CN103875935A - Probiotics-containing special complex enzyme for piglets and preparation method of complex enzyme - Google Patents
Probiotics-containing special complex enzyme for piglets and preparation method of complex enzyme Download PDFInfo
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Abstract
The invention relates to a probiotics-containing special enzyme for piglets and a preparation method of the special enzyme, and belongs to the field of feed enzyme preparations. The probiotics-containing special enzyme for the piglets comprises the following components in parts by weight: 5-20 parts of Chinese herbal medicine powder, 800-1200 parts of xylanase, 80-120 parts of cellulase, 250-300 parts of bacillus licheniformis culture, 15-25 parts of mannose, 600-1000 parts of protease and 50-80 parts of medium-temperature alpha-amylase, wherein the bacillus licheniformis culture is bacillus licheniformis beta-10-25; the preservation number of the bacillus licheniformis culture is CCTCCNO.M2013538. The preparation method of the probiotics-containing special enzyme for the piglets comprises the following steps: fermenting bacillus licheniformis, then filtering, concentrating and drying the bacillus licheniformis to obtain a culture, compounding the culture with other components. The complex enzyme is capable of improving the feed conversion rate, reducing the feed cost and improving preservation stability.
Description
Technical field
The invention belongs to enzyme preparation field, be specifically related to a kind of newborn piglet special composite enzyme containing profitable probliotics and preparation method thereof.
Background technology
In animal husbandry, newborn feeding piglet is the most complicated in all stages pig.The irrational consequence of nutrition supply is pig poor growth, and diarrhea rate is high, and the death rate is high, and then in making, large pig growth period is slow, extends marketing time.
But, in order to cut off the vertical transmission route of source of parents disease, shorten the sow breeding cycle simultaneously, extend the kind time limit, people are widely used in early weaned technology in the large-scale farming industry of pig.But because newborn piglet lacks congenital immunity, digestive system development is not perfect, a little less than digestive ferment secretion capacity, be directly that main feed is easy to cause the problems such as porkling is low to the digestibility of feed, feed intake is low, the speed of growth is slow, diarrhea rate is high for giving corn etc.
The variety of problems occurring after wean in order to overcome sucking pig, the many aspects such as supplementary feeding, food calling, the daily ration acidifying from weaning of majority enterprises, the feedstuff that uses good palatability and absorption easy to digest, the antigenicity of adding exogenous enzymes, reduction daily ration, the crude protein content that reduces daily ration and use promoter are improved and are optimized, but actual effect is uneven.
Consider that piglet is in when wean, the enzyme such as protease, the amylase work of himself stress and 1/3rd while declining to a great extent wean, and these enzymes obtain nutrition for piglet and energy plays vital effect.Meanwhile, fodder enzyme has original effect in Production of Livestock and Poultry, such as 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%.
Add enzyme preparation and can supplement protease and amylase, auxiliary albumen and the starch decomposing in daily ration, the ANFs of degraded in daily ration, and can effectively decompose the plant cell wall in corn/bean pulp type daily ration, makes intracellular nutriment discharge as starch, protein etc.Decomposing soluble fiber simultaneously, reduces the water absorbing capacity of string material and the viscosity of intestinal contents.In addition, a large amount of production practices and scientific experiment have also proved 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 official association have 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, we are through the experiment of repeatedly feeding, develop a kind of newborn piglet special composite enzyme that is added with the bacillus licheniformis culture that can produce high activity and stability 1,4 beta-glucanase, it can improve utilization rate, reduction diarrhea rate, the promotion growth of animal of feed effectively, with strong points, effective, remarkable in economical benefits.
Summary of the invention
The object of the present invention is to provide that a kind of enzyme activity is high, Antidiarrheic effect is significantly and containing newborn piglet special composite enzyme of profitable probliotics and preparation method thereof.
Object of the present invention can be achieved through the following technical solutions:
A kind of newborn piglet special composite enzyme containing profitable probliotics, it is characterized in that, described newborn piglet special composite enzyme is made up of the component of following parts by weight: Chinese herbal medicine powder 5-20 part, zytase 800-1200 part, cellulase 80-120 part, bacillus licheniformis culture 250-300 part, mannase 15-25, protease 600-1000, mesophilicα-diastase 50-80 part.
Described zytase, cellulase, mannonase protease and mesophilicα-diastase are 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 5-15 part; Radix Codonopsis 10-18 part; Radix Isatidis 10-15 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;
The preparation method of described bacillus licheniformis culture 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%, peptone 1%, soluble starch 1%, NaCl 1%, pH4.5-7.0,121-123 ℃ of sterilizing 30-40min.
(2) ferment tank
First class seed pot zymotic fluid is accessed in the fermentation tank that contains 3L fermentation medium to cultivation temperature 35-45 ℃, mixing speed 200-300r/min, ventilation (V/V) 1:1-3, incubation time 10-15h with 3% inoculum concentration; 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 in the time that 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 in the time that 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, herbal mediciment 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, 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 for subsequent use.
Dissolved oxygen control: 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: in the time that the content of reducing sugar in zymotic fluid is down to 3mg/ml-8mg/ml, start to add supplemented medium, feed supplement amount is to maintain 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 (be called for short CCTCC, address is: Wuchang District, Wuhan City, Hubei Province Luo Jia Shan Wuhan University Life Science College) on November 3rd, 2013, and preserving number is CCTCC NO:M 2013538.
Described bacterial strain is mutagenic obtained through physical chemistry, 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 newborn piglet special composite enzyme containing profitable probliotics forms by following parts by weight: 10 parts of Chinese herbal medicine powders, 1000 parts of zytases, 100 parts of cellulases, 275 parts of bacillus licheniformis cultures, 20 parts of mannases, 00 part of proteinase 8,65 parts of mesophilicα-diastases.
Described newborn piglet special composite enzyme is applicable to feed-processing plant and plant's autogamy feed, when use, should mix with other raw material in feed, can in advance the present invention be mixed with a small amount of feed, then be mixed in large quantities of feeds Direct-fed.Advise that complete diet pellet addition per ton is 60-120g.
Beneficial effect:
The present invention is according to newborn piglet digestive physiological characteristics, and through zoopery screening compound prescription, complex enzyme zymogram is set, effectively supplement the deficiency of the endogenous enzymes such as newborn piglet alimentary canal protease, amylase, effectively improve the digestive utilization ratio of newborn piglet to feed, prevent the generation of trophism diarrhea.In the bacillus licheniformis culture that the present invention adds, contain bacillus licheniformis and 1,4 beta-glucanase, not only simplified the purifying process of enzyme compared with adding pure enzymatic process, and add a large amount of probios---bacillus licheniformis in adding enzyme.The interpolation of bacillus licheniformis can produce protein and peptide antibacterial material on the one hand, antagonism enteron aisle pathogenetic bacteria, suppress 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 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.
In the present invention, added Chinese herbal medicine powder, it has the effect of natural antibacterial and enhancing immunity, can effectively put forward anti-stress ability, immunologic function and the Defense response function of newborn piglet.In addition, added the root of large-flowered skullcap, the Radix Isatidis 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; 15 parts of Radix Codonopsis; 12 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
A preparation method for bacillus licheniformis culture, comprises 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: secondary seed is accessed in 5000 milliliters of three grades of seed shaking flasks to 1000 milliliters of culture medium loading amounts, 100 revs/min of rotary shaking tables, 40 ℃ of cultivation temperature, incubation time 12h with 10% inoculum concentration;
4. first class seed pot is cultivated: the first class seed pot by three grades of seeds take 10% inoculum concentration access total measurement (volume) as 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 weight consists of: dusty yeast 0.5%, peptone 1%, soluble starch 1%, NaCl1%, pH5.5,121 ℃ of sterilizing 35min.
(2) ferment tank
First class seed pot zymotic fluid is accessed in the fermentation tank that contains 3L fermentation medium to 37 ℃ of cultivation temperature, mixing speed 250r/min, ventilation (V/V) 1:1-3, incubation time 12h with 3% inoculum concentration; Then by 1L through 121 ℃ of sterilizing 20min, temperature is that the fermentation medium of 10 ℃ fills into fermentation tank, constant temperature culture 18h in the time that 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 in the time that 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 for subsequent use.
The preparation method of Chinese herbal medicine powder is as follows:
Take 10 parts of the Radixs Astragali; 15 parts of Radix Codonopsis; 8 parts of Radix Isatidis, 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.
3 one kinds of preparation methods containing the newborn piglet special composite enzyme of profitable probliotics of embodiment
Containing a newborn piglet special composite enzyme for profitable probliotics, formed by the enzyme preparation of following parts by weight:
10 parts of Chinese herbal medicine powders, 1000 parts of zytases, 100 parts of cellulases, 275 parts of bacillus licheniformis cultures, 20 parts of mannases, 00 part of proteinase 8,65 parts of mesophilicα-diastases.
Get the zytase that originates from Hunan letter Wings Of The Eagle bioengineering limited company by above-mentioned parts by weight, cellulase, mannase, mesophilicα-diastase, protease, and the Chinese herbal medicine powder of embodiment 1 gained, the 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.
4 one kinds of preparation methods containing the newborn piglet special composite enzyme of profitable probliotics of embodiment
Containing a newborn piglet special composite enzyme for profitable probliotics, formed by the enzyme preparation of following parts by weight:
20 parts of Chinese herbal medicine powders, 1200 parts of zytases, 120 parts of cellulases, 300 parts of bacillus licheniformis cultures, 15 parts of mannases, 600 parts, protease, 80 parts of mesophilicα-diastases;
Chinese herbal medicine powder parts by weight composition: 15 parts of the Radixs Astragali; 18 parts of Radix Codonopsis; 10 parts of Radix Isatidis; 10 parts of Radix Angelicae Sinensis;
Get the zytase that originates from Hunan letter Wings Of The Eagle bioengineering limited company by above-mentioned parts by weight, cellulase, mannase, mesophilicα-diastase, protease, and Chinese herbal medicine powder, the bacillus licheniformis culture described in embodiment 2, under 5 ℃ of conditions, in mixer, stir, mixing time is 5 minutes.Stir vanning after in normal temperature or cold house, preserve.
5 one kinds of preparation methods containing the newborn piglet special composite enzyme of profitable probliotics of embodiment
Containing a newborn piglet special composite enzyme for profitable probliotics, formed by the enzyme preparation of following parts by weight:
5 parts of Chinese herbal medicine powders, 800 parts of zytases, 80 parts of cellulases, 200 parts of bacillus licheniformis cultures, 25 parts of mannases, 1200 parts, protease, 50 parts of mesophilicα-diastases;
Chinese herbal medicine powder parts by weight composition: 5 parts of the Radixs Astragali; 10 parts of Radix Codonopsis; 12 parts of Radix Isatidis; 12 parts of Radix Angelicae Sinensis;
Get the zytase that originates from Hunan letter Wings Of The Eagle bioengineering limited company by above-mentioned parts by weight, cellulase, mannase, mesophilicα-diastase, protease, and Chinese herbal medicine powder, the 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 6 Contrast on effect experiments
Choose 120 weanling pigs, be divided into experimental group and control group by body weight, blood lineage and no sex difference, every group of 3 repetitions, 20 pigs of each repetition.Experimental period is 28 days, and experimental group feed per ton adds complex enzyme formulation 120g described in the present embodiment 2, and control group does not add complex enzyme formulation, weighs on an empty stomach morning day at the whole story of experimental period, calculates feed conversion rate, daily gain, feedstuff-meat ratio, diarrhea rate and death rate.Experimental result is as shown in the table:
| ? | Daily gain | Feedstuff-meat ratio | Diarrhea rate | Death rate |
| Control group | 236.57g | 1.48 | 16.7% | 5% |
| Experimental group | 335.52g | 1.31 | 5.57% | 1.7% |
Embodiment 7 Contrast on effects
Choose 120 weanling pigs, be divided into experimental group and control group by body weight, blood lineage and no sex difference, every group of 3 repetitions, 20 pigs of each repetition.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 2, control group complex enzyme formulation used is market milk piglet specific enzyme, weigh morning day at the whole story in experimental period on an empty stomach, calculates feed conversion rate, daily gain, feedstuff-meat ratio diarrhea rate and death rate.Experimental result is as shown in the table:
| ? | Daily gain | Feedstuff-meat ratio | Diarrhea rate | Death rate |
| Control group | 278.57 | 1.43 | 10% | 2.5% |
| Experimental group | 335.52g | 1.31 | 5.57% | 1.7% |
Above experimental result shows, uses the newborn piglet special composite enzyme of the technology of the present invention production can effectively reduce the incidence of newborn Diarrhea after Piglets " Weaning, improves the speed of growth and the efficiency of feed utilization of newborn piglet, is a kind of desirable feed complex enzyme.
Embodiment 8 Chinese herbal medicine powders are preserved active impact to enzyme
The newborn piglet special composite enzyme obtaining take preparation method described in embodiment 2 is as sample 1, to remove Wheat Based Diet enzyme that the preparation method of Chinese herbal medicine powders all in embodiment 2 obtains as sample 2, the relatively preservation activity of two kinds of samples.At 45 ℃, preserve above-mentioned sample 18h, measure afterwards enzyme and live, result shows: the remnant enzyme activity of sample 1 is more than 73%, and the remnant enzyme activity of sample 2 remains on 60% left and right.
Claims (7)
1. the newborn piglet special composite enzyme containing profitable probliotics, it is characterized in that: the component by following parts by weight forms: Chinese herbal medicine powder 5-20 part, zytase 800-1200 part, cellulase 80-120 part, bacillus licheniformis culture 250-300 part, mannase 15-25, protease 600-1000, mesophilicα-diastase 50-80 part;
Described bacillus licheniformis is specially bacillus licheniformis (Β acillus licheniformis) β-10-25, and deposit number is CCTCC NO:M 2013538.
2. a kind of newborn piglet special composite 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 5-15 part; Radix Codonopsis 10-18 part; Radix Isatidis 10-15 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 newborn piglet special composite 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; First class seed pot zymotic fluid is accessed in the fermentation tank that contains 3L fermentation medium to cultivation temperature 35-45 ℃, mixing speed 200-300r/min, ventilation (V/V) 1:1-3, incubation time 10-15h with 3% inoculum concentration; 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 in the time that 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 in the time that 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 newborn piglet special composite enzyme containing profitable probliotics as claimed in claim 3, it is characterized in that, 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 preparation method of Chinese herbal medicine powder is as follows: take 10 parts of the Radixs Astragali; 15 parts of Radix Codonopsis; 8 parts of Radix Isatidis, respectively said herbal medicine being crushed to particle diameter is 2 millimeters of following mixing to obtain Chinese herbal medicine powder.
5. a method of preparing the newborn piglet special composite enzyme containing profitable probliotics as claimed in claim 1, comprises the steps:
Form zytase according to raw material, cellulase, mannase, protease, bacillus licheniformis culture, mesophilicα-diastase, Chinese herbal medicine powder mixes;
The preparation method of described Chinese herbal medicine powder is as follows:
Take Radix Astragali 5-15 part; Radix Codonopsis 10-18 part; Radix Isatidis 10-15 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%.
(2) ferment tank
First class seed pot zymotic fluid is accessed in the fermentation tank that contains 3L fermentation medium to cultivation temperature 35-45 ℃, mixing speed 200-300r/min, ventilation 1:1-3, incubation time 10-15h with 3% inoculum concentration; 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 in the time that 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 in the time that 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;
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 and alpha-amylase, ℃ 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 pH3-7,121-123 ℃ of sterilizing 30-40min is for subsequent use;
Dissolved oxygen control: 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: in the time that the content of reducing sugar in zymotic fluid is down to 3mg/ml-8mg/ml, start to add supplemented medium, feed supplement amount is to maintain 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 newborn piglet special composite enzyme containing profitable probliotics as claimed in claim 1, it is characterized in that: described complex enzyme is made up of the component of following parts by weight: 10 parts of Chinese herbal medicine powders, 1000 parts of zytases, 100 parts of cellulases, 275 parts of bacillus licheniformis cultures, 20 parts of mannases, 00 part of proteinase 8,65 parts of mesophilicα-diastases.
7. a kind of newborn piglet special composite enzyme containing profitable probliotics as claimed in claim application in newborn feeding piglet process.
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