CN104054914A - Wheat daily ration enzyme containing probiotics and preparation method thereof - Google Patents
Wheat daily ration enzyme containing probiotics and preparation method thereof Download PDFInfo
- Publication number
- CN104054914A CN104054914A CN201310739603.9A CN201310739603A CN104054914A CN 104054914 A CN104054914 A CN 104054914A CN 201310739603 A CN201310739603 A CN 201310739603A CN 104054914 A CN104054914 A CN 104054914A
- Authority
- CN
- China
- Prior art keywords
- culture
- parts
- bacillus licheniformis
- herbal medicine
- chinese herbal
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Abstract
The invention relates to a wheat daily ration enzyme containing probiotics and a preparation method thereof and belongs to the technical field of feed enzyme preparation. The wheat daily ration enzyme containing probiotics comprises following components, by weight, 100-140 parts of xylanase, 6-10 parts of cellulase, 20-30 parts of a bacillus licheniformis culture, 5-10 parts of a medium-temperature [alpha]-amylase and 0.5-2 parts of a Chinese herbal medicine powder, wherein the bacillus licheniformis [beta]-10-25 is assigned the accession number CCTCC NO:M2013538. The preparation method includes following steps: fermenting the bacillus licheniformis, performing a filtering process, a concentrating process and a drying process to the fermented bacillus licheniformis to obtain the culture, and compounding the culture with other components. The composite enzyme can increase feed conversion rate and reduce feed cost.
Description
Technical field
The invention belongs to enzyme preparation field, be specifically related to a kind of Wheat Based Diet enzyme and manufacture method thereof containing profitable probliotics.
Background technology
Development along with modern age zymotechnic and biotechnology, high-effect bioactivator---enzyme preparation is suitability for industrialized production on a large scale, and be applied in feed industry, many experiments and result of practical application all show, fodder enzyme preparation as a kind of feed addictive can effectively improve feed utilization rate, promote growth of animal and prevent and treat the generation of Animal diseases, compare with hormonal substance with antibiotic, there is remarkable security, caused the great attention of feedstuff industry in global range.
Feed enzyme preparation is in order to improve animal to the digestion of feed, utilization or to improve the metabolism usefulness in animal body and add the enzyme material in feed.The enzyme preparation that can add in feed at present comprises amylase, alpha-galactosidase, cellulase, 1,4 beta-glucanase, glucose oxidase, lipase, maltose, mannonase pectase, phytase, protease, keratinase, zytase etc.Due to the complexity of feedstuff structure, in feed industry production at present, more uses is complex enzyme formulation.Wheat Based Diet enzyme is a kind of novel feed addictive, and its application starts from 20 century 70s abroad, and the feed industry of China is just carried out the application of enzyme preparation in the nineties in 20th century.Therefore, to the research of feed complexing agent, the aquaculture development for China has very important significance in increasing.
Feeding enzyme is of a great variety, has all obtained goodish effect in Production of Livestock and Poultry.The scientist of the former Soviet Union adds amylodextrin enzyme in weanling pig daily ration in early days, makes daily gain improve 14.6%, and carbohydrate digestion rate improves 77.2%.According to U.S.'s interrelated data, Collier supplements enzyme preparation (containing protease, amylase, dextranase) in weanling pig daily ration in early days, and result experimental group has improved 25% than control group daily gain, and feed conversion rate has improved 15.5%.West Germany scholar once repeatedly carried out the experimental study that complex enzyme formulation is added in growing and fattening pigs, take barley in main daily ration, added 0.1% complex enzyme formulation, and result is: experimental group improves 5.1% than the control group speed of growth, and efficiency of feed utilization improves 4.1%.Visible, feed enzyme can significantly improve livestock and poultry to the digesting and assimilating of feed, and promotes growth, reduce feed cost, and as a kind of green feed additive, its use has security, is conducive to animal husbandry and produces the road for development of going on economy type.
For a long time, people's custom is usingd corn as energy feed, but along with the variation of aquaculture development to corn market on the increasing of corn demand and international market, cause corn buying difficulty and price to raise up, especially in non-main maize area, the shortage of corn becomes the key factor of the local animal husbandry development of restriction.And often the source of wheat is wider in non-main maize area, its price is also comparatively stable, and in harvesting wheat season, its price is compared with corn often has certain advantage, particularly along with the increase gradually of China's wheat yield, and the adding of WTO, import wheat pours in the impact bringing, and makes Wheat price on a declining curve.Simultaneously, animal husbandry development is rapid, and feed consumption constantly increases, and manufactures starch and lysine and also consumes a large amount of corns, the advantages such as the main corn price as energy feed rose steadily in the past, and wheat is wide with its cultivated area, output is high, price is low 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, the animal husbandry development road of the resource of selecting to be conducive to save food, improves efficiency of feed utilization, makes full use of limited grain resource and obtains livestock products as much as possible.The use of the enzyme preparations such as 1,4 beta-glucanase, can effectively address this problem, but the 1,4 beta-glucanase enzyme activity of the practical application in producing, testing is mostly lower at present, and poor heat stability.In addition, scientific experiment has confirmed that bacillus licheniformis has promotion growth of animals or poultry, reduces material anharmonic ratio; Increase intestinal beneficial bacterium quantity, improve animal and bird intestines micro-ecological environment; Prevention and treatment intestines problem, reduce the effect of the death rate.1989, U.S. FDA and U.S. feed control that official association announced can Direct-fed and be commonly considered as safe microorganism fungus kind list, comprising bacillus licheniformis.No. 318 bulletins of The Ministry of Agriculture of the People's Republic of China, MOA have also been listed feed addictive catalogue in bacillus licheniformis.
In view of above reason, the present invention will provide a kind of feed complex enzyme that is added with the bacillus licheniformis culture that can produce high activity and stability 1,4 beta-glucanase.
Summary of the invention
The object of the present invention is to provide composite Wheat Based Diet enzyme containing profitable probliotics of one kind of multiple enzymes and preparation method thereof.
Object of the present invention can be achieved through the following technical solutions:
A Wheat Based Diet enzyme containing profitable probliotics, is characterized in that, described Wheat Based Diet enzyme is comprised of the component of following parts by weight: Chinese herbal medicine powder 0.5-2 part, zytase 100-140 part, cellulase 6-10 part, bacillus licheniformis culture 20-30 part, mesophilicα-diastase 5-10 part.
Described zytase, cellulase and mesophilicα-diastase are Hunan letter Wings Of The Eagle bioengineering limited company or other company's products;
The preparation method of described Chinese herbal medicine powder is as follows:
Take Radix Astragali 5-12 part; Radix Codonopsis 8-15 part; Radix Isatidis 5-10 part; Radix Angelicae Sinensis 10-15 part; Respectively said herbal medicine being crushed to particle diameter is 2 millimeters of following mixing to 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: dusty yeast 0.5%, and peptone 1%, soluble starch 1%, NaCl 1%,, pH4.5-7.0,121-123 ℃ of sterilizing 30-40min.
(2) ferment tank
In the fermentation tank that first class seed pot zymotic fluid is contained to 3L fermentation medium with 3% inoculum concentration access, cultivation temperature 35-45 ℃, mixing speed 200-300r/min, ventilation (V/V) 1:1-3, incubation time 10-15h; Then by 1L through 121 ℃ of sterilizing 20min, temperature is that the fermentation medium of 10 ℃ fills into fermentation tank, constant temperature culture 15-20h when temperature rises to 35-45 ℃; Now, first class seed pot zymotic fluid is appended to access fermentation tank, constant temperature culture 10-15h with 2% inoculum concentration; Continuation is by 1L through 121 ℃ of sterilizing 20min, and temperature is that the fermentation medium of 10 ℃ fills into fermentation tank, constant temperature culture 10-15h when temperature rises to 35-45 ℃;
Described fermentation medium consists of: wheat bran 75g, corn flour 55g, rice bran 25g, beancake powder 20g, beta glucan 2g, Chinese herbal medicine powder 40g, trehalose 35g, dusty yeast 6g, corn steep liquor 3g, ammonium sulfate 2g, dipotassium hydrogen phosphate 2g, potassium dihydrogen phosphate 2g, natrium citricum 3g, defoamer 0.5g, pure water l000mL, pH value 4.5-7.0,121 ℃ of sterilizing 20min;
Chinese herbal medicine powder forms: get 10 parts of the Radixs Astragali; 9 parts of Radix Codonopsis; 8 parts of Radix Isatidis; 12 parts of Radix Angelicae Sinensis; Respectively said herbal medicine being crushed to particle diameter is 2 millimeters of following mixing to obtain Chinese herbal medicine powder;
The concocting method of described fermentation medium is: accurately take in proportion raw material, pure water in raw material, corn flour, beancake powder are dropped in material-compound tank, regulate pH value 3-7, add middle temperature amylase (3u/g corn flour) and alpha-amylase (30u/g corn flour), ℃ insulation 15-30min of warming while stirring to 70 simultaneously, is then slowly warming up to 90 ℃ of insulation 15-30min and liquefies, finally add other raw material, stir, adjust initial p H3-7,121-123 ℃ of sterilizing 30-40min is standby.
Dissolved oxygen is controlled: by adjusting speed of agitator and ventilation, control dissolved oxygen 15-30%;
PH controls: by mending ammoniacal liquor or phosphoric acid,diluted, in controlled fermentation process, pH value remains on 3-7;
Control of additive raw material: when the content of reducing sugar in zymotic fluid is down to 3mg/ml-8mg/ml, start to add supplemented medium, feed supplement amount be take and maintained zymotic fluid content of reducing sugar as 2mg/ml-5mg/ml;
Put tank standard: the old and feeble self-dissolving of 60-80% thalline, enzyme activity increasess slowly.
(3) zymotic fluid via hole diameter 10-1000 μ m coarse filtration, then obtains solid content 20-40% concentrate in the concentrated moisture of removing of 10-20 ℃ of loop ultrafiltration; Concentrate obtains bacillus licheniformis culture through vacuum freeze drying, ultramicro grinding.
Described bacillus licheniformis is specially bacillus licheniformis (Β acillus licheniformis) β-10-25.This bacterial strain is preserved in Chinese Typical Representative culture collection center on November 3rd, 2013, and (be 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 process physical chemistry mutagenesis 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 bacterium, and cellular morphology is rod-short, and raw in gemma, ellipse, does not expand.
Described bacterial strain physiological and biochemical property: V-P tests (+), Starch Hydrolysis (+), casein hydrolysis (+), gelatin hydrolysis (+), nitrate reduction (+), indole test (-), utilizes citrate (+), nitrate reductase (+), sweet mellow wine (+), wood sugar (+).
Described bacillus licheniformis β-10-25 is the bacterial strain of a plant height heat production stability 1,4 beta-glucanase, produces the 1,4 beta-glucanase enzyme activity of gained up to 8000-10000u/ml; At room temperature preserving after 6 months the enzyme loss of living is less than 25%, 4 ℃ and preserves after 12 months the enzyme loss of living and be less than 20%; Under 60 ℃ of conditions, preserve after 1h and still can keep 80% above enzyme to live, keep 20% above enzyme work after preserving 10min under 70 ℃ of conditions.Higher than existing 1,4 beta-glucanase enzyme activity, heat endurance and storage stability are all higher, are particularly suitable for the demand of feed industry.
Preferably, a kind of Wheat Based Diet enzyme containing profitable probliotics forms by following parts by weight: 120 parts of zytases, 8 parts of cellulases, 25 parts of bacillus licheniformis cultures, middle temperature amylase 8 part, 1 part of Chinese herbal medicine powder.
Described Wheat Based Diet enzyme is applicable to feed-processing plant and plant's autogamy feed, during use, should mix with other raw material in feed.Advise that complete diet pellet addition per ton is that in 80-120g(feed, wheat consumption is lower than 30%, addition is 80g/t, and wheat consumption surpasses 30%, and addition is 100-120g/t).
Beneficial effect:
The bacillus licheniformis culture that the present invention adds, compares with adding pure enzymatic process the purifying process of not only having simplified enzyme, and add a large amount of probios---bacillus licheniformis when adding enzyme.The interpolation of bacillus licheniformis can produce protein and peptide antibacterial material on the one hand, and antagonism enteron aisle pathogenetic bacteria suppresses the particularly breeding of the pathogen such as Escherichia coli of aerobic bacteria and facultative anaerobe; Can also produce the multiple enzyme except 1,4 beta-glucanase on the one hand, as protease, amylase, lipase and cellulase etc., thereby further improve the utilization rate of livestock and poultry to protein and energy, promote growth of animal; Finally, bacillus licheniformis also has enhancing Immune Function In Animals, resists the function that infects and improve animal physiological state.The present invention in addition adds 1,4 beta-glucanase enzyme activity in bacillus licheniformis culture up to 8000-10000u/ml; This enzyme still can keep 80% above enzyme to live preserve 1h under 60 ℃ of conditions after, keep 20% above enzyme work after preserving 10min under 70 ℃ of conditions; At room temperature preserving after 6 months the enzyme loss of living is less than 25%, 4 ℃ and preserves after 12 months the enzyme loss of living and be less than 20%.Higher than existing 1,4 beta-glucanase enzyme activity, enzyme effect optimum pH wide scope, heat endurance and storage stability are all higher.Compared to commercially available Wheat Based Diet enzyme, under same consumption the present invention higher to the enzymatic hydrolyzation of starch, thereby improve food conversion ratio, reduce feed cost.
In the present invention, added Chinese herbal medicine powder, it has natural antibacterial and the effect that strengthens immunity, can effectively improve anti-stress ability, immunologic function and the Defense response function of livestock and poultry animal.In addition, added the root of large-flowered skullcap, the radix bupleuri with the former effect of anti-heat stress in Chinese herbal medicine powder, made product of the present invention have better stability and preserve actively, the preservation activity of the product that does not more add Chinese herbal medicine powder under long-time hot conditions improved 15%.
The specific embodiment
The preparation method of embodiment 1 Chinese herbal medicine powder
A kind of preparation method of Chinese herbal medicine powder is as follows:
Take 10 parts of the Radixs Astragali; 12 parts of Radix Codonopsis; 8 parts of Radix Isatidis; 12 parts of Radix Angelicae Sinensis; Respectively said herbal medicine being crushed to particle diameter is 2 millimeters of following mixing to obtain Chinese herbal medicine powder.
The preparation method of 2 one kinds of bacillus licheniformis cultures of embodiment
Comprise the steps:
(1) liquid seeds expands cultivation
1. first order seed is cultivated: slant strains 2 articulatings after activation are entered in 500 ml shake flasks to 100 milliliters of culture medium loading amounts, 180 revs/min of rotary shaking tables, 40 ℃ of cultivation temperature, incubation time 10h;
2. secondary seed is cultivated: by first order seed, according in 500 milliliters of secondary seed shaking flasks of inoculum concentration access of 10%, condition of culture is identical with first order seed;
3. three grades of seed culture: by secondary seed with in 5000 milliliters of three grades of seed shaking flasks of 10% inoculum concentration access, 1000 milliliters of culture medium loading amounts, 100 revs/min of rotary shaking tables, 40 ℃ of cultivation temperature, incubation time 12h;
4. first class seed pot is cultivated: three grades of seeds be take to the first class seed pot that 10% inoculum concentration access total measurement (volume) is 150L, fermentation medium loading amount 100L, 41 ℃ of cultivation temperature, mixing speed 300rpm, ventilation (V/V) 1:1.5, tank pressure 0.05Mpa, incubation time 15h;
Described seed culture medium consists of: dusty yeast 0.5%, and peptone 1%, soluble starch 1%, NaCl 1%, pH5.5,121 ℃ of sterilizing 35min.
(2) ferment tank
In the fermentation tank that first class seed pot zymotic fluid is contained to 3L fermentation medium with 3% inoculum concentration access, 37 ℃ of cultivation temperature, mixing speed 250r/min, ventilation (V/V) 1:1-3, incubation time 12h; Then by 1L through 121 ℃ of sterilizing 20min, temperature is that the fermentation medium of 10 ℃ fills into fermentation tank, constant temperature culture 18h when temperature rises to 37 ℃; Now, first class seed pot zymotic fluid is appended to access fermentation tank, constant temperature culture 12h with 2% inoculum concentration; Continuation is by 1L through 121 ℃ of sterilizing 20min, and temperature is that the fermentation medium of 10 ℃ fills into fermentation tank, constant temperature culture 12h when temperature rises to 37 ℃;
Described fermentation medium consists of: wheat bran 75g, corn flour 55g, rice bran 25g, beancake powder 20g, beta glucan 2g, Chinese herbal medicine powder 40g, trehalose 35g, dusty yeast 6g, corn steep liquor 3g, ammonium sulfate 2g, dipotassium hydrogen phosphate 2g, potassium dihydrogen phosphate 2g, natrium citricum 3g, defoamer 0.5g, pure water l000mL, 5.5,121 ℃ of sterilizing 20min of pH value;
The concocting method of described fermentation medium is: accurately take in proportion raw material, pure water in raw material, corn flour, beancake powder are dropped in material-compound tank, regulate pH value 5.5, add middle temperature amylase (3u/g corn flour) and alpha-amylase (30u/g corn flour), ℃ insulation 20min of warming while stirring to 70 simultaneously, is then slowly warming up to 90 ℃ of insulation 20min and liquefies, finally add other raw material, stir, adjust initial p H5.5,121 ℃ of sterilizing 35min are standby.
Described Chinese herbal medicine powder forms: 10 parts of the Radixs Astragali; 9 parts of Radix Codonopsis; 8 parts of Radix Isatidis; 12 parts of Radix Angelicae Sinensis; Respectively said herbal medicine being crushed to particle diameter is 2 millimeters of following mixing to obtain Chinese herbal medicine powder;
(3) zymotic fluid via hole diameter 500 μ m coarse filtration, then obtain solid content 20% concentrate in the concentrated moisture of removing of 15 ℃ of loop ultrafiltrations; Concentrate obtains bacillus licheniformis culture through vacuum freeze drying, ultramicro grinding.
The preparation method of 3 one kinds of complex enzyme preparation for feeding of embodiment
Containing the Wheat Based Diet enzyme of profitable probliotics, by the enzyme preparation of following parts by weight, formed:
120 parts of zytases, 8 parts of cellulases, 25 parts of bacillus licheniformis cultures, 8 parts of mesophilicα-diastases, 1 part of Chinese herbal medicine powder.
Get the zytase that originates from Hunan letter Wings Of The Eagle bioengineering limited company or other companies, cellulase, mesophilicα-diastase, and 1 part of the Chinese herbal medicine powder of embodiment 1 gained, bacillus licheniformis culture described in embodiment 2, under 25 ℃ of conditions, in mixer, stir, mixing time is 5 minutes.Stir vanning after in normal temperature or cold house, preserve.
The preparation method of 4 one kinds of complex enzyme preparation for feeding of embodiment
Containing the Wheat Based Diet enzyme of profitable probliotics, by the enzyme preparation of following parts by weight, formed:
100 parts of zytases, 6 parts of cellulases, 30 parts of bacillus licheniformis cultures, 10 parts of mesophilicα-diastases, 0.5 part of Chinese herbal medicine powder;
The parts by weight of each component of Chinese herbal medicine powder consist of: 5 parts of the Radixs Astragali; 13 parts of Radix Codonopsis; 10 parts of Radix Isatidis; 15 parts of Radix Angelicae Sinensis;
By above-mentioned parts by weight, get the zytase that originates from Hunan letter Wings Of The Eagle bioengineering limited company, cellulase, mesophilicα-diastase, Chinese herbal medicine powder, bacillus licheniformis culture described in embodiment 2 stirs under 25 ℃ of conditions in mixer, and mixing time is 5 minutes.Stir vanning after in normal temperature or cold house, preserve.
The preparation method of 5 one kinds of complex enzyme preparation for feeding of embodiment
Containing the Wheat Based Diet enzyme of profitable probliotics, by the enzyme preparation of following parts by weight, formed:
140 parts of zytases, 10 parts of cellulases, 20 parts of bacillus licheniformis cultures, 5 parts of mesophilicα-diastases, 2 parts of Chinese herbal medicine powders;
The parts by weight of each component of Chinese herbal medicine powder consist of: 12 parts of the Radixs Astragali; 15 parts of Radix Codonopsis; 6 parts of Radix Isatidis; 10 parts of Radix Angelicae Sinensis;
By above-mentioned parts by weight, get the zytase that originates from Hunan letter Wings Of The Eagle bioengineering limited company, cellulase, mesophilicα-diastase, Chinese herbal medicine powder, bacillus licheniformis culture described in embodiment 2 stirs under 25 ℃ of conditions in mixer, and mixing time is 5 minutes.Stir vanning after in normal temperature or cold house, preserve.
Embodiment 5 Contrast on effect experiments
Choose 120 and fatten pig in early stage, by body weight, blood lineage and no sex difference, be divided into experimental group and control group, every group of 3 repetitions, each repeats 20 pigs.Experimental period is 28 days, and experimental group feed per ton adds complex enzyme formulation 120g described in the present embodiment 3, and control group does not add complex enzyme formulation, morning day at the whole story of experimental period, weighs on an empty stomach, calculates daily gain and feedstuff-meat ratio, and gives a mark to hair color.Experimental result is as shown in the table:
| Control group | Test group | |
| Average daily gain (g/d) | 655.20 | 713.27 |
| Feed/weightening finish | 2.45 | 2.15 |
| Hair color marking (ten point system) | 5.5 | 8.5 |
Embodiment 6 Contrast on effects
Choose 120 and fatten pig in early stage, by body weight, blood lineage and no sex difference, be divided into experimental group and control group, every group of 3 repetitions, each repeats 20 pigs.Experimental period is 28 days, experimental group and control group feed per ton all adds complex enzyme formulation 120g, experimental group complex enzyme formulation used is complex enzyme formulation described in embodiment 3, control group complex enzyme formulation used is in embodiment 3, to add commercially available Wheat Based Diet enzyme, weigh morning day at the whole story in experimental period on an empty stomach, calculate daily gain and feedstuff-meat ratio, and give a mark to hair color.Experimental result is as shown in the table:
| Control group | Test group | |
| Average daily gain (g/d) | 675.26 | 713.27 |
| Feed/weightening finish | 2.37 | 2.15 |
| Hair color marking (ten point system) | 6.5 | 8.5 |
Embodiment 7 Chinese herbal medicine powders are preserved active impact to enzyme
The Wheat Based Diet enzyme that the preparation method described in embodiment 2 of take obtains is sample 1, take that to remove the Wheat Based Diet enzyme that the preparation method of Chinese herbal medicine powders all in embodiment 2 obtains be sample 2, and relatively the preservation of two kinds of samples is active.At 45 ℃, preserve above-mentioned sample 18h, measure afterwards enzyme and live, result shows: the remnant enzyme activity of sample 1 is more than 74%, and the remnant enzyme activity of sample 2 remains on 60% left and right.
Claims (8)
1. the Wheat Based Diet enzyme containing profitable probliotics, it is characterized in that: the component by following parts by weight forms: zytase 100-140 part, cellulase 6-10 part, bacillus licheniformis culture 20-30 part, mesophilicα-diastase 5-10 part, Chinese herbal medicine powder 0.5-2 part;
Described bacillus licheniformis is bacillus licheniformis (Β acillus licheniformis) β-10-25, and deposit number is CCTCC NO:M2013538.
2. a kind of Wheat Based Diet enzyme containing profitable probliotics as claimed in claim 1, is characterized in that, the preparation method of described Chinese herbal medicine powder is as follows:
Take Radix Astragali 5-12 part; Radix Codonopsis 8-15 part; Radix Isatidis 5-10 part; Radix Angelicae Sinensis 10-15 part; Respectively said herbal medicine being crushed to particle diameter is 2 millimeters of following mixing to obtain Chinese herbal medicine powder.
3. a kind of Wheat Based Diet enzyme containing profitable probliotics as claimed in claim 1, is characterized in that, the preparation method of bacillus licheniformis culture, comprises the steps:
Bacillus licheniformis β-10-25 carries out one, two, three liquid seeds and seed tank culture after slant strains activation culture; In the fermentation tank that first class seed pot zymotic fluid is contained to 3L fermentation medium with 3% inoculum concentration access, cultivation temperature 35-45 ℃, mixing speed 200-300r/min, ventilation (V/V) 1:1-3, incubation time 10-15h; Then by 1L through 121 ℃ of sterilizing 20min, temperature is that the fermentation medium of 10 ℃ fills into fermentation tank, constant temperature culture 15-20h when temperature rises to 35-45 ℃; Now, first class seed pot zymotic fluid is appended to access fermentation tank, constant temperature culture 10-15h with 2% inoculum concentration; Continuation is by 1L through 121 ℃ of sterilizing 20min, and temperature is that the fermentation medium of 10 ℃ fills into fermentation tank, constant temperature culture 10-15h when temperature rises to 35-45 ℃;
Zymotic fluid via hole diameter 10-1000 μ m coarse filtration, then obtains solid content 20-40% concentrate in the concentrated moisture of removing of 10-20 ℃ of loop ultrafiltration; Concentrate obtains bacillus licheniformis culture through vacuum freeze drying, ultramicro grinding.
4. a kind of Wheat Based Diet enzyme containing profitable probliotics as claimed in claim 3, is characterized in that, the culture medium of described seed culture is: dusty yeast 0.5%, peptone 1%, soluble starch 1%, NaCl1%, pH4.5-7.0,121-123 ℃ of sterilizing 30-40min.
Described fermentation medium consists of: wheat bran 75g, corn flour 55g, rice bran 25g, beancake powder 20g, beta glucan 2g, Chinese herbal medicine powder 40g, trehalose 35g, dusty yeast 6g, corn steep liquor 3g, ammonium sulfate 2g, dipotassium hydrogen phosphate 2g, potassium dihydrogen phosphate 2g, natrium citricum 3g, defoamer 0.5g, pure water l000mL, pH value 4.5-7.0,121 ℃ of sterilizing 20min;
The concocting method of described fermentation medium is: accurately take in proportion raw material, pure water in raw material, corn flour, beancake powder are dropped in material-compound tank, regulate pH value 3-7, add middle temperature amylase (3u/g corn flour) and alpha-amylase (30u/g corn flour), ℃ insulation 15-30min of warming while stirring to 70 simultaneously, is then slowly warming up to 90 ℃ of insulation 15-30min and liquefies, finally add other raw material, stir, pH3-7,121-123 ℃ of sterilizing 30-40min is standby.
5. a method of preparing the Wheat Based Diet enzyme containing profitable probliotics as described in claim 1-5, comprises the steps:
According to raw material, form zytase, cellulase, bacillus licheniformis culture, mesophilicα-diastase, Chinese herbal medicine powder mixes;
Described zytase, cellulase, mesophilicα-diastase is Hunan letter Wings Of The Eagle bioengineering limited company or other existing products;
The preparation method of described Chinese herbal medicine powder is as follows:
Take Radix Astragali 5-12 part; Radix Codonopsis 8-15 part; Radix Isatidis 5-10 part; Radix Angelicae Sinensis 10-15 part; Respectively said herbal medicine being crushed to particle diameter is 2 millimeters of following mixing to 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: dusty yeast 0.5%, and peptone 1%, soluble starch 1%, NaCl1%,, pH4.5-7.0,121-123 ℃ of sterilizing 30-40min.
(2) ferment tank
In the fermentation tank that first class seed pot zymotic fluid is contained to 3L fermentation medium with 3% inoculum concentration access, cultivation temperature 35-45 ℃, mixing speed 200-300r/min, ventilation (V/V) 1:1-3, incubation time 10-15h; Then by 1L through 121 ℃ of sterilizing 20min, temperature is that the fermentation medium of 10 ℃ fills into fermentation tank, constant temperature culture 15-20h when temperature rises to 35-45 ℃; Now, first class seed pot zymotic fluid is appended to access fermentation tank, constant temperature culture 10-15h with 2% inoculum concentration; Continuation is by 1L through 121 ℃ of sterilizing 20min, and temperature is that the fermentation medium of 10 ℃ fills into fermentation tank, constant temperature culture 10-15h when temperature rises to 35-45 ℃;
The concocting method of described fermentation medium is: accurately take in proportion raw material, pure water in raw material, corn flour, beancake powder are dropped in material-compound tank, regulate pH value 3-7, add middle temperature amylase (3u/g corn flour) and alpha-amylase (30u/g corn flour), ℃ insulation 15-30min of warming while stirring to 70 simultaneously, then being slowly warming up to 90 ℃ of insulation 15-30min liquefies, finally add other raw material, stir, adjust initial pH3-7,121-123 ℃ of sterilizing 30-40min is standby;
Dissolved oxygen is controlled: by adjusting speed of agitator and ventilation, control dissolved oxygen 15-30%;
PH controls: by mending ammoniacal liquor or phosphoric acid,diluted, in controlled fermentation process, pH value remains on 3-7;
Control of additive raw material: when the content of reducing sugar in zymotic fluid is down to 3mg/ml-8mg/ml, start to add supplemented medium, feed supplement amount be take and maintained zymotic fluid content of reducing sugar as 2mg/ml-5mg/ml;
Put tank standard: the old and feeble self-dissolving of 60-80% thalline, enzyme activity increasess slowly;
(3) zymotic fluid via hole diameter 10-1000 μ m coarse filtration, then obtains solid content 20-40% concentrate in the concentrated moisture of removing of 10-20 ℃ of loop ultrafiltration; Concentrate obtains bacillus licheniformis culture through vacuum freeze drying, ultramicro grinding.
6. a kind of Wheat Based Diet enzyme containing profitable probliotics as claimed in claim 1, it is characterized in that: the component by following parts by weight forms: 120 parts of zytases, 8 parts of cellulases, 25 parts of bacillus licheniformis cultures, 8 parts of mesophilicα-diastases, 1 part of Chinese herbal medicine powder.
7. the application of a kind of Wheat Based Diet enzyme containing profitable probliotics as claimed in claim 1 in livestock and poultry Wheat Based Diet.
8. a kind of Wheat Based Diet enzyme containing profitable probliotics as claimed in claim 1, is characterized in that: described Chinese herbal medicine powder forms: 10 parts of the Radixs Astragali; 9 parts of Radix Codonopsis; 8 parts of Radix Isatidis; 12 parts of Radix Angelicae Sinensis.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310739603.9A CN104054914B (en) | 2013-12-27 | 2013-12-27 | A kind of Wheat ration enzyme containing profitable probliotics and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310739603.9A CN104054914B (en) | 2013-12-27 | 2013-12-27 | A kind of Wheat ration enzyme containing profitable probliotics and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104054914A true CN104054914A (en) | 2014-09-24 |
| CN104054914B CN104054914B (en) | 2016-12-07 |
Family
ID=51543072
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310739603.9A Active CN104054914B (en) | 2013-12-27 | 2013-12-27 | A kind of Wheat ration enzyme containing profitable probliotics and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104054914B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106318396A (en) * | 2016-08-15 | 2017-01-11 | 北京中科圣浩健康产业科技有限公司 | Compound enzyme soil modifier and preparation method, use method and applications thereof |
| 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 |
Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102100311A (en) * | 2009-12-21 | 2011-06-22 | 沈阳爱特杰牧业有限公司 | Additive for predigestion fermented feed for piglet |
| CN102119768A (en) * | 2011-01-25 | 2011-07-13 | 北京挑战农业科技有限公司 | Complex enzyme preparation for feeding piglets |
| CN102239967A (en) * | 2010-05-11 | 2011-11-16 | 河北农业大学 | Chinese herbal feed additive for improving meat quality of fattening pigs |
| CN102488099A (en) * | 2011-12-27 | 2012-06-13 | 庄晓曦 | Microbial enzyme feed additive, its feed formula and preparation method |
| CN102754753A (en) * | 2012-08-07 | 2012-10-31 | 山东新希望六和集团有限公司 | Feed for improving growth performances of broiler chickens and preparation method thereof |
| CN102771627A (en) * | 2011-05-09 | 2012-11-14 | 北京奕农顺丰生物技术有限公司 | Feed additive containing compound enzyme |
| CN102919565A (en) * | 2012-09-18 | 2013-02-13 | 沈阳巨源源生物科技有限公司 | Meat chick granulated feed containing active bacterium fermentation ingredient and traditional Chinese medicine ingredient |
| CN102987098A (en) * | 2012-12-07 | 2013-03-27 | 青岛根源生物技术集团有限公司 | Special complex enzyme preparation for wheat for livestock and poultry and application thereof |
| CN103250877A (en) * | 2013-05-17 | 2013-08-21 | 兰州天峰生物科技有限责任公司 | Natural Chinese herbal medicine animal feed additive and application thereof |
| CN103409347A (en) * | 2013-07-25 | 2013-11-27 | 山东隆科特酶制剂有限公司 | Bacterial strain capable of producing alkali protease and industrialized liquid fermentation method of bacterial strain |
-
2013
- 2013-12-27 CN CN201310739603.9A patent/CN104054914B/en active Active
Patent Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102100311A (en) * | 2009-12-21 | 2011-06-22 | 沈阳爱特杰牧业有限公司 | Additive for predigestion fermented feed for piglet |
| CN102239967A (en) * | 2010-05-11 | 2011-11-16 | 河北农业大学 | Chinese herbal feed additive for improving meat quality of fattening pigs |
| CN102119768A (en) * | 2011-01-25 | 2011-07-13 | 北京挑战农业科技有限公司 | Complex enzyme preparation for feeding piglets |
| CN102771627A (en) * | 2011-05-09 | 2012-11-14 | 北京奕农顺丰生物技术有限公司 | Feed additive containing compound enzyme |
| CN102488099A (en) * | 2011-12-27 | 2012-06-13 | 庄晓曦 | Microbial enzyme feed additive, its feed formula and preparation method |
| CN102754753A (en) * | 2012-08-07 | 2012-10-31 | 山东新希望六和集团有限公司 | Feed for improving growth performances of broiler chickens and preparation method thereof |
| CN102919565A (en) * | 2012-09-18 | 2013-02-13 | 沈阳巨源源生物科技有限公司 | Meat chick granulated feed containing active bacterium fermentation ingredient and traditional Chinese medicine ingredient |
| CN102987098A (en) * | 2012-12-07 | 2013-03-27 | 青岛根源生物技术集团有限公司 | Special complex enzyme preparation for wheat for livestock and poultry and application thereof |
| CN103250877A (en) * | 2013-05-17 | 2013-08-21 | 兰州天峰生物科技有限责任公司 | Natural Chinese herbal medicine animal feed additive and application thereof |
| CN103409347A (en) * | 2013-07-25 | 2013-11-27 | 山东隆科特酶制剂有限公司 | Bacterial strain capable of producing alkali protease and industrialized liquid fermentation method of bacterial strain |
Non-Patent Citations (5)
| Title |
|---|
| 冯焱等: "小麦型日粮添加益生菌与酶制剂对肉鸡生产性能及肉色的影响", 《粮食与饲料工业》 * |
| 牛竹叶等: "复合酶制剂在蛋鸡小麦型日粮中的应用", 《中国农学通报》 * |
| 王慧军等: "中草药添加剂在我国养殖业的应用现状", 《山东畜牧兽医》 * |
| 王继成主编: "《饲料添加剂配制与应用新技术》", 30 April 1996, 湖南科学技术出版社 * |
| 袁青杉等: "纤维素复合酶对小麦日粮酶解效果的研究", 《草业与畜牧》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106318396A (en) * | 2016-08-15 | 2017-01-11 | 北京中科圣浩健康产业科技有限公司 | Compound enzyme soil modifier and preparation method, use method and applications thereof |
| 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 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104054914B (en) | 2016-12-07 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103667222B (en) | Feed compound enzyme-containing dedicated enzyme for growing pigs and preparation method of feed compound enzyme-containing dedicated enzyme | |
| CN103805582B (en) | Beta-glucanase containing special enzyme for growing pigs and preparation method thereof | |
| CN110037165A (en) | A kind of Heisui River gadfly slurry solid state fermentation concentrated feed and preparation method | |
| CN106222114A (en) | The bacillus cereus of efficient degradation bean cake antigen protein and the method for bacterium enzyme mixed fermentation | |
| CN103667220B (en) | Neutral protease-containing growing pig dedicated enzyme and preparation method thereof | |
| CN103740601B (en) | A kind of aspergillus niger of high yield complex enzyme for feed and application thereof | |
| CN103749987B (en) | A kind of coarse cereals are mixed dregs of rice daily ration enzyme preparation and preparation method thereof | |
| CN103504123A (en) | Fermented soybean meal with function of complex enzymes and preparation method for fermented soybean meal | |
| CN105379947A (en) | Animal feed and preparation method thereof | |
| CN103766611A (en) | Neutral protease-containing coarse grain daily ration enzyme and preparation method thereof | |
| CN103156059B (en) | Preparation method of high-protein and low-fiber biological nutrition maize straw forage grass | |
| CN106260589A (en) | A kind of method that recycling wheat bran prepares albumen feedstuff | |
| CN103875935B (en) | A kind of suckling piglet special composite enzyme containing profitable probliotics and preparation method thereof | |
| CN103750023B (en) | Special beta-dextranase-containing complex enzyme for piglet and preparation method thereof | |
| CN109486724A (en) | A kind of synchronization preparation process and its feed applications mixing probiotics leaven | |
| CN103725661B (en) | A kind of suckling piglet specific enzyme containing complex enzyme for feed and preparation method thereof | |
| CN102643747A (en) | Method for preparing probiotics mixed preparation for processing straws | |
| CN103156091B (en) | Method for manufacturing biologically-fermented straw nutrition forage grass at low temperature for beef | |
| CN103865904B (en) | A kind of growing swine specific enzyme containing profitable probliotics and preparation method thereof | |
| CN104054914B (en) | A kind of Wheat ration enzyme containing profitable probliotics and preparation method thereof | |
| CN103392920A (en) | Fermentation method of soybean hulls | |
| CN103695396B (en) | A kind of high vigor complex enzyme for feed | |
| CN110150457A (en) | A kind of antibacterial type yeast culture and its application | |
| CN103805583B (en) | Wheat diet enzyme containing beta-glucanase and preparation method thereof | |
| CN103740683B (en) | A kind of Wheat ration enzyme containing neutral protease and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |