CN101273749B - Method of mixed fermentation treatment using vinasse as main feed raw material - Google Patents
Method of mixed fermentation treatment using vinasse as main feed raw material Download PDFInfo
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- CN101273749B CN101273749B CN2008100442162A CN200810044216A CN101273749B CN 101273749 B CN101273749 B CN 101273749B CN 2008100442162 A CN2008100442162 A CN 2008100442162A CN 200810044216 A CN200810044216 A CN 200810044216A CN 101273749 B CN101273749 B CN 101273749B
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
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Abstract
The invention discloses a method for processing feedstuff such as distilled grains, etc. by mixed fermentation, and the strain is brevibacterium lactofermentus and candida utilis; the technique is that: strain liquid cultured by a third class seed is mixed and then inoculated in a mixed fermentation medium; the mixed seed liquid is inoculated in the mixed fermentation medium with 50 to 60 percent of humidity according to 1:10 of inoculum concentration; the obtained mixture is fermented by sectional fermentation at the temperature of 30 to 37 DEG C, and the obtained leavening is dried at the temperature of 40 to 60 DEG C, and packed so as to obtain a finished product; the invention adopts the sectional fermentation mode which is aerobic firstly and then is anaerobic, thus eliminating rape seed cakes or thioglucoside and gossypol in cottonseed crumb, increasing the total content of protein in the distilled grains, degrading the alcohol substances and decomposing non-starch polysaccharide substances such as cellulose, etc.; the content of the cellulose of the obtained feedstuff finished product is reduced from 25 to 11 percent, and the protein content is increased from 15 percent to 30 to 40 percent; the feeding effect of animals is good.
Description
Technical field
The present invention relates to a kind of microbial fermentation and handle the method for mixed fodder raw materials such as vinasse, particularly, the present invention relates to a kind of method that adopts lactic acid bacteria and saccharomycete mixed culture fermentation to handle vinasse and albumen raw mixs such as dregs of beans, dish cake, cotton dregs or other assorted dregs of rice.
Background technology
Not only contain abundant thick protein (about 20%) in the distillers ' grains, also contain inorganic elements such as abundant crude fibre (about 25%), crude fat (about 10%), B family vitamin and phosphorus, potassium, this shows that the nutritive value of distillers ' grains and value of exploiting and utilizing are all high.In addition, also may have the micro-beneficiating ingredients such as amino acid, ribonucleic acid and purine that produce by microbial cells in the distillers ' grains, all these be cereal can not compare.Therefore, continuous always to the research of vinasse both at home and abroad, the research report of various relevant vinasse emerges in an endless stream.
China's research has in this regard at present obtained certain achievement, for example, utilize distillers ' grains to cultivate edible mushroom, extract compound amino acid and trace element, extract phytic acid and phytic acid calcium, extract protein, produce amylase and cellulase, utilize the vinasse anaerobic fermentation to reclaim biogas, produce forage protein, feed is produced in ensiling, produces GABA etc., but wherein except being used as feed, agricultural fertilizer, additive method all can not fundamentally solve the final whereabouts problem of distillers ' grains for this.
As the byproduct of liquor, distillers ' grains contains very abundant nutrition composition, helps growth of animals or poultry, can be directly as feed or feed addictive.For example, with solid state white vinasse separation rice husk drying or after drying, cooperate the feed that gets final product the production different cultivars again with other raw materials.Though the nutritional labeling of grains such as vinasse and wheat, corn, Chinese sorghum has certain similitude, but ANFs content such as crude fibre wherein and phytic acid are all higher, unless method is handled through shelling etc., otherwise be difficult to be applied in the feed, therefore seek more rational processing method, just can make vinasse more give full play to wherein inherent nutritive value.
Though it is commonplace to use physico-chemical method to handle distillers ' grains manufacturing extraction chemicals in the past, but this processing method can not obviously be improved the distillers ' grains nutritional labeling and be used for feed industry, and the vinasse waste liquid of handling with physico-chemical method still contains a large amount of organic matters, the COD value is also about 1500mg/L, if directly discharging will have a strong impact on the environment generation.Comparatively speaking, adopt the method for microbial fermentation that very big advantage is just arranged, both can prevent to pollute, can farthest utilize resource again, working condition is also fairly simple.It is reported that the at present domestic microorganism that is mainly used in the production mycoprotein has Aspergillus, rhizopus, candidiasis, Bacillus acidi lactici, streptococcus lactis, hay bacillus, lysine to produce bacterium, intends endomyces, geotrichum candidum etc.The bacterial classification that selects at present has:
(1) saccharomycete: the acidproof ability of yeast is strong, and is individual big, easily reclaims, and is suitable for vinasse fermentation, and saccharomycete commonly used has candida tropicalis bacterium, candida utili bacterium, skin shape trichosporon cutaneum bacterium, tree-shaped yeast.
(2) actinomyces: actinomyces have very strong cellulolytic ability, can directly utilize fibrous raw material manufacture order cell protein, and actinomyces commonly used have Nocard's bacillus, thermophilic actinomycete.
(3) mould: mould can produce highly active cellulase, the cellulose in the degraded vinasse, and mould commonly used has geotrichum candidum, aspergillus niger, mould, the Trichoderma viride of healthy and free from worry wood.
(4) basidiomycete etc.Be mainly used in and produce edible fungus etc.
Studies show that the effect of mixed fermentation is more obvious, adopt many bacterial classifications to mix the revolution fermentation, floor space is little, and fermentation 36～48h can be increased to the crude protein content of vinasse more than 30%, and amino acid whose content is more than 22%, and crude fibre has reduced about 2%.At fermentation stage, the bacterial strain metabolic enzyme has destroyed the tight structure between cellulose and the lignin, is easier to the animal digestion absorption, thereby has improved the biological value of vinasse protein feeds greatly.
Existing at present research with chicken manure and some additives, mixes the report of making feed as crude fibres such as maize straws with vinasse, has all obtained the effect of increasing production that increases weight preferably by the pig of feeding, ox, sheep, duck etc.With vinasse is that base stock adds rumen content or fermented by mixed bacterium, can obtain animal feeding-stuff containing somatic protein, like this, not only can turn waste into wealth, reduce and pollute, but also the vinasse that add as roughage originally can be become elaboration, be high nutrient content additive, feeding effect is relatively success also.
Summary of the invention
The object of the present invention is to provide a kind of method that adopts lactic acid bacteria and saccharomycete mixed culture fermentation to handle vinasse and albumen raw mixs such as dregs of beans, dish cake, cotton dregs or other assorted dregs of rice.Through the feed that this method obtains, its protein content height does not contain alcohols and other noxious material, thereby the animal good palatability, safe in utilization, feeding effect is good, not only make the nutritive value of vinasse inherence obtain giving full play to, but also solved vinasse pollution on the environment problem.
The present invention realizes the object of the invention by implementing following technical scheme:
It is the method for main feedstuff that mixed culture fermentation is handled with vinasse, it is characterized in that: employed bacterial classification is fermentation lactic acid brevibacterium and candida utili, processing step is: will be inoculated in the seed mixture culture medium after the fermentation lactic acid brevibacterium of three grades of seed culture and candida utili mix, obtain the seed mixture liquid of fermentation lactic acid brevibacterium and candida utili, again this seed mixture liquid being inoculated in humidity by 1: 10 inoculum concentration is in 50～60% the mixed fermentive culture medium, postvaccinal mixture behind stepwise fermentation, places the fermentate of gained again under 40～60 ℃ the temperature and dries under 30～37 ℃ temperature, obtain finished product after the packing.
The deposit number of described fermentation lactic acid brevibacterium (Brevibacterium lactofermentum) is CICC 10227, and the deposit number of candida utili (Candida tails) is CICC 1314, and bacterial classification is stored on the medium slant respectively.
Three grades of seed culture steps of described fermentation lactic acid brevibacterium (Brevibacterium lactofermentum) are:
A, 2～3 oese thalline of picking are inoculated in the 5ml lactic acid bacteria liquid culture medium, and 220rpm cultivated 16 hours for 37 ℃, got the first order seed nutrient solution;
B, the first order seed nutrient solution of 5mL incubated overnight is inoculated in the 100mL lactic acid bacteria liquid culture medium, 220rpm cultivated 16 hours, and was cultured to cell concentration＞10 for 37 ℃
9CFU/mL gets the secondary seed nutrient solution;
C, 100mL secondary seed nutrient solution is inoculated in the 10L lactic acid bacteria liquid culture medium, 220rpm cultivated 16 hours, and was cultured to cell concentration＞10 for 37 ℃
9CFU/mL, three grades of seed culture fluids of fermentation lactic acid brevibacterium.
Described lactic acid bacteria liquid culture medium is formulated as follows: yeast extract 7.5g, peptone 7.5g, glucose 10g, KH
2PO
42g, the 100mL of Tomato juice, Tween-80 0.5mL, distilled water 900mL, 7.0,15 pounds of sterilizations of pH 30 minutes.
Three grades of seed culture steps of described candida utili (Candida tails) are:
A, 2～3 oese thalline of picking are inoculated in the 5ml malt juice liquid medium, and 220rpm cultivated 16 hours for 28 ℃, got the first order seed nutrient solution;
B, the first order seed nutrient solution of 5mL incubated overnight is inoculated in the 100mL malt juice liquid medium, 220rpm cultivated 16 hours, and was cultured to cell concentration＞10 for 28 ℃
9CFU/mL gets the secondary seed nutrient solution;
C, 100mL secondary seed nutrient solution is inoculated in the 10L malt juice liquid medium, 220rpm cultivated 16 hours, and was cultured to cell concentration＞10 for 28 ℃
9CFU/mL, three grades of seed culture fluids of candida utili.
Described malt extract medium is formulated as follows: the raw material (not hopping) with fermentation beer, be diluted to 12 Berlin, and sterilized 30 minutes for 15 pounds.
Described lactic acid bacteria or malt juice liquid medium can obtain solid medium by the agar powder that adds gross weight 1.5%.
The culture process of the seed mixture liquid of described fermentation lactic acid brevibacterium and candida utili is:
With the fermentation lactic acid brevibacterium with after candida utili mixes according to 1: 0.01～1 ratio, be inoculated in the seed culture medium 28～30 ℃ of cultivation temperature, speed of agitator 220～300r/min by 1: 10 inoculum concentration again, cultivated 48 hours, and be cultured to cell concentration＞10
9CFU/mL.
The optimum mixture ratio example of described fermentation lactic acid brevibacterium and candida utili is 1: 0.1.
Described seed culture medium is formulated as: cornstarch 5%, urea: CaHPO 1%,
40.5%, natural pH value was sterilized 30 minutes for 15 pounds.
The manufacture craft of described mixed fermentive culture medium is: with fresh vinasse in dry or squeeze under 45～55 ℃ be 25～35% to water content after, by weight percentage, be 80～90% according to spirit stillage with the albumen raw material earlier: 10～20% mix, mix according to 97.3～98.9%% mixture and 1～2.5% urea again, add 0.1%CaCO
3, selectivity adds 0.5% CaHPO again
4, 0.1% feeding mineral additive or 0.1% feeding vitamin preparation fermentation medium, natural pH value, 105 ℃ of steam batch (-type)s sterilizations 1 hour, 2～3 times, each 3～4 hours intermittences.
The humidity of described mixed fermentive culture medium is by adding entry or an amount of wheat bran is controlled.
Described albumen raw material is selected from dregs of beans, cotton dregs, rape cake or the coconut dregs of rice.
The raw material composition and the weight proportion of the described feeding mineral additive of preparation 1000Kg are as follows:
CuSO
4·5H
2O 24Kg
FeSO
4·7H
2O 250Kg
ZnSO
4·H
2O 200Kg
MnSO
4 200Kg
Mass percent concentration is 5% KI 5Kg
Mass percent concentration is 1% Na
2SeO
318Kg
Carrier: zeolite powder 303Kg
Compound method is: elder generation joins 70% zeolite powder 212.1Kg in the blender, adds CuSO by above-mentioned weight proportion then
4.5H
2O, FeSO
4.7H
2O, ZnSO
4.H
2O, MnSO
4, 5% KI and 1% Na
2SeO
3, the last 87.9Kg zeolite powder that adds remainder again mixes and promptly obtains the feeding mineral additive of 1000Kg.
Described feeding vitamin is selected from " the general multivitamin of pig " produced by Beijing company of monarch's moral Tontru.
Described stepwise fermentation is meant: postvaccinal mixture is sub-packed in the closed bag, carries out fermentation process then in two stages, first aerobic fermentation 12 hours, 30～32 ℃ of fermentation temperatures; Back anaerobic fermentation 48～72 hours, 35～37 ℃ of fermentation temperatures, when mixture pH drops to 3.5～4.0 stable state, and the thalline number reaches 10
7～10
8During CFU/g, stop fermentation.
The invention has the advantages that:
1, the present invention adopts fermentation lactic acid brevibacterium and two kinds of microorganism mixed culture fermentations of candida utili to handle vinasse and albumen raw mixs such as dregs of beans, dish cake, cotton dregs or other assorted dregs of rice, thereby obtained protein content up to 30～40%, no alcohols material, and good palatability, no noxious material, the feedstuff that safety in utilization is good;
2, the present invention is in the sweat to feedstuff, adopt stepwise fermentation mode aerobic earlier, the back anaerobism, thereby sloughed the glucosinolate in the albumen raw materials such as dish cake effectively, in the final finished feed that obtains, the content of glucosinolate is lower than 0.01%, meets the feed safety requirement: promptly isothiocyanic acid content less than 0.045%, oxazolidine thioketones content less than 0.025%;
3, the present invention passes through acting in conjunction aerobic earlier, the back anaerobic fermentation, thereby sloughed the gossypol in the albumen raw materials such as cotton dregs effectively, in the final finished feed that obtains, the content of gossypol is lower than 0.01%, meets the feed safety requirement: promptly the content of gossypol is less than 0.02%;
4, two kinds of microorganisms of fermentation lactic acid brevibacterium and candida utili are aerobic earlier by adopting, back anaerobism co-fermentation vinasse in the present invention, thereby increased the total protein content in the vinasse, the alcohols material of having degraded, SNSP class materials such as cellulose have been decomposed, make the content of cellulose of the finished feed of final acquisition drop to 11% by original 25%, protein content is increased to 30～40% by original 15%.
The specific embodiment
Embodiment 1
It is the method for main feedstuff that mixed culture fermentation is handled with vinasse, one of employed bacterial classification is fermentation lactic acid brevibacterium (Brevibacterium lactofermentum), its deposit number is CICC 10227, two of bacterial classification is candida utili (Candida tails), its deposit number is CICC 1314, and bacterial classification is stored on the medium slant respectively.
Described fermentation lactic acid brevibacterium and candida utili are respectively through three grades of seed culture, be inoculated in after the mixing in the seed mixture culture medium, obtain the seed mixture liquid of fermentation lactic acid brevibacterium and candida utili, again this seed mixture liquid being inoculated in humidity by 1: 10 inoculum concentration is in 50% the mixed fermentive culture medium, postvaccinal mixture behind stepwise fermentation, obtains finished product after the fermentate of gained being placed again oven dry under 50 ℃ the temperature, packing under 30～37 ℃ temperature.
Three grades of seed culture steps of described fermentation lactic acid brevibacterium (Brevibacterium lactofermentum) are:
A, 2～3 oese thalline of picking are inoculated in the 5ml lactic acid bacteria liquid culture medium, and 220rpm cultivated 16 hours for 37 ℃, got the first order seed nutrient solution;
B, the first order seed nutrient solution of 5mL incubated overnight is inoculated in the 100mL lactic acid bacteria liquid culture medium, 220rpm cultivated 16 hours, and was cultured to cell concentration＞10 for 37 ℃
9CFU/mL gets the secondary seed nutrient solution;
C, 100mL secondary seed nutrient solution is inoculated in the 10L lactic acid bacteria liquid culture medium, 220rpm cultivated 16 hours, and was cultured to cell concentration＞10 for 37 ℃
9CFU/mL, three grades of seed culture fluids of fermentation lactic acid brevibacterium.
Described lactic acid bacteria liquid culture medium is formulated as follows: yeast extract 7.5g, peptone 7.5g, glucose 10g, KH
2PO
42g, the 100mL of Tomato juice, Tween-80 0.5mL, distilled water 900mL, 7.0,15 pounds of sterilizations of pH 30 minutes.
Three grades of seed culture steps of described candida utili (Candida tails) are:
A, 2～3 oese thalline of picking are inoculated in the 5ml malt juice liquid medium, and 220rpm cultivated 16 hours for 28 ℃, got the first order seed nutrient solution;
B, the first order seed nutrient solution of 5mL incubated overnight is inoculated in the 100mL malt juice liquid medium, 220rpm cultivated 16 hours, and was cultured to cell concentration＞10 for 28 ℃
9CFU/mL gets the secondary seed nutrient solution;
C, 100mL secondary seed nutrient solution is inoculated in the 10L malt juice liquid medium, 220rpm cultivated 16 hours, and was cultured to cell concentration＞10 for 28 ℃
9CFU/mL, three grades of seed culture fluids of candida utili.
Described malt extract medium is formulated as follows: the raw material (not hopping) with fermentation beer, be diluted to 12 Berlin, and sterilized 30 minutes for 15 pounds.
The culture process of the seed mixture liquid of described fermentation lactic acid brevibacterium and candida utili is:
The fermentation lactic acid brevibacterium with after candida utili mixes according to 1: 0.5 ratio, is inoculated in the seed culture medium by 1: 10 inoculum concentration again, 30 ℃ of cultivation temperature, speed of agitator 220r/min cultivated 48 hours, was cultured to cell concentration＞10
9CFU/mL.
Described seed culture medium is formulated as: cornstarch 5%, urea: CaHPO 1%,
40.5%, natural pH value was sterilized 30 minutes for 15 pounds.
The manufacture craft of described mixed fermentive culture medium is: with fresh vinasse in dry under 45 ℃ be 30% to water content after, by weight percentage, earlier 80% spirit stillage is mixed with 20% rape cake, after getting 97.4% vinasse rape cake mixture again and 2% urea mixing, add 0.1%CaCO
3, 0.5%CaHPO
4, natural pH value, 105 ℃ of steam batch (-type)s were sterilized 1 hour, and 2 times, intermittently 4 hours.
Described stepwise fermentation is meant: postvaccinal mixture is sub-packed in the closed bag, carries out fermentation process then in two stages, first aerobic fermentation 12 hours, nature fermentation, 30 ℃ of temperature; Anaerobic fermentation 60 hours, 35 ℃ of fermentation temperatures, when mixture pH drops to 4.0 stable state, and the thalline number reaches 10
7～10
8During CFU/g, stop fermentation.
The key technical indexes of this product is as follows:
Content of microorganisms 10
7～10
8CFU/g, pH＜4.0, protein content is more than 30%, and glucosinolate content is less than 0.01%.
Embodiment 2
Fermentation lactic acid brevibacterium and candida utili are carried out three grades of seed culture respectively, three grades of seed liquor are inoculated in the seed mixture culture medium after mixing, obtain the seed mixture liquid of fermentation lactic acid brevibacterium and candida utili, again this seed mixture liquid being inoculated in humidity by 1: 10 inoculum concentration is in 60% the mixed fermentive culture medium, postvaccinal mixture behind stepwise fermentation, obtains finished product after the fermentate of gained being placed again oven dry under 40 ℃ the temperature, packing under 30～37 ℃ temperature.
The culture process of the seed mixture liquid of described fermentation lactic acid brevibacterium and candida utili is:
The fermentation lactic acid brevibacterium with after candida utili mixes according to 1: 1 ratio, is inoculated in the seed culture medium by 1: 10 inoculum concentration again, 28 ℃ of cultivation temperature, speed of agitator 220r/min cultivated 50 hours, was cultured to cell concentration＞10
9CFU/mL.
The manufacture craft of described mixed fermentive culture medium is: with fresh vinasse squeeze be 35% to water content after, by weight percentage, earlier 80% spirit stillage is mixed with 20% cotton dregs, after getting 97.3% vinasse cotton dregs mixture again and 2.5% urea mixing, adding 0.1%CaCO
3, 0.1% feeding mineral additive, natural pH value, 105 ℃ of steam batch (-type)s sterilization 1 hour, 3 times, each intermittently 3～4 hours.
All the other method steps are with embodiment 1.
The key technical indexes of this product is as follows:
Content of microorganisms 10
7～10
8CFU/g, pH＜4.0, protein content is more than 30%.
Embodiment 3
Fermentation lactic acid brevibacterium and candida utili are carried out three grades of seed culture respectively, three grades of seed liquor are inoculated in the seed mixture culture medium after mixing, obtain the seed mixture liquid of fermentation lactic acid brevibacterium and candida utili, again the inoculum concentration of this seed mixture liquid by 1: 10 is inoculated in the mixed fermentive culture medium of humidity 55%, postvaccinal mixture behind stepwise fermentation, obtains finished product after the fermentate of gained being placed again oven dry under 60 ℃ the temperature, packing under 30～37 ℃ temperature.
The culture process of the seed mixture liquid of described fermentation lactic acid brevibacterium and candida utili is:
The fermentation lactic acid brevibacterium with after candida utili mixes according to 1: 0.01 ratio, is inoculated in the seed culture medium by 1: 10 inoculum concentration again, 30 ℃ of cultivation temperature, speed of agitator 300r/min cultivated 72 hours, was cultured to cell concentration＞10
9CFU/mL.
The manufacture craft of described mixed fermentive culture medium is: with fresh vinasse in dry under 55 ℃ be 25% to water content after, by weight percentage, earlier 90% spirit stillage is mixed with 10% dregs of beans, after getting 98.9% vinasse dregs of beans mixture again and 1% urea mixing, adding 0.1%CaCO
3, natural pH value, 105 ℃ of steam batch (-type)s were sterilized 1 hour, and 3 times, each intermittently 3～4 hours.
All the other method steps are with embodiment 1.
The key technical indexes of this product is as follows:
Content of microorganisms 10
7～10
8CFU/g, pH＜4.0, protein content is more than 30%.
Embodiment 4
Fermentation lactic acid brevibacterium and candida utili are carried out three grades of seed culture respectively, three grades of seed liquor are inoculated in the seed mixture culture medium after mixing, obtain the seed mixture liquid of fermentation lactic acid brevibacterium and candida utili, again the inoculum concentration of this seed mixture liquid by 1: 10 is inoculated in the mixed fermentive culture medium of humidity 58%, postvaccinal mixture behind stepwise fermentation, obtains finished product after the fermentate of gained being placed again oven dry under 40～50 ℃ the temperature, packing under 30～37 ℃ temperature.
The culture process of the seed mixture liquid of described fermentation lactic acid brevibacterium and candida utili is:
The fermentation lactic acid brevibacterium with after candida utili mixes according to 1: 0.1 ratio, is inoculated in the seed culture medium by 1: 10 inoculum concentration again, 30 ℃ of cultivation temperature, speed of agitator 220r/min cultivated 55 hours, was cultured to cell concentration＞10
9CFU/mL.
The manufacture craft of described mixed fermentive culture medium is: with fresh vinasse in dry under 45 ℃ be 32% to water content after, by weight percentage, earlier 80% spirit stillage is mixed with 20% the coconut dregs of rice, after getting 97.8% vinasse coconut dregs of rice mixture again and 2% urea mixing, add 0.1%CaCO
3, 0.1% feeding vitamin, natural pH value, 105 ℃ of steam batch (-type)s sterilization 1 hour, 2 times, intermittently 3 hours.
All the other method steps are with embodiment 1.
The key technical indexes of this product is as follows:
Content of microorganisms 10
7～10
8CFU/g, pH＜4.0, protein content is more than 30%.
Claims (7)
1. a mixed culture fermentation processing is the method for main feedstuff with vinasse, it is characterized in that employed bacterial classification is fermentation lactic acid brevibacterium and candida utili, processing step is: will be inoculated in the seed mixture culture medium after the fermentation lactic acid brevibacterium of three grades of seed culture and candida utili mix, obtain the seed mixture liquid of fermentation lactic acid brevibacterium and candida utili, again this seed mixture liquid being inoculated in humidity by 1: 10 inoculum concentration is in 50～60% the mixed fermentive culture medium, postvaccinal mixture behind stepwise fermentation, places the fermentate of gained again under 40～60 ℃ the temperature and dries under 30～37 ℃ temperature, obtain finished product after the packing;
The culture process of described fermentation lactic acid brevibacterium and candida utili seed mixture liquid is:
With the fermentation lactic acid brevibacterium with after candida utili mixes according to 1: 0.01～1 ratio, be inoculated in the seed culture medium 28～30 ℃ of cultivation temperature, speed of agitator 220～300r/min by 1: 10 inoculum concentration again, cultivated 48 hours, and be cultured to cell concentration＞10
9CFU/mL;
Described seed culture medium is formulated as: cornstarch 5%, urea: CaHPO 1%,
40.5%, natural pH value was sterilized 30 minutes for 15 pounds;
The manufacture craft of described mixed fermentive culture medium is:
With fresh vinasse in dry or squeeze under 45～55 ℃ be 25～35% to water content after, by weight percentage, be 80～90% according to spirit stillage with the albumen raw material earlier: 10～20% mix, mix according to 97.3～98.9% mixture and 1～2.5% urea again, add 0.1%CaCO
3, selectivity adds 0.5% CaHPO again
4, 0.1% feeding mineral additive or 0.1% feeding vitamin preparation fermentation medium, natural pH value, 105 ℃ of steam batch (-type)s sterilizations 1 hour, 2～3 times, each 3～4 hours intermittences;
Described stepwise fermentation is meant: postvaccinal mixture is sub-packed in the closed bag, carries out fermentation process then in two stages, first aerobic fermentation 12 hours, 30～32 ℃ of fermentation temperatures; Back anaerobic fermentation 48～72 hours, 35～37 ℃ of fermentation temperatures, when mixture pH drops to 3.5～4.0 stable state, and the thalline number reaches 10
7～10
8During CFU/g, stop fermentation.
2. it is the method for main feedstuff that mixed culture fermentation as claimed in claim 1 is handled with vinasse, it is characterized in that three grades of seed culture steps of described fermentation lactic acid brevibacterium are:
A, 2～3 oese thalline of picking are inoculated in the 5ml lactic acid bacteria liquid culture medium, and 220rpm cultivated 16 hours for 37 ℃, got the first order seed nutrient solution;
B, the first order seed nutrient solution of 5mL incubated overnight is inoculated in the 100mL lactic acid bacteria liquid culture medium, 220rpm cultivated 16 hours, and was cultured to cell concentration＞10 for 37 ℃
9CFU/mL gets the secondary seed nutrient solution;
C, 100mL secondary seed nutrient solution is inoculated in the 10L lactic acid bacteria liquid culture medium, 220rpm cultivated 16 hours, and was cultured to cell concentration＞10 for 37 ℃
9CFU/mL, three grades of seed culture fluids of fermentation lactic acid brevibacterium.
3. it is the method for main feedstuff that mixed culture fermentation as claimed in claim 2 is handled with vinasse, it is characterized in that described lactic acid bacteria liquid culture medium is formulated as follows: yeast extract 7.5g, peptone 7.5g, glucose 10g, KH
2PO
42g, the 100mL of Tomato juice, Tween-80 0.5mL, distilled water 900mL, 7.0,15 pounds of sterilizations of pH 30 minutes.
4. it is the method for main feedstuff that mixed culture fermentation as claimed in claim 1 is handled with vinasse, it is characterized in that three grades of seed culture steps of described candida utili (Candida tails) are:
A, 2～3 oese thalline of picking are inoculated in the 5ml malt juice liquid medium, and 220rpm cultivated 16 hours for 28 ℃, got the first order seed nutrient solution;
B, the first order seed nutrient solution of 5mL incubated overnight is inoculated in the sub-100mL malt juice liquid medium, 220rpm cultivated 16 hours, and was cultured to cell concentration＞10 for 28 ℃
9CFU/mL gets the secondary seed nutrient solution;
C, 100mL secondary seed nutrient solution is inoculated in the 10L malt juice liquid medium, 220rpm cultivated 16 hours, and was cultured to cell concentration＞10 for 28 ℃
9CFU/mL, three grades of seed culture fluids of candida utili.
5. it is the method for main feedstuff that mixed culture fermentation as claimed in claim 4 is handled with vinasse, it is characterized in that described malt extract medium is formulated as follows: the fermentation beer raw material of hopping is not diluted to 12 Berlin, sterilizes 30 minutes for 15 pounds.
6. it is the method for main feedstuff that mixed culture fermentation as claimed in claim 1 is handled with vinasse, and the mixed proportion that it is characterized in that described fermentation lactic acid brevibacterium and candida utili is 1: 0.1.
7. it is the method for main feedstuff that mixed culture fermentation as claimed in claim 1 is handled with vinasse, it is characterized in that described albumen raw material is selected from dregs of beans, cotton dregs, rape cake or the coconut dregs of rice.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2008100442162A CN101273749B (en) | 2008-04-15 | 2008-04-15 | Method of mixed fermentation treatment using vinasse as main feed raw material |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2008100442162A CN101273749B (en) | 2008-04-15 | 2008-04-15 | Method of mixed fermentation treatment using vinasse as main feed raw material |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101273749A CN101273749A (en) | 2008-10-01 |
| CN101273749B true CN101273749B (en) | 2011-09-07 |
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| CN108552408A (en) * | 2018-05-15 | 2018-09-21 | 湖北工业大学 | A kind of cattle feed and preparation method thereof made of vinasse |
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| CN111357882A (en) * | 2020-05-11 | 2020-07-03 | 哈工大机器人（山东）智能装备研究院 | Preparation method of pet rice wine lees probiotic food |
| CN111780497A (en) * | 2020-07-06 | 2020-10-16 | 宿州市皖神面制品有限公司 | Vinasse drying process |
| CN112617041A (en) * | 2020-12-21 | 2021-04-09 | 青岛镜川食品有限公司 | High-protein easily-digestible pet feed and preparation method thereof |
| CN113180149B (en) * | 2021-05-11 | 2023-10-13 | 江南大学 | Method for producing probiotics feed by continuously fermenting vinasse serving as raw material |
| CN114886133B (en) * | 2022-03-26 | 2023-10-17 | 刘文治 | Complete set of equipment for producing feed raw material by using fermentation tank containing large quantity of rice husk |
| CN115812844B (en) * | 2022-12-28 | 2024-05-24 | 青岛啤酒股份有限公司 | Fermented feed based on whole spent grains and preparation method thereof |
| CN116268207A (en) * | 2023-02-24 | 2023-06-23 | 山东泰山生力源集团股份有限公司 | Method for preparing high-protein feed by improving ammonia nitrogen assimilation rate of saccharomycetes |
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