CN104054914B - A kind of Wheat ration enzyme containing profitable probliotics and preparation method thereof - Google Patents
A kind of Wheat ration enzyme containing profitable probliotics and preparation method thereof Download PDFInfo
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Abstract
The present invention relates to a kind of Wheat ration enzyme containing profitable probliotics and preparation method thereof, belong to feed enzyme preparation field.The described Wheat ration enzyme containing profitable probliotics is made up of the component of following parts by weight: xylanase 100 140 parts, cellulase 6 10 parts, Bacillus licheniformis culture 20 30 parts, middle temperature alpha amylase 5 10 parts, Chinese herbal medicine powder 0.5 2 parts.Wherein Bacillus licheniformis (Β acillus licheniformis) β 10 25, deposit number CCTCC NO:M2013538.Preparation method is as follows: the culture filtering, concentrate, be dried acquisition after Bacillus licheniformis is fermented compounds with other compositions, and gained compound enzyme can be improved food conversion ratio, and reduces feed cost.
Description
Technical field
The invention belongs to enzymic preparation field, be specifically related to a kind of Wheat ration enzyme containing profitable probliotics and manufacture method thereof.
Background technology
Along with zymotechnic in modern age and the development of biotechnology, high-effect bioactive substance enzyme preparation can industry on a large scale
Metaplasia is produced, and is applied in feed industry, and many experiments and result of practical application all show, fodder enzyme preparation is raised as one
Feed additives can be effectively improved the utilization rate of feedstuff, promotes growth of animal and prevent and treat the generation of Animal diseases, with antibiotic and swash
Element class material is compared, and has the safety of brilliance, causes the great attention of feedstuff industry in global range.
Feed enzyme preparation be in order to improve animal to the digestion of feedstuff, utilize or the metabolism usefulness of improving in animal body and add feedstuff
In enzyme material.The enzyme preparation can added in feedstuff at present include amylase, alpha-galactosidase, cellulase, β-
Glucanase, glucoseoxidase, lipase, maltase, mannonase pectase, phytase, protease, angle
Protease, xylanase etc..Due to the complexity of feedstuff structure, current feed industry in producing more use compound
Enzyme preparation.Wheat ration enzyme is a kind of novel feed additive, and it is applied and starts from 20 century 70s abroad, and China
Feed industry just carry out the application of enzyme preparation in the nineties in 20th century.Therefore, the research to feedstuff complexing agent is strengthened for me
The aquaculture development of state has very important significance.
Feeding enzyme is of a great variety, all achieves goodish effect in Production of Livestock and Poultry.Former Soviet Union scientist is broken in early days
Adding amylodextrin enzyme in milk piglet diet so that daily gain improves 14.6%, carbohydrate digestion rate improves 77.2%.
According to U.S.'s relevant information, in Collier ablactational baby pig daily ration in early days supplement enzyme preparation (containing protease, amylase,
Glucanase), result experimental group improves 25% than matched group daily gain, and feed conversion rate improves 15.5%.West Germany
Scholar the most repeatedly carries out growing and fattening pigs and adds the experimentation of compound enzymic preparation, in the daily ration based on Fructus Hordei Vulgaris, adds
The compound enzymic preparation of 0.1%, result is: experimental group improves 5.1% than the matched group speed of growth, and efficiency of feed utilization improves
4.1%.Visible, feed enzyme can significantly improve poultry and digest and assimilate feedstuff, promotes growth, reduces feed cost,
And as a kind of green feed additive, its use has safety, beneficially Animal husbandry production goes on economic joint
The about road for development of type.
For a long time, people are accustomed to using Semen Maydis as energy feed, but along with Semen Maydis demand is strengthened by aquaculture development
With the change of Semen Maydis market on international market, Semen Maydis buying difficulty and price is caused to raise up, especially in non-main maize area,
The shortage of Semen Maydis becomes the key factor limiting local animal husbandry development.And in the source of non-main maize area often Semen Tritici aestivi
Relatively wide, its price is the most relatively stable, and in harvesting wheat season, its price compare with Semen Maydis often have certain
Advantage, in particular with being gradually increased of China's wheat yield, and the addition of WTO, import wheat pours in and brings
Impact so that Wheat price is on a declining curve.Meanwhile, animal husbandry development is rapid, and feedstuff consumption constantly increases, manufacture starch and
Lysine also consumes substantial amounts of Semen Maydis, and the corn price being mainly used as energy feed in the past rises steadily, and Semen Tritici aestivi is with its kind
Plant the advantages such as area is wide, yield is high, price is low and become the first-selection of replacement of corn.
At present, the shortage of grain is the principal element of puzzlement China feed industrial development.Therefore, joint to be advantageously selected for
The about animal husbandry development road of grain resource, improves efficiency of feed utilization, makes full use of limited grain resource acquisition and to the greatest extent may be used
The livestock products that energy is many.The use of the enzyme preparations such as 1,4 beta-glucanase, can effectively solve the problem that this problem, but at present producing,
The 1,4 beta-glucanase enzyme activity of the actual application in experiment is the most relatively low, and poor heat stability.Additionally, scientific experiments card
Real Bacillus licheniformis has promotion growth of animals or poultry, reduces feed-weight ratio；Increase intestinal beneficial bacterium quantity, improve animal and bird intestines micro-
Ecological environment；Prevention and treatment intestinal tract disease, reduce the effect of mortality rate.1989, U.S. FDA and U.S. feed control official
Member association discloses can be with Direct-fed and be commonly considered as safe microorganism fungus kind list, including Bacillus licheniformis.
The Ministry of Agriculture of the People's Republic of China, MOA 318 bulletin has also listed feed additive catalogue in Bacillus licheniformis.
For these reasons, the present invention will provide a kind of being added with can produce the lichens bud of high activity and stability 1,4 beta-glucanase
The feed complex enzyme of spore alphacterium culture.
Summary of the invention
It is an object of the invention to provide compounding Wheat ration enzyme containing profitable probliotics of one kind of multiple enzymes and preparation method thereof.
The purpose of the present invention can be achieved through the following technical solutions:
A kind of Wheat ration enzyme containing profitable probliotics, it is characterised in that described Wheat ration enzyme is by the component group of following parts by weight
Become: Chinese herbal medicine powder 0.5-2 part, xylanase 100-140 part, cellulase 6-10 part, Bacillus licheniformis culture 20-30
Part, mesophilicα-diastase 5-10 part.
Described xylanase, cellulase and mesophilicα-diastase be Hunan letter Wings Of The Eagle biological engineering limited company or other
Products；
The preparation method of described Chinese herbal medicine powder is as follows:
Weigh Radix Astragali 5-12 part；Radix Codonopsis 8-15 part；Radix Isatidis 5-10 part；Radix Angelicae Sinensis 10-15 part；Respectively by said herbal medicine powder
Being broken to particle diameter is that less than 2 millimeters mixing obtain Chinese herbal medicine powder；
Described Bacillus licheniformis culture preparation method is as follows:
(1) seed culture
Bacillus licheniformis slant strains is fermented through three grades of shake flask fermentation agent first class seed pots；
The culture medium of described seed culture is: yeast powder 0.5%, peptone 1%, soluble starch 1%, NaCl 1%,
PH4.5-7.0,121-123 DEG C of sterilizing 30-40min.
(2) ferment tank
First class seed pot fermentation liquid is accessed in the fermentation tank containing 3L fermentation medium with 3% inoculum concentration, cultivation temperature
35-45 DEG C, mixing speed 200-300r/min, ventilation (V/V) 1:1-3, incubation time 10-15h；Then by 1L through 121 DEG C
Sterilizing 20min, temperature is that the fermentation medium of 10 DEG C fills into fermentation tank, constant temperature culture 15-20h when temperature rises to 35-45 DEG C；
Now, first class seed pot fermentation liquid is added access fermentation tank, constant temperature culture 10-15h with 2% inoculum concentration；Continue to pass through 1L
121 DEG C of sterilizing 20min, temperature is that the fermentation medium of 10 DEG C fills into fermentation tank, constant temperature culture when temperature rises to 35-45 DEG C
10-15h；
Described fermentation medium consists of: Testa Tritici 75g, Semen Maydis powder 55g, Testa oryzae 25g, soybean cake powder 20g, beta glucan 2g,
Chinese herbal medicine powder 40g, trehalose 35g, yeast powder 6g, Semen Maydis pulp 3g, ammonium sulfate 2g, dipotassium hydrogen phosphate 2g, potassium dihydrogen phosphate
2g, sodium citrate 3g, defoamer 0.5g, pure water l000mL, pH value 4.5-7.0,121 DEG C of sterilizing 20min；
Chinese herbal medicine powder forms: take the Radix Astragali 10 parts；Radix Codonopsis 9 parts；Radix Isatidis 8 parts；Radix Angelicae Sinensis 12 parts；Respectively by said herbal medicine
Being crushed to particle diameter is that less than 2 millimeters mixing obtain Chinese herbal medicine powder；
The concocting method of described fermentation medium is: the most accurately weigh raw material, by the pure water in raw material, Semen Maydis powder, bean
Cake powder puts in material-compound tank, regulates pH value 3-7, adds midrange thermal stable amylase (3u/g Semen Maydis powder) and alpha-amylase (30u/g
Semen Maydis powder), warming while stirring simultaneously to 70 DEG C insulation 15-30min, then it is to slowly warm up to 90 DEG C of insulation 15-30min and carries out
Liquefaction, is eventually adding other raw material, stirs, and adjusts initial p H3-7, and 121-123 DEG C of sterilizing 30-40min is standby.
Dissolved oxygen controls: by adjusting speed of agitator and ventilation, control dissolved oxygen 15-30%；
PH controls: by mending ammonia or phosphoric acid,diluted, controls pH value in sweat and is maintained at 3-7；
Control of additive raw material: when the content of reducing sugar in fermentation liquid is down to 3mg/ml-8mg/ml, starts to add supplemented medium, mends
Doses is to maintain fermentation liquid content of reducing sugar as 2mg/ml-5mg/ml；
Put tank standard: 60-80% thalline aging self-dissolving, enzyme activity increasess slowly.
(3) fermentation liquor aperture 10-1000 μm coarse filtration, then goes moisture removal to obtain solid content in 10-20 DEG C of loop ultrafiltration concentration and contains
Amount 20-40% concentrated solution；Concentrated solution obtains Bacillus licheniformis culture through vacuum lyophilization, micronizing.
Described Bacillus licheniformis is specially Bacillus licheniformis (Β acillus licheniformis) β-10-25.This bacterial strain is in 2013
On November 3, in is preserved in China typical culture collection center, and (being called for short CCTCC, address is: Wuchang District, Wuhan City, Hubei Province Luo
Jia Shan Wuhan University Life Science College), preserving number is CCTCC NO:M2013538.
This bacterial strain is through physics and chemistry behavior gained, and feature is as follows:
Described bacterial strain bacterium colony on solid plate is white, and edge is irregular, and dry tack free is opaque, and microscopy is Gram-positive
Antibacterial, cellular morphology is rod-short, raw in spore, oval, does not expands.
Described bacterial strain physiological and biochemical property: V-P test (+), Starch Hydrolysis (+), casein hydrolysis (+), gelatin hydrolysis (+),
Nitrate reduction (+), indole test (-), utilize citrate (+), nitrate reductase (+), mannitol (+), xylose (+).
Described Bacillus licheniformis β-10-25 is the bacterial strain of a plant height heat production stability 1,4 beta-glucanase, and the β-Portugal producing gained gathers
Carbohydrase enzyme activity is up to 8000-10000u/ml；After at room temperature preserving 6 months, enzyme is lived and is lost less than 25%, and 4 DEG C preserve 12
After Yue, enzyme lives loss less than 20%；Remain to after preserving 1h under the conditions of 60 DEG C keep more than 80% enzyme to live, preserve under the conditions of 70 DEG C
More than 20% enzyme is kept to live after 10min.Higher than existing 1,4 beta-glucanase enzyme activity, heat stability and storage stability are the highest,
It is particularly suitable for the demand of feed industry.
Preferably, a kind of Wheat ration enzyme containing profitable probliotics is formed by following parts by weight: xylanase 120 parts, cellulose
Enzyme 8 parts, Bacillus licheniformis culture 25 parts, midrange thermal stable amylase 8 parts, Chinese herbal medicine powder 1 part.
Described Wheat ration enzyme is applicable to feed-processing plant and plant's autogamy feedstuff, should mix with other raw material in feedstuff during use
Close uniformly.Advising that complete feedstuff addition per ton is that in 80-120g(feedstuff, Semen Tritici aestivi consumption is less than 30%, addition is 80g/t, little
Wheat consumption is more than 30%, and addition is 100-120g/t).
Beneficial effect:
The Bacillus licheniformis culture that the present invention is added, not only simplify the purifying process of enzyme compared with adding pure enzymatic process,
And add a large amount of probiotic bacteria Bacillus licheniformis while adding enzyme.On the one hand the interpolation of Bacillus licheniformis can produce
Protein and peptide antibiotic substance, antagonism enteropathogenic antibacterial, suppresses the cause of disease such as aerobe and facultative anaerobe particularly escherichia coli
The breeding of bacterium；On the one hand the multiple enzyme in addition to 1,4 beta-glucanase can also be produced, such as protease, amylase, lipase and fiber
Element enzyme etc., thus improve poultry further to protein and the utilization rate of energy, promotion growth of animal；Finally, lichens brood cell bar
Bacterium also has enhancing Immune Function In Animals, resists the function infecting and improving animal physiological state.Additionally the added lichens of the present invention
1,4 beta-glucanase enzyme activity in bacillus culture is up to 8000-10000u/ml；After this enzyme preserves 1h under the conditions of 60 DEG C
Remain to keep more than 80% enzyme to live, after preserving 10min under the conditions of 70 DEG C, keep more than 20% enzyme to live；At room temperature preserve 6 months
The loss of living of rear enzyme less than 25%, 4 DEG C preserve 12 months after enzyme live loss less than 20%.It is higher than existing 1,4 beta-glucanase enzyme activity,
Enzyme effect optimum pH wide scope, heat stability and storage stability are the highest.Compared to commercially available Wheat ration enzyme,
Under same consumption, the present invention is higher to the enzymatic hydrolyzation of starch, thus improves food conversion ratio, and reduces feed cost.
With the addition of Chinese herbal medicine powder in the present invention, it has the effect of natural antibacterial and enhancing immunity, can effectively improve poultry
The anti-stress ability of animal, immunologic function and Defense response function.It addition, with the addition of in Chinese herbal medicine powder, to have anti-heat stress former
The Radix Scutellariae of effect, Radix Bupleuri so that product of the present invention has more preferable stability and preserves activity, less adds Chinese herbal medicine powder
Product preservation activity under long-time hot conditions improves 15%.
Detailed description of the invention
The preparation method of embodiment 1 Chinese herbal medicine powder
The preparation method of a kind of Chinese herbal medicine powder is as follows:
Weigh the Radix Astragali 10 parts；Radix Codonopsis 12 parts；Radix Isatidis 8 parts；Radix Angelicae Sinensis 12 parts；Said herbal medicine is crushed to particle diameter respectively is
Less than 2 millimeters mixing obtain Chinese herbal medicine powder.
The preparation method of 2 one kinds of Bacillus licheniformis cultures of embodiment
Comprise the steps:
(1) liquid seeds amplification culture
1. first order seed is cultivated: access in 500 milliliters of shaking flasks by slant strains 2 ring after activation, culture medium loading amount 100 milliliters,
Rotary shaker 180 revs/min, cultivation temperature 40 DEG C, incubation time 10h；
2. secondary seed is cultivated: accesses in 500 milliliters of secondary seed shaking flasks by first order seed according to the inoculum concentration of 10%, cultivates bar
Part is identical with first order seed；
3. three grades of seed culture: accessing in 5000 milliliters of three grades of seed flask by secondary seed with 10% inoculum concentration, culture medium fills
Measure 1000 milliliters, rotary shaker 100 revs/min, cultivation temperature 40 DEG C, incubation time 12h；
4. first class seed pot is cultivated: with 10% inoculum concentration, three grades of seeds are accessed the total measurement (volume) first class seed pot as 150L, fermentation
Culture medium loading amount 100L, cultivation temperature 41 DEG C, mixing speed 300rpm, ventilation (V/V) 1:1.5, tank pressure 0.05Mpa,
Incubation time 15h；
Described seed culture medium consists of: yeast powder 0.5%, peptone 1%, soluble starch 1%, NaCl 1%, pH5.5,
121 DEG C of sterilizing 35min.
(2) ferment tank
First class seed pot fermentation liquid is accessed in the fermentation tank containing 3L fermentation medium with 3% inoculum concentration, cultivation temperature 37 DEG C,
Mixing speed 250r/min, ventilation (V/V) 1:1-3, incubation time 12h；Then by 1L through 121 DEG C of sterilizing 20min,
Temperature is that the fermentation medium of 10 DEG C fills into fermentation tank, constant temperature culture 18h when temperature rises to 37 DEG C；Now, by first order seed
Tank fermentation liquid adds with 2% inoculum concentration and accesses fermentation tank, constant temperature culture 12h；Continue 1L through 121 DEG C of sterilizing 20min,
Temperature is that the fermentation medium of 10 DEG C fills into fermentation tank, constant temperature culture 12h when temperature rises to 37 DEG C；
Described fermentation medium consists of: Testa Tritici 75g, Semen Maydis powder 55g, Testa oryzae 25g, soybean cake powder 20g, beta glucan 2g,
Chinese herbal medicine powder 40g, trehalose 35g, yeast powder 6g, Semen Maydis pulp 3g, ammonium sulfate 2g, dipotassium hydrogen phosphate 2g, potassium dihydrogen phosphate
2g, sodium citrate 3g, defoamer 0.5g, pure water l000mL, pH value 5.5,121 DEG C of sterilizing 20min；
The concocting method of described fermentation medium is: the most accurately weigh raw material, by the pure water in raw material, Semen Maydis powder, bean
Cake powder puts in material-compound tank, regulates pH value 5.5, adds midrange thermal stable amylase (3u/g Semen Maydis powder) and alpha-amylase (30u/g
Semen Maydis powder), warming while stirring simultaneously to 70 DEG C insulation 20min, then it is to slowly warm up to 90 DEG C of insulation 20min and liquefies,
Being eventually adding other raw material, stir, adjust initial p H5.5,121 DEG C of sterilizing 35min are standby.
Described Chinese herbal medicine powder forms: the Radix Astragali 10 parts；Radix Codonopsis 9 parts；Radix Isatidis 8 parts；Radix Angelicae Sinensis 12 parts；Respectively by above-mentioned medium-height grass
It is that less than 2 millimeters mixing obtain Chinese herbal medicine powder that medicated powder is broken to particle diameter；
(3) fermentation liquor aperture 500 μm coarse filtration, then concentrates in 15 DEG C of loop ultrafiltrations and goes moisture removal to obtain solid content 20%
Concentrated solution；Concentrated solution obtains Bacillus licheniformis culture through vacuum lyophilization, micronizing.
The preparation method of 3 one kinds of complex enzyme preparation for feeding of embodiment
A kind of Wheat ration enzyme containing profitable probliotics, is made up of the enzyme preparation of following parts by weight:
Xylanase 120 parts, cellulase 8 parts, Bacillus licheniformis culture 25 parts, mesophilicα-diastase 8 parts, medium-height grass
1 part of medicated powder.
Take and originate from Hunan letter Wings Of The Eagle biological engineering limited company or the xylanase of other companies, cellulase, middle temperature α-shallow lake
Powder enzyme, and the Chinese herbal medicine powder of embodiment 1 gained 1 part, the Bacillus licheniformis culture described in embodiment 2, in 25 DEG C of bars
Stirring in mixer under part, mixing time is 5 minutes.Stir and preserve in room temperature or cold house after casing.
The preparation method of 4 one kinds of complex enzyme preparation for feeding of embodiment
A kind of Wheat ration enzyme containing profitable probliotics, is made up of the enzyme preparation of following parts by weight:
Xylanase 100 parts, cellulase 6 parts, Bacillus licheniformis culture 30 parts, mesophilicα-diastase 10 parts, in
0.5 part of medical herbs powder；
The parts by weight of each component of Chinese herbal medicine powder consist of: the Radix Astragali 5 parts；Radix Codonopsis 13 parts；Radix Isatidis 10 parts；Radix Angelicae Sinensis 15 parts；
The xylanase originating from Hunan letter Wings Of The Eagle biological engineering limited company, cellulase, middle temperature is taken by above-mentioned parts by weight
α-amylase, Chinese herbal medicine powder, the Bacillus licheniformis culture described in embodiment 2, stirs under the conditions of 25 DEG C in mixer
Uniformly, mixing time is 5 minutes.Stir and preserve in room temperature or cold house after casing.
The preparation method of 5 one kinds of complex enzyme preparation for feeding of embodiment
A kind of Wheat ration enzyme containing profitable probliotics, is made up of the enzyme preparation of following parts by weight:
Xylanase 140 parts, cellulase 10 parts, Bacillus licheniformis culture 20 parts, mesophilicα-diastase 5 parts, in
2 parts of medical herbs powder；
The parts by weight of each component of Chinese herbal medicine powder consist of: the Radix Astragali 12 parts；Radix Codonopsis 15 parts；Radix Isatidis 6 parts；Radix Angelicae Sinensis 10 parts；
The xylanase originating from Hunan letter Wings Of The Eagle biological engineering limited company, cellulase, middle temperature is taken by above-mentioned parts by weight
α-amylase, Chinese herbal medicine powder, the Bacillus licheniformis culture described in embodiment 2 stirs all under the conditions of 25 DEG C in mixer
Even, mixing time is 5 minutes.Stir and preserve in room temperature or cold house after casing.
Embodiment 5 Contrast on effect is tested
Choose 120 and fatten early stage pig, be divided into experimental group and matched group, often 3 repetitions of group by body weight, blood lineage and no sex difference,
20 pigs of each repetition.Experiment periods is 28 days, and experimental group feedstuff per ton adds compound enzymic preparation 120g described in the present embodiment 3,
Matched group, without compound enzymic preparation, is weighed on an empty stomach morning day at the whole story of experimental period, calculates daily gain and feedstuff-meat ratio, and gives hair color
Give a mark.Experimental result is as shown in the table:
| Matched group | Test group | |
| Average daily gain (g/d) | 655.20 | 713.27 |
| Feedstuff/weightening finish | 2.45 | 2.15 |
| Hair color marking (ten point system) | 5.5 | 8.5 |
Embodiment 6 Contrast on effect
Choose 120 and fatten early stage pig, be divided into experimental group and matched group, often 3 repetitions of group by body weight, blood lineage and no sex difference,
20 pigs of each repetition.Experiment periods is 28 days, and experimental group and matched group feedstuff per ton all add compound enzymic preparation 120g, experiment
Group compound enzymic preparation used is compound enzymic preparation described in embodiment 3, and compound enzymic preparation used by matched group is will to be added in embodiment 3
Add Commercial wheat daily ration enzyme, weigh on an empty stomach morning day at the whole story of experimental period, calculate daily gain and feedstuff-meat ratio, and carry out to hair color
Marking.Experimental result is as shown in the table:
| Matched group | Test group | |
| Average daily gain (g/d) | 675.26 | 713.27 |
| Feedstuff/weightening finish | 2.37 | 2.15 |
| Hair color marking (ten point system) | 6.5 | 8.5 |
Embodiment 7 Chinese herbal medicine powder preserves the impact of activity to enzyme
With described in embodiment 2 preparation method obtain Wheat ration enzyme for sample 1, with remove in embodiment 2 all of in
The Wheat ration enzyme that the preparation method of medical herbs powder obtains is sample 2, compares the preservation activity of two kinds of samples.On preserving at 45 DEG C
Stating sample 18h, measure enzyme afterwards and live, result shows: the remnant enzyme activity of sample 1 is more than 74%, and the remnant enzyme activity of sample 2 is protected
Hold about 60%.
Claims (8)
1. the Wheat ration enzyme containing profitable probliotics, it is characterised in that: it is made up of the component of following parts by weight: xylanase
100-140 part, cellulase 6-10 part, Bacillus licheniformis culture 20-30 part, mesophilicα-diastase 5-10 part, medium-height grass
Medicated powder 0.5-2 part；
Described Bacillus licheniformis is Bacillus licheniformis (Β acillus licheniformis) β-10-25, and deposit number is CCTCC
NO:M 2013538。
A kind of Wheat ration enzyme containing profitable probliotics, it is characterised in that the preparation method of described Chinese herbal medicine powder
As follows:
Weigh Radix Astragali 5-12 part；Radix Codonopsis 8-15 part；Radix Isatidis 5-10 part；Radix Angelicae Sinensis 10-15 part；Respectively by said herbal medicine powder
Being broken to particle diameter is that less than 2 millimeters mixing obtain Chinese herbal medicine powder.
A kind of Wheat ration enzyme containing profitable probliotics, it is characterised in that the preparation of Bacillus licheniformis culture
Method, comprises the steps:
Bacillus licheniformis β-10-25 successively carries out one, two, three liquid seeds and one-level kind after slant strains activation culture
Sub-tank is cultivated；Afterwards the fermentation liquid of first class seed pot is accessed in the fermentation tank containing 3L fermentation medium with 3% inoculum concentration, training
Support temperature 35-45 DEG C, mixing speed 200-300r/min, ventilation (V/V) 1:1-3, incubation time 10-15h；Then by 1L
Through 121 DEG C of sterilizing 20min, temperature is that the fermentation medium of 10 DEG C fills into fermentation tank, constant temperature when temperature rises to 35-45 DEG C
Cultivate 15-20h；Now, first class seed pot fermentation liquid is added access fermentation tank, constant temperature culture 10-15h with 2% inoculum concentration；Continue
Continuing 1L through 121 DEG C of sterilizing 20min, temperature is that the fermentation medium of 10 DEG C fills into fermentation tank, treats that temperature rises to 35-45 DEG C
Time constant temperature culture 10-15h；
Fermentation liquor aperture 10-1000 μm coarse filtration, then concentrates in 10-20 DEG C of loop ultrafiltration and goes moisture removal to obtain solid content
20-40% concentrated solution；Concentrated solution obtains Bacillus licheniformis culture through vacuum lyophilization, micronizing.
A kind of Wheat ration enzyme containing profitable probliotics, it is characterised in that one, two, three liquid described
The culture medium that seed and first class seed pot are cultivated is: yeast powder 0.5%, peptone 1%, soluble starch 1%, NaCl 1%,
PH4.5-7.0,121-123 DEG C of sterilizing 30-40min；
Described fermentation culture based raw material consists of: Testa Tritici 75g, Semen Maydis powder 55g, Testa oryzae 25g, soybean cake powder 20g, beta glucan
2g, Chinese herbal medicine powder 40g, trehalose 35g, yeast powder 6g, Semen Maydis pulp 3g, ammonium sulfate 2g, dipotassium hydrogen phosphate 2g, di(2-ethylhexyl)phosphate
Hydrogen potassium 2g, sodium citrate 3g, defoamer 0.5g, pure water 1000mL；
The concocting method of described fermentation medium is: the most accurately weigh raw material, by the pure water in raw material, Semen Maydis powder, bean
Cake powder puts in material-compound tank, regulates pH value 3-7, adds midrange thermal stable amylase and alpha-amylase, and midrange thermal stable amylase addition is 3u/g
Semen Maydis powder, alpha-amylase addition is 30u/g Semen Maydis powder, warming while stirring simultaneously to 70 DEG C insulation 15-30min, then
It is to slowly warm up to 90 DEG C of insulation 15-30min liquefy, is eventually adding other raw material, stirs, pH3-7,121-123 DEG C
Sterilizing 30-40min is standby.
A kind of Wheat ration enzyme containing profitable probliotics, it is characterised in that: by the component of following parts by weight
Composition: xylanase 120 parts, cellulase 8 parts, Bacillus licheniformis culture 25 parts, mesophilicα-diastase 8 parts,
Chinese herbal medicine powder 1 part.
A kind of Wheat ration enzyme containing profitable probliotics, it is characterised in that: described Chinese herbal medicine powder forms: yellow
Stilbene 10 parts；Radix Codonopsis 9 parts；Radix Isatidis 8 parts；Radix Angelicae Sinensis 12 parts.
7. the method preparing the Wheat ration enzyme containing profitable probliotics as described in claim 1-4 any one, comprises the steps:
Form xylanase, cellulase, Bacillus licheniformis culture, mesophilicα-diastase, Chinese herbal medicine powder according to raw material
Mix homogeneously；
Described xylanase, cellulase, mesophilicα-diastase is Hunan letter Wings Of The Eagle biological engineering limited company or other are existing
There is product；
The preparation method of described Chinese herbal medicine powder is as follows:
Weigh Radix Astragali 5-12 part；Radix Codonopsis 8-15 part；Radix Isatidis 5-10 part；Radix Angelicae Sinensis 10-15 part；Respectively by said herbal medicine powder
Being broken to particle diameter is that less than 2 millimeters mixing obtain Chinese herbal medicine powder；
Described Bacillus licheniformis culture preparation method is as follows:
(1) seed culture
Bacillus licheniformis β-10-25 slant strains is fermented through three grades of shake flask fermentation agent first class seed pots；
The culture medium of described seed culture is: yeast powder 0.5%, peptone 1%, soluble starch 1%, NaCl 1%,
PH4.5-7.0,121-123 DEG C of sterilizing 30-40min；
(2) ferment tank
First class seed pot fermentation liquid is accessed in the fermentation tank containing 3L fermentation medium with 3% inoculum concentration, cultivation temperature
35-45 DEG C, mixing speed 200-300r/min, ventilation (V/V) 1:1-3, incubation time 10-15h；Then by 1L through 121 DEG C
Sterilizing 20min, temperature is that the fermentation medium of 10 DEG C fills into fermentation tank, constant temperature culture 15-20h when temperature rises to 35-45 DEG C；
Now, first class seed pot fermentation liquid is added access fermentation tank, constant temperature culture 10-15h with 2% inoculum concentration；Continue to pass through 1L
121 DEG C of sterilizing 20min, temperature is that the fermentation medium of 10 DEG C fills into fermentation tank, constant temperature culture when temperature rises to 35-45 DEG C
10-15h；
The concocting method of described fermentation medium is: the most accurately weigh raw material: Testa Tritici 75g, Semen Maydis powder 55g, Testa oryzae 25g,
Soybean cake powder 20g, beta glucan 2g, Chinese herbal medicine powder 40g, trehalose 35g, yeast powder 6g, Semen Maydis pulp 3g, ammonium sulfate 2g,
Dipotassium hydrogen phosphate 2g, potassium dihydrogen phosphate 2g, sodium citrate 3g, defoamer 0.5g, pure water 1000mL, by raw material
Pure water, Semen Maydis powder, soybean cake powder put in material-compound tank, regulate pH value 3-7, add midrange thermal stable amylase and alpha-amylase, in
Temperature amylase addition is 3u/g Semen Maydis powder, and alpha-amylase addition is 30u/g Semen Maydis powder, warming while stirring simultaneously to 70 DEG C
Insulation 15-30min, is then to slowly warm up to 90 DEG C of insulation 15-30min and liquefies, be eventually adding other raw material, and stirring is all
Even, adjust initial pH3-7,121-123 DEG C of sterilizing 30-40min is standby；
Dissolved oxygen controls: by adjusting speed of agitator and ventilation, control dissolved oxygen 15-30%；
PH controls: by mending ammonia or phosphoric acid,diluted, controls pH value in sweat and is maintained at 3-7；
Control of additive raw material: when the content of reducing sugar in fermentation liquid is down to 3mg/ml-8mg/ml, starts to add supplemented medium, mends
Doses is to maintain fermentation liquid content of reducing sugar as 2mg/ml-5mg/ml；
Put tank standard: 60-80% thalline aging self-dissolving, enzyme activity increasess slowly；
(3) fermentation liquor aperture 10-1000 μm coarse filtration, then goes moisture removal to obtain solid content in 10-20 DEG C of loop ultrafiltration concentration and contains
Amount 20-40% concentrated solution；Concentrated solution obtains Bacillus licheniformis culture through vacuum lyophilization, micronizing.
The application in poultry Wheat Based Diet of a kind of Wheat ration enzyme containing profitable probliotics.
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| CN114214247A (en) * | 2021-12-27 | 2022-03-22 | 威海市世代海洋生物科技股份有限公司 | Bacillus licheniformis and application thereof |
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