CN105533140A - Making method of lactic acid bacteria and compound enzyme solid state fermented corn germ meal - Google Patents
Making method of lactic acid bacteria and compound enzyme solid state fermented corn germ meal Download PDFInfo
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Abstract
Belonging to the technical field of biological feed, the invention relates to a making method of lactic acid bacteria and compound enzyme solid state fermented corn germ meal. The method includes the steps of: crushing soybean meal into 40-mesh corn germ meal powder; introducing 5% of a lactic acid bacteria fermented seed solution with a viable count of 2.0*10<8>-2.0*10<9>cfu/ml into the corn germ meal powder, adding 0.5% of compound enzyme, adjusting the water content to 40% and the pH value to 5.0-7.0, mixing the substances evenly to obtain a fermentation base material; spreading the fermentation base material at a fermentation tank bottom at 30-45DEG C, carrying out anaerobic fermentation for 36-48h to obtain fermented corn germ meal, transferring the fermented corn germ meal into a fluidized bed to perform drying, and conducting cooling to room temperature so as to obtain a finished product. The corn germ meal is subjected co-fermentation by lactic acid bacteria and compound enzyme, the generated various enzymes, organic acids, amino acids and probiotics greatly improve the nutritional value of corn germ meal, and the protein content of the fermented corn germ meal is significantly increased, and the content of cellulose and other antinutritional factors is significantly decreased. As high quality protein feed, the lactic acid bacteria and compound enzyme solid state fermented corn germ meal can significantly increase the feed intake and digestibility of animals and enhance the antioxidant capacity of animal bodies.
Description
Technical field
The invention belongs to biological feedstuff technical field, be specifically related to the preparation method of lactic acid bacteria and complex enzyme solid state fermentation corn germ cake.
Background technology
China is the country that lacks the most of protide feed resource in the world, the external dependence degree of protide feed is more than 70%, the annual Soybean Meal import volume for feed is up to 5,000 ten thousand tons, and fish meal import volume 1,200,000 tons, annual total import value is 1500-1800 hundred million yuan.Therefore, develop unconventional protein feedstuff resource and seem particularly important.Corn germ cake take corn as raw material, before production starch or alcohol, carried out by corn seed soaking, pulverize and extracting oil after germ separation or remaining byproduct after lixiviate grease.Corn germ cake extracts after grease through squeezing or lixiviate, except fat content reduces, and all the other nutrient content unchanged.The crude protein content of corn germ cake is between 20-27%, it is 2-3 times of corn seed, but its lysine content is lower is only 27% of Soybean Meal, and amino acid composition is also uneven, and crude fat content is about 2%, close with Soybean Meal content, but crude fiber content is higher, and due to mouthfeel slant acidity, animal palatability is poor, make to use in growing-finishing pigs diets very restricted, general addition is at 5-10%.At present, conventional solution has amino acid balance method, adds combined-enzyme method etc., and the difference on effect of different disposal method is a lot, and these methods fundamentally do not change the auxotrophy of corn germ cake.Microbial fermentation technology process is considered to the most safe and effective processing method.
The phytoprotein that large molecule is difficult to digest is in fermentation process, the proteasome degradation of secreted generation is little peptide and free amino acid, significantly add the utilization rate of protein, the a large amount of beneficial bacteriums produced during the fermentation, and the secondary metabolite such as organic acid, bacteriocin, can effectively to degrade ANFs, improve animal body immunity, and make corn germ cake have special sour fragrance, increase palatability, there is prebiotic activity simultaneously, the generation of enteron aisle oxidative stress and diarrhoea can be alleviated.Fermentable corn germ cake as the animal feed protein source of high-quality, and can increase probio quantity, improves peptic ulcer bleeding, reduces feed waste, ensures animal health g and D.
Summary of the invention
The object of the present invention is to provide the preparation method of a kind of lactic acid bacteria and complex enzyme solid state fermentation corn germ cake.
The preparation method of a kind of lactic acid bacteria of the present invention and complex enzyme solid state fermentation corn germ cake, comprises the following steps:
Dregs of beans is pulverized by 1.1, forms 40 object corn germ cake powder;
1.2 to prepare viable count be 2.0 × 10
8-2.0 × 10
9the lactobacillus-fermented seed liquor of cfu/ml, comprises the following steps:
1.2.1 by glucose 20g, peptone 10g, yeast extract 5g, potassium dihydrogen sulfate 2g, water 1000ml, pH value 7.0,121 DEG C of condition sterilizings, as fluid nutrient medium;
1.2.2 use the fluid nutrient medium of aseptic straw aspiration step 1.2.1, instillation is equipped with in the bottle of lactic acid bacteria freeze drying powder, and vibration (100 times/min), makes freeze-drying thalline be suspension; The ratio of fluid nutrient medium and lactic acid bacteria freeze drying powder is 1:10;
Whole suspension of the freeze-drying thalline 1.2.3 obtained with aseptic straw aspiration step 1.2.2, be transplanted in MRS solid medium, 36-48h is cultivated at 37 DEG C, then be inoculated on MRS slant medium, 36-48h is cultivated on inclined-plane, inoculate in the fluid nutrient medium of step 1.2.1, carry out the Shaking culture 12-24h that frequency of oscillation is 160 times/min, obtaining viable count is 1.0 × 10
8-1.0 × 10
9the lactobacillus-fermented seed liquor of cfu/ml;
1.2.4 the lactobacillus-fermented seed liquor obtained by step 1.2.3 is inoculated in 100L seeding tank with the bacterium amount that connects of 5-10%, cultivate 12-24h, described lactobacillus-fermented seed liquor is inoculated in 100L seeding tank with the bacterium amount that connects of 5-10%, the liquid amount referring to fermentation tank is 70% of fermenter volume, connects bacterium amount and refers to this 5-10% of 70%;
1.2.5 the lactobacillus-fermented seed liquor obtained by step 1.2.4 connects with the inoculum concentration kind of 5-10% in 1t fermentation tank, and cultivate 24-36h, obtaining viable count is 2.0 × 10
8-2.0 × 10
9the lactobacillus-fermented seed liquor of cfu/ml, described lactobacillus-fermented seed liquor is inoculated in 1t fermentation tank with the bacterium amount that connects of 5-10%, refers to that the liquid amount of fermentation tank is 70% of fermenter volume, connects bacterium amount and refers to this 5-10% of 70%.；
1.3 lactobacillus-fermented seed liquor step 1.2 prepared access in the corn germ cake powder that step 1.1 obtains by 5% (v/w), and add the complex enzyme of 0.5% (w/w), adjustment water content is 40%, and pH value is 5.0-7.0, obtains the basestocks that ferments after mixing;
The fermentation basestocks obtained in step 1.3 stirs by 1.4, and paves at the bottom of fermentation vat by the thickness of 60-70cm, and fermentation vat temperature remains on 30-45 DEG C, anaerobic fermentation 36-48h, obtains the fermented maize germ dregs of rice;
1.5 dry in the fermented maize germ dregs of rice obtained in step 1.4 immigration fluid bed, and bake out temperature is 50-55 DEG C;
1.6 to be dried after the fermented maize germ dregs of rice be down to room temperature, namely obtain lactic acid bacteria and complex enzyme solid state fermentation corn germ cake finished product.
Complex enzyme composition described in step 1.3 is: acid protease 50000IU/g, cellulase 36000IU/g, 1,4 beta-glucanase 3000IU/g, zytase 10000IU/g.
Compared with prior art, the present invention has the following advantages:
1. corn germ cake is under the combined ferment of lactic acid bacteria and complex enzyme, the various enzymes, organic acid, amino acid, probio etc. that produce substantially increase the nutritive value of corn germ cake, in the fermented maize germ dregs of rice, protein content significantly increases, the ANFs content such as cellulose significantly reduce, decreasing the harm to animal, is a kind of protein feed resource of high-quality;
2. in animal, add feed intake and the digestibility that significantly can increase animal, improve antioxidant ability of organism.
Detailed description of the invention
Below in conjunction with embodiment, the preparation method of lactic acid bacteria provided by the invention and complex enzyme solid state fermentation corn germ cake is described in detail.
Embodiment 1
With reference to the lactic acid bacteria of the present embodiment and the method for production of complex enzyme solid state fermentation corn germ cake, it comprises following concrete steps:
(1) preparation of fermentation raw material
By fermentation raw material---corn germ cake is crushed to the powdery of 40 orders (0.42mm).
(2) preparation of lactic acid bacterial liquid
Suitable fluid nutrient medium is drawn with aseptic straw, instillation lactic acid bacteria freeze drying powder is preserved in bottle, vibrates gently, makes freeze-drying thalline be suspension, draw whole bacteria suspension, transplant in MRS solid medium, after cultivating 36h at 37 DEG C, be inoculated on MRS slant medium, after 36h is cultivated on inclined-plane, be inoculated in fluid nutrient medium, Shaking culture 12h, obtaining viable count is 1.0 × 10
8seed liquor, by seed liquor with 5% the bacterium amount that connects be inoculated in 100L seeding tank, cultivate after 12h that culture transferring is in 1t fermentation tank, obtaining viable count after cultivating 24h is 2.0 × 10
8fermentation seed liquid.
(3) ferment
The lactobacillus-fermented seed liquor of above-mentioned preparation is linked in corn germ cake by 5% (v/w), and (complex enzyme composition is: acid protease 50000IU/g to add the complex enzyme of 0.5% (w/w), cellulase 36000IU/g, 1,4 beta-glucanase 3000IU/g, with zytase 10000IU/g), adjustment water content is 40%, and pH value 5.0 obtains the basestocks that ferments after mixing; Postvaccinal fermentation basestocks presses thickness 60cm, paves uniformly at the bottom of fermentation vat, and fermentation vat temperature remains on 30 DEG C, after stirring, anaerobic fermentation 36h, obtains fermentation enzymolysis corn germ cake.
(4) dry
Product after fermentation all moves in fluid bed and dries, and bake out temperature controls at 50 DEG C, after the product after to be dried is down to room temperature, obtains fermented maize germ dregs of rice finished product.
Embodiment 2
With reference to the lactic acid bacteria of the present embodiment and the method for production of complex enzyme solid state fermentation corn germ cake, it comprises following concrete steps:
(1) preparation of fermentation raw material
By fermentation raw material---corn germ cake is crushed to the powdery of 40 orders (0.42mm).
(2) preparation of lactic acid bacterial liquid
Suitable fluid nutrient medium is drawn with aseptic straw, instillation lactic acid bacteria freeze drying powder is preserved in bottle, vibrates gently, makes freeze-drying thalline be suspension, draw whole bacteria suspension, transplant in MRS solid medium, after cultivating 42h at 37 DEG C, be inoculated on MRS slant medium, after 42h is cultivated on inclined-plane, be inoculated in fluid nutrient medium, Shaking culture 18h, obtaining viable count is 3.0 × 10
8seed liquor, by seed liquor with 7% the bacterium amount that connects be inoculated in 100L seeding tank, cultivate after 18h that culture transferring is in 1t fermentation tank, obtaining viable count after cultivating 24h is 1.0 × 10
9fermentation seed liquid.
(3) ferment
The lactobacillus-fermented seed liquor of above-mentioned preparation is linked in corn germ cake by 5% (v/w), and (complex enzyme composition is: acid protease 50000IU/g to add 0.5% (w/w) complex enzyme, cellulase 36000IU/g, 1,4 beta-glucanase 3000IU/g, with zytase 10000IU/g), adjustment water content is 40%, and pH value 6.0 obtains the basestocks that ferments after mixing; Postvaccinal fermentation basestocks presses thickness 65cm, paves uniformly at the bottom of fermentation vat, and fermentation vat temperature remains on 35 DEG C, after stirring, anaerobic fermentation 42h, obtains fermentation enzymolysis corn germ cake.
(4) dry
Product after fermentation all moves in fluid bed and dries, and bake out temperature controls at 53 DEG C, after the product after to be dried is down to room temperature, obtains fermented maize germ dregs of rice finished product.
Embodiment 3
With reference to the lactic acid bacteria of the present embodiment and the method for production of complex enzyme solid state fermentation corn germ cake, comprise following concrete steps:
(1) preparation of fermentation raw material
By fermentation raw material---corn germ cake is crushed to the powdery of 40 orders (0.42mm).
(2) preparation of lactic acid bacterial liquid
Suitable fluid nutrient medium is drawn with aseptic straw, instillation lactic acid bacteria freeze drying powder is preserved in bottle, vibrates gently, makes freeze-drying thalline be suspension, draw whole bacteria suspension, transplant in MRS solid medium, after cultivating 48h at 37 DEG C, be inoculated on MRS slant medium, after 48h is cultivated on inclined-plane, be inoculated in fluid nutrient medium, Shaking culture 24h, obtaining viable count is 2.0 × 10
8seed liquor, by seed liquor with 10% the bacterium amount that connects be inoculated in 100L seeding tank, cultivate after 24h that culture transferring is in 1t fermentation tank, obtaining viable count after cultivating 36h is 2.0 × 10
9fermentation seed liquid.
(3) ferment
The lactobacillus-fermented seed liquor of above-mentioned preparation is linked in corn germ cake by 5% (v/w), and (complex enzyme composition is: acid protease 50000IU/g to add 0.5% (w/w) complex enzyme, cellulase 36000IU/g, 1,4 beta-glucanase 3000IU/g, with zytase 10000IU/g), adjustment water content is 40%, and pH value 7.0 obtains the basestocks that ferments after mixing; Postvaccinal fermentation basestocks presses thickness 70cm, paves uniformly at the bottom of fermentation vat, and fermentation vat temperature remains on 40 DEG C, after stirring, anaerobic fermentation 48h, obtains fermentation enzymolysis corn germ cake.
(4) dry
Product after fermentation all moves in fluid bed and dries, and bake out temperature controls at 55 DEG C, after the product after to be dried is down to room temperature, obtains fermented maize germ dregs of rice finished product.
By following test data, the practical application effect of lactic acid bacteria and the complex enzyme solid state fermentation corn germ cake adopting the present invention to make can be embodied:
One, lactic acid bacteria and complex enzyme solid state fermentation are on the impact of corn germ cake nutritional labeling and ANFs content
1, materials and methods
Corn germ cake (commercially available, CP is 21.05%), lactic acid bacteria is the autonomous preservation of Jilin Academy of Agricultural Science Animal nutrition research institute of herding branch.MRS solid medium: peptone 10g, beef extract 10g, yeast extract 5g, dibasic ammonium citrate 2g, sodium acetate 5g, K
2hPO42g, MnSO
44H
2o0.25g, MgSO
47H
2o0.58g, glucose 20g, Tween-80 1mL, agar 25g, water 1000mL, adjust pH6.2 ~ 6.4,121 DEG C of sterilizing 20min.Fluid nutrient medium: glucose 20g, peptone 10g, yeast extract 5g, K
2hPO
42g, water 1000mL, pH7.0,121 DEG C of sterilizing 20min.
Acid protease (50000IU/g) cellulase (36000IU/g), 1,4 beta-glucanase (3000IU/g), and zytase (10000IU/g) is purchased from Zhaodong, Heilongjiang Province Cheng enzyme preparation Co., Ltd.
2. fermented sample preparation and testing index
According to fermentation process of the present invention, by corn germ cake sealed fermenting 48h.Sample after fermentation ends, dry standby survey in 50 ~ 55 DEG C.Testing index has: crude protein, crude fibre, crude fat, lysine, cellulose, xylan and beta glucan.
3. results and analysis
Nutritional labeling before and after corn germ cake fermentation is in table 1.
Before and after the fermentation of table 1 corn germ cake, nutritional labeling compares
As can be seen from Table 1, corn germ cake is after lactic acid bacteria and complex enzyme solid state fermentation, crude protein and lysine content significantly increase, crude protein content increases by 19%, lysine increases by 44%, crude fibre, cellulose, xylan and beta glucan content significantly reduce, crude fibre declines 66%, cellulose declines 82%, and xylan declines 78%, and beta glucan is almost completely degraded, therefore, corn germ cake, after lactic acid bacteria and complex enzyme solid state fermentation, can significantly improve its nutritional labeling, reduces the ANFs such as crude fibre.
Two, lactic acid bacteria and complex enzyme solid state fermentation corn germ cake are tested the feeding effect of growing-finishing pig
In order to prove the feeding effect of lactic acid bacteria and complex enzyme solid state fermentation corn germ cake further, carry out the feeding experiment to growing-finishing pig.
1. materials and methods
The preparation of lactic acid bacteria and complex enzyme solid state fermentation corn germ cake, according to fermentation process of the present invention, by corn germ cake sealed fermenting 48h.In 50 ~ 55 DEG C of dry for standby after fermentation ends.
Experimental design: 60 55 ages in days, the healthy commercial generation PIC pig of first starting weight 24 ± 1.1kg, male and female half and half, is divided into 3 groups.Test adopts single factor design, often organizes 10 repetitions, each repetition 2 pigs, free choice feeding and drinking-water.First group of ordinary maize dregs of beans daily ration of feeding is control group, second group of 15% untreated corn germ cake of feeding, the 3rd group of 15% fermented maize germ dregs of rice of feeding.Preliminary trial period 7d, 27 days experimental periods, amounts to 34 days.
Test daily ration:
Control group daily ration take corn-soybean meal as primary raw material, test in 1 group and add 15% untreated corn germ cake, test the fermented maize germ dregs of rice that 2 groups are added 15%, Diet Formula is with reference to China " lean meat species growing-finishing pig feeding standard NY/T65-2004 " design.Daily ration composition and trophic level are in table 2.
Table 2 daily ration composition and trophic level
Premix: pig multidimensional, 200g; Copper sulphate, 380g; Ferrous sulfate, 400g; Zinc sulfate, 200g; Manganese sulfate, 45g; 1% sodium selenite, 50g; 1% KI, 40g; 1% cobalt chloride, 20g; Sodium bicarbonate, 500g, choline, 1000g.
Testing index:
Growth indexes: just trying the 1st day phase and the 27th day respectively, weighs on an empty stomach before raising morning, and calculates its average daily ingestion amount, daily gain and feedstuff-meat ratio.
Blood parameters: just trying the 27th day phase, raise front empty stomach morning, each repetition adopts 1, often organizes 10, vena cava anterior blood sampling 10ml, and under room temperature, leave standstill, naturally separate out serum, serum is divided in 1.5ml centrifuge tube, in-20 DEG C of preservations, and standby survey.Testing index has total protein, albumin, urea nitrogen.
Nutriment apparent digestibility: adopt endogenous indicator method (acid insoluble ash) to start to collect excrement sample the 21st day experimental period, all collect 4d continuously at 9:00 and 16:00, the excrement sample that every day collects fully stirs and evenly mixs, by the 10% reduction excrement sample collecting total excrement amount, every 100g excrement sample adds 20ml10%H
2sO
4, standby survey crude protein, separately gets 500g excrement sample and dries under 65 DEG C of conditions, make air-dry sample, pulverized 40 mesh sieves, standby survey crude fibre, crude fat and coarse ash.
Assay method in feed and excrement sample: crude protein adopts full-automatic Kjeldahl determination device to measure; Crude fat adopts surname extraction instrument; Crude fibre, ADF and NDF adopt fibre analyser to measure; Acid insoluble ash adopts the method for GB/T23742-2009.
Certain Nutrient Apparent Digestibility (%)=[1-(and in excrement certain nutrient content/feed in certain nutrient content) × (in feed acid insoluble ash content/excrement in the content of acid insoluble ash)] × 100%.
Enteron aisle major microorganisms measures: just trying the last day of phase, often organize the grower pigs of random choose 4 basic uniformities of body condition, put to death, open abdominal cavity rapidly, cecal ligation two ends, cut caecum and be placed on aseptic operating platform, get cecal content 1g wherein, put into the triangular flask of the sterile saline filling 99ml, fully vibrate, get dilution and carry out 10 times of dilutions.Draw 0.1ml to be inoculated on Selective agar medium.Escherichia coli, with EMB agar medium 37 DEG C cultivation, carry out colony counting after 48h; Lactic acid bacteria, with LBS culture medium 37 DEG C cultivation, carries out colony counting after 48h; Bifidobacterium, with BBL culture medium 37 DEG C of Anaerobic culturel, carries out colony counting after 48h.All microorganism count results represent with logarithmic form carries out variance analysis.
2. results and analysis
2.1 impacts on pig growth performance
Feed fermentation and the non-fermented maize germ dregs of rice the results are shown in Table 2 to growth of growing-finishing pigs performance impact.
Table 2 is fed the impact of the fermented maize germ dregs of rice on grower pigs production performance
Note: colleague's shoulder notes not marking-up mother person for difference remarkable (P>0.05), different lowercase is significant difference (P<0.05), and different capitalization person is difference extremely remarkable (P<0.01).
As can be seen from the table, counterpoise at the beginning of between each processed group and last counterpoise difference are all not significantly (P>0.05).Average daily gain and the average daily ingestion amount of testing 1 group are all remarkable in control group and test 2 groups (P<0.05), test 2 groups and control group difference remarkable (P>0.05); Feed-weight ratio is tested 1 group and is significantly higher than control group and test 2 groups, illustrate that the corn germ cake of interpolation 15% in grower pigs daily ration can have a certain impact to the growth of pig, but add the fermented maize germ dregs of rice, growth performance does not have difference compared with conventional corn-soybean meal daily ration of feeding.
2.2 impacts on grower pigs biochemical indicator
Feed fermentation and the non-fermented maize germ dregs of rice on the impact of growing-finishing pig blood parameters in table 3.
Table 3 is fed the impact of the fermented maize germ dregs of rice on grower pigs blood parameters
Note: colleague's shoulder notes not marking-up mother person for difference remarkable (P>0.05), different lowercase is significant difference (P<0.05), and different capitalization person is difference extremely remarkable (P<0.01).
As can be seen from Table 3, between each processed group, seralbumin and total protein content difference are all not significantly (P>0.05), test the serum urea nitrogen content of 2 groups significantly lower than control group and test 1 group (P<0.05).
2.3 impacts on nutritive digestibility
On the impact of nutritive digestibility in table 4.
Table 4 is fed the impact of the fermented maize germ dregs of rice on grower pigs nutritive digestibility
Note: colleague's shoulder notes not marking-up mother person for difference remarkable (P>0.05), different lowercase is significant difference (P<0.05), and different capitalization person is difference extremely remarkable (P<0.01).
As can be seen from Table 4, between each processed group, dry digestibility difference is not significantly (P>0.05), the crude protein digestibility testing 2 groups is significantly higher than control group and test 1 group (P<0.05), and test 1 group is significantly lower than control group (P<0.05), test 1 group of crude fiber digestibility significantly lower than control group and test 2 groups (P<0.05), and control group and test 2 groups of differences are not significantly (P>0.05).Illustrate, the fermented maize germ dregs of rice of feeding can significantly improve thick protein and coarse-fibred digestibility.
2.4 impacts on cecum microorganisms
What affect cecum microorganisms the results are shown in Table 5.
Table 5 is fed the impact of the fermented maize germ dregs of rice on grower pigs cecum microorganisms
Note: colleague's shoulder notes not marking-up mother person for difference remarkable (P>0.05), different lowercase is significant difference (P<0.05), and different capitalization person is difference extremely remarkable (P<0.01).
As can be seen from Table 5, the fermented maize germ dregs of rice of feeding (testing 2 groups) can significantly reduce colibacillary quantity, improve the quantity of Bacillus acidi lactici and Bifidobacterium, illustrate that the fermented maize germ dregs of rice have prebiotic effect.
3 conclusions
Corn germ cake, after lactic acid bacteria and complex enzyme solid state fermentation, can significantly improve its nutritional labeling, reduces the ANFs such as crude fibre.Proved by the test at growing-finishing pig, the fermented maize germ dregs of rice adding 15% can remarkable daily gain, effectively improves feed-weight ratio, and crude protein, coarse-fibred digestibility can be significantly improved, harmful intestinal tract bacteria is significantly reduced, and beneficial bacterium significantly increases, and has prebiotic function.
Claims (2)
1. a preparation method for lactic acid bacteria and complex enzyme solid state fermentation corn germ cake, is characterized in that comprising the following steps:
Dregs of beans is pulverized by 1.1, forms 40 object corn germ cake powder;
1.2 to prepare viable count be 2.0 × 10
8-2.0 × 10
9the lactobacillus-fermented seed liquor of cfu/ml, comprises the following steps:
1.2.1 by glucose 20g, peptone 10g, yeast extract 5g, potassium dihydrogen sulfate 2g, water 1000ml, pH value 7.0,121 DEG C of condition sterilizings, as fluid nutrient medium;
1.2.2 use the fluid nutrient medium of aseptic straw aspiration step 1.2.1, instillation is equipped with in the bottle of lactic acid bacteria freeze drying powder, and vibration (100 times/min), makes freeze-drying thalline be suspension; The ratio of fluid nutrient medium and lactic acid bacteria freeze drying powder is 1:10;
Whole suspension of the freeze-drying thalline 1.2.3 obtained with aseptic straw aspiration step 1.2.2, be transplanted in MRS solid medium, 36-48h is cultivated at 37 DEG C, then be inoculated on MRS slant medium, 36-48h is cultivated on inclined-plane, inoculate in the fluid nutrient medium of step 1.2.1, carry out the Shaking culture 12-24h that frequency of oscillation is 160 times/min, obtaining viable count is 1.0 × 10
8-1.0 × 10
9the lactobacillus-fermented seed liquor of cfu/ml;
1.2.4 the lactobacillus-fermented seed liquor obtained by step 1.2.3 is inoculated in 100L seeding tank with the bacterium amount that connects of 5-10%, cultivate 12-24h, described lactobacillus-fermented seed liquor is inoculated in 100L seeding tank with the bacterium amount that connects of 5-10%, the liquid amount referring to fermentation tank is 70% of fermenter volume, connects bacterium amount and refers to this 5-10% of 70%;
1.2.5 the lactobacillus-fermented seed liquor obtained by step 1.2.4 connects with the inoculum concentration kind of 5-10% in 1t fermentation tank, and cultivate 24-36h, obtaining viable count is 2.0 × 10
8-2.0 × 10
9the lactobacillus-fermented seed liquor of cfu/ml, described lactobacillus-fermented seed liquor is inoculated in 1t fermentation tank with the bacterium amount that connects of 5-10%, refers to that the liquid amount of fermentation tank is 70% of fermenter volume, connects bacterium amount and refers to this 5-10% of 70%;
1.3 lactobacillus-fermented seed liquor step 1.2 prepared access in the corn germ cake powder that step 1.1 obtains by 5% (v/w), and add the complex enzyme of 0.5% (w/w), adjustment water content is 40%, and pH value is 5.0-7.0, obtains the basestocks that ferments after mixing;
The fermentation basestocks obtained in step 1.3 stirs by 1.4, and paves at the bottom of fermentation vat by the thickness of 60-70cm, and fermentation vat temperature remains on 30-45 DEG C, anaerobic fermentation 36-48h, obtains the fermented maize germ dregs of rice;
1.5 dry in the fermented maize germ dregs of rice obtained in step 1.4 immigration fluid bed, and bake out temperature is 50-55 DEG C;
1.6 to be dried after the fermented maize germ dregs of rice be down to room temperature, namely obtain lactic acid bacteria and complex enzyme solid state fermentation corn germ cake finished product.
2. press the method for lactic acid bacteria according to claim 1 and complex enzyme solid state fermentation corn germ cake, it is characterized in that the complex enzyme composition described in step 1.3 is: acid protease 50000IU/g, cellulase 36000IU/g, 1,4 beta-glucanase 3000IU/g, zytase 10000IU/g.
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| CN107047987A (en) * | 2017-03-29 | 2017-08-18 | 内蒙古河套农牧业技术研究院 | A kind of fermented composite feed for feeding Fattening Sheep and its application |
| CN109874940A (en) * | 2019-04-28 | 2019-06-14 | 新昌县大船畈生物科技有限公司 | Pig fattening period mixed feed and preparation method thereof |
| CN110250324A (en) * | 2019-07-29 | 2019-09-20 | 宁夏大学 | A kind of caragana microphylla fermented type fully mixed diet preparation method and application |
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