CN101411389B - Method for producing feed from detoxified corn residue after producing alcohol by mixed bacterial micro-ecological formulation - Google Patents
Method for producing feed from detoxified corn residue after producing alcohol by mixed bacterial micro-ecological formulation Download PDFInfo
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Abstract
The invention discloses a method for producing feedstuff by utilization of mixed flora microecological preparation for detoxication of corn alcohol residue, wherein candida utili, Bacillus subtilis and rhamnose lactobacillus are subjected to high-density fermentation, so that the viable count of the candida utili is more than 4.0*10<8>CFU/g, the viable count of the Bacillus subtilis is more than 3.8*10<8>CFU/g, and the viable count of the rhamnose lactobacillus is more than 3.0*10<8>CFU/g; and the preparation is directly mixed with the corn residue after production of alcohol or other toxic feedstuff polluted by mildew according to the proportion of between 5 and 10 grams per kilogram, and the mixture is sprayed with sterile water to maintain a humidity of more than 40 percent, reacts for 2 to 10 hours at a temperature of between 30 and 35 DEG C, is dried at a temperature of between 45 and 50 DEG C, and can degrade ZEA in the corn alcohol residue or the other toxic feedstuff, wherein the detoxication rate reaches more than 94 percent, so that the corn alcohol residue or the other toxic feedstuff becomes safe animal feedstuff.
Description
Technical field
The present invention relates to a kind of method of producing feed, particularly relate to the method for mixed bacterial micro-ecological formulation producing feed from detoxified corn residue after producing alcohol.
Background technology
It is the ubiquitous problem in the whole world that feed and food go mouldy, according to FAO (Food and Agriculture Organization of the United Nation) (FAO) data in 2002, there is 25% crops grain every year in the whole world approximately by mycotoxin contamination, and China is every year on average because the caused feed loss of going mouldy accounts for more than 10% of total output, topmost going mouldy derives from a kind of estrogens mycotoxin that sickle-like bacteria produces---and zearalenone (zearalenone, ZEA).Find that from various places feed factory sampling Detection the ZEA recall rate reaches 100% in corn, protein feeds and the mixed feed, mean concentration is 104.99 μ g/kg, and it is serious to exceed standard.
At present, the detoxification technology of mycotoxin is mainly taked physics detoxicity method (heating, ultraviolet ray irradiation, organic clay, active carbon, mannan-oligosaccharides etc.) and chemical detoxication method (soda acid is handled, nitrogen treatment, organic solvent etc.).
Chinese invention patent CN99101577.0 discloses a kind of fermentation method for removing toxic substance from rapeseed cake, the same leavening of rape cake (starch based byproduct or pseudo-ginseng chaff or corn flour or wheat bran etc.) is stirred by a certain percentage, at normal temperature more than 20 ℃, place fermentation vat (leakage-proof and seepage-proof) Nei Jiashui, the water surface overflows about 5 centimetres of charge levels, the fermentation detoxification, thus high-quality feed obtained.
The physics detoxicity method is mainly removed mycotoxin by adsorbents adsorb, and the detoxification amount is limited, and the adsorbent major part that adds can not be digested and assimilated by animal, has the adverse effect of nutriment in the adsorption feed.Mycotoxin is removed in effect by chemical reagent, be applicable to the go mouldy processing of feed of seed class bulky grains such as corn, but time-consuming taking a lot of work, the detoxification of feed is not suitable for going mouldy in a large number, and nutritional labeling caused great destruction, cause handling the uncertainty of back, and bring environmental pollution health hazard.
Trichosporon bacteria (having a liking for) mycotoxin (Trichosporon mycotoxinivorans) that Austria Biomin GmbH company screens from a kind of candida microorganisms is by biology conversion effet degradable ochracin and ZEA.
Many microorganisms have antagonism to fungi, and for example the bacillus subtilis of food-grade and lactic acid bacteria can produce organic acid or produce some protein analogues such as cyclic peptide, chitinase etc. and suppress conk by metabolism.The interpolation bacillus subtilis can be fermented and be produced amylase and protease in the feed, helps directly to decompose in animal intestinal, digest feed nutrient, improves the alimentary canal chyme.Can give the organoleptic properties of product fine and add lactic acid bacteria, promote the security of fermented feed.
Summary of the invention
The objective of the invention is to overcome the shortcoming of prior art, a kind of method that ZEA toxin in the corn alcohol residue reaches the mixed bacterial micro-ecological formulation producing feed from detoxified corn residue after producing alcohol more than 94% of removing is provided.
Technical scheme of the present invention is as follows:
The method of mixed bacterial micro-ecological formulation producing feed from detoxified corn residue after producing alcohol comprises the steps:
(1) probiotics of mixed bacterial: adopt candida utili, bacillus subtilis and Lactobacillus rhamnosus, make a kind of probiotics of mixed bacterial by the composite back of high density fermentation, centrifugal treating and interpolation carrier; Wherein the candida utili viable count is 4.0 * 10
8More than the CFU/g, the bacillus subtilis viable count is 3.8 * 10
8More than the CFU/g, the Lactobacillus rhamnosus viable count is 3.0 * 10
8More than the CFU/g;
(2) preparation of feed: with the probiotics spice of mixed bacterial in the corn residue of producing behind the alcohol, per kilogram is produced the probiotics of the corn residue interpolation 5～10g mixed bacterial behind the alcohol, spraying sterilized water keeps the humidity of compound more than 40%, 30～35 ℃ of reaction 5～10h, more than 45～50 ℃ of oven dry 2h, make the detoxification of corn alcohol residue become safe feed.
Described high density fermentation comprises the steps:
A, with dull and stereotyped activation yeast, bacillus subtilis and Lactobacillus rhamnosus 1～3 day;
B, picking list colony inoculation are in liquid seed culture medium, and 28～37 ℃, 220～250r/min jolting overnight incubation obtains kind of a liquid;
C, get kind of liquid and insert in the liquid enriched medium by 5～10% inoculum concentration, 28～37 ℃, 250r/min jolting overnight incubation obtains enrichment culture liquid;
D, prepared enrichment culture liquid is inserted fermentation tank according to 5～10% inoculum concentration, candida utili and bacillus subtilis are adopted and continuously ferment, fermentation temperature is controlled at 28～37 ℃, and Lactobacillus rhamnosus is adopted fed-batch fermentation, 30～37 ℃ of fermentation temperature controls; The fermentation medium initial glucose concentration is 30～60g/L, prepares glucose solution in advance and is used for feed supplement; When feed supplement finishes, guarantee that the sweat total sugar concentration is 40～120g/L.
The rotating speed of described centrifugal treating is 3000～5000r/min.
Described carrier is zeolite powder and CaCO
3Composite carrier, the percentage by weight of zeolite powder are 25～75%, CaCO
3Percentage by weight be 25～75%, adopt drum mixer to mix.
The present invention adopts candida utili, bacillus subtilis and the Lactobacillus rhamnosus main component as mixed bacterial, by high density fermentation barm cell concentration is reached more than the 30g/L, when the thalline addition reaches 2.0 * 10
7During CFU/mL, 1mg/L ZEA fully can degrade in 1h.Candida utili, bacillus subtilis and Lactobacillus rhamnosus can be bought from Chinese Research for Industrial Microbial Germ preservation center among the present invention, cultivate by high density fermentation, can be made into mixed bacterial micro-ecological formulation, it is handled the corn alcohol residue, but efficient degradation zearalenone toxin not only, and add bacillus subtilis and help in animal intestinal, directly to decompose, digest feed nutrient, improve the alimentary canal chyme; Add lactic acid bacteria and can give feed good organoleptic properties, strengthen the animal intestinal resistance, prevention and treatment diarrhoea are regulated microecological balance, promote the security of fermented feed.
The present invention has following beneficial effect:
Adopt this mixed bacterial micro-ecological formulation and corn alcohol residue or the direct spice of other poisonous feeds, can remove the ZEA toxin and reach more than 94%.Simultaneously, owing to added lactobacillus and bacillus subtilis with prebiotic effect, can promote the digestion of animal to feed, improve and strengthen the animal intestinal resistance, prevention and treatment diarrhoea, regulate microecological balance, make corn alcohol residue or other poisonous feeds become safe feed, and realized the higher value application of industrial residu.
Description of drawings
Fig. 1 is the mixed bacterial micro-ecological formulation degraded ZEA efficiency curve diagram of embodiment 2.
The specific embodiment
For better understanding the present invention, below in conjunction with embodiment the present invention is done detailed description further, but the scope of protection of present invention is not limited to the scope that embodiment represents.
1. the candida utili Candida utilis CLY01 with preservation (purchases in Guangdong Microbes Inst DSMZ, GIM2.9), bacillus subtilis Bacillus subtilis H008 (purchases in Guangdong Microbes Inst DSMZ, GIM1.135) and Lactobacillus rhamnosus Lactobacillus casei subsp.rhamnosus6013 (purchase in Chinese industrial microorganism fungus kind preservation center, 6013) use brewer's wort solid agar medium (purchase encircle triumphant microorganism Co., Ltd) respectively in Guangzhou, LB solid medium (tryptone 10g/L, yeast extract 5g/L, NaCl 10g/L, agar 20g/L, pH7.0) and lactobacillus solid medium (glucose 20g/L, peptone 10g/L, beef extract 10g/L, dusty yeast 5g/L, sodium acetate 2g/L, ammonium citrate 2g/L, Tween 80 1mL/L, MgSO
47H
2O0.2g/L, K
2HPO
42g/L, MnSO
4H
2O0.05g/L, pH6.5) activation culture 1 day separately.
2. (seed culture medium is identical with the activation medium composition for the 10mL seed culture medium of picking list colony inoculation in test tube, do not contain agar) in, candida utili CLY01 is 28 ℃ of cultivations, bacillus subtilis H008 and Lactobacillus rhamnosus 6013 are 37 ℃ of cultivations, 220r/min jolting overnight incubation obtains seed culture fluid.
3. get kind of liquid 5mL and insert in the 100mL liquid enriched medium (enriched medium is identical with the activation medium composition, does not contain agar), 28～37 ℃, 220r/min jolting overnight incubation obtains enrichment culture liquid.
4. enrichment culture liquid is inserted fermentation tank according to about 5% inoculum concentration, candida utili CLY01 and bacillus subtilis H008 adopt and continuously ferment, fermentation temperature is controlled at 28 ℃ and 37 ℃ respectively, and Lactobacillus rhamnosus 6013 is adopted fed-batch fermentation, 37 ℃ of fermentation temperature controls.The fermentation medium initial glucose concentration is 30g/L, prepares the 500g/L glucose solution in advance and is used for feed supplement.When feed supplement finished, detecting the sweat total sugar concentration was 118.6g/L.
5. detect barm cell concentration and degraded ZEA efficient behind high density fermentation, the dry ferment amount reaches more than the 30g in every liter of zymotic fluid, and adds 2.0 * 10
7The CFU/mL thalline stops fermentation in the time of can degrading 1mg/L ZEA fully in 1h; Bacillus subtilis reaches 8.08 * 10 at cell concentration
9Stop fermentation when CFU/mL is above; Lactobacillus rhamnosus reaches 6.50 * 10 at cell concentration
8Stop fermentation when CFU/mL is above, receive bacterium.
6. candida utili CLY01, bacillus subtilis H008 and Lactobacillus rhamnosus 6,013 three fermented liquids are mixed, 4000r/min centrifugal treating 5min, the viable bacteria yield in the final bacterium mud is 84.6%, wherein the content of candida utili about 2.43 * 10
9CFU/g, bacillus subtilis is about 2.29 * 10
9CFU/g, Lactobacillus rhamnosus is about 1.84 * 10
9CFU/g.
7. with carrier (zeolite powder and CaCO
3The mass percent example be 2:1) composite after, mix with the ratio of 2:1 with wet thallus, product is through 55 ℃ of dry 2h, water content is 1.96%, obtains mixed bacterial micro-ecological formulation, wherein the viable count of three bacterium detects as follows: candida utili is about 4.05 * 10
8CFU/g, bacillus subtilis is about 3.82 * 10
8CFU/g, Lactobacillus rhamnosus is about 3.05 * 10
8CFU/g.
8. get corn alcohol residue sample 1kg and pulverize, sampling is soaked 2h with the 10mL sterilized water, and the centrifuging and taking supernatant detects ZEA content with ELISA, and the ZEA content that calculates corn residue sample is 83.96 μ g/kg.
9. get the corn alcohol residue spice of mixed bacterial micro-ecological formulation 5g and pulverizing and mix, spraying sterilized water keeps the humidity of mixing residue 40%, stir with mixer 30r/min average rate, 30 ℃ of reaction 5h, 45 ℃ of oven dry 2h, it is 2.96 μ g/kg that sampling detects its residual ZEA content, and virus elimination rate is 96.5%, makes the corn alcohol residue become safe animal feed.
Detect in fluid sample, the degrade efficient of ZEA of mixed bacterial micro-ecological formulation.Get standard items ZEA and be mixed with the solution 10mL that content is 5.23 μ g/mL, get the made mixed bacterial micro-ecological formulation of 0.5g embodiment 1 step 7 and add and make that saccharomycete concentration is about 2.0 * 10 in the reaction system
7CFU/mL, get simultaneously do not add probiotics ZEA solution in contrast.Reaction condition: 30 ℃, 200r/min, vibration 5h, respectively at 0.5h, 1h, 1.5h, 3h, 5h sampling, centrifugal 20min under the 5000r/min condition, get supernatant and detect its ZEA content with ELISA, experimental group that different time is surveyed and the mapping of the ZEA content of control group, as shown in Figure 1, can clearly see when adding mixed bacterial micro-ecological formulation reaction 0～1h, most ZEA is by efficient degradation in the system, and during reaction 1～5h, the ZEA degradation rate slows down.Behind reaction 1h, experimental group ZEA content is 0.26 μ g/mL, and virus elimination rate is 95.1%.
Getting corn alcohol residue sample 1kg pulverizes, sampling is soaked 2h with the 10mL sterilized water, the centrifuging and taking supernatant, detect ZEA content with ELISA, the ZEA content that calculates the mixed feed sample is about 107.15 μ g/kg, the mixed bacterial micro-ecological formulation spice that the feed sample pulverized and 10g embodiment 1 step 7 is made also mixes, spraying sterilized water keeps the humidity of mixture more than 40%, stir with mixer 30r/min average rate, 30 ℃ of reaction 10h, it is 6.43 μ g/kg that 50 ℃ of oven dry, sample thief detect its residual ZEA content, and virus elimination rate is 94.0%.
The protein feeds sample 1kg that gets mould contamination pulverizes, sampling is soaked 2h with the 10mL sterilized water, the centrifuging and taking supernatant, detect ZEA content with ELISA, the ZEA content that calculates the protein feeds sample is about 110.0 μ g/kg, the mixed bacterial micro-ecological formulation spice that the feed sample pulverized and 10g embodiment 1 step 7 is made also mixes, spraying sterilized water keeps the humidity of mixture more than 40%, stir with mixer 30r/min average rate, 30 ℃ of reaction 7h, it is 5.67 μ g/kg that 50 ℃ of oven dry, sample thief detect its residual ZEA content, and virus elimination rate is 94.8%.
1. the candida utili Candida utilis CLY01 with preservation (purchases in Guangdong Microbes Inst DSMZ, GIM2.9), bacillus subtilis Bacillus subtilis H008 (purchases in Guangdong Microbes Inst DSMZ, GIM1.135) and Lactobacillus rhamnosus Lactobacillus casei subsp.rhamnosus6013 (purchase in Chinese industrial microorganism fungus kind preservation center, 6013) use brewer's wort solid agar medium (purchase encircle triumphant microorganism Co., Ltd) respectively in Guangzhou, LB solid medium (tryptone 10g/L, yeast extract 5g/L, NaCl10g/L, agar 20g/L, pH7.0) and lactobacillus solid medium (glucose 20g/L, peptone 10g/L, beef extract 10g/L, dusty yeast 5g/L, sodium acetate 2g/L, ammonium citrate 2g/L, Tween 80 1mL/L, MgSO
47H
2O0.2g/L, K
2HPO
42g/L, MnSO
4H
2O0.05g/L, pH6.5) activation culture 2 days separately.
2. (seed culture medium is identical with the activation medium composition for the 10mL seed culture medium of picking list colony inoculation in test tube, do not contain agar) in, candida utili CLY01 is 28 ℃ of cultivations, bacillus subtilis H008 and Lactobacillus rhamnosus 6013 are 35 ℃ of cultivations, 230r/min jolting overnight incubation obtains seed culture fluid.
3. get kind of liquid 7mL and insert in the 100mL liquid enriched medium (enriched medium is identical with the activation medium composition, does not contain agar), 28～35 ℃, 230r/min jolting overnight incubation obtains enrichment culture liquid.
4. enrichment culture liquid is inserted fermentation tank according to about 7% inoculum concentration, candida utili CLY01 and bacillus subtilis H008 adopt and continuously ferment, fermentation temperature is controlled at 28 ℃ and 35 ℃ respectively, and Lactobacillus rhamnosus 6013 is adopted fed-batch fermentation, 35 ℃ of fermentation temperature controls.The fermentation medium initial glucose concentration is 30g/L, prepares the 500g/L glucose solution in advance and is used for feed supplement.When feed supplement finished, detecting the sweat total sugar concentration was 111.4g/L.
5. detect barm cell concentration and degraded ZEA efficient behind high density fermentation, the dry ferment amount reaches more than the 30g in every liter of zymotic fluid, and adds 2.0 * 10
7The CFU/mL thalline stops fermentation in the time of can degrading 1mg/L ZEA fully in 1h; Bacillus subtilis reaches 8.08 * 10 at cell concentration
9Stop fermentation when CFU/mL is above; Lactobacillus rhamnosus reaches 6.50 * 10 at cell concentration
8Stop fermentation when CFU/mL is above, receive bacterium.
6. candida utili CLY01, bacillus subtilis H008 and Lactobacillus rhamnosus 6,013 three fermented liquids are mixed, 3000r/min centrifugal treating 5min, the viable bacteria yield in the final bacterium mud is 89.3%, wherein the content of candida utili about 2.47 * 10
9CFU/g, bacillus subtilis is about 2.29 * 10
9CFU/g, Lactobacillus rhamnosus is about 1.97 * 10
9CFU/g.
7. with carrier (zeolite powder and CaCO
3The mass percent example be 2:1) composite after, mix with the ratio of 2:1 with wet thallus, product is through 55 ℃ of dry 2h, water content is 1.92%, obtains mixed bacterial micro-ecological formulation, wherein the viable count of three bacterium detects as follows: candida utili is about 4.12 * 10
8CFU/g, bacillus subtilis is about 3.81 * 10
8CFU/g, Lactobacillus rhamnosus is about 3.28 * 10
8CFU/g.
8. get corn alcohol residue sample 1kg and pulverize, sampling is soaked 2h with the 10mL sterilized water, and the centrifuging and taking supernatant detects ZEA content with ELISA, and the ZEA content that calculates corn residue sample is 101.53 μ g/kg.
9. get the corn alcohol residue spice of mixed bacterial micro-ecological formulation 7g and pulverizing and mix, spraying sterilized water keeps the humidity of mixing residue 40%, stir with mixer 30r/min average rate, 35 ℃ of reaction 7h, 48 ℃ of oven dry 2h, it is 4.55 μ g/kg that sampling detects its residual ZEA content, and virus elimination rate is 95.52%, makes the corn alcohol residue become safe animal feed.
Embodiment 6
1. the candida utili Candida utilis CLY01 with preservation (purchases in Guangdong Microbes Inst DSMZ, GIM2.9), bacillus subtilis Bacillus subtilis H008 (purchases in Guangdong Microbes Inst DSMZ, GIM1.135) and Lactobacillus rhamnosus Lactobacillus casei subsp.rhamnosus6013 (purchase in Chinese industrial microorganism fungus kind preservation center, 6013) use brewer's wort solid agar medium (purchase encircle triumphant microorganism Co., Ltd) respectively in Guangzhou, LB solid medium (tryptone 10g/L, yeast extract 5g/L, NaCl10g/L, agar 20g/L, pH7.0) and lactobacillus solid medium (glucose 20g/L, peptone 10g/L, beef extract 10g/L, dusty yeast 5g/L, sodium acetate 2g/L, ammonium citrate 2g/L, Tween 80 1mL/L, MgSO
47H
2O0.2g/L, K
2HPO
42g/L, MnSO
4H
2O0.05g/L, pH6.5) activation culture 3 days separately.
2. (seed culture medium is identical with the activation medium composition for the 10mL seed culture medium of picking list colony inoculation in test tube, do not contain agar) in, candida utili CLY01 is 30 ℃ of cultivations, bacillus subtilis H008 and Lactobacillus rhamnosus 6013 are 35 ℃ of cultivations, 250r/min jolting overnight incubation obtains seed culture fluid.
3. get kind of liquid 10mL and insert in the 100mL liquid enriched medium (enriched medium is identical with the activation medium composition, does not contain agar), 30～35 ℃, 250r/min jolting overnight incubation obtains enrichment culture liquid.
4. enrichment culture liquid is inserted fermentation tank according to about 10% inoculum concentration, candida utili CLY01 and bacillus subtilis H008 adopt and continuously ferment, fermentation temperature is controlled at 30 ℃ and 35 ℃ respectively, and Lactobacillus rhamnosus 6013 is adopted fed-batch fermentation, 35 ℃ of fermentation temperature controls.The fermentation medium initial glucose concentration is 30g/L, prepares the 500g/L glucose solution in advance and is used for feed supplement.When feed supplement finished, detecting the sweat total sugar concentration was 107.3g/L.
5. detect barm cell concentration and degraded ZEA efficient behind high density fermentation, the dry ferment amount reaches more than the 30g in every liter of zymotic fluid, and adds 2.0 * 10
7The CFU/mL thalline stops fermentation in the time of can degrading 1mg/L ZEA fully in 1h; Bacillus subtilis reaches 8.08 * 10 at cell concentration
9Stop fermentation when CFU/mL is above; Lactobacillus rhamnosus reaches 6.50 * 10 at cell concentration
8Stop fermentation when CFU/mL is above, receive bacterium.
6. candida utili CLY01, bacillus subtilis H008 and Lactobacillus rhamnosus 6,013 three fermented liquids are mixed, 5000r/min centrifugal treating 5min, the viable bacteria yield in the final bacterium mud is 86.8%, wherein the content of candida utili about 2.53 * 10
9CFU/g, bacillus subtilis is about 2.32 * 10
9CFU/g, Lactobacillus rhamnosus is about 2.04 * 10
9CFU/g.
7. with carrier (zeolite powder and CaCO
3The mass percent example be 2:1) composite after, mix with the ratio of 2:1 with wet thallus, product is through 55 ℃ of dry 2h, water content is 1.91%, obtains mixed bacterial micro-ecological formulation, wherein the viable count of three bacterium detects as follows: candida utili is about 4.22 * 10
8CFU/g, bacillus subtilis is about 3.87 * 10
8CFU/g, Lactobacillus rhamnosus is about 3.40 * 10
8CFU/g.
8. get corn alcohol residue sample 1kg and pulverize, sampling is soaked 2h with the 10mL sterilized water, and the centrifuging and taking supernatant detects ZEA content with ELISA, and the ZEA content that calculates corn residue sample is 104.28 μ g/kg.
9. get the corn alcohol residue spice of mixed bacterial micro-ecological formulation 10g and pulverizing and mix, spraying sterilized water keeps the humidity of mixing residue 40%, stir with mixer 30r/min average rate, 35 ℃ of reaction 10h, 50 ℃ of oven dry 2h, it is 3.07 μ g/kg that sampling detects its residual ZEA content, and virus elimination rate is 97.01%, makes the corn alcohol residue become safe animal feed.
Claims (4)
1. the method for mixed bacterial micro-ecological formulation producing feed from detoxified corn residue after producing alcohol is characterized in that comprising the steps:
(1) probiotics of mixed bacterial: adopt candida utili, bacillus subtilis and Lactobacillus rhamnosus, make a kind of probiotics of mixed bacterial by the composite back of high density fermentation, centrifugal treating and interpolation carrier; Wherein the candida utili viable count is 4.0 * 10
8More than the CFU/g, the bacillus subtilis viable count is 3.8 * 10
8More than the CFU/g, the Lactobacillus rhamnosus viable count is 3.0 * 10
8More than the CFU/g;
(2) preparation of feed: with the probiotics spice of mixed bacterial in the corn residue of producing behind the alcohol, per kilogram is produced the probiotics of the corn residue interpolation 5～10g mixed bacterial behind the alcohol, spraying sterilized water keeps the humidity of compound more than 40%, 30～35 ℃ of reaction 5～10h, more than 45～50 ℃ of oven dry 2h, make the detoxification of corn alcohol residue become safe feed.
2. the method for mixed bacterial micro-ecological formulation producing feed from detoxified corn residue after producing alcohol according to claim 1 is characterized in that: described high density fermentation comprises the steps:
A, with dull and stereotyped activation candida utili, bacillus subtilis and Lactobacillus rhamnosus 1～3 day;
B, picking list colony inoculation are in liquid seed culture medium, and 28～37 ℃, 220～250r/min jolting overnight incubation obtains kind of a liquid;
C, get kind of liquid and insert in the liquid enriched medium by 5～10% inoculum concentration, 28～37 ℃, 250r/min jolting overnight incubation obtains enrichment culture liquid;
D, prepared enrichment culture liquid is inserted fermentation tank according to 5～10% inoculum concentration, candida utili and bacillus subtilis are adopted and continuously ferment, fermentation temperature is controlled at 28～37 ℃, and Lactobacillus rhamnosus is adopted fed-batch fermentation, 30～37 ℃ of fermentation temperature controls; The fermentation medium initial glucose concentration is 30～60gL, prepares glucose solution in advance and is used for feed supplement; When feed supplement finishes, guarantee that the sweat total sugar concentration is 40～120g/L.
3. the method for mixed bacterial micro-ecological formulation producing feed from detoxified corn residue after producing alcohol according to claim 1 is characterized in that: the rotating speed of described centrifugal treating is 3000～5000r/min.
4. the method for mixed bacterial micro-ecological formulation producing feed from detoxified corn residue after producing alcohol according to claim 1 is characterized in that: described carrier is zeolite powder and CaCO
3Composite carrier, the percentage by weight of zeolite powder are 25～75%, CaCO
3Percentage by weight be 25～75%.
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| CN1242162A (en) * | 1999-03-19 | 2000-01-26 | 徐太树 | Fermentation method for removing toxic substance from rapeseed cake |
| CN1843151A (en) * | 2006-03-16 | 2006-10-11 | 上海交通大学 | Composite mold toxin sorbent |
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