryzae from the test tube slant being stored in 0-4 DEG C of refrigerator, be seeded on potato glucose slant tube culture medium, 28 ~ 30 DEG C of constant temperature culture 24 ~ 48h, the newborn mycelium of picking, be transferred on potato glucose slant tube culture medium, 28 ~ 30 DEG C of constant temperature culture 24 ~ 48h.
C. the activation of bacillus subtilis: from the test tube slant picking one ring bacillus subtilis of 0-4 DEG C of refrigerators, be seeded on LB slant tube culture medium, 37 DEG C of constant temperature culture 20 ~ 24h, then be transferred on LB slant tube culture medium, 37 DEG C of constant temperature culture 20 ~ 24h.
D. the activation of Lactobacillus plantarum: from the test tube slant picking one ring Lactobacillus plantarum of 0-4 DEG C of refrigerators, be seeded on MRS fluid nutrient medium, 37 DEG C of constant temperature culture 20 ~ 24h, then transfer once on MRS fluid nutrient medium, 37 DEG C of constant temperature culture 20 ~ 24h.
E. the activation of brewer's yeast: from the test tube slant picking one ring brewer's yeast of 0 ~ 4 DEG C of refrigerator, be seeded on YPD fluid nutrient medium, 37 DEG C of constant temperature culture 20 ~ 24h, then transfer once on YPD fluid nutrient medium, 37 DEG C of constant temperature culture 20 ~ 24h.
B. the preparation of bacterial classification pure culture
A. the preparation of head mold seed pure culture: inoculate the rhizopus filament after activation on dregs of beans bran mass solid medium, 28 ~ 30 DEG C of constant temperature culture 3 ~ 5d.
B. the preparation of aspergillus oryzae seed pure culture: inoculate the rhizopus filament after activation on dregs of beans bran mass solid medium, 28 ~ 30 DEG C of constant temperature culture 3 ~ 5d.
C. the preparation of bacillus subtilis seed pure culture: switching 1ml ~ 2ml activation after brewer's yeast liquid nutrient solution to 50mlLB seed culture fluid, 37 DEG C of constant-temperature shaking culture 10 ~ 18h.
D. the preparation of Lactobacillus plantarum seed pure culture: switching 1ml ~ 2ml activation after brewer's yeast liquid nutrient solution to 50mlMRS seed culture fluid, 37 DEG C of quiescent culture 10 ~ 18h.
E. the preparation of brewer's yeast seed pure culture: switching 1ml ~ 2ml activation after brewer's yeast liquid nutrient solution to 50mlYPD seed culture fluid, 28 ~ 30 DEG C of quiescent culture 12 ~ 18h.
C. the pulverizing of fermentation raw material
Get corn 10 ~ 15 parts, 10 ~ 15 parts, wheat bran, dregs of beans 15 ~ 25 parts, palm kernel meal 35 ~ 45 parts, 10 ~ 15 parts, secondary powder, 1 ~ 2 part, urea, glucose 0.5 ~ 1 part, mixing, pulverizing, sieve, fineness is 20 ~ 40 orders.
D. fermentation raw material mix water
Material-water ratio is 1:0.6 ~ 1.0, mixes thoroughly, sabot.
E. the sterilizing of fermentation raw material
Carry out high pressure steam sterilization to fermentation raw material, the condition of sterilizing is 0.1Mpa ~ 0.2Mpa, 15 ~ 30min.
F. aerobic fermentation
Inoculation head mold, aspergillus oryzae, bacillus subtilis, Lactobacillus plantarum, brewer's yeast inoculum are in fermentation raw material, inoculation quality ratio is as follows, rhizopus: raw material 0.25 ~ 1.5:100, aspergillus oryzae: raw material 0.25 ~ 1.5:100, bacillus subtilis: raw material 0.5 ~ 2:100, Lactobacillus plantarum: raw material 0.5 ~ 2:100: brewer's yeast: raw material 2 ~ 8:100.Mix thoroughly after inoculation, be placed in 28 ~ 30 DEG C of incubator moisturizing aerobic cultivation 48h, temperature raises and is placed on 37 DEG C of constant incubator aerlbic cultures, controls relative humidity 85% ~ 95%.
G. anaerobic fermentation
After aerobic stage terminates, break up, mix fermented feed thoroughly, dress one-way exhaust sealing bag, be placed in Anaerobic culturel under room temperature.
After anaerobic fermentation 6d, measure the content of protein in feed, α-diastatic content, the content of acid protease and the viable count of lactic acid bacteria.
Embodiment 4 external enzyme preparation anaerobic fermentation feed method
Concrete implementation step is as follows:
A. the preparation of seed liquor
A. the preparation of bacillus subtilis seed pure culture: switching 1ml ~ 2ml activation after bacillus subtilis liquid medium to 50mlLB seed culture fluid, 37 DEG C of constant-temperature shaking culture 10 ~ 18h.
B. the preparation of Lactobacillus plantarum seed pure culture: switching 1ml ~ 2ml activation after Lactobacillus plantarum liquid medium to 50mlMRS seed culture fluid, 37 DEG C of quiescent culture 10 ~ 18h.
C. the preparation of brewer's yeast seed pure culture: switching 1ml ~ 2ml activation after brewer's yeast liquid nutrient solution to 50mlYPD seed culture fluid, 28 ~ 30 DEG C of quiescent culture 12 ~ 18h.
B. the pulverizing of fermentation raw material
Get corn 10 ~ 15 parts, 10 ~ 15 parts, wheat bran, dregs of beans 15 ~ 25 parts, palm kernel meal 35 ~ 45 parts, 10 ~ 15 parts, secondary powder, 1 ~ 2 part, urea, glucose 0.5 ~ 1 part, mixing, pulverizing, sieve, fineness is 20 ~ 40 orders.
C. fermentation raw material mix water
Material-water ratio is 1:0.6 ~ 1.0, mixes thoroughly, sabot.
D. the sterilizing of fermentation raw material
Carry out high pressure steam sterilization to fermentation raw material, the condition of sterilizing is 0.1Mpa ~ 0.2Mpa, 15 ~ 30min.
E. inoculate
Inoculation bacillus subtilis, Lactobacillus plantarum, brewer's yeast inoculum are in fermentation raw material, inoculation quality ratio is as follows, bacillus subtilis: raw material 0.5 ~ 2:100, Lactobacillus plantarum: raw material 0.5 ~ 2:100: brewer's yeast: raw material 2 ~ 8:100.
F. external enzyme preparation
Postvaccinal fermented feed inoculation α-amylase 100U/g, acid protease 100U/g, mixes thoroughly, loads one-way exhaust sealing bag, room temperature bottom fermentation.
After fermentation 6d, measure the viable count of the content of thick protein in feed, α-diastatic content, the content of acid protease and saccharomycete, lactic acid bacteria.
Above in each example fermentation raw material kind and proportioning identical, material protein content is 20.11% (in over dry material), and the viable count of α-amylase, acid protease content and lactic acid bacteria is all 0.
Carry out correlation analysis to the content of the fermented feed gross protein value in embodiment 1, embodiment 2, embodiment 3, embodiment 4, α-amylase, acid protease, the viable count of lactic acid bacteria, analysis result is listed in Fig. 1, Fig. 2, Fig. 3 and Fig. 4.As can be seen from Figure 1, in 4 kinds of embodiments, in the final tunning of embodiment 3, protein content is the highest, be 32.00%, improve 59.12% than protein content in former feed, the final fermented feed protein content of embodiment 4 is secondary high, be 27.10%, improve 34.26%.Visible embodiment 3 zymotechnique is conducive to the raising of protein content in fermentation raw material most.As can be seen from Figure 2, in 4 in embodiment, in the final fermented feed of embodiment 3, α-amylase activity is the highest, is 103U/g, and embodiment 4 times is high, and be 70U/g, embodiment 2 is minimum, is 24U/g.As can be seen from Figure 3, in 4 kinds of embodiments, in the final fermented feed of embodiment 3, acid protease activity is the highest, is 123U/g, and embodiment 4 times is high, and be 71U/g, embodiment 2 is minimum, is 47U/g.As can be seen from Figure 4, in the final fermented feed of embodiment 3, live lactobacillus number is the highest, and its logarithm value is 12.1.
In addition, mould extracellular proteinase and carbohydrase in sweat, mould inoculum concentration is too low, can not provide enough hydrolases, and then can not obtain enough hydrolysates, affects the growth of saccharomycete, bacterium; Mould inoculum concentration is too much, and fungus growth is too vigorous, on the one hand because competitive relation can affect the growth of saccharomycete and bacterium, can cause on the other hand for hypoxgia.
In sweat, lactic acid bacteria, bacillus subtilis and saccharomycete play different effects, and the proportioning of each bacterial classification directly has influence on the quality of fermented feed, specifically in table 1.
Table 1 fermented feed bacterium, saccharomycete inoculum concentration result of the test
As shown in Table 1, when bacillus subtilis inoculum concentration 1%, saccharomycete inoculum concentration 6%, lactobacillus inoculum amount 1%, fermented feed protein matter content is the highest.
Aerobic stage can produce a large amount of hydrolases, is also that single cell protein produces the topmost stage simultaneously.The length of aerobic stage again with temperature correlation.Continue heat-preservation fermentation 24h to be warming up to 37 DEG C after 28-30 DEG C of heat-preservation fermentation 36-48h, the protein content and the enzymatic activity that obtain fermented feed are the highest.
Although the present invention with preferred embodiment openly as above; but it is also not used to limit the present invention, any person skilled in the art, without departing from the spirit and scope of the present invention; all can do various changes and modification, what therefore protection scope of the present invention should define with claims is as the criterion.
Claims (10)
1. a method for multi-cultur es two step combined ferment feed, is characterized in that, mainly comprise the following steps:
(1) activated spawn, and the pure culture preparing head mold, aspergillus oryzae, bacillus subtilis, Lactobacillus plantarum, brewer's yeast respectively;
(2) fermentation raw material pretreatment, comprises raw material pulverizing, mixes the rear sterilizing of water mixing;
(3) aerobic fermentation: the pure culture of inoculation head mold, aspergillus oryzae, bacillus subtilis, Lactobacillus plantarum, brewer's yeast is in fermentation raw material, mix thoroughly after inoculation, be placed in 28 ~ 30 DEG C of aerobic cultivation 36 ~ 48h of moisturizing, temperature raises and is placed on 37 DEG C of constant incubator aerlbic culture 24h, controls relative humidity 85% ~ 95%;
(4) anaerobic fermentation: after aerobic stage terminates, breaks up, mixes fermented feed thoroughly, loads one-way exhaust sealing bag, is placed in Anaerobic culturel 4 ~ 8d under room temperature.
2. method according to claim 1, it is characterized in that, example inoculation in mass ratio, material after every 100 parts of sterilizings, inoculation rhizopus wheat bran seed pure culture 0.2 ~ 0.7 part, aspergillus oryzae wheat bran seed pure culture 0.2 ~ 0.7 part, bacillus subtilis pure culture 0.8 ~ 1.2 part, Lactobacillus plantarum pure culture 0.5 ~ 1.5 part, brewer's yeast pure culture 2 ~ 8 parts.
3. method according to claim 1, it is characterized in that, material after every 100 parts of sterilizings, inoculation rhizopus wheat bran seed pure culture 0.2 ~ 0.7 part, aspergillus oryzae wheat bran seed pure culture 0.2 ~ 0.7 part, the inoculum concentration of bacillus subtilis pure culture is 1 part, and the inoculum concentration of Lactobacillus plantarum pure culture is 1 part, and the inoculum concentration of brewer's yeast pure culture is 6 parts.
4. method according to claim 1, it is characterized in that, described fermentation raw material pretreatment comprises: the pulverizing of (1) fermentation raw material: get corn 10 ~ 15 parts in mass ratio, 10 ~ 15 parts, wheat bran, dregs of beans 15 ~ 25 parts, palm kernel meal 35 ~ 45 parts, 10 ~ 15 parts, secondary powder, 1 ~ 2 part, urea, glucose 0.5 ~ 1 part, mixing, pulverizing, sieve, fineness is 20 ~ 40 orders; (2) fermentation raw material mix water, material-water ratio is 1:0.6 ~ 1.0, mixes thoroughly, sabot; (3) sterilizing of fermentation raw material: carry out high pressure steam sterilization to fermentation raw material, the condition of sterilizing is 0.1Mpa ~ 0.2Mpa, 15 ~ 30min.
5. method according to claim 1, is characterized in that, the preparation of head mold seed pure culture: inoculate the rhizopus filament after activation on dregs of beans bran mass solid medium, 28 ~ 30 DEG C of constant temperature culture 3 ~ 5d.
6. method according to claim 1, is characterized in that, the preparation of aspergillus oryzae seed pure culture: inoculate the aspergillus oryzae filament after activation on dregs of beans bran mass solid medium, 28 ~ 30 DEG C of constant temperature culture 3 ~ 5d.
7. method according to claim 1, is characterized in that, the preparation of bacillus subtilis seed pure culture: inoculate the bacillus subtilis after slant activation to 50ml LB seed culture fluid, 37 DEG C of constant-temperature shaking culture 10 ~ 18h.
8. method according to claim 1, is characterized in that, the preparation of Lactobacillus plantarum seed pure culture: switching 1ml ~ 2ml activation after Lactobacillus plantarum liquid medium to 50ml MRS seed culture fluid, 37 DEG C of quiescent culture 10 ~ 18h.
9. method according to claim 1, is characterized in that, the preparation of brewer's yeast seed pure culture: switching 1ml ~ 2ml activation after brewer's yeast liquid nutrient solution to 50ml YPD seed culture fluid, 28 ~ 30 DEG C of quiescent culture 12 ~ 18h.
10. according to the feed that the arbitrary described method of claim 1-9 prepares.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510498368.XA CN104996722B (en) | 2015-08-13 | 2015-08-13 | A kind of method of the step combined ferment feed of multi-cultur es two |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510498368.XA CN104996722B (en) | 2015-08-13 | 2015-08-13 | A kind of method of the step combined ferment feed of multi-cultur es two |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104996722A true CN104996722A (en) | 2015-10-28 |
| CN104996722B CN104996722B (en) | 2018-04-06 |
Family
ID=54369832
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510498368.XA Active CN104996722B (en) | 2015-08-13 | 2015-08-13 | A kind of method of the step combined ferment feed of multi-cultur es two |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104996722B (en) |
Cited By (25)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105876197A (en) * | 2016-04-15 | 2016-08-24 | 南宁学院 | Preparation method of compound sow feed in lactation period |
| CN106173204A (en) * | 2016-07-22 | 2016-12-07 | 日照金禾博源生化有限公司 | A kind of method preparing high protein feed for base material fermentation with citric acid corn starch residue and mycelia slag |
| CN106376725A (en) * | 2016-08-25 | 2017-02-08 | 北京康缘益生生物科技有限公司 | Biological fermented feed and preparation method thereof |
| CN106538862A (en) * | 2016-11-02 | 2017-03-29 | 南京宝辉生物饲料有限公司 | A kind of biofermentation pig starter feed |
| CN106615702A (en) * | 2016-09-19 | 2017-05-10 | 湛江五洲生物工程有限公司 | Wet process deep fermentation method of fermented bean pulp |
| CN107568415A (en) * | 2017-10-24 | 2018-01-12 | 山西大禹生物工程股份有限公司 | A kind of livestock and poultry biological fermentation feed |
| CN108148779A (en) * | 2018-01-16 | 2018-06-12 | 天津生机集团股份有限公司 | A kind of organic matter decomposing inoculant and preparation method thereof |
| CN108477395A (en) * | 2018-03-23 | 2018-09-04 | 哈尔滨伟平科技开发有限公司 | A kind of production method of feed additive for ruminant |
| CN108576373A (en) * | 2018-03-29 | 2018-09-28 | 湖北吾尔利生物工程股份有限公司 | A kind of novel biological fermentation feed production technology |
| CN108795820A (en) * | 2018-06-29 | 2018-11-13 | 江苏盐城源耀生物科技有限公司 | It is a kind of for the mixing probiotics of bean pulp fermentation and its fermentation process |
| CN108850692A (en) * | 2018-06-20 | 2018-11-23 | 西南大学 | A kind of composite bait for fish culture |
| CN109221616A (en) * | 2018-09-30 | 2019-01-18 | 南宁学院 | A kind of feed and preparation method thereof improving boar reproductive performance |
| CN109486724A (en) * | 2018-12-26 | 2019-03-19 | 江苏牧之歌生态农业科技有限公司 | A kind of synchronization preparation process and its feed applications mixing probiotics leaven |
| CN109793097A (en) * | 2018-12-26 | 2019-05-24 | 河南巨龙生物工程股份有限公司 | A kind of fermentation process using sugaring tailing and tryptophan mother liquor production biological feedstuff |
| CN110100963A (en) * | 2019-05-05 | 2019-08-09 | 广西壮族自治区畜牧研究所 | A kind of Silaging method of sugarcane caudal lobe |
| CN110331104A (en) * | 2019-07-05 | 2019-10-15 | 四川大学 | A kind of lactobacillus plantarum CV10D1 and its application |
| CN110408553A (en) * | 2019-08-06 | 2019-11-05 | 苏州吉态来胺生物科技有限公司 | A method of single cell protein is produced by raw material of palm waste |
| CN110463828A (en) * | 2019-09-10 | 2019-11-19 | 宁夏健力肽生物科技有限公司 | A kind of method of palm kernel meal production biological feedstuff |
| CN110583964A (en) * | 2019-09-23 | 2019-12-20 | 江南大学 | Biological removal method for efficiently removing four aflatoxins in peanut meal |
| CN111436525A (en) * | 2020-03-18 | 2020-07-24 | 广州智特奇生物科技股份有限公司 | Method for processing feed by integrating fermentation and enzymolysis |
| CN111557377A (en) * | 2020-05-12 | 2020-08-21 | 江南大学 | Method for preparing fruit and vegetable waste fermented feed |
| CN112205541A (en) * | 2020-10-23 | 2021-01-12 | 安徽天邦饲料科技有限公司 | Fermented aquatic animal-keeping feed and production process thereof |
| CN113100330A (en) * | 2021-05-20 | 2021-07-13 | 安徽农业大学 | Method for preparing feed and fertilizer by using livestock slaughtering leftovers |
| CN114052121A (en) * | 2021-09-30 | 2022-02-18 | 山东泰山生力源集团股份有限公司 | Large-scale preparation method of palm meal biological feed |
| CN115553380A (en) * | 2022-10-13 | 2023-01-03 | 徐州工程学院 | Fermented feed and fermentation process thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101401614A (en) * | 2008-10-30 | 2009-04-08 | 河南省龙腾高科实业有限公司 | Multi-bacterial fermentation production of protein feedstuff with steam exploration of vegetables dregs and cotton dregs and method thereof |
| CN101449739A (en) * | 2008-12-25 | 2009-06-10 | 浙江大学 | Micro-ecology fermentation protein feedstuff and preparation method thereof |
| CN101632411A (en) * | 2009-08-20 | 2010-01-27 | 陈志敏 | Method for preparing high-quality rapeseed protein containing conjugated linoleic acid by multi-strain fermentation |
| CN102318732A (en) * | 2011-08-15 | 2012-01-18 | 中国农业科学院油料作物研究所 | Method for preparing oil cake protein feedstuff through low moisture solid state fermentation |
| CN102754732A (en) * | 2011-04-29 | 2012-10-31 | 弗曼燊生物科技(上海)有限公司 | Microbial fermented feed production method adopting multi-fungus joint transformation |
| CN103829036A (en) * | 2014-02-17 | 2014-06-04 | 郑州市中食农产品加工研究院 | Preparation method of microecological fermented feed by utilizing byproducts of corn deep-processing as raw materials |
-
2015
- 2015-08-13 CN CN201510498368.XA patent/CN104996722B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101401614A (en) * | 2008-10-30 | 2009-04-08 | 河南省龙腾高科实业有限公司 | Multi-bacterial fermentation production of protein feedstuff with steam exploration of vegetables dregs and cotton dregs and method thereof |
| CN101449739A (en) * | 2008-12-25 | 2009-06-10 | 浙江大学 | Micro-ecology fermentation protein feedstuff and preparation method thereof |
| CN101632411A (en) * | 2009-08-20 | 2010-01-27 | 陈志敏 | Method for preparing high-quality rapeseed protein containing conjugated linoleic acid by multi-strain fermentation |
| CN102754732A (en) * | 2011-04-29 | 2012-10-31 | 弗曼燊生物科技(上海)有限公司 | Microbial fermented feed production method adopting multi-fungus joint transformation |
| CN102318732A (en) * | 2011-08-15 | 2012-01-18 | 中国农业科学院油料作物研究所 | Method for preparing oil cake protein feedstuff through low moisture solid state fermentation |
| CN103829036A (en) * | 2014-02-17 | 2014-06-04 | 郑州市中食农产品加工研究院 | Preparation method of microecological fermented feed by utilizing byproducts of corn deep-processing as raw materials |
Non-Patent Citations (2)
| Title |
|---|
| 谢邦祥: "《农家小曲酒酿造实用技术》", 31 March 2003, 金盾出版社 * |
| 颜方贵: "《发酵微生物学》", 31 January 1993, 中国农业大学出版社 * |
Cited By (29)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105876197A (en) * | 2016-04-15 | 2016-08-24 | 南宁学院 | Preparation method of compound sow feed in lactation period |
| CN106173204B (en) * | 2016-07-22 | 2020-10-02 | 日照金禾博源生化有限公司 | Method for preparing high-protein feed by fermenting citric acid corn starch residues and hypha residues serving as base materials |
| CN106173204A (en) * | 2016-07-22 | 2016-12-07 | 日照金禾博源生化有限公司 | A kind of method preparing high protein feed for base material fermentation with citric acid corn starch residue and mycelia slag |
| CN106376725A (en) * | 2016-08-25 | 2017-02-08 | 北京康缘益生生物科技有限公司 | Biological fermented feed and preparation method thereof |
| CN106615702A (en) * | 2016-09-19 | 2017-05-10 | 湛江五洲生物工程有限公司 | Wet process deep fermentation method of fermented bean pulp |
| CN106538862A (en) * | 2016-11-02 | 2017-03-29 | 南京宝辉生物饲料有限公司 | A kind of biofermentation pig starter feed |
| CN107568415A (en) * | 2017-10-24 | 2018-01-12 | 山西大禹生物工程股份有限公司 | A kind of livestock and poultry biological fermentation feed |
| CN108148779A (en) * | 2018-01-16 | 2018-06-12 | 天津生机集团股份有限公司 | A kind of organic matter decomposing inoculant and preparation method thereof |
| CN108477395A (en) * | 2018-03-23 | 2018-09-04 | 哈尔滨伟平科技开发有限公司 | A kind of production method of feed additive for ruminant |
| CN108576373A (en) * | 2018-03-29 | 2018-09-28 | 湖北吾尔利生物工程股份有限公司 | A kind of novel biological fermentation feed production technology |
| CN108850692A (en) * | 2018-06-20 | 2018-11-23 | 西南大学 | A kind of composite bait for fish culture |
| CN108795820A (en) * | 2018-06-29 | 2018-11-13 | 江苏盐城源耀生物科技有限公司 | It is a kind of for the mixing probiotics of bean pulp fermentation and its fermentation process |
| CN109221616A (en) * | 2018-09-30 | 2019-01-18 | 南宁学院 | A kind of feed and preparation method thereof improving boar reproductive performance |
| CN109486724A (en) * | 2018-12-26 | 2019-03-19 | 江苏牧之歌生态农业科技有限公司 | A kind of synchronization preparation process and its feed applications mixing probiotics leaven |
| CN109793097A (en) * | 2018-12-26 | 2019-05-24 | 河南巨龙生物工程股份有限公司 | A kind of fermentation process using sugaring tailing and tryptophan mother liquor production biological feedstuff |
| CN110100963A (en) * | 2019-05-05 | 2019-08-09 | 广西壮族自治区畜牧研究所 | A kind of Silaging method of sugarcane caudal lobe |
| CN110100963B (en) * | 2019-05-05 | 2022-08-30 | 广西壮族自治区畜牧研究所 | Silage method of sugarcane tail leaves |
| CN110331104A (en) * | 2019-07-05 | 2019-10-15 | 四川大学 | A kind of lactobacillus plantarum CV10D1 and its application |
| CN110408553A (en) * | 2019-08-06 | 2019-11-05 | 苏州吉态来胺生物科技有限公司 | A method of single cell protein is produced by raw material of palm waste |
| CN110463828A (en) * | 2019-09-10 | 2019-11-19 | 宁夏健力肽生物科技有限公司 | A kind of method of palm kernel meal production biological feedstuff |
| CN110583964A (en) * | 2019-09-23 | 2019-12-20 | 江南大学 | Biological removal method for efficiently removing four aflatoxins in peanut meal |
| CN111436525A (en) * | 2020-03-18 | 2020-07-24 | 广州智特奇生物科技股份有限公司 | Method for processing feed by integrating fermentation and enzymolysis |
| CN111557377A (en) * | 2020-05-12 | 2020-08-21 | 江南大学 | Method for preparing fruit and vegetable waste fermented feed |
| CN111557377B (en) * | 2020-05-12 | 2022-09-27 | 江南大学 | Method for preparing fruit and vegetable waste fermented feed |
| CN112205541A (en) * | 2020-10-23 | 2021-01-12 | 安徽天邦饲料科技有限公司 | Fermented aquatic animal-keeping feed and production process thereof |
| CN113100330A (en) * | 2021-05-20 | 2021-07-13 | 安徽农业大学 | Method for preparing feed and fertilizer by using livestock slaughtering leftovers |
| CN113100330B (en) * | 2021-05-20 | 2024-01-16 | 安徽农业大学 | Method for preparing feed and fertilizer by utilizing livestock slaughtering offal |
| CN114052121A (en) * | 2021-09-30 | 2022-02-18 | 山东泰山生力源集团股份有限公司 | Large-scale preparation method of palm meal biological feed |
| CN115553380A (en) * | 2022-10-13 | 2023-01-03 | 徐州工程学院 | Fermented feed and fermentation process thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104996722B (en) | 2018-04-06 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104996722B (en) | A kind of method of the step combined ferment feed of multi-cultur es two | |
| CN101433270B (en) | Preparation method of vegetable seed protein feed | |
| CN102715342B (en) | Method for processing microbiological feed based on spirit vinasse and miscellaneous meal | |
| CN102687792B (en) | Feeding microecologic preparation based on beer grains and rice bran meal | |
| CN101215535B (en) | Solid fermentation process for preparing bacillus natto microecological preparation | |
| CN102934736B (en) | Method for preparing sweet potato skin/ sweet potato powder dreg fermented feed | |
| CN104824337A (en) | Preparation method of fermented soybean meal for feeding | |
| CN102696860B (en) | Highly efficient and low-cost microbiological feed proteins based on vinegar residue and miscellaneous meal | |
| CN108157673A (en) | A kind of preparation method for the prawn mixed feed that ferments | |
| CN110384175A (en) | The method of yeast culture and the application of yeast culture are prepared using vinasse | |
| CN104186957A (en) | Method for improving forage nutritional value of cottonseed meal through microbial fermentation | |
| CN104026331B (en) | The preparation method of mature vinegar vinegar grain feed | |
| CN102334611A (en) | Solid-state fermentation method for bacillus natto-saccharomycete composite viable bacteria preparation with rice bran as matrix | |
| CN103555598A (en) | Edible ferment complex inoculant as well as preparation method thereof | |
| CN103156051B (en) | Method using composite bacterium to ferment bean dregs to manufacture protein feed | |
| CN108378223A (en) | A kind of preparation method of the feeding fermented dregs of beans of low antigen | |
| CN106173204A (en) | A kind of method preparing high protein feed for base material fermentation with citric acid corn starch residue and mycelia slag | |
| CN104855674A (en) | Production method for microbial fermentation complete feed by combining strain joint transformations | |
| CN102960541A (en) | Method for producing environment-friendly biological feed employing distiller grains and glycoprotein | |
| CN110301526A (en) | Complex micro organism fungicide and its method for preparing bioactive feed | |
| CN108795812A (en) | A kind of compost decomposing agent and the preparation method and application thereof | |
| CN101491289A (en) | Rape seed protein feedstuff and preparation method thereof | |
| CN104232547A (en) | Microbial flora additive used for sheep feed, and preparation method thereof | |
| CN102943049B (en) | Process for producing cryytococcus neoformans culture through solid fermentation | |
| CN103060206B (en) | Fermentation bacteria agent and preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |