CN101449739A - Micro-ecology fermentation protein feedstuff and preparation method thereof - Google Patents
Micro-ecology fermentation protein feedstuff and preparation method thereof Download PDFInfo
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- CN101449739A CN101449739A CNA2008101638438A CN200810163843A CN101449739A CN 101449739 A CN101449739 A CN 101449739A CN A2008101638438 A CNA2008101638438 A CN A2008101638438A CN 200810163843 A CN200810163843 A CN 200810163843A CN 101449739 A CN101449739 A CN 101449739A
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Abstract
The invention provides a micro-ecological fermented protein feed and a preparation method, wherein the feed is prepared by mixing bean pulp and silkworms with quality ratio 90-98:2-10, then cooking to obtain solid state fermention medium, then inoculating mixed strains of bacillus subtilis, Saccharomyces cerevisiae, lactobacillus and Rhizopusoligosporus on solid fermention medium, then co-mixing for 30-45 hours at 33-37 DEG C, drying fermentation products after completing fermenting, crushing and obtaining the micro-ecological fermented protein feed. The inventive micro-ecological fermented protein feed contains a plurality of biological active peptides, content of small peptide reach more than 22%, which is favourable for digestion and development; contains lactobacillus, microzyme, which is favourable for controlling useful microbiological population in intestinal tracts of animals; has good delicious flavor and good luring effect, advances young animals for ingestion; contains soybean isoflavone more than 0.05%, markedly increases immunity of bodies, increases producibility of animals; relieving odmour of fecal waste; fully takes place of animal protein feed of fish meal and plasma protein flour and the like.
Description
(1) technical field
The present invention relates to a kind of micro-ecology fermentation protein feedstuff and preparation method thereof.
(2) background technology
The problem of relevant " promoting the young animal growth " is to culture the focus that the scientific worker pays close attention to always, in order to satisfy the special physiological growth needs of young animal, the feed makers-up often adds a large amount of or raw materials of being easy to digest and assimilate close with maternal source in feed, as fish meal, meat meal tankage, SDPP, whey powder etc.But the application of these raw materials makes that the price of prescription feed is higher; influence the benefit of aquaculture; especially because fish meal as a kind of inferior scarce resource, is subjected to strict limited production protection, SDPP etc. also owing to the use that is under an embargo of bio-safety problem in each main state of origin.The restriction of animal protein feed is used, and the demand of high-quality feed protein raw materials increases day by day, the trying situation that present just feed high-quality protein raw material faces.
In order to seek the protein raw materials of high-quality, people transfer to sight on the vegetable raw material dregs of beans again.Dregs of beans is as a kind of important vegetable protein source, and wherein gross protein value is 43～48%, and is made up of the amino acid than balance, and especially lysine content reaches 2.5～2.8%.But feeding dregs of beans contains the factor of CKIs matter digestibility and utilization such as antitrypsin, anti-elk protease, agglutinin etc.; Contain the factor of influential carbohydrate digestion such as oligosaccharides, aldehydes matter, tannin etc.; Contain the factor that reduces the mineral element utilization rate such as phytic acid etc.; Contain the antivitamin factor such as antivitamin A, E, B
12Etc. the factor; Contain the factor that increases food elk viscosity, disturbs Nutrients Digestion to absorb SNSP is arranged; Other factors also have goitrogen, saponin, isoflavones, rise cyanogen glucoside etc.Therefore dregs of beans being improved, improved its digestibility, reduce its ANFs, will be to make dregs of beans become a kind of effective way of high-quality feed albumen.Many people have made extensive work in this respect, eliminate or reduce ANFs in the dregs of beans as waiting with physics method, chemical method, biological method of breeding, though these methods have certain effect, but exist or cost height or residual chemical agents or destroy some nutrition or still residual more ANFs shortcoming, in addition, also lack animal protein potential distinctive biologically active peptide after digesting, therefore fail fine application.
(3) summary of the invention
At the problem of above-mentioned existence, the present invention adopts many bacterial classifications combined ferment to produce a kind of micro-ecology fermentation protein feedstuff dregs of beans and silkworm chrysalis combination.Not only eliminated the ANFs in the dregs of beans, and dregs of beans and silkworm chrysalis contain the various active peptide after fermentation, give full play to the combined effect of dregs of beans and silkworm chrysalis active peptide, can substitute animal protein feeds such as fish meal, SDPP fully.
The technical solution used in the present invention is:
A kind of micro-ecology fermentation protein feedstuff, described feed is to add the water boiling after being mixed with 90～98:2～10 mass ratioes by dregs of beans and silkworm chrysalis to make solid-state fermentation culture medium, and on solid-state fermentation culture medium, inoculate bacillus subtilis (Bacillus subtilis), saccharomyces cerevisiae (Saccharomycescerevisiae), Bacillus acidi lactici (Lactobacillus sp.) and Rhizopus oligosporus (Rhizopus oligosporus) mixed bacteria, again in 33～37 ℃ of following blend fermented and cultured 30～45 hours, after the fermentation ends with the tunning drying, pulverizing makes described micro-ecology fermentation protein feedstuff.
Preferred described solid-state fermentation culture medium makes as follows: after dregs of beans and silkworm chrysalis mix with 90～98:2～10 mass ratioes, add quality and be the water of 0.65～1.0 times of dregs of beans and silkworm chrysalis quality sum, boiling 30～40min obtains solid-state fermentation culture medium under 0.12～0.15MPa pressure.
A kind of preparation method of micro-ecology fermentation protein feedstuff, described method is that bacillus subtilis (Bacillus subtilis), saccharomyces cerevisiae (Saccharomyces cerevisiae), Bacillus acidi lactici (Lactobacillus sp.) and Rhizopus oligosporus (Rhizopus oligosporus) mixed bacteria are seeded to solid-state fermentation culture medium, in 33～37 ℃ of following blend fermented and cultured 30～45 hours, after the fermentation ends with tunning drying, pulverizing, obtain described micro-ecology fermentation protein feedstuff (crude protein 〉=48%, Bacillus acidi lactici 〉=0.5 * 10
8Individual/g, yeast count 〉=6 * 10
8Individual/g); Adding the water boiling after described solid-state fermentation culture medium is mixed with 90～98:2～10 mass ratioes by dregs of beans and silkworm chrysalis obtains.
Each bacterial classification inoculation amount can be unrestricted, selects its conventional inoculum concentration to get final product usually.Preferably, described mixed bacteria inoculum concentration is: account for solid-state fermentation culture medium dry weight mass percent in seed liquor or kind Qu Zhiliang, the bacillus subtilis seed liquor is 0.1%～2%, saccharomyces cerevisiae kind song is 5%～12%, Bacillus acidi lactici seed liquid is 0.4%～1.2%, Rhizopus oligosporus kind song 1～2%.Described bacillus subtilis seed liquor is prepared by following method: bacillus subtilis is seeded to the liquid seed culture medium that this area routine is applicable to bacillus subtilis, 35～37 ℃, 220r/min shaking table were cultivated 24～26 hours, obtained the bacillus subtilis seed liquor; Described saccharomyces cerevisiae kind song is prepared by following method: saccharomyces cerevisiae is seeded to the liquid seed culture medium that this area routine is applicable to saccharomyces cerevisiae, cultivates 18～20 hours, and obtains saccharomyces cerevisiae kind song for 30～32 ℃; Described Bacillus acidi lactici seed liquid is prepared by following method: Bacillus acidi lactici is seeded to the liquid seed culture medium that this area routine is applicable to Bacillus acidi lactici, and 25～27 ℃, 220r/min were cultivated 24～26 hours, obtained Bacillus acidi lactici seed liquid; Described Rhizopus oligosporus kind song is prepared by following method: Rhizopus oligosporus is seeded to the liquid seed culture medium that this area routine is applicable to Rhizopus oligosporus, 35～37 ℃, 220r/min were cultivated 24～26 hours, and the results mycelium is pulverized, sterilization obtains Rhizopus oligosporus kind song.
Preferably, described bacillus subtilis is Bacillus subtilis ACCC01185, and administrative center provides by the preservation of Chinese agriculture microorganism fungus kind.
Preferably, described saccharomyces cerevisiae is Saccharomyces cerevisiae ACCC20042, and administrative center provides by the preservation of Chinese agriculture microorganism fungus kind.
Preferably, described Bacillus acidi lactici is Lactobacillus sp.ACCC11026, and administrative center provides by the preservation of Chinese agriculture microorganism fungus kind.
Preferably, described Rhizopus oligosporus is Rhizopus oligosporus ACCC30496, and administrative center provides by the preservation of Chinese agriculture microorganism fungus kind.
Preferably, after described solid-state fermentation culture medium is mixed with 90～98:2～10 mass ratioes by dregs of beans and silkworm chrysalis, add quality and be the water of 0.65～1.0 times of dregs of beans and silkworm chrysalis quality sum, boiling 30～40min obtains under 0.12～0.15MPa pressure.
Concrete, described method is as follows:
(1) Bacillus subtilis ACCC01185 is seeded to the liquid seed culture medium that routine is applicable to bacillus subtilis, and 35～37 ℃, 220r/min shaking table were cultivated 24～26 hours, obtained the bacillus subtilis seed liquor;
(2) Saccharomyces cerevisiae ACCC20042 is seeded to the seed culture medium that routine is applicable to saccharomyces cerevisiae, cultivates 18～20 hours, and obtains saccharomyces cerevisiae kind song for 30～32 ℃;
(3) Lactobacillus sp.ACCC11026 is seeded to the liquid seed culture medium that routine is applicable to Bacillus acidi lactici, and 25～27 ℃, 220r/min were cultivated 24～26 hours, obtained Bacillus acidi lactici seed liquid;
(4) Rhizopus oligosporus ACCC30496 is seeded to the liquid seed culture medium that routine is applicable to Rhizopus oligosporus, and 35～37 ℃, 220r/min were cultivated 24～26 hours, and the results mycelium is pulverized, and sterilization obtains Rhizopus oligosporus kind song;
(5) after dregs of beans and silkworm chrysalis mix with 90～98:2～10 mass ratioes, add quality and be the water of 0.65～1.0 times of dregs of beans and silkworm chrysalis quality sum, boiling 30～40min obtains solid-state fermentation culture medium under 0.12～0.15MPa pressure, described solid-state fermentation culture medium is cooled to 33～35 ℃, inoculation quality is respectively the bacillus subtilis seed liquor of solid-state fermentation culture medium dry weight 0.1%～2%, 5%～12% saccharomyces cerevisiae kind song, 0.4%～1.2% Bacillus acidi lactici seed liquid, 1%～2% Rhizopus oligosporus kind song, 33～37 ℃ of blend fermented and cultured 30～45 hours;
(6) after fermented and cultured finishes, be dried to water content less than 10%, oven dry, pulverizing promptly obtain described micro-ecology fermentation protein feedstuff.
Beneficial effect of the present invention is mainly reflected in: micro-ecology fermentation protein feedstuff ANFs of the present invention is degraded more than 95%, digests and assimilates efficient and reaches more than 90%, contains multiple biologically active peptide, and little peptide content reaches more than 22%, has aid digestion and promote to grow; Through the multiple beneficial microbial fermentation, form through low temperature drying, be rich in Bacillus acidi lactici, saccharomycete, help regulating and control the animal intestinal beneficial microbe colony; Have natural fermenting aroma, good palatability has good food calling effect, promotes young animal to search for food; Contain isoflavones more than 0.05%, can significantly improve immunity of organisms, improve breeding performonce fo animals; Also can alleviate the stink of excrement; Can substitute animal protein feeds such as fish meal, SDPP fully.
(4) specific embodiment
The present invention is described further below in conjunction with specific embodiment, but protection scope of the present invention is not limited in this:
Embodiment 1:
1, the bent preparation of the kind of bacterial classification
(1) bacillus subtilis (Bacillus subtilis ACCC01185, Chinese agriculture microorganism fungus kind preservation administrative center provides):
Slant medium is: peptone 1g, beef extract 0.3g, NaCl 0.5g, agar 2.0g, distilled water 100ml, pH value 7.0.Inoculation Bacillus subtilis ACCC01185 cultivates 24h, obtains slant strains for 37 ℃;
Liquid seed culture medium is: glucose 0.2g, NaCl 0.5g, yeast extract 0.5g, peptone 1g, distilled water 100mL, pH7.0.Liquid seeds is cultivated every 300ml triangular flask loading amount 50ml liquid seed culture medium, the 30min that under 120 ℃ of conditions, sterilizes, and the cooling back connects a ring lawn to seed culture medium from the inclined-plane, putting 37 ℃ of temperature control shaking tables cultivates, rotating speed 220r/min cultivates 24h, viable bacteria metric density 〉=2.5 * 10
10Cfu/mL is the bacillus subtilis seed liquor.
(2) Rhizopus oligosporus RT-3 (Rhizopus oligosporus ACCC30496, Chinese agriculture microorganism fungus kind preservation center provides):
PDA medium slant preservation bacterial classification.(bean sprouts 1000g adds 1000ml water boil 30min, filtered through gauze in the basic culture solution base with spore inoculating.Filtered juice adds glucose 30g, (NH
4)
2SO
44g, MgSO
47H
2O 2.5g and KH
2PO
41g, dissolving and moisturizing are to 1000ml, transfer pH5～5.5 with lactic acid, packing, the 0.1MPa 30min that sterilizes), the inoculum concentration of Rhizopus oligosporus is 100ml basic culture solution inoculation 1.0g mycelium (weight in wet base), 37 ℃ of shaken cultivation 24h, the results mycelium is with the pulverizer rubbing of the bacterium of going out, be Rhizopus oligosporus bacterial classification song, 4～8 ℃ of preservations are standby.
(3) saccharomyces cerevisiae (Saccharomyces cerevisiae ACCC20042, Chinese agriculture microorganism fungus kind preservation administrative center provides):
Former bacterium test tube---little triangular flask seed (maltose liquid glucose 100ml, 7～8 ° of Be of pol, 30 ℃ of heat insulating culture 12h---change big triangular flask (maltose liquid glucose 600ml over to, 7～8 ° of Be of pol), 30 ℃ of heat insulating culture 12h---12h---Ka Shi jar yeast starter is spread cultivation at the distiller's yeast cylinder, and (culture medium is that every cylinder is put clear water 300kg, flour 30kg to change 30 ℃ of cultivations of Ka Shi jar interior (syrup 12kg, 7～8 ° of Be of pol enlarge cultivation) over to, wheat bran 10kg) 20h is yeast kind song.
(4) Bacillus acidi lactici (Lactobacillus sp.ACCC11026, Chinese agriculture microorganism fungus kind preservation administrative center provides):
Strain inclined plane storage medium: yeast extraction 10g, glucose 15g, calcium carbonate 15g, agar 15g, distilled water 1000mL.
Fermentation seed culture medium: peptone 5g, tryptone 5g, yeast extraction 10g, distilled water 1000mL, pH6.8.
Seed culture method: slant strains is inoculated in respectively in the liquid seed culture medium, and in 25 ℃, 220r/min cultivates 24h, is Bacillus acidi lactici seed liquid.
2, fermentation raw material is handled
Dregs of beans 900g and silkworm chrysalis 100g mix, and add 1000g water, and boiling 35min sterilizes under 0.12Mpa pressure, closes inlet valve after the sterilization, and shelving 35min discharging obtains solid-state fermentation culture medium.
3, inoculation
Above-mentioned through pretreated solid-state fermentation culture medium, ventilation blast-cold to 33～35 ℃ are inoculated bacillus subtilis seed liquor 1g respectively, saccharomyces cerevisiae seed liquor 50g, and Bacillus acidi lactici seed liquid 4g, the bent 10g of Rhizopus oligosporus RT-3 kind is at 35～37 ℃ of fermented and cultured 40h.
4, temperature controlled fermentation
Fermentation temperature has mainly been managed in fermentation.Open temperature-controlled instrument, connect fan power, the temperature upper control limit is 37 ℃, is limited to 33 ℃ down.The diligent observation, diligent test.Temperature controller probe drop point wants correct, the degree of depth will reach 10cm (doing contrast but plug in thermometer on the limit), mainly prevent probe, temperature controller and connection instrument fault, begin behind the general 6h to heat up, wind is touring in the early stage used chamber, and is higher or expect that the temperature rise temperature is too quickly when weather temperature, but air door outside the opening chamber reaches the purpose of regulating growth temperature.Finish through the 40h fermentation, can strengthen air door, open door, do not make the material temperature surpass 40 ℃, wait for out the room drying.
5, drying
Adopt low temperature drying, expect temperature control early stage in 38 ℃, and later stage material temperature control is in 45 ℃, and moisture can go out the room less than 10%.Be crushed to after the oven dry more than fineness 60 orders, obtain moisture≤10%,, crude protein 〉=48%, Bacillus acidi lactici 〉=0.5 * 10
8Individual/g, yeast count 〉=6 * 10
8The micro-ecology fermentation protein feedstuff of individual/g.
6, effect
Select 120 of 28 ages in days wean three way cross piglets, by nest not, sex, principle that body weight is close be divided into 4 groups, every group of 3 repetitions, each repeats 10 pigs.The control group group basal diet of feeding, test I, test II, test III group are respectively with the dregs of beans of the alternative basal diet moderate of micro-ecology fermentation protein feedstuff of 5%, 10% and 15% present embodiment.Experimental period from 28 ages in days to 56 ages in days.Effect sees Table 1.
Table 1: different gradient micro-ecology fermentation protein feedstuffs are to the influence of weanling pig production performance
As can be seen from Table 1, the micro-ecology fermentation protein feedstuff addition is that 5%, 10% and 15% test group daily gain improves 6.48% (P＜0.05), 10.5% (P＜0.05) and 11.9% (P＜0.05) than control group respectively.
Embodiment 2:
1, the bent preparation of the kind of bacterial classification
(1) bacillus subtilis (Bacillus subtilis ACCC01185, Chinese agriculture microorganism fungus kind preservation administrative center provides):
Slant medium is: peptone 1g, beef extract 0.3g, NaCl 0.5g, agar 2.0g, distilled water 100ml, pH value 7.0.Inoculation Bacillus subtilis ACCC01185 cultivates 24h, obtains slant strains for 37 ℃;
Liquid seed culture medium is: glucose 0.2g, NaCl 0.5g, yeast extract 0.5g, peptone 1g, distilled water 100mL, pH7.0.Liquid seeds is cultivated every 300ml triangular flask loading amount 50ml liquid seed culture medium, 30min sterilizes under 120 ℃ of conditions, the cooling back connects a ring lawn to seed culture medium from the inclined-plane, putting 37 ℃ of temperature control shaking tables cultivates, rotating speed 220r/min, reaching to the gemma rate (needs 24h approximately) more than 90% time stops, and is the bacillus subtilis seed liquor.
(2) Rhizopus oligosporus (Rhizopus oligosporus ACCC30496, Chinese agriculture microorganism fungus kind preservation center provides):
PDA medium slant preservation bacterial classification.(bean sprouts 1000g adds 1000ml water boil 30min, filtered through gauze in the basic culture solution base with spore inoculating.Filtered juice adds glucose 30g, (NH
4)
2SO
44g, MgSO
47H
2O 2.5g and KH
2PO
41g.Dissolving and moisturizing are to 1000ml, transfer pH5～5.5 with lactic acid, packing, 0.1MPa sterilization 30min), the inoculum concentration of Rhizopus oligosporus is 100ml basic culture solution inoculation 2.0g mycelium (weight in wet base), 37 ℃ of shaken cultivation 24h, the results mycelium, pulverizer with the bacterium of going out rubs, and is Rhizopus oligosporus bacterial classification song, and 4～8 ℃ of preservations are standby.
(3) saccharomyces cerevisiae (Saccharomyces cerevisiae ACCC20042, Chinese agriculture microorganism fungus kind preservation administrative center provides):
Former bacterium test tube---little triangular flask seed (maltose liquid glucose 100ml, 7～8 ° of Be of pol) 30 ℃ of heat insulating culture 10h---change big triangular flask (maltose liquid glucose 600ml over to, 7～8 ° of Be of pol), 30 ℃ of heat insulating culture 10h---change (syrup 13kg in the Ka Shi jar over to, 7～8 ° of Be expansion of pol cultivation) 10h---Ka Shi jar yeast starter is spread cultivation at the distiller's yeast cylinder, and (culture medium is that every cylinder is put clear water 200kg in 30 ℃ of cultivations, flour 30kg, wheat bran 10kg) total incubation time 20h, be yeast starter liquid.
(4) Bacillus acidi lactici (Lactobacillus sp.ACCC11026, Chinese agriculture microorganism fungus kind preservation administrative center provides):
Strain inclined plane storage medium: yeast extraction 10g, glucose 15g, calcium carbonate 15g, agar 15g, distilled water 1000mL.
Fermentation seed culture medium: peptone 5g, tryptone 5g, yeast extraction 10g, distilled water 1000mL, pH6.8.
Seed culture method: slant strains is inoculated in respectively in the liquid seed culture medium, and in 25 ℃, 220r/min cultivates 24h, is Bacillus acidi lactici seed liquid.
2, fermentation raw material is handled
Dregs of beans 980g and silkworm chrysalis 20g mix, and add 650g water, and boiling 35min sterilizes under 0.12Mpa pressure, closes inlet valve after the sterilization, shelving 35min discharging.
3, inoculation
Through pretreated solid-state fermentation culture medium, ventilation blast-cold to 33～35 ℃ are inoculated bacillus subtilis seed liquor 20g respectively, S. cervisiae seed liquor 120g, and Bacillus acidi lactici seed liquid 12g, the bent 20g of Rhizopus oligosporus RT-3 kind is at 33～37 ℃ of fermented and cultured 45h.
4. temperature controlled fermentation
Fermentation temperature has mainly been managed in fermentation.Open temperature-controlled instrument, connect fan power, the temperature upper control limit is 37 ℃, is limited to 33 ℃ down.The diligent observation, diligent test.Temperature controller probe drop point wants correct, the degree of depth will reach 10cm (doing contrast but plug in thermometer on the limit), mainly prevent probe, temperature controller and connection instrument fault, begin behind the general 6h to heat up, wind is touring in the early stage used chamber, and is higher or expect that the temperature rise temperature is too quickly when weather temperature, but air door outside the opening chamber reaches the purpose of regulating growth temperature.Finish through the 45h fermentation, can strengthen air door, open door, do not make the material temperature surpass 40 ℃, wait for out the room drying.
5, drying
Adopt low temperature drying, expect temperature control early stage in 38 ℃, and later stage material temperature control is in 45 ℃, and moisture can go out the room less than 10%.Be crushed to after the oven dry more than fineness 60 orders, obtain moisture≤10%,, crude protein 〉=48%, Bacillus acidi lactici 〉=0.5 * 10
8Individual/g, yeast count 〉=6 * 10
8The micro-ecology fermentation protein feedstuff of individual/g.
6, effect
Choose 108 of 21 ages in days wean three way cross piglets, by nest not, sex, principle that body weight is close be divided into 3 groups, every group of 3 repetitions, each repeats 12 pigs.The control group group basal diet of feeding, test I, test II group substitute the fish meal of basal diet moderate respectively with 2.5% and 5% micro-ecology fermentation protein feedstuff of present embodiment.Influence to the diarrhea of weaned piglets rate during the 21-35 age in days sees Table-2.
Table 2: micro-ecology fermentation protein feedstuff is to the influence of diarrhea of weaned piglets rate
Table 2 as can be known, micro-ecology fermentation protein feedstuff can obviously reduce the diarrhea rate of weanling pig, test I and test II reduce by 44.5% (P＜0.05) and 48.7% (P＜0.05) than control group respectively.
Embodiment 3:
1, the bent preparation of the kind of bacterial classification
(1) bacillus subtilis (Bacillus subtilis ACCC01185, Chinese agriculture microorganism fungus kind preservation administrative center provides):
Slant medium is: peptone 1g, beef extract 0.3g, NaCl 0.5g, agar 2.0g, distilled water 100ml, pH value 7.0.Inoculation Bacillus subtilis ACCC01185 cultivates 24h, obtains slant strains for 37 ℃;
Liquid seed culture medium is: glucose 0.2%, NaCl 0.5%, yeast extract 0.5%, peptone 1%, pH7.0.Liquid seeds is cultivated every 300ml triangular flask loading amount 50ml liquid seed culture medium, 30min sterilizes under 120 ℃ of conditions, the cooling back connects a ring lawn to seed culture medium from the inclined-plane, putting 37 ℃ of temperature control shaking tables cultivates, rotating speed 220r/min, reaching to the gemma rate (needs 24h approximately) more than 90% time stops, and is the bacillus subtilis seed liquor.
(2) Rhizopus oligosporus (Rhizopus oligosporus ACCC30496, Chinese agriculture microorganism fungus kind preservation center provides):
PDA medium slant preservation bacterial classification.(bean sprouts 1000g adds 1000ml water boil 30min, filtered through gauze in the basic culture solution base with spore inoculating.Filtered juice adds glucose 30g, (NH
4)
2SO
44g, MgSO
47H
2O 2.5g and KH
2PO
41g, dissolving and moisturizing are to 1000ml, transfer pH5～5.5 with lactic acid, packing, the 0.1MPa 30min that sterilizes), the inoculum concentration of Rhizopus oligosporus is 100ml basic culture solution inoculation 2.0g mycelium (weight in wet base), 37 ℃ of shaken cultivation 24h, the results mycelium is with the pulverizer rubbing of the bacterium of going out, be Rhizopus oligosporus bacterial classification song, 4～8 ℃ of preservations.
(3) saccharomyces cerevisiae (Saccharomyces cerevisiae ACCC20042, Chinese agriculture microorganism fungus kind preservation administrative center provides):
Former bacterium test tube---little triangular flask seed (glucose liquid glucose 100ml, 7～8 ° of Be of pol) 30 ℃ of heat insulating culture 12h---change big triangular flask (maltose liquid glucose 600ml over to, 7～8 ° of Be of pol), 30 ℃ of heat insulating culture 12h---change (syrup 13kg in the Ka Shi jar over to, 7～8 ° of Be expansion of pol cultivation) 112h---Ka Shi jar yeast starter is spread cultivation at the distiller's yeast cylinder, and (culture medium is that every cylinder is put clear water 300kg in 30 ℃ of cultivations, flour 30kg, wheat bran 10kg) total incubation time 20h, be yeast starter liquid.
(4) Bacillus acidi lactici (Lactobacillus sp.ACCC11026, Chinese agriculture microorganism fungus kind preservation administrative center provides):
Strain inclined plane storage medium: yeast extraction 10g, glucose 15g, calcium carbonate 15g, agar 15g, distilled water 1000mL.
Fermentation seed culture medium: peptone 5g, tryptone 5g, yeast extraction 10g, distilled water 1000mL, pH6.8.
Seed culture method: slant strains is inoculated in respectively in the liquid seed culture medium, and in 25 ℃, 220r/min cultivates 24h, is Bacillus acidi lactici seed liquid.
2, fermentation raw material is handled
Dregs of beans 9.4kg and silkworm chrysalis 0.6kg mix, and add entry 8.25kg and mix, and boiling 35min sterilizes under 0.12Mpa pressure, closes inlet valve after the sterilization, shelving 35min discharging.
3, inoculation
Through pretreated solid-state fermentation culture medium, ventilation blast-cold to 33～35 ℃ are inoculated bacillus subtilis seed liquor 105g respectively, the bent 850g of saccharomyces cerevisiae bacterial classification, and Bacillus acidi lactici seed liquid 80g, the bent 150g of Rhizopus oligosporus RT-3 kind is at 33～37 ℃ of fermented and cultured 40h.
4. temperature controlled fermentation
Fermentation temperature has mainly been managed in fermentation.Open temperature-controlled instrument, connect fan power, the temperature upper control limit is 37 ℃, is limited to 33 ℃ down.The diligent observation, diligent test.Temperature controller probe drop point wants correct, the degree of depth will reach 10cm (doing contrast but plug in thermometer on the limit), mainly prevent probe, temperature controller and connection instrument fault, begin behind the general 6h to heat up, wind is touring in the early stage used chamber, and is higher or expect that the temperature rise temperature is too quickly when weather temperature, but air door outside the opening chamber reaches the purpose of regulating growth temperature.Finish through the 40h fermentation, can strengthen air door, open door, do not make the material temperature surpass 40 ℃, wait for out the room drying.
5, drying
Adopt low temperature drying, expect temperature control early stage in 38 ℃, and later stage material temperature control is in 45 ℃, and moisture can go out the room less than 10%.Be crushed to after the oven dry more than fineness 60 orders, obtain moisture≤10%,, crude protein 〉=48%, Bacillus acidi lactici 〉=0.5 * 10
8Individual/g, yeast count 〉=6 * 10
8The micro-ecology fermentation protein feedstuff of individual/g.
6, effect
Select 1 age in days AA meat chick, 360 plumages, be divided into control group at random and organize 4 processed group with test I, II, III.Every group of 3 repetitions.Each repeats 30.The control group group basal diet of feeding, test I, test II, test III group are respectively with the dregs of beans of the alternative basal diet moderate of micro-ecology fermentation protein feedstuff of 5%, 10% and 15% present embodiment.Influence to fryer average weight, average daily gain and material anharmonic ratio sees Table-3.
Table 3: micro-ecology fermentation protein feedstuff is to the broiler growth Effect on Performance
Table 3 shows: add micro-ecology fermentation protein feedstuff in the daily ration and can significantly improve the growth of meat chicken performance, average daily gain test I, test II, the test III group in 1～21 age in days and 1～42 age in days stage improve 5.0% (P＜0.05) than control group respectively, 9.6% (P＜0.05), 8.7% (P＜0.05) and 7.0% (P＜0.05), 11.5% (P＜0.05), 9.3% (P＜0.05); Material anharmonic ratio test I, test II, the test III group in 1～21 age in days and 1～42 age in days stage reduce by 9.0% (P＜0.05) than control group respectively, 10.2% (P＜0.05), 13.3% (P＜0.05) and 8.1% (P＜0.05), 12.1% (P＜0.05), 11.1% (P＜0.05).
Claims (6)
1. micro-ecology fermentation protein feedstuff, described feed is to add the water boiling after being mixed with 90～98:2～10 mass ratioes by dregs of beans and silkworm chrysalis to make solid-state fermentation culture medium, and on solid-state fermentation culture medium, inoculate bacillus subtilis (Bacillus subtilis), saccharomyces cerevisiae (Saccharomycescerevisiae), Bacillus acidi lactici (Lactobacillus sp.) and Rhizopus oligosporus (Rhizopusoligosporus) mixed bacteria, again in 33～37 ℃ of following blend fermented and cultured 30～45 hours, after the fermentation ends with the tunning drying, pulverizing makes described micro-ecology fermentation protein feedstuff.
2. micro-ecology fermentation protein feedstuff as claimed in claim, it is characterized in that described solid-state fermentation culture medium makes as follows: after dregs of beans and silkworm chrysalis mix with 90～98:2～10 mass ratioes, add quality and be the water of 0.65～1.0 times of dregs of beans and silkworm chrysalis quality sum, boiling 30～40min obtains solid-state fermentation culture medium under 0.12～0.15MPa pressure.
3. the preparation method of a micro-ecology fermentation protein feedstuff, described method is that bacillus subtilis (Bacillus subtilis), saccharomyces cerevisiae (Saccharomyces cerevisiae), Bacillus acidi lactici (Lactobacillus sp.) and Rhizopus oligosporus (Rhizopus oligosporus) mixed bacteria are seeded to solid-state fermentation culture medium, in 33～37 ℃ of following blend fermented and cultured 30～45 hours, after the fermentation ends tunning is dried to water content less than 10%, is crushed to 40～60 orders and promptly obtains described micro-ecology fermentation protein feedstuff; Adding the water boiling after described solid-state fermentation culture medium is mixed with 90～98:2～10 mass ratioes by dregs of beans and silkworm chrysalis obtains.
4. the method for claim 1, it is characterized in that described mixed bacteria inoculum concentration is: account for solid-state fermentation culture medium dry weight mass percent in seed liquor or kind Qu Zhiliang, the bacillus subtilis seed liquor is 0.1%～2%, saccharomyces cerevisiae kind song is 5%～12%, Bacillus acidi lactici seed liquid is 0.4%～1.2%, Rhizopus oligosporus kind song 1～2%;
Described bacillus subtilis seed liquor is prepared by following method: bacillus subtilis is seeded to the liquid seed culture medium that is applicable to bacillus subtilis, and 35～37 ℃, 220r/min shaking table were cultivated 24～26 hours, obtained the bacillus subtilis seed liquor;
Described saccharomyces cerevisiae kind song is prepared by following method: saccharomyces cerevisiae is seeded to the seed culture medium that is applicable to saccharomyces cerevisiae, cultivates 18～20 hours, and obtains saccharomyces cerevisiae kind song for 30～32 ℃;
Described Bacillus acidi lactici seed liquid is prepared by following method: Bacillus acidi lactici is seeded to the liquid seed culture medium that is applicable to Bacillus acidi lactici, and 25～27 ℃, 220r/min were cultivated 24～26 hours, obtained Bacillus acidi lactici seed liquid;
Described Rhizopus oligosporus kind song is prepared by following method: Rhizopus oligosporus is seeded to the liquid seed culture medium that is applicable to Rhizopus oligosporus, and 35～37 ℃, 220r/min were cultivated 24～26 hours, and the results mycelium is pulverized, and sterilization obtains Rhizopus oligosporus kind song.
5. as claim 3 or 4 described methods, it is characterized in that described bacillus subtilis is Bacillussubtilis ACCC01185, saccharomyces cerevisiae is Saccharomyces cerevisiaeACCC20042, Bacillus acidi lactici is Lactobacillus sp.ACCC11026, and Rhizopus oligosporus is Rhizopus oligosporus ACCC30496.
6. as claim 3 or 4 described methods, after it is characterized in that described solid-state fermentation culture medium is mixed with 90～98:2～10 mass ratioes by dregs of beans and silkworm chrysalis, add quality and be the water of 0.65～1.0 times of dregs of beans and silkworm chrysalis quality sum, boiling 30～40min obtains under 0.12～0.15MPa pressure.
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